Handbook of Environmental Engineering 26

Lawrence K. Wang
Mu-Hao Sung Wang
Yung-Tse Hung Editors

Waste Treatment in
the Biotechnology,
Agricultural and
Food Industries
Volume 1
Handbook of Environmental Engineering 26

Series Editors
Lawrence K. Wang
PhD, Rutgers University, New Brunswick, NJ, USA
MS, University of Rhode Island, Kingston, RI, USA
MSCE, Missouri University of Science and Technology, Rolla, MO, USA
BSCE, National Cheng Kung University, Tainan, Taiwan, ROC

Mu-Hao Sung Wang

PhD, Rutgers University, New Brunswick, NJ, USA
MS, University of Rhode Island, Kingston, RI, USA
BSCE, National Cheng Kung University, Tainan, Taiwan, ROC
The past 75 years have seen the emergence of a growing desire worldwide to take
positive actions to restore and protect the environment from the degrading effects of
all forms of pollution: air, noise, solid waste, and water. The principle intention of
the Handbook of Environmental Engineering (HEE) series is to help readers
formulate answers to the fundamental questions facing pollution in the modern era,
mainly how serious is pollution and is the technology needed to abate it not only
available, but feasible. In a highly practical manner, HEE offers educators, students,
and engineers a strong grounding in the principles of Environmental Engineering,
as well as providing effective methods for developing optimal abatement
technologies at costs that are fully justified by the degree of abatement achieved.
With an emphasis on using the Best Available Technologies, the authors of these
volumes present the necessary engineering protocols derived from the fundamental
principles of chemistry, physics, and mathematics, making these volumes a must
have for environmental pollution researchers.

More information about this series at http://www.springer.com/bookseries/7645

Lawrence K. Wang
Mu-Hao Sung Wang • Yung-Tse Hung
Editors

Waste Treatment
in the Biotechnology,
Agricultural and Food
Industries
Volume 1
Editors
Lawrence K. Wang Mu-Hao Sung Wang
Lenox Institute of Water Technology Lenox Institute of Water Technology
Latham, NY, USA Latham, NY, USA
Agricultural Engineering Department
University of Illinois
Urbana-Champaign, IL, USA

Yung-Tse Hung
Department of Civil and Environmental
Engineering, Cleveland State University
Cleveland, OH, USA

ISSN 2512-1359     ISSN 2512-1472 (electronic)

Handbook of Environmental Engineering
ISBN 978-3-031-03589-0    ISBN 978-3-031-03591-3 (eBook)
https://doi.org/10.1007/978-3-031-03591-3

© Springer Nature Switzerland AG 2022

This work is subject to copyright. All rights are solely and exclusively licensed by the Publisher, whether
the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of
illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and
transmission or information storage and retrieval, electronic adaptation, computer software, or by similar
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The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication
does not imply, even in the absence of a specific statement, that such names are exempt from the relevant
protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in this book
are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the
editors give a warranty, expressed or implied, with respect to the material contained herein or for any
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claims in published maps and institutional affiliations.

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The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland
Preface

The past 75 years have seen the emergence of a growing desire worldwide that posi-
tive actions be taken to restore and protect the environment from the degrading
effects of all forms of pollution—air, water, soil, thermal, radioactive, and noise.
Since pollution is a direct or indirect consequence of waste, the seemingly idealistic
demand for “zero discharge” can be construed as an unrealistic demand for zero
waste. However, as long as waste continues to exist, we can only attempt to abate
the subsequent pollution by converting it to a less noxious form, or reusable form.
Three major questions usually arise when a particular type of pollution has been
identified: (1) How serious are the environmental pollution and natural resources
crisis? (2) Is the technology to abate them or recycle them available? and (3) Do the
costs of abatement justify the degree of treatment achieved for environmental pro-
tection and resources conservation? This book is one of the volumes of the Handbook
of Environmental Engineering series. The principal intention of this series is to help
readers formulate answers to the above three questions.
The traditional approach of applying tried-and-true solutions to specific environ-
mental and natural resources problems has been a major contributing factor to the
success of environmental engineering and has accounted in large measure for the
establishment of a “methodology of pollution control.” However, the realization of
the ever-increasing complexity and interrelated nature of current environmental
problems renders it imperative that intelligent planning of pollution abatement sys-
tems be undertaken. A prerequisite to such planning is an understanding of the per-
formance, potential, and limitations of the various methods of environmental
protection and resources recovery available for environmental scientists and engi-
neers. In this series of handbooks, we will review at a tutorial level a broad spectrum
of engineering systems (natural environment, processes, operations, and methods)
currently being utilized, or of potential utility, for pollution abatement, environmen-
tal protection, and natural resources conservation. We believe that the unified inter-
disciplinary approach presented in these handbooks is a logical step in the evolution
of environmental engineering.

v
vi Preface

Treatment of the various engineering systems presented will show how an engi-
neering formulation of the subject flows naturally from the fundamental principles
and theories of chemistry, microbiology, physics, and mathematics. This emphasis
on fundamental science recognizes that engineering practice has in recent years
become more firmly based on scientific principles rather than on its earlier depen-
dency on an empirical accumulation of facts. It is not intended, though, to neglect
empiricism where such data lead quickly to the most economical design. Certain
engineering systems are not readily amenable to fundamental scientific analysis,
and in these instances we have resorted to less science in favor of more art and
empiricism.
Since a bio-environmental engineer must understand science within the context
of applications, we first present the development of the scientific basis of a particu-
lar subject, followed by exposition of the pertinent design concepts and operations,
and detailed explanations of their applications to natural resources conservation or
environmental protection. Throughout the series, methods of mathematical model-
ing, system analysis, practical design, and calculation are illustrated by numerical
examples. These examples clearly demonstrate how organized analytical reasoning
leads to the most direct and clear solutions. Wherever possible, pertinent cost data
or models have been provided.
Our treatment of wastes from biotechnology, agricultural, and food industries is
offered in the belief that the trained engineer should more firmly understand funda-
mental principles, be more aware of the similarities and/or differences among many
of the bio-environmental engineering systems, and exhibit greater flexibility and
originality in the definition and innovative solution of bio-environmental system
problems. In short, the bio-environmental engineers should, by conviction and prac-
tice, be more readily adaptable to change and progress.
Coverage of the unusually broad field of environmental science, technology,
engineering, and mathematics (STEM) has demanded expertise that could only be
provided through multiple authorships. Each author (or group of authors) was
permitted to employ, within reasonable limits, the customary personal style in
organizing and presenting a particular subject area; consequently, it has been dif-
ficult to treat all subject materials in a homogeneous manner. Moreover, owing to
limitations of space, some of the authors’ favored topics could not be treated in
great detail, and many less important topics had to be merely mentioned or com-
mented on briefly. All authors have provided an excellent list of references at the
end of each chapter for the benefit of the interested readers. As each chapter is
meant to be self-­contained, some mild repetition among the various texts was
unavoidable. In each case, all omissions or repetitions are the responsibility of the
editors and not the individual authors. With the current trend toward metrication,
the question of using a consistent system of units has been a problem. Wherever
possible, the authors have used the British system (fps) along with the metric
equivalent (mks, cgs, or SIU) or vice versa. The editors sincerely hope that this
redundancy of units’ usage will prove to be useful rather than being disruptive to
the readers.
Preface vii

The goals of the Handbook of Environmental Engineering (HEE) series are: (1)
to cover entire environmental fields, including air, land, water, and noise pollution
control, solid waste processing and resource recovery, physicochemical treatment
processes, biological treatment processes, biotechnology, biosolids management,
flotation technology, membrane technology, desalination technology, water
resources, natural control processes, radioactive waste disposal, hazardous waste
management, and thermal pollution control; and (2) to employ a multimedia
approach to environmental conservation and protection since air, water, soil, and
energy are all interrelated.
This book (Waste Treatment in the Biotechnology, Agricultural and Food
Industries, Volume 1) and its sister books of the Handbook of Environmental
Engineering (HEE) series have been designed to serve as a mini-series of bio-­
environmental engineering and management textbooks as well as supplemental ref-
erence books. We hope and expect they will prove of equally high value to advanced
undergraduate and graduate students, to designers of sustainable biological resources
systems, and to scientists and researchers. The editors welcome comments from
readers in all of these categories. It is our hope that the bio-environmental engineer-
ing and management books will not only provide information on bio-resources
engineering but will also serve as a basis for advanced study or specialized investi-
gation of the theory and analysis of various biological systems.
This book, Waste Treatment in the Biotechnology, Agricultural and Food
Industries, Volume 1, covers the topics on: treatment and management of livestock
wastes; waste treatment in the pharmaceutical biotechnology industry using green
environmental technologies; vermicomposting process for processing agricultural
and food industry wastes; the impacts of climate change on agricultural, food, and
public utility industries; innovative PACT-activated sludge, CAPTOR-activated
sludge, activated bio-filter, vertical loop reactor, and PHOSTRIP processes; agricul-
tural waste treatment by water hyacinth aquaculture, wetland aquaculture, evapo-
transpiration, rapid rate land treatment, slow rate land treatment, and subsurface
infiltration; production and applications of crude polyhydroxyalkanoate-containing
bioplastic from the agricultural and food-processing wastes; optimization processes
of biodiesel production from pig and neem (Azadirachta indica A. juss) seeds blend
oil using alternative catalysts from waste biomass; making castor oil a promising
source for the production of flavor and fragrance through lipase-mediated biotrans-
formation; and treatment and minimization of waste in baker’s yeast industry.
The editors are pleased to acknowledge the encouragement and support received
from Mr. Aaron Schiller, Executive Editor of the Springer Nature Switzerland AG,
and his colleagues, during the conceptual stages of this endeavor. We wish to thank
the contributing authors for their time and effort, and for having patiently borne our
reviews and numerous queries and comments. We are very grateful to our respective
families for their patience and understanding during some rather trying times.

Auburndale, MA, USA; Latham, NY, USA Lawrence K. Wang

Auburndale, MA, USA; Latham, NY, USA  Mu-Hao Sung Wang
Cleveland, OH, USA  Yung-Tse Hung
Contents

1 
Management and Treatment of Livestock Wastes��������������������������������    1
Dale H. Vanderholm, Donald L. Day, Arthur J. Muehling, Lawrence
K. Wang, Yung-Tse Hung, Erick Butler, Mu-Hao Sung Wang,
and Haneen Yehya
2 
Waste Treatment in the Pharmaceutical Biotechnology
Industry Using Green Environmental Technologies ����������������������������   79
Lawrence K. Wang, Mu-Hao Sung Wang, Nazih K. Shammas,
and Ping Wang
3 
Vermicomposting Process for Treating Agricultural and Food
Wastes�������������������������������������������������������������������������������������������������������� 173
Lawrence K. Wang, Mu-Hao Sung Wang, Yung-Tse Hung,
Kathleen Hung Li, Hamidi Abdul Aziz, Mohd Suffian Yusoff, and
Puganeshwary Palaniandy
4 
The Impacts of Climate Change on Agricultural, Food,
and Public Utility Industries ������������������������������������������������������������������ 205
Josephine O. Wong, Erick Butler, Nai-Yi Wang, Mu-Hao Sung
Wang, and Lawrence K. Wang
5 
Innovative PACT Activated Sludge, CAPTOR Activated Sludge,
Activated Bio-­Filter, Vertical Loop Reactor,
and PhoStrip Processes���������������������������������������������������������������������������� 241
Lawrence K. Wang, Mu-Hao Sung Wang, and Nazih K. Shammas
6 
Agricultural Waste Treatment by Water Hyacinth Aquaculture,
Wetland Aquaculture, Evapotranspiration, Rapid Rate
Land Treatment, Slow Rate Land Treatment,
and Subsurface Infiltration �������������������������������������������������������������������� 277
Lawrence K. Wang, Mu-Hao Sung Wang, and Nazih K. Shammas

ix
x Contents

7 
Production and Applications of Crude
Polyhydroxyalkanoate-Containing Bioplastic
from the Agricultural and Food-­Processing Wastes ���������������������������� 317
Volodymyr Ivanov, Yung-Tse Hung, Viktor Stabnikov,
Robert Lee-­Kong Tiong, and Anatoliy Salyuk
8 
Optimization Processes of Biodiesel Production from Pig
and Neem (Azadirachta indica A. Juss) Seeds Blend Oil Using
Alternative Catalysts from Waste Biomass�������������������������������������������� 341
T. F. Adepoju and Yung-Tse Hung
9 
Castor Oil: A Promising Source for the Production of Flavor
and Fragrance Through Lipase-Mediated Biotransformation������������ 363
Suman Singh, Naziya Syed, Shivani Chaturvedi,
Ashween Deepak Nannaware, Prashant Kumar Rout,
and Yung-Tse Hung
10 
Treatment and Minimization of Waste in Baker’s Yeast Industry������ 391
Esra Can Doğan, Ayla Arslan, Nevim Genç, Levent Dağaşan,
and Yung-Tse Hung

Index������������������������������������������������������������������������������������������������������������������ 471
About the Editors

Lawrence K. Wang has served the society as a pro-

fessor, inventor, chief engineer, chief editor, and public
servant (UN, USEPA, New York State) for 50+ years,
with experience in entire field of environmental sci-
ence, technology, engineering, and mathematics
(STEM). He is a licensed NY-MA-NJ-PA-OH
Professional Engineer, a certified NY-MA-RI
Laboratory Director, a licensed MA-NY Water
Operator, and an OSHA Instructor. He has special pas-
sion and expertise in developing various innovative
technologies, educational programs, licensing courses,
international projects, academic publications, and
humanitarian organizations, all for his dream goal of
promoting world peace. He is a retired Acting President/
Professor of the Lenox Institute of Water Technology,
USA, a Senior Advisor of the United Nations Industrial
Development Organization (UNIDO), Vienna, Austria,
and a former professor/visiting professor of Rensselaer
Polytechnic Institute, Stevens Institute of Technology,
University of Illinois, National Cheng-Kung University,
Zhejiang University, and Tongji University. Dr. Wang is
the author of 750+ papers and 50+ books and is cred-
ited with 29 invention patents. He holds a BSCE degree
from National Cheng-Kung University, Taiwan, ROC;
an MSCE degree from the University of Missouri; an
MS degree from the University of Rhode Island; and a
PhD degree from Rutgers University, USA. Currently,
he is the book series editor of CRC Press, Springer
Nature Switzerland, Lenox Institute Press, World
Scientific Singapore, and John Wiley. Dr. Wang has
been a Delegate of the People to People International

xi
xii About the Editors

Foundation; a Diplomate of the American Academy of

Environmental Engineers; a member of ASCE, AIChE,
ASPE, WEF, AWWA, CIE, and OCEESA; and a recipi-
ent of many US and international engineering and sci-
ence awards.

Mu-Hao Sung Wang has been an engineer of the

New York State Department of Environmental
Conservation, an editor of CRC Press, Springer Nature
Switzerland, and Lenox Institute Press, and a university
professor of the Stevens Institute of Technology,
National Cheng-Kung University, and the Lenox
Institute of Water Technology. Totally, she has been a
government official and an educator in the USA and
Taiwan for over 50 years. Dr. Wang is a licensed
Professional Engineer, and a Diplomate of the American
Academy of Environmental Engineers (AAEE). Her
publications have been in the areas of water quality,
modeling, environmental sustainability, solid and haz-
ardous waste management, NPDES, flotation technol-
ogy, industrial waste treatment, and analytical methods.
Dr. Wang is the author of over 50 publications and an
inventor of 14 US and foreign patents. She holds a
BSCE degree from National Cheng-Kung University,
Taiwan, ROC; an MS degree from the University of
Rhode Island, RI, USA; and a PhD degree from Rutgers
University, NJ, USA. She is the co-series editor of the
Handbook of Environmental Engineering series
(Springer Nature Switzerland), Coeditor of the
Advances in Industrial and Hazardous Wastes
Treatment series (CRC Press of Taylor & Francis
Group) and the Coeditor of the Environmental Science,
Technology, Engineering and Mathematics series
(Lenox Institute Press). She is a member of AWWA,
NYWWA, NEWWA, WEF, NEWEA, CIE, and
OCEESA.
About the Editors xiii

Yung-Tse Hung has been Professor of Civil

Engineering at Cleveland State University, Cleveland,
Ohio, USA, since 1981. He is a Fellow of the American
Society of Civil Engineers, a licensed Professional
Engineer in Ohio and North Dakota, and a Diplomate
of American Academy of Environmental Engineers. He
has taught at 16 universities in 8 countries. His research
interests and publications have been involved with bio-
logical treatment processes, solid wastes, hazardous
waste management, and industrial waste treatment. Dr.
Hung is credited with over 470 publications and pre-
sentations, 28 books, 159 book chapters, in water and
wastewater treatment. He received his BSCE and
MSCE degrees from National Cheng-Kung University,
Taiwan, ROC, and his PhD degree from the University
of Texas at Austin, USA. He is the Editor-in-Chief of
International Journal of Environment and Waste
Management, International Journal of Environmental
Engineering, and International Journal of
Environmental Engineering Science, and Coeditor of
the Advances in Industrial and Hazardous Wastes
Treatment series (CRC Press of Taylor & Francis
Group), and the Handbook of Environmental
Engineering series (Springer). Dr. Hung is also the
Chief Editor of the Handbook of Environment and
Waste Management series (World Scientific Singapore)
and the Permanent Executive Director and Ex-President
of OCEESA (Overseas Chinese Environmental
Engineers and Scientists Association).
Contributors

T. F. Adepoju Chemical/Petrochemical Engineering Department, Akwa-Ibom

State University, Ikot Akpaden, Nigeria
Ayla Arslan Environmental Engineering Department, Kocaeli University,
Kocaeli, Turkey
Hamidi Abdul Aziz School of Civil Engineering and Solid Waste Management
Cluster, Universiti Sains Malaysia, Pulau Pinang, Malaysia
Erick Butler School of Engineering and Computer Science, West Texas A&M
University, Canyon, TX, USA
Shivani Chaturvedi Enzyme and Microbial Biochemistry Laboratory, Department
of Chemistry, Indian institute of Technology Delhi, New Delhi, India
Maddison Cole Department of Civil and Environmental Engineering, Cleveland
State University, Cleveland, OH, USA
Levent Dağaşan SVL Food Biotechnology Consultancy, Istanbul, Turkey
Donald L. Day Agricultural Engineering Department, University of Illinois,
Urbana-Champaign, IL, USA
Esra Can Doğan Environmental Engineering Department, Kocaeli University,
Kocaeli, Turkey
Nevim Genç Environmental Engineering Department, Kocaeli University,
Kocaeli, Turkey
Christopher R. Huhnke Department of Civil and Environmental Engineering,
Cleveland State University, Cleveland, OH, USA
Yung-Tse Hung Department of Civil and Environmental Engineering, Cleveland
State University, Cleveland, OH, USA
Volodymyr Ivanov School of Civil and Environmental Engineering, Nanyang
Technological University, Singapore

xv
xvi Contributors

Kathleen Hung Li NEC Business Network Solutions, Inc., Irving, TX, USA
Arthur J. Muehling Agricultural Engineering Department, University of Illinois,
Urbana-Champaign, IL, USA
Ashween Deepak Nannaware Phytochemistry Division, CSIR-Central Institute
of Medicinal and Aromatic Plants, Lucknow, Uttar Pradesh, India
Puganeshwary Palaniandy School of Civil Engineering, Universiti Sains
Malaysia, Pulau Pinang, Malaysia
Howard H. Paul Department of Information Systems, Cleveland State University,
Cleveland, OH, USA
Prashant Kumar Rout Phytochemistry Division, CSIR-Central Institute of
Medicinal and Aromatic Plants, Lucknow, Uttar Pradesh, India
Anatoliy Salyuk Department of Biochemistry and Ecological Control, National
University of Food Technologies, Kiev, Ukraine
Nazih K. Shammas Lenox Institute of Water Technology, Latham, NY, USA
Suman Singh Phytochemistry Division, CSIR-Central Institute of Medicinal and
Aromatic Plants, Lucknow, Uttar Pradesh, India
Viktor Stabnikov Department of Microbiology and Biotechnology, National
University of Food Technologies, Kiev, Ukraine
Naziya Syed Phytochemistry Division, CSIR-Central Institute of Medicinal and
Aromatic Plants, Lucknow, Uttar Pradesh, India
Robert Lee-Kong Tiong School of Civil and Environmental Engineering,
Nanyang Technological University, Singapore, Singapore
Dale H. Vanderholm Institute of Agricultural and Natural Resources, University
of Nebraska, Castle Rock, CO, USA
Lawrence K. Wang Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-­
Champaign, IL, USA
Mu-Hao Sung Wang Lenox Institute of Water Technology, Latham, NY, USA
Nai-Yi Wang Chemistry Department, University of Buffalo, Buffalo, NY, USA
Ping Wang Civil and Environmental Engineering Department, Rensselaer
Polytechnic Institute, Troy, NY, USA
Chaya Wilks Department of Civil and Environmental Engineering, Cleveland
State University, Cleveland, OH, USA
Contributors xvii

Josephine O. Wong City of San Francisco, San Francisco, CA, USA

Haneen Yehya Department of Civil and Environmental Engineering, Cleveland
State University, Cleveland, OH, USA
Mohd Suffian Yusoff School of Civil Engineering, Universiti Sains Malaysia,
Pulau Pinang, Malaysia
Chapter 1
Management and Treatment of Livestock
Wastes

Dale H. Vanderholm, Donald L. Day, Arthur J. Muehling,

Lawrence K. Wang, Yung-Tse Hung, Erick Butler, Mu-Hao Sung Wang,
and Haneen Yehya

Nomenclature

AU Number of 1000 lb animal units per animal type

BOD5 Five-day biochemical oxygen demand
BUW Bedding unit weight, lb/ft3
Ca+2 Calcium cation
C Targeted rate concentration
C* Background rate concentration
Co Initial concentration of conditions

D. H. Vanderholm
Agricultural Research Division, Institute of Agricultural and Natural Resources, University of
Nebraska, Castle Rock, CO, USA
e-mail: [email protected]
D. L. Day · A. J. Muehling
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
L. K. Wang (*)
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]
Y.-T. Hung · H. Yehya
Department of Civil and Environmental Engineering, Cleveland State University,
Cleveland, OH, USA
e-mail: [email protected], [email protected]
E. Butler
School of Engineering and Computer Science, West Texas A & M University,
Canyon, TX, USA
e-mail: [email protected]
M.-H. S. Wang
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 1

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_1
2 D. H. Vanderholm et al.

COD Chemical oxygen demand

CH3COOH Acetic acid
CO Carbon monoxide
CO2 Carbon dioxide
D Number of days in storage period
DS Dissolved solids
DVM Daily volume of manure production for animal type, ft3/AU/day
FR Volumetric void ratio
FS Fixed solids
H2 Diatomic hydrogen
HLR Hydraulic loading rate
k First-order rate constant (cm/day)
Mg+2 Magnesium cation
MMCTCO2e Million metric tons of CO2 equivalent
N2 Diatomic nitrogen
NH3-N Ammonia-nitrogen
NH4-N Ammonium-nitrogen
NO Nitrous oxide
OLR Organic loading rate
PO4−3 Phosphate ion
q Hydraulic loading rate (cm/day)
SS Suspended solids
TKN Total Kjeldahl nitrogen
TP Total phosphorus
TS Total solids
TBV Total bedding volume stored, ft3
TVM Total volume of stored manure, ft3
TWW Total wastewater stored, ft3
TVS Total volatile solids
VMD Volume of manure production for animal type for storage period, ft3
WB Weight of bedding used for animal type, lb/AU/day
WV Volume of waste stored, ft3

1.1 Introduction

In recent years, livestock waste management has been a rapidly changing technol-
ogy. It is subject to government regulation and sensitive to population growth pat-
terns, community attitudes, and land-use changes. It is influenced by variables such
as soil type, topography, climate, crops, and livestock production practices. The
evolution of larger and more concentrated livestock operations has accentuated the
problems of waste management. Better management methods are necessary not
only to hold down labor requirements and expense but also to minimize detrimental
effects on the environment. Where animals are allowed to roam freely on pastures,
such as is still done in many areas of the state, the manure from the livestock is
1 Management and Treatment of Livestock Wastes 3

deposited directly on the land and recycled with a minimum hazard to the environ-
ment. Even pasture production of livestock, however, requires management to pre-
vent overgrazing, overcrowding, loss of vegetative cover, and the development of
potential nonpoint sources of pollution. The facilities that cause the greatest envi-
ronmental threat, however, are those in which the livestock are confined perma-
nently or frequently on a regular basis. Figure 1.1 provides the consequences of
infiltrated livestock waste.
In general, the regulations do not stipulate how waste must be handled but rather
delineate the unsatisfactory practices and acceptable methods for correcting unsat-
isfactory situations. The decision-making process, when a farmer has to deal with
correcting a problem situation, is essentially left to the farmer as to the selection of
the system or combination of systems to correct the problems.
The frequent use of the term “waste” in this chapter is not intended to imply that
we are dealing with a material of no value. The intent is to convey the understanding
that the material consists of more than just the feces and urine excreted by the ani-
mals, for example, hair, soil, spilled feed, and other materials. In actuality, there is
much that can be recovered and reused from this material for supplying plant

Fig. 1.1 Consequences of infiltrated livestock waste [1]

4 D. H. Vanderholm et al.

fertilizers, livestock feed additives, and conversion to energy. Practical management

practices to realize these and other benefits are encouraged whenever possible.
The manual has components grouped together by function, and systems are com-
posed of components with different functions. For this reason, some skipping
around in the manual will be necessary when using it for planning purposes. The
important thing is to insure that the components selected for the system are compat-
ible and adequate for their purpose as well as to insure that the entire system accom-
plishes its management objective. English units of measurement are used in
examples, although metric units are included in many tables.
Another point to consider in consistent planning is whether the failure of one
component will result in the failure of the entire system or if adequate flexibility is
provided to permit continued operation without disastrous effects when unforeseen
events happen. Often simple emergency or contingency measures can be planned
into a system at various points, thereby preventing difficult situations later.
Data presented on waste production and characteristics are values generated
from different parts of the United States, making it nearly impossible to define con-
sistent values. Where specific values for an individual system can be obtained, these
should be used in preference to the manual values. The values found in this chapter
are deemed to provide perspective on what occurs in livestock operations across the
country.
Selecting a system and the individual components involved is a process that
includes engineering, economics, regulatory considerations, personal preferences,
and other factors. There is no single system which is best. Each component, facility,
or process has advantages and disadvantages. Each of these factors mentioned in the
previous sentence needs to be given consideration in order to develop the most suit-
able waste management system for a given situation.
The information provided in this chapter is intended to create a frame for plan-
ning and sizing waste management system components. If systems require further
explanation, the reader should consult the resources for further direction on deter-
mining what constituents are necessary to create a more adequate design. It may
also be necessary to obtain professional design assistance.

1.1.1 Federal Regulations

Federal regulations have been mandated by the US Environmental Protection

Agency (USEPA) since its establishment in 1970. For the purpose of livestock
waste treatment, legislation is applicable for both air and water. Air pollution
research began in 1955 prior to the formation of the USEPA when the Air Pollution
Act was passed to support funding and research. In 1970, the Clean Air Act required
air quality standards for existing facilities and the refusal of building new infrastruc-
ture if not compliance with current legislation [2]. In addition, legislation has the
USEPA control air emissions from mobile and stationary sources and establishes
the National Ambient Air Quality Standards (NAAQs). NAAQs regulate hazardous
1 Management and Treatment of Livestock Wastes 5

air pollutants for the purpose of protecting the public health and environment and
are incorporated with State Implementation Plans [3].
Nevertheless, agriculture persists with odor problems, and further mandates
were added later through the Clean Air Act Amendments of 1990. In the amend-
ments, the legislation headed by the USEPA and Secrecies of Agriculture and
Energy required reduction emissions that produce acid rain and for the protection of
ozone, ammonia volatilization from animal and other agricultural operations for
water and soil acidification, and methane emissions from rice and livestock produc-
tion for ozone depletion [2]. Figure 1.2 provides the various methods in which air
pollution can be caused by the livestock industry.
Water legislation began as early as 1886 with the River and Harbors Act of 1886
and 1889. Following the induction of the USEPA, the passing of the Federal
Pollution Control Act of 1972 placed federal government responsible for creating
and enforcing standards for water pollution control and maintaining the integrity of
the water supplies, where a goal of having 0% discharge by 1985 was set. However,
the biggest impact to water treatment in livestock wastes was the Clean Water Act
of 1977. The Clean Water Act of 1977 introduces stringent legislation on feedlots
and also required National Pollution Discharge Elimination System (NPDES) per-
mits [2].
The National Pollution Discharge Elimination System (NDPES) regulates the
quantity of waste entering navigable waters and also point sources [5]. In regard to

Fig. 1.2 Pathways for manure contaminants in the air [4]

6 D. H. Vanderholm et al.

livestock wastes, the NPDES require permits when discharging in the following
conditions [2]:
1. Feeding operations consisting of 1000 animals confined for a time greater than
45 days per year and pollution less than 25 year, 24 storm events
2. Feeding operations with 300 animals discharge through a manmade device into
navigable waters either from a feed lot of a manmade device
3. Hatcheries and fish farm cold-water ponds that have a total of 20,000 lbs animal
production with 5000 lbs of food discharging 30 days per year, or warm-water
ponds discharging 30 days per year
There have been several revisions made to NPDES permit involving concentrated
animal feeding operations (CAFOs) or feedlots. The 2003 revision makes permits
necessary for both open lots and CAFOs, refines the definition of CAFO require-
ments, and incorporates a nutrition management plan that considers faculty and land
application issues where the lack of compliance can require CAFOs to point source.
Proposed revisions have been suggested in 2008 and 2011 from outcomes of law-
suits submitted by both the industry and environmental interest groups. For exam-
ple, in 2011, a proposal was made where it would have been required for a CAFO
or its affiliated state to release information. The proposal was not mandated as the
USEPA decided to make additional measures to ascertain existing techniques to
collect necessary information [6, 7].

1.1.2 State Regulations

Regulations imposed by the state will vary. There are many resources available to
the user to determine which regulations are appropriate for a given state. An inves-
tigation of specific state investigation will be up to the user. A list of each state’s
environmental agency with associated links is in Table 1.1.

1.2 Wastewater Characteristics

1.2.1 General Characteristics of Wastewater

1.2.1.1 Terminology

Prior to evaluating the properties of wastewater, it is important to understand the

general terminology related to quantifying the characteristics of wastewater. Overall,
wastes can be evaluated based on their physical and chemical properties. Tables 1.2
and 1.3 summarize the physical and chemical properties along with characteristics
from excreted beef. The most important physical properties within waste include
the weight, volume, and moisture content. These properties quantify the amount of
 1

Table 1.1 List of state environmental agencies with associated links

State State agency Website
Alabama Alabama Department of Environmental Management http://www.adem.state.al.us/default.cnt
Alaska Alaska Department of Environmental Conservation http://dec.alaska.gov/
Arizona Arizona Department of Environmental Quality http://www.azdeq.gov/
Arkansas Arkansas Department of Environmental Quality http://www.adeq.state.ar.us/
California California Environmental Protection Agency http://www.calepa.ca.gov/
Colorado Colorado Department of Public Health and Environment https://www.colorado.gov/cdphe/
Connecticut Connecticut Department of Energy and Environmental Protection http://www.ct.gov/deep/site/default.asp
Delaware Delaware Department of Natural Resources and Environmental http://www.dnrec.delaware.gov/Pages/Portal.aspx
Control
Florida Florida Department of Environmental Protection http://www.dep.state.fl.us/
Georgia Georgia Environmental Protection Division http://epd.georgia.gov/
Hawaii Hawaii Office of Environmental Quality Control http://health.hawaii.gov/oeqc/
Idaho Idaho Department of Environmental Quality http://www.deq.idaho.gov/
Management and Treatment of Livestock Wastes

Illinois Illinois Environmental Protection Agency http://www.epa.illinois.gov/index

Indiana Indiana Department of Environmental Management https://secure.in.gov/idem/index.htm
Iowa Iowa Department of Natural Resources http://www.iowadnr.gov/Environment.aspx
Kansas Kansas Department of Health and Environment: Division of http://www.kdheks.gov/environment/
Environment
Kentucky Kentucky Department for Environmental Protection http://dep.ky.gov/Pages/default.aspx
Louisiana Louisiana Department of Environmental Quality http://www.deq.louisiana.gov/portal/
Maine Maine Department of Environmental Protection http://www.maine.gov/dep/
Maryland Maryland Department of the Environment http://www.mde.state.md.us/Pages/Home.aspx
Massachusetts Massachusetts Department of Environmental Protection http://www.mass.gov/eea/agencies/massdep/
Michigan Michigan Department of Environmental Quality http://www.michigan.gov/deq
Montana Montana Department of Environmental Quality http://www.deq.mt.gov/default.mcpx
(continued)
7
 8

Table 1.1 (continued)

State State agency Website
Minnesota Minnesota Pollution Control Agency http://www.pca.state.mn.us/
Mississippi Mississippi Department of Environmental Quality http://www.deq.state.ms.us/
Missouri Missouri Department of Environmental Quality http://dnr.mo.gov/env/index.html
Nebraska Nebraska Department of Environmental Quality http://www.deq.state.ne.us/
Nevada Nevada Division of Environmental Protection http://ndep.nv.gov/
New New Hampshire Department of Environmental Services http://des.nh.gov/index.htm
Hampshire
New Mexico New Mexico Environmental Department http://www.nmenv.state.nm.us/
New York New York Department of Environmental Conservation http://www.dec.ny.gov/
North Carolina North Carolina Department of Environment and Natural Resources http://www.ncdenr.gov/web/guest
Noth Dakota North Dakota Environmental Health http://www.ndhealth.gov/EHS/
Ohio Ohio Environmental Protection Agency http://www.epa.state.oh.us/
Oklahoma Oklahoma Department of Environmental Quality http://www.deq.state.ok.us/
Oregon Oregon Department of Environmental Quality http://www.oregon.gov/deq/pages/index.aspx
Pennsylvania Pennsylvania Department of Environmental Protection http://
www.depweb.state.pa.us/portal/server.pt/community/dep_home/5968
Rhode Island Rhose Island Department of Environmental Management http://www.dem.ri.gov/
South Carolina South Carolina Department of Health and Environmental Control http://www.scdhec.gov/HomeAndEnvironment/
South Dakota South Dakota Department of Environment and Natural Resources http://denr.sd.gov/
Tennessee Tennessee Department of Environment and Conservation http://www.state.tn.us/environment/
Texas Texas Commission of Environmental Quality http://www.tceq.state.tx.us/
Utah Utah Department of Environmental Quality http://deq.utah.gov/
Vermont Vermont Department of Environmental Conservation http://www.anr.state.vt.us/dec/dec.htm
Virginia Virginia Department of Environmental Quality http://deq.state.va.us/
D. H. Vanderholm et al.
 1

State State agency Website

Washington Washington Department of Ecology http://www.ecy.wa.gov/
West Virginia West Virginia Department of Environmental Protection http://www.dep.wv.gov/Pages/default.aspx
Wisconsin Wisconsin Department of Natural Resources http://dnr.wi.gov/
Wyoming Wyoming Department of Environmental Quality http://deq.wyoming.gov/
Management and Treatment of Livestock Wastes
9
10 D. H. Vanderholm et al.

Table 1.2 Physical and chemical properties of waste [2]

Physical properties
Moisture content Component of a waste that can be removed by evaporation and
drying
Total solids Component of a waste that is left after evaporation
Volatile solids Component of a waste that has been removed when a waste
sample is placed in a muffle furnace at 1112 °F
Fixed solids Component of a waste that remains after a waste sample is heated
in a muffle furnace at 1111 °F
Suspended solids Component of a waste removed by means of filtration
Chemical properties
Five-day biological oxygen Water quality index that measures the amount of oxygen needed
demand (BOD5) for microorganisms to degrade material
Chemical oxygen demand Water quality index that determines the amount of oxygen
(COD) consumed by organic material

Table 1.3 Excreted beef waste characteristics [8]

Components Units Beef cow in confinement Growing calf confined (450–750 lb)
Weight lb/da-a 125 50
Volume ft3/d-a 2.0 0.8
Moisture %wet basis 88 88
TS lb/d-a 15 6.0
VS lb/d-a 13 5.0
BOD lb/d-a 3.0 1.1
N lb/d-a 0.42 0.29
P lb/d-a 0.097 0.055
K lb/d-a 0.30 0.19

waste that must be handled and subsequently treated. Secondary physical properties
evaluate categories that are found within a given waste. These secondary properties
include total solids (TS), volatile solids (VS), fixed solids (FS), dissolved solids
(DS), and suspended solids (SS) [2].
On the other hand, chemical properties are represented as nutrients or wastewater
quality indices. Nitrogen (N), phosphorus (P), and potassium are the elements
mainly considered as nutrients. These nutrients are further subdivided into subse-
quent forms that can be beneficial or detrimental to the handling of livestock.
Figures 1.3 and 1.4 summarize nitrogen and phosphorus processes that occur within
livestock waste. Five-day biochemical oxygen demand (BOD5) and chemical oxy-
gen demand (COD) are two of the many wastewater quality indices. These indices
are evaluated within a laboratory and are important in determining the nature of the
wastewater present. BOD5 relates the amount of oxygen required to degrade waste
1 Management and Treatment of Livestock Wastes 11

Fig. 1.3 Nitrogen processes involved in manure management (from top to bottom: mineralization,
nitrification, denitrification (bottom left), volatilization (bottom right)). (Adapted from [4])

by microorganisms in 5 days at 20 °C, while COD involves the consumption of

oxygen by organic and inorganic constituents [2].

1.2.1.2 Wastewater Characteristics

It can be said that the type of manure in wastewater produced varies not only on
characteristics but also on the time of year. Based on the data collected between
summer and winter for cattle manure and bedding, Loehr (1974) found that the
ranges for parameters are different between summer and winter. For example, per-
cent total solids (%TS) in winter have an average of 2.8% versus 2.3% in summer.
In regard to biochemical and chemical oxygen demands (BOD5 and COD), winter
indicates higher values of BOD at 13,800 mg/L versus only 10,300 mg/L in sum-
mer. Nutrient presence is higher at 2350 mg/L as N for total nitrogen in summer, as
compared with 1800 mg/L ion summer, and total phosphorus is 280 mg/L in winter,
while only 190 mg/L in summer. These results can be reflected based on conditions
such as precipitation and temperature [9].
In addition, having considered swine lagoon analysis in Missouri, liquid wastes
are significantly higher in total solids, total nitrogen and ammonia, salts, and miner-
als as compared to sludge. In particular, liquid wastes contained 3091 mg/L, as
12 D. H. Vanderholm et al.

Fig. 1.4 Phosphorus cycle in relation to waste application and transformation of phosphorus in the
soil profile [1]

compared to only 203.843 mg/L in solids. This trend is also noticed in terms of salts
(Na 470 mg/L, Ca 257 mg/L, and Mg 64 mg/L versus 4.627 mg/L, 6.176 mg/L, and
1.514 mg/L in liquid, respectively) [10].
Also, the waste characteristics of different industries vary. The supernatant for
different animal wastes sampled from a lagoon and municipal waste treatment was
compared. Poultry lagoons contained the highest concentration of wastes. The mean
COD for poultry was 3700 mg/L, compared with 2050 mg/L and 1672 for the swine
and dairy lagoons, respectively. This trend can be highly seen in BOD5, TS, total
volatile solids (TVS), suspended solids (SS), and ammonia nitrogen (NH3-N),
where the poultry lagoon contained the highest amounts of all three. Nevertheless,
untreated municipal wastewater has significantly lower values for every category; in
some cases such as COD values, the lowest animal waste value (1672 mg/L for
dairy lagoons) was four times the COD than in municipal waste and almost ten
times less than the highest (poultry) [11]. Tables 1.4 and 1.5 present characteristics
of manure based on various livestock types. Table 1.6 presents wastewater charac-
teristics of swine waste.
On the other hand, while waste constituents were higher in the animal waste, the
untreated municipal wastewater contained higher amounts of trace metals, specifi-
cally cadmium, chromium, copper, and lead. In fact, examining copper, the range
for copper was between 190 and 440 mg/L for poultry lagoons; however, in untreated
1 Management and Treatment of Livestock Wastes 13

Table 1.4 Total manure, nitrogen, phosphorus, and potassium excreted by different livestock
species [12]
Fresh manure N (lb/ P2O5 (lb/ P (lb/ K2O (lb/ K (lb/
Livestock type (gal/day) day) day) day) day) day)
Beef cattle (1000 lb 7.5 0.34 0.25 0.11 0.29 0.24
body weight)
Dairy cow (1000 lb 11 0.41 0.17 0.074 0.32 0.27
body weight)
Swine (100 lb body 1 0.045 0.034 0.015 0.036 0.030
weight)
Poultry (4 lb body 0.028 0.0029 0.0026 0.0011 0.0015 0.0012
weight)
Note: Livestock type is based on 1000 lb body weight

Table 1.5 Manure characteristics per animal [13]

Average Total manure Total solids Volatile solids
Animal type weight (lb) production (ft3/day) production (lb/day) production (lb/day)
Swine
 Nursery 35 0.04 0.39 0.30
pig
 Growing 65 0.07 0.72 0.55
pig
 Finishing 150 0.16 1.65 1.28
pig
 Gestation 275 0.15 0.82 0.66
sow
 Sow and 375 0.36 2.05 1.64
litter
 Boar 350 0.19 1.04 0.84
Cattle
 Dairy 1000 1.39 12.00 10.00
 Beef 1000 0.95 8.50 7.20
Poultry
 Layers 4 0.0035 0.064 0.048
 Broilers 2 0.0022 0.044 0.034

municipal wastewater, it was found that the range of copper was between 20 and
3360 mg/L, almost four times as much for the averages of these ranges. With the
exception of arsenic and cadmium, poultry lagoons consistently had higher amounts
of trace elements [11].
14 D. H. Vanderholm et al.

Table 1.6 Swine waste characteristics [2]

Sow Nursing/
Grower Replacement nursery pig
Component Units 40–220 lb gilt Gestation Lactation Boar 0–40 lb
Weight lb/d/1000# 63.40 32.80 27.20 60.00 20.50 106.00
Volume ft3/d/1000# 1.00 0.53 0.44 0.96 0.33 1.70
Moisture % 90.00 90.00 90.80 90. 00 90.70 90.00
TS % w.b. 10.00 10.00 9.20 10. 00 9.30 10.00
lb/d/1000# 6.34 3.28 2.50 6.00 1.90 10.60
VS " 5.40 2.92 2.13 5.40 1.70 8.80
FS " 0.94 0.36 0.37 0.60 0.30 1.80
COD " 6.06 3.12 2.37 5.73 1.37 9.80
BOD5 " 2.08 1.08 0.83 2.00 0.65 3.40
N " 0.42 0.24 0.19 0.47 0.15 0.60
P " 0.16 0.08 0.06 0.15 0.05 0.25
K " 0.22 0.13 0.12 0.30 0.10 0.35
TDS 1.29
C:N ratio 7 7 6 6 6 8
Average daily production for weight range noted. Increase solids and nutrients by 4% for each 1%
feed waste more than 5%

1.2.2 Milk House Wastewater Characteristics

Milk house wastewater is generated from various sources within the dairy industry.
These sources include but are not limited to [14]:
1. Wash water from cleaning bulk tanks
2. Cleaning of milk pipelines
3. Cleaning of milking units
4. Cleaning equipment
5. Cleaning of milk house floor
6. Remnant within the milk pipelines, receiver, and bulk tanks
7. Chemicals
8. Water softener recharge
9. Manure
10. Bedding
11. Floor dirt and grit
12. Washing the udders of the cows
Typical milk house and diary wastewater characteristics are listed in Tables 1.7
and 1.8.
The Wisconsin National Resource Conservation Service (NRCS) describes three
constituents within milk house wastewater—solids, phosphorus, and ammonia
nitrogen and chlorides. Solids contain manure, primarily made of lignin and cellu-
lose. These are a major producer of milk house wastewater. Solids usually have a
1 Management and Treatment of Livestock Wastes 15

Table 1.7 Characteristics of milk house wastewater [14]

Parameter Final effluent tank (mg/L) Design (mg/L)

BOD5 500–2600 1200
Total Solids (TS) 200–1000 450
Fats, Oils, Grease 90–500 225
30–100 65
Total Phosphorus 21–100 55
pH 6.2–8.0 7.5
Temperature 53–70 °C –

Table 1.8 Dairy waste characterization; milking center [15, 16]

Milk
house Milk house Milk house, parlor, Milk house, parlor,
Component Units only and parlor and holding area and holding area
Volume ft3/day/1000 0.22 0.60 1.40 1.60
head
Water gal/ 2.3 6.3 14.7 16.8
volume day/1400 lb
cow
Moisture % 99.72 99.40 99.70 98.50
COD lb/1000 gal 25.30 41.70 – –
BOD5 lb/1000 gal – 8.37 – –
N lb/1000 gal
P lb/1000 gal 0.58 0.83 0.23 0.83
K lb/1000 gal 1.50 2.50 0.57 3.33

concentration range between 1600 and 7000 mg/L. Depending on the source, some
solids can be comprised of high-concentration BOD. For example, it has been deter-
mined that raw waste milk can have a BOD concentration of 100,000 mg/L [15].
The presence of phosphorus has been attributed to daily cleaning operations such
as pipeline washing or the presence of cleaning chemicals such as detergents and
acid rinses, many of which can have 3.1–10.6% phosphorus by weight. Phosphorus
in milking house centers is usually soluble and can cause eutrophication [15].
Ammonia is found in manure, urine, and decomposed milk. The discharge of
milk house wastewater with substantial concentrations of ammonia can be toxic to
fish. On the other hand, chlorides are also found in urine, milking system cleaners
and sanitation, and water softening generation. The presence of chlorides can have
an impact on the salinity of the wastewater being treated [15].
The daily operations within a milk house require daily cleaning of equipment
and pipelines. The University of Minnesota Extension describes a four-stage clean-
ing process. Cleaning begins with rinsing the transfer lines to remove any raw milk
16 D. H. Vanderholm et al.

that may remain. Next, organic material is removed by a detergent with an active
chlorine concentration of 100 mg/L. This detergent raises the pH above 11. Then, an
acid rise is completed to reduce inorganic material. The pH is lowered to around 3.5
to prevent bacteria formation and neutralize any detergent residue that may remain.
Finally, chlorine with a concentration of 200 mg/L is added to kill microorganisms
in the line. The process of cleaning equipment and pipelines accounts for an addi-
tional source of wastewater that needs to be treated prior to any discharge [14].

1.2.2.1 Treatment of Milk House Wastewater

There are several treatment methods for milk house wastewater. Table 1.9 lists sev-
eral treatment methods that are being used in the state of Minnesota. For example,
a viable option of treating milk house wastewater is through a two-stage septic
system. It is important to note that wastewater entering into the tank does not include
waste milk from cows. Waste milk will be disposed with manure. Treatment by the
septic system is contingent on the strength of the wastewater, leaving the parlor and
also time spent in the septic tanks [17, 18].
Wastewater is pretreated using two septic tanks consisting of inlet and outlet
baffles. The tanks remove settable solids, fats, and grease and inhibit contamination
throughout the remaining sections of the treatment plant. In the state of Minnesota,
tank sizing is based on either a hydraulic retention time of 3 days or a volume of
1000 gallons, whichever is greater. In addition, Minnesota requires 4 ft of soil cover.
Prior to exiting the septic tank, the wastewater passes through an effluent filter. The
effluent filter prevents suspended solids from leaving the septic tank [17, 18].
Next, wastewater moves through a bark bed. The bark bed combines soil with
bark and shredded wood. The depth of the bark bed is between 18 and 24 inches.
The purpose of the mixture is to prevent the soil in colder climates and allows for
more oxygen transfer, which in turn increases the rate of degradation at the soil-­
effluent interface. The sizing and application within the bark bed is determined by
the soil type. Typical bark beds consist of a depth of 2 ft of soil to the bedrock or
groundwater. Sizing of the bed is computed by taking the loading rate of the soil
(contingent on soil type) and dividing it by the total wastewater volume. The load-
ing rate is read from a table based on soil type. Presented values consider a BOD5
concentration of 750 mg/L, flow rate of 5 gallons per day, and a BOD5 loading rate
of 0.0062 lbs/gallon. Bark beds can also be sized using hydraulic loading as
well [19].
Another treatment method that can be employed is the use of constructed wet-
lands. Because constructed wetlands are not unique to milk house waste treatment,
they will be discussed in Sect. 1.3.
Nevertheless, literature has discussed the efficiency of constructed wetlands for
treating dairy wastewater. A three-celled surface wetland was used to treat dairy
wastewater. The study compared the performance of the summer and winter sea-
sons. The results found that total suspended solids (TSS), total phosphorus (TP),
and total Kjeldahl nitrogen (TKN) were reduced in the summer as compared to the
1 Management and Treatment of Livestock Wastes 17

Table 1.9 Treatment methods for milk house wastewater treatment [17, 18]
Treatment
method Description Requirements
Chemical Coagulation and Effluent BOD ≤205 mg/L
batch reactor flocculation Discharge into infiltration/filtration system
Bark bed Soil infiltration with Requirement of soil texture to a minimum of 3 ft
18–24 inches of barkwood bedrock. Treatment consists of three processes:
Pressure distribution  1. Primary treatment is completed by two septic
system disperses effluent tanks. Tanks are designated based on an HRT
of 3 days or the volume whichever is greater
 2. Infiltration area
 3. Distribution system: The system consists of a
pump, transferring pipe. Effluent traveling to
the pipe must have a minimum velocity of
2 ft/s. The transferring pipe must have a
diameter of 2 inches with a drainage slope of
1%. Distribution is done through gravel bed
or a chamber system
Aeration and Aerobic treatment or Treatment will consist of three processes:
media recirculating media filter  1. Primary treatment will use two septic tanks.
filtration Design requirements similar to bark bed
primary treatment
 2. Aerobic treatment follows primary treatment
where the goal must be less than 200 mg/L
effluent BOD
 3. Following aerobic treatment the discharge
will enter an infiltration/filtration system
Irrigation Treatment consists of  1. A proper site for irrigation consists of a
water filled within the tank location where 20% of materials from 2 ft
that will be dispersed onto below the buffer zone pass through a #200
crops sieve
 2. The irrigation area must have a minimum 3%
slope, where the down gradient should be
50 ft away karst, surface water, or any private
wells
 3. Treatment consists of using a septic tank.
Design requirements are similar to bark bed
primary treatment
 4. Wastewater moves to a 3-day holding dosing
tank with piping for distribution and pumping
Vegetated Wastewater from a septic  1. Both siting and primary treatment use similar
treatment tank is distributed onto design criteria as previously mentioned
dosing vegetation by a sloping  2. Treated waste from a septic system will travel
system elevated pipe where the through a distribution system to a dosing tank
upslope side of the pipe is by a perforated pipe with perforations
enclosed between 1/2 and 1 inch diameter. The pipe is
elevated 1–1.5 ft above the ground
 3. Determination of vegetated area is based on
either a flow depth no greater than 0.5 ft using
a treatment time of 15 min and a Manning
constant of 0.24, or the smallest area that can
handle a design loading rate no greater than
0.9 inches/week
18 D. H. Vanderholm et al.

winter. In addition, BOD5 removal was lower than 30 mg/L during the summer
months as compared to the winter months. Finally, fecal coliform removal was
approximately 31% [20].
To avoid eutrophication in a local surface water body, a three-celled parallel free
water surface wetland was used to treat dairy wastewater. The treatment process
began with the concrete settling pad for the purpose of eliminating solids prior to
entry into the wetland. Following treatment into the constructed wetland, a three-­
sump pump transfers the wastewater into a holding pond. The authors concluded
that BOD5, conductivity, total dissolved solids (TDS), TSS, TKN, TP, phosphate,
ammonia, nitrate, nitrite, and fecal coliform bacteria were generally reduced by the
wetland. In addition, all parameters with the exception of nitrate and nitrate were
diminished from the settling pad to the holding pond. Fecal coliform was reduced
provided that cows were kept from grazing in the constructed wetlands [21].

1.2.2.2 Conservation

Along with dairy wastewater treatment, water conservation is another important

facet to properly handle wastewater. Water conservation is important because it pro-
vides the dairy plant owners an opportunity to reduce the cost for treatment. In
general, wastewaters with high BOD5 concentration discharged into a municipal
wastewater treatment system incur high costs. It can also become expensive for
onsite treatment as well; therefore, water conservation efforts provide owners an
opportunity to save funds. In addition, methods have a positive impact on areas
where water resources are currently being depleted and can also reduce the potential
of stringent legislation. In the dairy industry, water reuse can reduce freshwater
demand to 1 gal of water/1 gal of milk produced if proper management of goals is
provided and maintenance is regularly scheduled [22].

1.3 Waste Treatment

1.3.1 Anaerobic Digestion

Anaerobic digestion is the fermentation of organic waste by hydrolytic microorgan-

isms into fatty acid chains, carbon dioxide (CO2), and hydrogen (H2). Short fatty
acids are then converted into acetic acid (CH3COOH), H2, CO2, and microorgan-
isms. Acetic acid forms biogas, a combination of methane (CH4), CO2, and trace
elements by means of methanogenic bacteria. Occasionally, biogas can form hydro-
gen sulfide by sulfate-reducing bacteria. In general, CH4 in biogas produces between
55 and 80%, while approximately 65% is found in animal manure [23].
The processes in anaerobic digestion are driven by temperature, moisture, and
solid content. There are three major temperature ranges defined—psychrophilic
1 Management and Treatment of Livestock Wastes 19

(<20 °C), mesophilic (35–40 °C), and thermophilic (51–57 °C). Ideally an anaero-
bic digester should operate at temperatures greater than 35 °C. A moisture content
of 60–99% is ideal, while solid content in the digester should be less than 15% [24].
Recently, there has been a big interest in anaerobic digestion for the purpose of
energy conversion [25]. Since 1996, the Environmental Protection Agency has part-
nered with the US Department of Agriculture, the National Resource Conservation
Service (NCRS), and the US Department of Energy to develop a program known as
AgStar, an opportunity for monetary support in projects related to anaerobic diges-
tive systems. In 1998, the program began by promoting seven farm digesters across
the country [26].
There have been reports of profit being made on the energy that has been cap-
tured through the use of livestock manure. These values have greatly depended upon
the monetary cost of electricity. For example, if one were to sell electricity in
Wisconsin and California, a 1000-head dairy farm with manure production would
be worth about $56,000 and $77,500, respectively [25]. Statistically speaking, it
was found that in 2009, approximately 151 biogas systems that have been installed
within the state of Wisconsin produced about 11.6 megawatts of electricity, enough
for use by 10,000 homes. Within January 2007 and June 2008 alone, 150,000 kilo-
watt hours (kWh) of electricity were produced by farms that had 2000-head of ani-
mals and 440,000 kWh of electricity for those between 2000 and 4500 [27].
Figure 1.5 indicates the net value of dollars based on the digester per number of
head of cattle. Figure 1.6 indicates the number of dairies operating at a given carbon
price per operation size.
There are a plethora of reasons why AgStar has become a popular consideration
for the development of biogas. Consider that the state of Wisconsin has spent
between $16 and 18 billion each year for coal energy imports whereas about $853
million for transportation [27]. If the state of Wisconsin, rich in manure and crop
remains and waste components from the dairy processing, fats, and greases can
transport this material into fuels, it would create an infrastructure that would be
safer and easier to be controlled as compared to the current energy options on the
market today and additional revenue for farmers [27].
A recent 2013 study conducted by the US Environmental Protection Agency
(USEPA) evaluating the AgStar program found that anaerobic digesters reduce
greenhouse gases by 1.73 million metric tons of CO2 equivalent (MMCTCO2e). This
is because methane is captured and burned before entering into the atmosphere. On
the other hand, anaerobic digesters produced 840 million kWh in 2013. These ben-
efits were contingent on the type of anaerobic digester applied. For example, the
most commonly used digesters in the United States were complete mixed and mixed
plug flow [28]. Biogas production is also dependent upon the type of livestock.
Table 1.10 provides information concerning the daily production of biogas per ani-
mal type.
20 D. H. Vanderholm et al.

Fig. 1.5 Net value in dollars of digesters per head vs. number of head [25]

Fig. 1.6 Number of dairies operating at a given carbon price vs. operation size [25]
1 Management and Treatment of Livestock Wastes 21

Table 1.10 Biogas production by animal [23]

Animal type Average weight (kg) Biogas/animal/d (m3)

Dairy 625 1.3
Beef 447 0.32
Swine 70 0.14
Poultry 1.2 0.0092

Table 1.11 Characteristics of various anaerobic digester types [23]

Anaerobic digestion system OLR COD/m3/kg HRT (d)

Covered anaerobic lagoon 0.05–0.2 60–360
Plug flow digester 1–6 18–20
Mixed 1–10 5–20

Fig. 1.7 Covered lagoon digester [29]

1.3.1.1 Types of Anaerobic Digesters

There are six types of anaerobic digesters—covered anaerobic lagoons, plug flow,
continually stirred tank reactor, fixed film, induced blanket reactor, and anaerobic
sequencing batch reactors. Table 1.11 reports the characteristics of three of the six
anaerobic digesters (covered anaerobic lagoons, plug flow digester, and mixed). The
selection of the appropriate anaerobic digester is determined by appropriate param-
eters such as the geographic location. Covered anaerobic lagoons form biogas from
manure stored in structures and are low cost, simplistic in design, and manageable.
There are two types of covers—full and partial. Production of biogas by a covered
anaerobic lagoon depends on the temperature. Therefore, covered lagoons are more
appropriate in areas of warmer climate. Biogas production in a covered lagoon is
collected in pipes at the top of the digester and then transported by using a low
vacuum. From there, the remaining biogas is then flared. Additional characteristics
of a covered anaerobic lagoon include high total solid (TS) concentration, organic
loading rate (OLR) of 0.2–0.5 kg chemical oxygen demand (COD)/m3 day, and a
22 D. H. Vanderholm et al.

Fig. 1.8 Complete mix digester [29]

Fig. 1.9 Plug flow digester [29]

hydraulic retention time (HRT) of 60–360 days [23]. Figures 1.7 and 1.8 are dia-
grams of a covered lagoon digester and a completely mixed digester.
On the other hand, manure in a plug flow digester enters undigested and leaves
digested. A typical plug flow digester includes concrete and geosynthetic material
for gas collection. Manure enters into the digester and is limited to 11–14% total
solid concentration, 1–6 kg COD/m3 day OLR, and an HRT between 20 and 30 days.
In a continually stirred tank reactor, manure enters into a tank and is mixed to main-
tain a consistent concentration throughout the reactor. Unlike a plug flow digester
which is limited to 6 kg COD/m3 OLR, the maximum allowable organic loading rate
for total solids entering into a continually stirred tank reactor is 10 kg COD/m3 day.
In addition, the hydraulic retention time is shorter than a plug flow reactor ranging
between 5 and 20 days [23]. Figure 1.9 is a diagram of a plug flow digester.
A fixed film digester is an attached growth reactor with fixed film media. When
waste enters into the fixed film digester, anaerobic biomass attaches to the fixed film
media. Typical fixed film digesters have a low HRT between 0.5 and 4 days. Influent
manure in a fixed film digester has an OLR between 5 and 10 kg COD/m3 day with
a solid concentration less than 1% [23].
Finally, an induced blanket reactor forms a sludge blanket by digesting the waste.
Manure moves upward from the bottom of the reactor to the top. Inside the blanket,
1 Management and Treatment of Livestock Wastes 23

manure moves upward contacting with anaerobic biomass to become digested. At

the top of the tank, the biogas is created while the sludge blanket moves back to the
bottom of the reactor. There are two types of blanket reactors—upflow anaerobic
sludge blanket (UASB) digester and induced blanket reactor (IBR). UASB involves
low concentration of solids, while IBR usually handles high solid concentra-
tions [30].
The cost of an anaerobic digester application has been contingent on the type. In
the design and construction of a system, the price involves the initial cost of the
system and its operation and maintenance (O & M). The US Department of
Agriculture (USDA) reported values on 38 different digesters. The overall cost of an
anaerobic digester has been estimated to be between $114,000 and 326,000.
Operation and maintenance (O & M) was found to be contingent on the type of
waste. The O & M for swine waste was 2.3% of the initial cost for the system, while
dairy was 7% [23].
Within the last 5 years, other anaerobic digestion processes have been tested. A
specific type of anaerobic digestion design is known as a temperature-phased anaer-
obic digestion reactor. Temperature-phased anaerobic digestion (TPAD) is a system
that completes treatment in two stages at two temperatures—during the first stage,
the digester operates at a temperature at the highest thermophilic temperatures,
approximately 55 °C while the second stage at the lower ended mesophilic condi-
tions or approximately 35 °C. When using a TPAD for livestock waste, the advan-
tages are significant as the digester is capable of increasing a higher probability of
bioconversion and methane production, with lower hydraulic retention times (HRT)
and also size reduction [31]. Harikishan and Sung (2003) used a TPAD process to
treat livestock wastewater for the purpose of analyzing dairy cattle manure. Having
organic loadings of 1.87–5.82 g VS/L/day, 36–41% of volatile solids were removed,
converting 0.52–0.62 L methane/g VS. In addition, fecal coliform and Salmonella
counts meet USEPA Class A standards [31].
Other authors have researched and found results under different conditions. King
et al. (2011) used a 3-year pilot in-storage psychrophilic anaerobic digester (ISPAD)
to consider swine manure and if it is able to handle psychrophilic conditions and be
able to complete anaerobic digestion and successfully produce methane. Results
based on the microbial community analysis were able to produce methane, provided
that volatile solids (VS) had a rate of 44.6 dm3/kg day at 35°, 9.8 dm3/kg day at 18°,
and 8.5 dm3/kg day at 8° and an organic matter content of 24% [32]. Rao et al.
(2010) used a self-mixed anaerobic digester (SMAD) combined with a multistage
high-rate biomethanation process where the authors were capable of reducing vola-
tile solids (VS) by 58% and producing a methane yield of 0.16 m3/kg, with a loading
rate of 3.5 kg VS/m3 day and a hydraulic retention time (HRT) of 13 days. The
authors considered using the opportunity to reduce the loading rate and the hydrau-
lic retention time and percent treatment [33].
24 D. H. Vanderholm et al.

1.3.2 Constructed Wetlands

1.3.2.1 Description

The purpose of a constructed wetland is to provide a low-maintenance treatment

system that creates a quality effluent for areas that have a high volume of wastewa-
ter. Constructed wetlands house wastewater within wide channels. These channels
also support plant life that grows by using the nutrients from the wastewater. There
are four major processes employed in constructed wetlands—sedimentation, filtra-
tion, plant uptake (oxygen is provided at the plant root for waste decomposition),
and biological decomposition (plants provide adequate binding sites for microor-
ganisms) [15].
The basic idea of a wetland is to maintain moist conditions for pollutants to be
trapped and broken down by the plant that are contained within them. In addition,
constructed wetlands take advantage of combining anaerobic and aerobic condi-
tions that persist through the wetland. The majority of constructed wetland design
consists of using either subsurface flow or surface flow. Surface flow wetlands con-
sist of having a “free water zone” about 30 cm deep on top of a soil layer where the
majority of plant growth would occur. The advantage of designing a wetland by this
manner is that it would place microbial growth in the best advantage to occur in the
areas where the water and its contaminants would be. Subsurface flow wetlands,
also known as “root zone method,” remove the “free water zone” for the purpose of
allowing direct contact between plant material and contaminants present [34]. There
are several design parameters that are necessary for treatment—hydraulic loading
rate, length-to-width ratio, bottom slope, water depth, and vegetation [35].
The water depth of a constructed wetland is usually between 20 and 40 cm deep.
The advantage of using surface constructed wetlands is the biological and physical
methods that are employed within the system. Microbial activity (biological)
degrades much of the organic materials, while colloids are either settled within the
wetland or can become filtered out (physical). Nitrogen is capable of being removed
by means of nitrification (the formation of nitrate from ammonium nitrogen) and
denitrification (the formation of atmospheric nitrogen from nitrates) [2], while
ammonia is volatilized by the use of algal photosynthesis. If any phosphorus is
removed, it is by means of wetland plants eventually by either absorption or precipi-
tation [36].

1.3.2.2 Constructed Wetland Types

Literature recognizes three major types of constructed wetlands—free water surface

(FWS), vegetated submerged or subsurface system, and floating aquatic plant (FAP)
systems [38]. Figures 1.10, 1.11, and 1.12 are drawings of each type of constructed
wetland.
1 Management and Treatment of Livestock Wastes 25

Fig. 1.10 Free water surface (FWS) constructed wetland [37]

Fig. 1.11 Subsurface constructed wetland [37]

Fig. 1.12 Floating aquatic plant (FAP) constructed wetland [37]

26 D. H. Vanderholm et al.

In a free water surface system, the wastewater depth is usually shallow, anywhere
between 6 and 18 inches with a flat-bottom slope. Because of their shallow depths,
FWS wetlands usually degrade wastewater under aerobic conditions. When waste-
water enters an FWS, it moves above the sediment, having direct contact with the
plants at the surface. However, the efficiency of FWS treatment is contingent upon
the presence of microorganism located throughout the surface. Nevertheless, micro-
organisms attach themselves to plant stems and/or litter below the water surface, or
at the soil/plant-root matrix, creating the proper environment for wastewater treat-
ment. Prior to entry of an FWS, a pretreatment system to remove settling and float-
ing solids is recommended or ammonia [38]. FWS-constructed wetlands have been
proven to reduce BOD5 and TSS to 30 mg/L, ammonia, and ammonium-nitrogen to
10 mg/L [39]. In addition, to the effluent quality, FWS wetlands are very common
in livestock operations because they are inexpensive and can be in operation year
round [38].
Under the National Resource Conservation Service guidelines, an FWS is to be
designed based on a 25-year storm event depending on the state. Sometimes a deten-
tion pond downstream may be necessary to meet this requirement. The sizing of an
FWS is done by using one of the two methods—presumptive method or the field test
method. The presumptive method assumes a BOD5 concentration, while the field
test method is based on an actual daily measurement of BOD5 from the given live-
stock operation [39]. The presumptive method approximates a pollutant entering
into a wetland by reviewing the BOD5 or nitrogen concentration and then applies
the value to an areal loading rate (typically 65 lb BOD5/acre/day). The presumptive
method has been well-known since the Tennessee Valley Authority (TVA) intro-
duced it in 1989 [38].
The field test method requires a collection of samples and analysis based on
BOD5 and total nitrogen (TN). Some of the important factors examined include
average daily flow, temperature, and decay rate constant. The data collection from
the field test is used to determine the size of the wetland. The purpose of the field
test method is to ensure that the design of the wetland does not exceed discharge
limits [38].
On the other hand, in vegetated submerged systems, wastewater flows within the
sediment bed, having more contact with the plant roots. The sediment bed is usually
made of rock, gravel, and soils. Vegetation is usually planted at the top of the wet-
land [38]. Because wastewater flows at lower depths, wastewater is usually degraded
at anaerobic conditions. The slope of this wetland ranges between 2 and 6%. Sizing
of submerged systems is contingent on flow rate and influent and desired outflow
BOD5 [39]. Vegetated submerged systems are not as prolific as surface flow wet-
lands. This is because the sediment beds can easily accumulate solids. Also, the
beds can be very expensive to construct. Nevertheless, vegetated submerged sys-
tems can be used to treat wastewater with low flows and solids [38].
Finally, floating aquatic plant systems comprise of one or more ponds. The ponds
are designed for plants to grow and float at the top of the ponds. Each pond is
designed for a depth between 3 and 5 ft for the purpose of avoiding non-desired
plant species to grow and become prominent within the system and gives the plant
1 Management and Treatment of Livestock Wastes 27

access to nutrients within the wastewater. There are several factors for appropriately
harvesting. These include the number, size, and arrangement of ponds and the tech-
nique for harvesting. There are two major plant species in FAP systems—water
hyacinths and duckweed [38].

1.3.2.3 Constructed Wetland Design

Constructed wetland design usually consists of first-order models under plug flow
conditions, alternating between looking for values of BOD, TSS, ammonium, and
fecal coliforms [34]:

k
   
ln  C  C   Co  C    
q
(1.1)
where
C0 = initial concentration of conditions
C = targeted rate concentration
C* = background rate concentration
k = first-order rate constant (cm/d)
q = hydraulic loading rate (cm/d)
An alternative method to designing a constructed wetland would be the use of
regression equations for one had the desire to consider looking at multiple compo-
nents at one time.
Stone et al. (2004) used constructed wetlands, particularly marsh-pond-marsh
wetland system at North Carolina A & T University. Six wetland systems with the
dimensions of 11 × 40 m treated nitrogen by removing % concentration of ammonia
nitrogen of 30% but only removing 8% phosphorus treatment. First-order kinetics
were 3.7–4.5 m/day for total N and 4.2–4.5 m/day for P, much lower than the typical
model rate constant [40].
In addition, the Environmental Protection Agency has tracked several constructed
wetlands that have been used for the purpose of waste treatment. Seven locations to
treat three different waste types—swine, dairy, and poultry—were constructed. For
swine wastewater, a project in Duplin County, North Carolina, was undertaken for
the purpose of removing Total Kjeldahl Nitrogen (TKN), as it was observed that a
major factor affecting treatment was loading rates of TKN (3 kg/ha/d TKN) and was
able to remove between 91 and 96% TKN, while 10 kg/ha/day only removed
approximately 73%. A wetland in Essex, Ontario, reduced TSS (97%), BOD5
(97%), and 99% fecal coliforms, and 95% E. coli from dairy farm milk house waste-
water. Auburn University used a constructed wetland for poultry lagoon that consid-
ered a series of five wetlands at 3.1 cm/day, a loading rate of 145 kg/ha-day for
chemical oxygen demand (COD), and 30 kg/ha-day total TKN at a maximum of
49.8% BOD6, 60.7% COD, and 36.8% PO4 [41].
28 D. H. Vanderholm et al.

1.3.3 Lagoons

A lagoon is an earthen basin that treats wastewater and stores both liquids and solids
[2]. Lagoons can store wastewater, manure, or rainfall runoff [42]. Lagoons are
capable of reducing BOD and chemical oxygen demand (COD), nitrogen, and odors
[2]. Lagoons can take a round, square, or rectangular shape with a typical length-to-­
width ratio of 3:1 [43]. In addition, lagoons can be situated as a single or multiple-­
stage lagoon system. A single lagoon is divided into three major volumes—sludge
storage, treatment, and effluent storage. Above the effluent storage is a freeboard for
the purpose of protecting the lagoon from storm situations [44]. Figure 1.13 pro-
vides a cross-sectional area of a lagoon.
In the sludge storage, sludge settles at the bottom of the lagoon and is digested at
the top of the layer. Over time, sludge will accumulate within this layer until it
becomes equal to the liquid present. The treatment volume is located above sludge
storage consisting of manure at the bottom. Biological degradation converts sludge
into organic acids and other compounds. The products of organic acids include
methane and carbon dioxide, hydrogen sulfide, ammonia, and volatile organics.
Treated wastewater not leaving the lagoon is stored in the effluent storage section.
Effluent is stored for the purpose of watering crops [44].
Lagoons are designed based on a 25-year, 24-h storm event. This value is contin-
gent on the location of the lagoon as the 25-year, 24-h storm event varies across the
country. The design loading into the lagoon is determined by the number, size, and
the species of animal, along with the geographical location of the lagoon. Prior to

Fig. 1.13 Cross-sectional area of lagoons [13]

1 Management and Treatment of Livestock Wastes 29

land application, dewatering the lagoon is very important. Frequency of dewatering

is contingent on the salt concentration and the soil type [45].
The sizing of a lagoon is based on the volume, depth, and pH. The volume of a
lagoon is contingent on the loading rate of volatile solids per 1000 cubic foot. This
is a function of temperature. The depth of a lagoon is predicated on the precipitation
and evaporation rates where the lagoon is located. A typical minimum depth is 6 ft
but can be 10 ft for colder climates. However, these values are general and are con-
tingent on the type of lagoon constructed. The optimum pH should be maintained at
6.5 to avoid inhibiting methane bacteria. Anytime the pH is below 6.5, lagoons will
experience a high organic loading [2].
Before construction of a lagoon, it is imperative that a soil and groundwater
study is done. This is to ensure that sensitive areas are protected from any dis-
charged from the lagoon. These areas would be any region that leads to surface
runoff. Avoid areas that are geologically unstable [42]. Pretreatment of wastewater
may be beneficial to reduce odor if the BOD5 loading rate is 50 lb BOD5/AC/day
and the depth of the pond is between 6 and 20 ft [43]. In addition, lagoons should be
in close proximity if manure is scraped into the lagoon or below the manure
source [42].
Lagoon maintenance is important for controlling odors. Lagoons should be ana-
lyzed for the presence of algal blooms. Algal blooms occur in basins that have high
loading of nutrients (nitrogen and phosphorus). If a lagoon is void of algal blooms,
ensure that aerobic lagoons do not become anaerobic. Anaerobic conditions can
produce products that can cause odors. The operator should also check and if neces-
sary provide adequate dilution of waste prior to entry into the lagoon and avoid
overloading [46]. This can be accomplished by using a combination of runoff and
wash water [45]. If odors still persist, lime addition to the lagoon can reduce the
presence of odors [46].
Lagoon operators should also evaluate the species of algae and check for the
presence of weeds and grasses and protect them from erosion and unauthorized
access. A healthy lagoon should have green algae. Blue-green and filamentous algae
can clump within a lagoon blocking the sun. Gray, black, or purple algae are very
unhealthy for a lagoon. The presence of weeds can cause a lagoon to short circuit,
thereby affecting the flow of wastewater within the unit. Grass covers on the slopes
and level surfaces of the lagoon can be beneficial but should be mowed and properly
fertilized and should be checked for food, trash, or scum on or near the premise.
These items should be discarded. Trees or any bushes should not be present near the
berm of a lagoon and should be removed [46]. This will also protect the embank-
ments [44]. In the event of erosion, operators should determine the source and make
necessary adjustments to the lagoon if necessary. Unauthorized activity can be
avoided by placing fences and warning signs adjacent to the lagoon [46].
Finally, operators should also monitor the sludge storage and sludge depth.
Remove excess sludge that has accumulated within the lagoon [44].
30 D. H. Vanderholm et al.

1.3.3.1 Anaerobic Lagoons

Anaerobic lagoons are the most common lagoon used for treatment of livestock
wastewater. One of the biggest reasons is because anaerobic bacteria have a higher
rate of organic decomposition as compared with aerobic bacteria [42]. This is
because anaerobic bacteria operate in environments without molecular oxygen a
condition that does not require constant maintenance. Generally, anaerobic lagoons
are usually very deep. Ranges for depth can vary on the region [46]. For example,
the University of Missouri Extension and the State of Mississippi state that lagoons
can have depths between 8 and 20 ft [42, 43]. Based on treatment desired, lagoons
can be designed to be completed as single stage with no secondary treatment, or in
multiple stages where further treatment is completed by additional lagoons [45].
Figure 1.14 is a diagram of a two-stage anaerobic lagoon system.
Anaerobic lagoon can be circular, square, or rectangular. A length-to-width ratio
of 3:1 for rectangular anaerobic is desired, with earthen dike and banks slopes
between 2:1 and 3:1 [42, 43, 45]. Anaerobic lagoons should have a 1 foot spillway
below the top of the berm where inlets should be located on the longest side of the
lagoon [42].
During the wastewater treatment process, anaerobic lagoons separate into top
and bottom layers. At the top of the lagoon, less dense materials such as oils float to
the top of the lagoon, while sludge settles the bottom. The presence of oils and other
materials prevents oxygen entry, maintaining anaerobic conditions within the sys-
tem [46].
Anaerobic lagoons are sized based on the volatile solid (VS) loading rate. These
values can be expressed in 1000 ft3/day or lb VS/1000 ft3-day. These numbers are
affected by the climate. For example, in South Carolina, the volatile solids’ loading
rate is 5 lb VS/1000 ft3-day, while Iowa has a VS loading rate of 3.5 lb VS/10003-­
day [48].
Nevertheless, anaerobic lagoons are problematic because of odors. These odors
are a product of hydrogen sulfide, ammonia, organic acids [49], and methane. Odors
can also be caused by winter to fall and summer to fall turnover within the lagoon
or during land application [42]. There are many solutions that can resolve persisting
odor problems in a pond. Anaerobic lagoons can be covered to prevent the release
of methane gas exiting the system. Anaerobic lagoons can also have induced aerobic

Fig. 1.14 Two-stage anaerobic lagoons for livestock manure treatment [50]
1 Management and Treatment of Livestock Wastes 31

Fig. 1.15 Floating aerator [50]

layers at the top of the lagoon. This can be done by including a floating cover or
aerating the top of the lagoon at very low rates [44]. Figure 1.15 is a floating aerator.

1.3.3.2 Aerobic Lagoons

Aerobic lagoons degrade organic matter by the application of dissolved oxygen

throughout the lagoon. Because dissolved oxygen persists throughout the lagoon,
odors are not present within the system. In order to maintain aerobic conditions,
aerobic ponds are shallow but require a large land requirement. These ponds are
more commonly found in warm and sunny climates. There two subcategories of
aerobic lagoons—naturally and mechanically aerated [46]. Figure 1.16 is a diagram
of an aerobic lagoon.
Naturally aerobic, oxidation ponds reduce organic materials within wastewater
by using either oxygen from the atmosphere or algae by means of photosynthesis
[46]. Wind on the pond surface also mixes with the water within the oxidation pond
[44]. These ponds are very shallow with a minimum depth between 1 and 5 ft with
a maximum of 5 and 6 ft [46]. The main design parameter is the organic loading
rate, which is typically 50 lbs BOD5/acre of surface area [49]. Nevertheless, natu-
rally aerobic lagoons are not often used for the treatment of livestock wastewater.
Mechanically aerated lagoons mix oxygen throughout the lagoon by mechanical
means. The need for supplying energy can make these lagoons expensive. In many
cases, solar or wind power supply the power to operate aeration equipment. Also the
lagoons can be designed to have anaerobic segments to reduce energy requirement
[46]. Compared with naturally aerobic, mechanically aerated lagoons do not have a
large area requirement but usually have a depth of 10 ft However, in addition to
32 D. H. Vanderholm et al.

Fig. 1.16 Two-cell aerobic lagoon to treat swine waste [51]

being more expensive, mechanically aerated lagoons tend to generate more sludge,
have a high tendency for foaming, and may require additional treatment such as a
septic tank to collect and remove solids [49].

1.3.3.3 Facultative Lagoons

Facultative lagoons are basins that operate in both aerobic and anaerobic conditions.
These lagoons can be arranged as a two-staged pond system where each pond has a
depth of 4 ft or as a single-pond system with a depth of 6 ft [43]. Facultative lagoons
usually have three layers. At the top is an aerobic layer. This layer receives sunlight
and wind, promoting the process of photosynthesis, and provides oxygen. The mid-
dle layer is a facultative layer. In this layer of the lagoon, anaerobic and aerobic
conditions exist. The extent as to which condition is prominent is contingent on the
geographical location of the lagoon [46]. Bacteria that can thrive in anaerobic or
aerobic conditions (facultative bacteria) are commonly found in this layer. The bot-
tom layer is anaerobic. This layer contains an accumulation of sludge from lagoon
activities [46]. Because of the layering of the lagoon, odors can be minimized [52].
1 Management and Treatment of Livestock Wastes 33

1.3.4 Thermal and Biological Chemical Treatment

for Biogas Producton

1.3.4.1 Description

Recent developments have occurred where there has been a call for the conversion
of livestock wastes that can be used for energy, specifically biofuels. To summarize,
biochemical processes are transforming organic materials to fuels by means of vari-
ous processes such as anaerobic and photosynthesis. Following a biochemical pro-
cess, the remaining solid and slurry within the reactor becomes viable as a reusable
resource such as fertilizer [53].
Thermochemical processes convert organic matter into gas, fuels, or other car-
bon residuals by the use of high temperatures to physically convert the bonds of
organic matter. Some of the major chemical conversion procedures include combus-
tion, pyrolysis, gasification, and liquefaction [53].

1.3.4.2 Pyrolysis

Pyrolysis ultimately transfers a given biomass into either char or a volatile gas that
can form bio-oil or combustible pyrolytic oils. Slow pyrolysis methods have been
used to form char, an entity that has the benefit of producing energy for coal com-
bustion plants, or as an addendum to soil. Some authors have found that chars from
various pyrolytic processes are capable of having better absorption than those made
from granular activation carbon [53].
There are two major types of pyrolysis—fast and slow/moderate. Fast pyrolysis
is a pyrolytic process that consists of using high heat rate and residence time. The
resultant products include low molecular weight or an insoluble organic compound
such as tar. Reactor examples include bubble fluidized bed, circulating fluidized
bed, and vacuum reactor. The requirements within fast pyrolysis includes a particle
size less than 1 mm. Slow/moderate pyrolysis is the antithesis of fast as it requires a
long vapor residence time and low heat rate. The resultant products are charcoal,
depending on the concentration of lignin and hemicelluloses. Examples include
rotary kiln and moving bed reactor [54].
Pyrolysis applications have been experimented with various manure types. It has
been determined that the effectiveness of char production was based upon manure
type and the conditions, as it was observed that organic materials differ between two
different waste types [55].
34 D. H. Vanderholm et al.

1.3.4.3 Direct Liquefaction

Direct liquefaction is another thermochemical process that converts organic mate-

rial, specifically lignin components, into various organic oils. Ideal conditions for
liquefaction would be having very high pressures (5–20 MPa) and low temperatures
(250–350 °C). Following the process, the remainders of direct liquefaction are non-­
reactive and stable, which are then converted into oil-based compounds with high
molecular weights [53].
The process of liquefaction begins when the bonds of organic material are bro-
ken into simpler compounds, resulting into the forms of chars, instead of the process
of oils. To prevent the formation of chars, solvents are typically added to slow down
higher-order solid-state reactions, reducing condensation and the subsequent char
formation. Examples of the solvents that are used include dioxane, MDSO, DMF,
acetone, and methyl alcohol [54].

1.3.4.4 Gasification

Gasification operates at high temperatures and atmospheric pressure within the

range of 800–1300 °C for the purpose of producing chars and a low energy fuel. The
gasification process has three components—first, pyrolysis, or the conversion of
organic materials into both tars and hydrogen-based combustible fuels. Second,
exothermic reactions with the presence of oxygen can occur to remove the bonds
within the organic material at high temperatures. Third, methanation or the forma-
tion of methane from hydrogen and carbon monoxide proceeds where the condi-
tions consist of lower temperatures [53].
A fixed bed 10 kW power, counter-current atmospheric pressure gasifier was
capable of achieving a gas product made from either high ash feedlot manure (HFB)
or poultry litter biomass (HLB) that consisted of the following product: H2,
5.8 ± 1.7%; CO, 27.6 ± 3.6%; CH4, 1.0 ± 0.5%; CO2, 6.7 ± 4.3%; and N2, 59.0 ± 7.1%.
Ideal processes included air-blown gasification for the purpose of having a higher
energy fuel [56]. If application of a catalyst such as nickel or aluminum would bet-
ter assist in the formation of gas production by preventing tar cracking, it would be
preferable [57].
Priyadarsana et al. (2005) completed gasification studies for the production of
both cattle manure and chicken litter biomass under batch mode where it was deter-
mined that the molar composition of gas was 27–30% CO, 7–10% H2, 1–3% CH4,
2–6% CO2, and 51–63% N2 based on the use of air mass flow rate of 1.48 and
1.97 kg/g, where particle sizes are 9.4 and 5.15 mm, respectively [58].
1 Management and Treatment of Livestock Wastes 35

1.3.5 Composting

There are many reasons to compost. Composting is done to reduce organic material,
degrade dead livestock, and reduce disease transmission at a low cost. There are
several factors that affect the quality of composting—carbon-to-nitrogen ratio,
moisture content, temperature, and the type of composting materials [59]. A proper
carbon-to-nitrogen ratio reduces the odors while the temperature affects the micro-
bial degradation [60]. The temperature affects degradation processes. During the
winter season, degradation can be reduced in some places by 20% [61]. Composting
materials include sawdust, wood chips, and litter. Composting consist of microor-
ganisms (bacteria and fungi) degrading organic materials within the compost pile to
simple products [59].
The general composting values are shown in Table 1.12 below. These values are
based on manure composting. Composting consist of primary and secondary pro-
cesses. In primary composting, the temperature is raised and the organic material is
degraded. As composting progresses, degradation begins to slow and the tempera-
ture is reduced. Eventually, degradation ends and the material is left idle [60] in a
process known as curing. Curing maintains the conditions within the pile. It also
allows items such as bones to be degraded [61].
There are two types of composting facilities—bins and piles or windrows. These
are contingent on the type of livestock industry. Bins are used in poultry and swine.
Beef and dairy cattle use piles or windrows [59]. Windrows or piles place materials
into rows at triangular cross-sections. They are usually combined with bulking
agents [62]. Aeration occurs by turning the piles by using frontend loader or com-
post turners [63]. Piles constructed in arid regions will need to receive outside mois-
ture. This can be done by using a high-pressure nozzle from holding ponds or lagoon
wastewater. On the other hand, piles in areas with precipitation may need to be
covered to prevent odor production [63]. Bins can be designed to have dimensions
of 6 ft by 8 ft with a wall height between 5 and 6 ft Bins can be made of 2 × 6 or
2 × 8 lumber or using plywood with a 2 × 6 to provide support behind the plywood
[64]. The foundation of bins can be made up of pallets, gravel, concrete, and bare
soil [65].
There are two entities that can be composted—manure and dead livestock. Dead
animal composting is an option to remove livestock carcasses without having

Table 1.12 Factors that affect composting [59]

Factor Value
C:N ratio 25–40:1 (optimum: 30:1)
Moisture 40–65% (optimum: 50%)
Temperature 43–66 (optimum: 54–60 C), >71 not ideal
Site selection 1–5% (2–3% account for runoff and erosion)
36 D. H. Vanderholm et al.

detrimental effects on the environment [59]. Dead animal composting maintains

aerobic conditions, provided gases and liquids are taken away from the system [60].
Livestock operators should consider state requirements to decide what the state
requirement of handling dead animals is. For example in the state of Kansas, com-
posting facilities of dead livestock require a roof and floor to sustain moisture and
avoid groundwater contamination with a fence surrounding the facility [59]. The
process of composting is contingent on the size of the carcass materials [60].
A dead animal composting pile begins with a layer of sawdust 1–2 ft in depth.
The dead livestock are then spread evenly across the sawdust layer [60]. Animals
are laid on the side in an attempt to maximize the space for livestock [61]. Another
layer of sawdust 2 ft in depth covers the dead animals. This second layer of sawdust
maintains heat, prevents odors from escaping, and collects liquids and air to encour-
age microbial activity within the pile [60]. The amount of sawdust needed is contin-
gent on the type of livestock to be composted. A rule of thumb for sawdust
application is that in every 1000 lb of carcass, apply 7.4 yd3 of sawdust in the dimen-
sions of 9 ft × 10 ft [61]. When livestock need to be added to the composting pile,
the top sawdust layer is removed, exposing the dead animal layer. At this point, the
new animals are added and then covered up with a new sawdust layer. To maintain
the quality of the pile, it advised that the pile is turned every 90 days. Once compost-
ing is complete, the products can be applied to the land or reused in other capacities
[60]. This will usually happen anywhere between 4 and 12 months of composting
time [61].
While composting dead livestock is advantageous, there are several concerns
involved. These can include leachate of fluids from the carcasses entering into the
surface and groundwater and disease-spreading pathogens [65]. Therefore, it is nec-
essary to consider the best place to site the place for composting dead livestock.
Changes can include placing the facilities away from the water table, away from low
permeable soils, and downwind from neighbors. Facilities should also be con-
structed away from livestock to suppress disease potential. Livestock operators
should also have an emergency plan in case of outbreak [61]. For additional protec-
tion, the livestock operator can create a barrier wall to prevent access to the com-
posting pile. The barrier can be 4 ft high using four steel t-posts with concrete floors,
wooden walls, and a metal roof [65].

1.3.6 Vermicomposting

An alternative method of treating wastes that has been used related to composting is
as vermicomposting. Vermicomposting is a method where earthworms digest a
small portion of organic matter where the majority becomes waste in a form known
as worm casts. The processes involved in earthworm digestion are typically physical
or mechanical, grinding and mixing, and biological or microbial decomposition in
nature. In vermicomposting, waste is added to the system. It must be added into the
system in thin layers for the purpose of increasing degradation. There is great
1 Management and Treatment of Livestock Wastes 37

competition between earthworms and microorganisms for the carbon sources.

Application of waste can change—it will either increase or decrease productiv-
ity [66].
Vermicomposting treatment technology has been used extensively in animal
excretion, sewage, and agroindustrial wastes but not animal manures. Therefore,
Loh et al. (2004) treated cattle and goat manures using the earthworm, Eisenia
foetida. The experiments found that total C, P, and K were high in goat manure
worm casts as compared to cattle, whereas cattle worm casts were richer in N con-
tent. In addition, cattle manure had a higher biomass and reproductive performance
as well along with a higher cocoon production per worm [66]. Other studies have
been compiled on cow, buffalo, horse, donkey, goat, and animal [67], dairy [68], and
pig [69, 70] to name a few. Within continuous feeding reactors, two different types
of pig slurry were compared with 500 earthworms (Eisenia foetida); microbial bio-
mass was specifically measured with 3 kg of pig slurry; loss of C was not related to
pig slurry rate; rate of manure-earthworm relationships was investigated [71].

1.3.7 Summary

There are many treatment methods that can be considered for the handling of wastes
that persist within the livestock industry. An operator must consider what is avail-
able in regard to space and the desired treatment needed in order to make an appro-
priate decision on selecting the proper treatment method.

1.4 Land Application of Livestock Wastes

1.4.1 Description

Land application is a waste management technique that involves recovery of nutri-

ents from manure by plants for the purpose of producing a crop [2]. The classifica-
tion of manure depends on the percent of dry matter present and the type of livestock
waste industry. Manure can be in liquid (less than 5% dry matter), semiliquid
(5–10% dry matter), or solid (greater than 15% dry matter) form. Generally, beef
and poultry industry handles solid manure, while dairy and swine manure is usually
in liquid form [72].
Regardless of the industry, the nutrient content is the primary focus for applica-
tion. Nutrient content within the manure is affected by the type of animal species,
the process for handling of manure, livestock housing, bedding system, diet, tem-
perature, and the nutrients present. The primary nutrients of concern are nitrogen,
phosphorus, and potassium. The nitrogen presence affects the type of plants and
quality of the produce. There are two important forms of nitrogen that must be
38 D. H. Vanderholm et al.

considered—organic nitrogen and ammonium nitrogen. When organic nitrogen

enters into soils, it is mineralized into inorganic nitrogen. Mineralization is contin-
gent on the temperature and time of year. Warm and moist soils are better for the
degradation of organic nitrogen as compared with cool and dry soils. Ammonium-­
nitrogen is converted to organic nitrogen by plants in a process known as nitrifica-
tion. Moreover, 25–50% of organic nitrogen is converted to ammonium-nitrogen.
However, improper application of manure can lead to volatilization or the conver-
sion of ammonium-nitrogen to ammonia-nitrogen. This becomes problematic
because ammonia-nitrogen dissipates into the atmosphere. On the other hand,
potassium and phosphorus must be converted to inorganic forms in order for it to be
of use by plants [73]. Manure can also be problematic because it can produce vari-
ous gases. These gases can have grave effects depending on the concentration.
Table 1.13 summarizes the major gases found in manure. Previous treatment meth-
ods can affect land application. Table 1.14 discusses the various treatment processes
and their effects on land application. Therefore, the type of handling equipment,
time, and rate of application should be considered if an operator is to consider land
application.

1.4.2 Manure Handling Equipment

The equipment necessary for handling manure depends on the type of manure. Each
operator must make a decision of handling manure that best distributes the nutrients
to the crops being planted. Depending on the type of manure handled, there are
unique pieces of equipment that are used in order to safely move the manure onto
the field.

1.4.2.1 Solid Manure

Solid manure is incorporated at the surface by using spreaders that are trucked-­
mounted or trailer-towed. Regardless of the type of spreader, manure can be spread
at the side or the rear. Nevertheless, rear manure spreaders are more likely used
today [72]. For example, livestock operators in the state of Missouri primarily use

Table 1.13 Manure gases [74]

Gas Effects (percent indicates percent or concentration in ppm)
Ammonia (NH3) Eye irritation (<1%)
Coughing, irritation of throat, eyes, lungs (3–5%)
Carbon dioxide (CO2) Difficulty breathing, drowsiness, headaches (3–6%)
Death (>30%)
Methane (CH4) Asphyxiation (5–15%)
Hydrogen sulfide (H2S) Dizziness irritation, headache (50 ppm)
Death
1 Management and Treatment of Livestock Wastes 39

Table 1.14 Various wastewater and biosolid treatment processes and methods and their effects on
land application processes [75]
Process/ Effect on land
Method Process definition Effects on biosolids application process
Wastewater treatment process
Thickening Low force separation of Increases solid content by Lowers transportation
water and solids by removing water costs
gravity, flotation, or
centrifugation
Stabilization method
Digestion Biological stabilization Reduces biological oxygen Reduces the quality
(anaerobic and/ through conversion of demand, pathogen density, of biosolids
or aerobic) organic matter to carbon and attractiveness of the
dioxide, water, and material to vectors
methane (disease-spreading
organisms)
Alkaline Stabilization through the Raises pH. Temporarily High pH immobilizes
stabilization addition of alkaline decreases biological activity. metals as long as pH
materials (e.g. lime, kiln Reduces pathogen density levels are maintained
dust) and attractiveness of the
material to vectors
Heat drying Drying of biosolids by Destroys pathogens, Greatly reduces
increasing temperature eliminates most of water sludge volume
of solids during
wastewater treatment
Chemical and physical processes that enhance the handling of stabilized biosolids
Conditioning Processes that cause Improves sludge dewatering The ease of spreading
biosolids to coagulate to characteristics. May increase may be reduced by
aid in the separation of dry solids mass and improve treating biosolids
water stabilization with polymers
Dewatering High force separation of Increases solids Reduces land
water and solids. concentration to 15–45%. requirements and
Methods include Lowers nitrogen (N) and lowers transportation
vacuum filters, potassium (K) costs
centrifuges, filter and concentrations. Improves
belt presses, etc. ease of handling
Advanced stabilization method
Composting Aerobic, thermophilic, Lowers biological activity, Excellent soil
biological stabilization destroys most pathogens, conditioning
in a windrow, aerated and degrades sludge to properties. Contains
static pile, or vessel humus-like material less plant available N
than other biosolids

rear-end box-type spreaders with beaters. These spreaders can consist of a conveyor
with chains to move manure from the front of the spreader to the beaters or a front
endgate that moves the manure to the beaters [76]. Once it is moved to the rear, the
beaters scatter the manure onto the ground [72].
Rear-end box-type spreaders can have single, horizontal, or double vertical beat-
ers. However, each beater type is limited in its ability to properly distribute
40 D. H. Vanderholm et al.

nutrients. Single beaters cannot spread manure homogenously onto the land.
Horizontal beaters only spread manure in areas of close proximity to the trailer.
Double vertical beaters spread manure very wide and thin. Overall, rear-end box-­
type spreaders have a problem with spreading manure homogenously onto the land.
They are also very heavy and have the potential to compact soils if land application
is done in the fall and spring. Similar to box-type spreaders, truck-mounted spread-
ers apply manure using double beaters in various horizontal or vertical configura-
tions. Regardless of application, solid manure handling should be applied within
24 h. This is to ensure the minimization of nutrient loss, the presence of odors,
nutrient runoff, and compaction [72].
Since the application of solid manure can generate odors, there are methods that
can be done to suppress odors in manure land application. These include placing a
cover over solid manure not being applied, using chemical treatment such as alum
(also advantageous for preventing ammonia volatilization), and considering the
wind direction when applying onto the surface. There are also mechanisms that can
be employed that can better spread the manure upon entry on the field. These include
a tandem disk or a field cultivator. Solid manure can also be pretreated by drying or
composting [77].

1.4.2.2 Semisolid Manure

Semisolid manure is handled by using spreaders with an endgate. The configuration

can range from side discharge or a V-shaped hopper. Each of these can be handled
by power takeoff (PTO) or ground wheel tractor spreaders or a truck-mounted
spreader. The process of application consists of moving the manure by augers to be
flung at the point of emission on the spreader. Manure is flung either by using a
rotating or flail-type expeller. A rotating expeller directly flings manure, while in a
flail-type expeller, manure travels from a hopper onto a rotating shaft with chain-­
suspended hammers. Once the manure is on the hammers, it is tossed onto land [76].

1.4.2.3 Liquid Manure

Liquid manure can be applied at or below the surface. Surface application of liquid
manure is completed by fixed sprinklers, hand-carried sprinkler, traveling guns, or
central pivot irrigators [76]. Factors that control application by irrigation equipment
are nozzle size and pressure. These affect the size of the drops applied to the surface.
Larger-sized drops are greatly preferred to control the loss of nitrogen and decrease
odors [77]. A recommended size is greater than 150 μm. Other ways include adding
dilution water or drop nozzles [78]. Surface application of manure is preferred in
areas where odors and nutrient loss are minimal [76]. Figures 1.17 and 1.18 provide
diagrams of irrigation systems.
Subsurface application injects liquid manure below the surface where it is then
applied below the soil surface by a self-propelled application. Manure can also be
1 Management and Treatment of Livestock Wastes 41

Fig. 1.17 Irrigation system to apply liquid manure [76]

Region of highest
Region of highest application rate if
application rate if nozzles are
smallest nozzles are correctly sized
oversized

Center of pivot
Pump

Mannure
lagoon
Sprinklers
Underground
main line
Lateral
Towers

Fig. 1.18 Center pivot irrigation system [79]

transferred by a drag hose or a tractor-drawn applicator. The method chosen is

determined by the size of the operation. Usually larger operations opt to use a drag
hose or a tractor-drawn applicator. When liquid waste is applied below the surface,
injectors have chisels that break up material or sweeps that uniformly apply the
liquid manure below the root surface to avoid leaching [76]. Chisel-type knives also
prohibit odors and volatilization, while sweep-knife injection reduces volatilization,
denitrification, and material degradation [73].
Subsurface is preferred to surface application for several reasons. First, subsur-
face reduces the potential of ammonia-nitrogen emissions [76], greenhouse gases,
and odors [72]. For example, research has shown odors to be reduced by 90% when
42 D. H. Vanderholm et al.

incorporating a subsurface method [78]. Second, subsurface application reduces

runoff potential, availing more nitrogen to plants. Third, subsurface injection
spreads the manure so it does not have an impact on the surface of the soil. Despite
its many advantages, subsurface application is energy intensive; requires more
maintenance, time, and management; has higher equipment costs; and is incapable
of being used on rocky soils. Therefore, assessment should be made to determine
whether or not subsurface injection is a more viable option than any surface applica-
tion method [80].

1.4.3 Time of Application

The time when manure is applied determines nutrient availability to plants. Spring
is the best season for manure application because nutrients are broken down into the
soils during the growing season. Organics are quickly broken down in the soils,
increasing nitrogen availability. Summer applications are appropriate if growing
hay, pasture, and warm-season grasses and if application is completed by travel
guns or the central pivot system. Applying manure during the fall is only appropri-
ate if temperature stays below 50 °F [80]. This is because manure is immobilized
and remains in the soil [73, 80], leading to more time for degradation. But when the
temperature is above 50°, nitrification, leaching, and denitrification occur [73].
Winter application of manure is never recommended as manure hardly enters the
soil and has a higher potential for runoff [80] into surface waters. If manure applica-
tion is a necessity in the winter, apply at low concentrations or during periods of
snow melt [73].

1.4.4 Rate of Application

The amount of nitrogen, type of manure, how manure is applied and used, and addi-
tional economic or environmental are the factors that determine how frequently
manure will be applied to a given crop. The University of Minnesota Extension
provides four steps to determine the process by properly determining the rate of
application [73]:
1. Determine the nutrient needs of the crop.
2. Analyze the nutrient content within the manure.
3. Uncover the nutrient available to the crop.
4. Compute the rate of application.
1 Management and Treatment of Livestock Wastes 43

1.4.5 Summary

In summary, the purpose of land application attempts to resolve the issue of losing
nutrients that are vital to the growth of crops. Manure should be applied uniformly
to land to avoid the volatilization of nitrogen into the atmosphere. It should also
maintain the potassium (K) and phosphorus (P) on the field. The time of application
should be considered in order to have nutrients maintained within the soils and
avoid any subsequent losses that occur during improper times of application [81].
Manure application should be done to avoid the presence of odors [76] and other
potential issues. The rate of application depends on the crop’s needs.

1.5 Storage of Livestock Wastes

1.5.1 Description

Most often, the treatment of livestock waste is done for the purpose of recycling
products back within the system. This can include land application for growing
plants. However, there may be times when the conditions are not conducive for
reusing treated wastes. Therefore, livestock wastes must be stored until the appro-
priate conditions take place. There are several factors that should be considered
when deciding whether or not to store manure: first, if the soil is saturated, wet,
frozen, or snow covered or if the soil will compact under the weight of manure han-
dling equipment; second, if the temperature and/or humidity create a proper envi-
ronment for the generation of odors; third, if a livestock operation may not have the
proper equipment or personnel available to apply manure at the present moment;
fourth, if the cropping schedule may require temporary storage; and finally, if there
is a higher volume of manure and wastewater than what can be handled [82]. There
are several methods for storing wastes. These methods are employed usually based
on the time of storage and type of waste treated—i.e., solid, semisolid, and liq-
uid wastes.

1.5.2 Storage Time

Livestock wastes can be stored either on a short-term or long-term basis. When

wastes are stored for 60–90 days or up to 180 days, it is termed as short-term stor-
age. Short-term storage is a viable option when poor weather conditions persist, or
when setup is not appropriate to properly handle manure. Short-term storage is also
used in mild climates or when growing crops [83]. However, it is very seldom for
operators to store liquid manure on a short-term basis. Dairy wastes are the most
appropriate to be stored short term.
44 D. H. Vanderholm et al.

There are many methods for storing manure on a short-term basis. These can
include stacking within a field, covered with a plastic sheet, or storage in a detention
pond. Manure can also be scraped into open lots in mounds or inside pole sheds.
Regardless of the method, the operator should choose to avoid any contamination of
water supplies or exposure to bacteria from the manure [84].
Long-term storage can last for approximately 180 days. Facilities are available to
hold solid, semisolid, or liquid wastes. For example, walls and slabs can stack solid
manure, while semisolid pumps or scrapers help transport waste into areas desig-
nated for storage. Liquid waste is usually transported by pumps or pipes [84].
Sometimes manure can be held for longer than 180 days. For example, waste is
stored for 6 months for the purpose of application on annual row or small grain
crops. In the center and upper Midwest, storage can happen for a full year if fall
applications are unsuccessful because of wet conditions [82].

1.5.3 Facilities to Store Livestock Waste

There are many facilities that can be used to store manure. However, the practicality
of each facility depends if the operator is storing solid, semisolid, or liquid manure.
Table 1.15 provides an estimated cost for manure storage facilities.

1.5.3.1 Solid Manure Storage

The objective in storing solid manure is to reduce the volume, odor, and potential
for runoff. Solid manure is stored based on climate and industry. Because the evapo-
ration rate is greater than precipitation, arid regions can store solid manure in a
different fashion as compared to regions that retain precipitation. Arid regions sim-
ply store manure in stacks or piles. In the beef and dairy industry, manure is com-
posted using windrows or piles, while in the poultry industry, the manure is
contained inside stack houses. On the other hand, non-arid regions require the solid

Table 1.15 Estimated costs for manure storage facilities. Numbers based on 500,000 gallon
capacity [85]
Cost
Storage type ($/1000 gallon)
Naturally lined earthen basin 25–36
Clay-lined earth basin using clay onsite 50–70
Clay-lined earth basin using clay from off-farm borrow site (depending on 80–100
hauling distance)
Earthen basin with plastic liner 100–140
Earthen basin with concrete 120–280
Aboveground pre-cast concrete tank 200–250
Aboveground concrete tank poured in place 230–270
1 Management and Treatment of Livestock Wastes 45

manure to be walled with a concrete bottom and covered with a roof. If solid manure
is not housed in this manner, it could also be composted [83]. However, there are
alternatives for non-arid region storage of solid manure. Purdue University
Extension states that if manure is dried and bedding is added to form a solid, it can
be stored on concrete pads. Concrete pad storage of manure reduces the potential of
groundwater leaching and runoff provided the operator constructs a roof [86].

1.5.3.2 Semisolid Manure Storage

Pits are a main way to store semisolid manure. Pits in general are a viable option for
waste storage because they can reduce waste volume and reduce the production of
odors provided they are properly maintained. Pits can be fabricated from concrete
or a coated metal or can be completely made of earth. Manure is transferred into
them by means of slated floors. Fabricated pits can be constructed for a location
completely above, partially above, or below the surface of the ground. The process
of transferring semisolid manure is by scraping or flushing the manure from its
source. Equipment used for transferring can include collection sump pumps or by
gravity, depending where the pit is located. Semisolid manure should be agitated
before transfer to ensure all suspended solids are relocated into the pit [83].
Pits made from earthen structures are capable of housing large quantities of
semisolid wastes. Therefore, operators will need to ensure ample space is available
if a pit from earth is to be used [83]. The incorporation of manure at the bottom of
the pit protects the pit from leaching nutrients. This is especially advantageous for
very clayey soils. Pits are also lined to protect leaching from the walls. The change
in fluid levels can alter the stability of the pit, leading to the formation of cracks
[86]. In addition, earthen structures require vegetative cover. Maintenance is then
necessary for its upkeep. As with fabricated pits, manure entering into an earth-­
structured pit also requires agitation. Transporting semisolid wastes into the pit is
easily done with the use of a built-in access ramp. This can make hauling and trans-
porting waste very time-consuming. Nevertheless, earthen pits can be a culprit for
odor production so proper maintenance is necessary. Despite the time-consuming
hauling and the high potential for odors, earthen pits are less expensive as compared
to fabricated pits [83].

1.5.3.3 Liquid Manure Storage

Facilities that can store liquid manure can include lagoons, runoff holding ponds,
and storage tanks. Table 1.16 provides a detailed description of the solid content
within liquid manure systems. Lagoons are a beneficial option for storing liquid
manure because they can house liquid manure for 6–24 months [86], can be cost-­
effective per animal, and reduce odors [83]. Lagoons provide a mechanism for liq-
uid waste to be treated prior to land application [86]. Lagoons require a higher
volume than treatment of semisolid manure and must consider the temperature,
46 D. H. Vanderholm et al.

Table 1.16 Solid content for liquid manure systems [76]

System Solid content

Manure pit
Swine 4–8%
Cattle 10–15%
Holding pond
Pit overflow 1–3%
Feedlot runoff <1%
Dairy bard wastewater <1%
Lagoon, single or first stage
Swine ½–1%
Cattle 1–2%
Lagoon, second stage <1/2%

climate, and volume of wastewater to be housed. Biological activities in the lagoon

are maintained by replenishing the lagoon with dilution water and prevent salt
buildup. This should be monitored during high rates of evaporation [86]. Lagoons
should also be monitored to avoid a buildup of settled solids [87]. More information
on lagoons can be found in Sect. 1.2.
Runoff holding ponds are typically used for storage during rainfall events. This
means that any liquid manure housed must be pumped out following the event [83].
Holding ponds are designed to be smaller than lagoons. This reduces the rate of
degradation within the pond. Erosion and overflow is controlled by installing a
12-inch spillway. To maintain liquids within the holding ponds, a settling basin is
set up to collect 60–75% of the solid manure. This allows waste removal to be com-
pleted by irrigation systems [86].
Storage tanks for liquid waste can be made from glass, concrete, or earth. Similar
to pits, storage tanks can be placed above, partially above, or underground. A stor-
age tank is divided into five major sections—residual volume, manure storage, wash
water, rainfall and evaporation, and safety volume depth. The residual volume com-
prises of 6–12 inches from the bottom of the tank. Above the residual volume houses
the manure. The manure is pumped into this section of the tank and can be stored
for 3–6 months. The wash water stores wash or freshwater. If the tank is open, the
net rainfall and evaporation section collects any rainfall that may occur. Finally, the
safety volume depth provides adequate space to handle a 25-year, 24-h storm event.
Depending on the type of material, storage tanks will have a different depth [88].
1 Management and Treatment of Livestock Wastes 47

1.5.4 Storage Area Design

The storage of manure has been published by the US Department of Agriculture

(USDA) and follows the following calculation based on storage volume [2]:

VMD  AU  DVM  D (1.2)

where.
VMD = volume of manure production for animal type for storage period, ft3
AU = number of 1000 lb animal units per animal type
DVM = daily volume of manure production for animal type, ft3/AU/day
D = number of days in storage period
The second equation calculates the bedding storing volume:

BV  (FR  WB  AU  D) /BUW (1.3)

wheres
FR = volumetric void ratio (values range from 0.3 to 0.5)
WB = weight of bedding used for animal type, lb/AU/day
BUW = bedding unit weight, lb/ft3
Sometimes this equation is multiplied by 0.5 as a volumetric void ratio.
Sizing for a liquid and slurry waste storage can be calculated from the following
equation:

WV = TVM + TWM + TBV (1.4)

where
WV = volume of waste stored, ft3
TVM = total volume of stored manure, ft3
TWW = total wastewater stored, ft3
TBV = total bedding volume stored, ft3

1.5.5 Summary

The type of manure affects the manure facility chosen. Within the types of manure,
there are various facilities that can house manure. Each facility should be analyzed
carefully before installation. This ensures that the proper facility is constructed
based on the needs of the operation.
48 D. H. Vanderholm et al.

1.6 Feedlot Runoff Control Systems

1.6.1 Description

Section 1.1 of this chapter indicates that feedlots are required to have NPDES per-
mits as defined in the Clean Water Act of 1977 [89]. This limits the amount of dis-
charge that can occur at a particular location. A major source of discharge from
feedlots is runoff. There are several different systems that properly contain runoff.
Many of systems have been discussed in prior sections, and therefore information
concerning the significance for runoff control will only be presented. Runoff control
protects a feedlot from the presence of weeds, odors, and insects. The collected
water provides an alternative source for fertilizers and irrigation water [90].

1.6.2 Runoff Control Systems

1.6.2.1 Descrtiption

The processes of a runoff control systems are multifaceted. A runoff control system
captures and reroutes rain or snowmelt. It can also provide a method to treat runoff
before it is to be discharged. There are two major subsets of runoff control sys-
tems—full containment and discharge runoff control systems. Full containment
systems (also known as clean water diversion systems) include the use of terraces,
channels, and roof gutters [89].

1.6.2.2 Clean Water Diversion

The purpose of diversion is to control runoff entry into holding ponds and settling
basins [94]. In addition, precipitation is prevented from invading manure storage
systems, preventing the potential for creating polluted runoff [90].

1.6.2.3 Discharge Runoff Control

Discharge runoff control systems include settling basins and runoff holding ponds.
Settling basins are a runoff control system that separate liquid from solids. The
separation of liquids from solids allows liquids to be further treated by methods
such as storage ponds. Solids settle to the bottom while the liquids remain at the top.
There are several processes that will cause solids to separate from liquids. These
include risers, slotted board, or porous dams. Settling basins consist of channels or
boxes made of concrete or earth. Cleaning the basin is necessary to allow for solid
1 Management and Treatment of Livestock Wastes 49

placement. The cleaning of the basin should be done if 50% of the basin is filled
with solids. Solids are taken from the basin and led away from the feedlot. If clean-
ing is not permissible, an alternative method is to increase the size of the basin by
25–50%. Scrapers, high-pressure water systems, and metal screens prevent the sys-
tem from being clogged. Figure 1.19 is a diagram of a solid-liquid separator [90].
Figure 1.20 depicts a system to handle runoff. Figure 1.21 provides a diagram of a
settling basin.
Runoff holding ponds receive and store liquid runoff from settling basins. This
process can happen 15–30 min before entry into a settling basin [92]. In general,
they are smaller than holding ponds. This means that when wastewater is collected,
it will only remain in the ponds for a short period of time. They must be dewatered
by using equipment such as a sprinkling systems or perforated pipes. However, if
holding ponds are constructed in arid regions, dewatering is not necessary as evapo-
ration will be sufficient. Water removed from the holding pond can be applied onto
crops [90]. Figure 1.22 is a diagram of a holding pond.
In general, holding ponds are designed based on a 25-year, 24-h storm [90].The
volume chosen for the pond is also contingent upon the time of storage permit-
ted [92].

Fig. 1.19 Solid-liquid separator [8]

50 D. H. Vanderholm et al.

Fig. 1.20 Lot runoff handling system for milking wastewater [91]

1.6.2.4 Vegetative Filter Strips

Another method to control feedlot runoff includes vegetative filter strips. Vegetative
filter strips (VFS) are a feedlot runoff control system consisting of vegetation. This
vegetation is grown in close proximity to the feedlot, reducing constituents such as
sediments, nutrients, and pesticides [93] and COD [94]. In a VFS system, vegetation
uptakes pollutants from runoff prohibiting transport beyond the feedlot. The removal
of these particulates from the runoff results in clean water [95]. Associated pro-
cesses include settling, filtration, dilution, pollution absorption, and infiltration [96].
VFS systems are capable of removing 60–70% suspended solids, 70–80% nitrogen
1 Management and Treatment of Livestock Wastes 51

Fig. 1.21 Settling basins for manure management [91]. (a) Earthen sidewall setting basin. (b)
Concrete setting basin

Fig. 1.22 Holding pond for storing milk house wastewater [91]

[94], 7–100% phosphorus, and 64–87% pathogen removal [97]. VFS systems create
a mechanism that can reduce non-point pollution runoff. Several factors affect the
efficiency of a VFS system. These include the type of pollutant, soil type, vegeta-
tion, state of flow, and current plant status [93].
52 D. H. Vanderholm et al.

The nature of the pollutant is important in determining its ability to be treated by

vegetative filter strips. Vegetative filter strips are capable of reducing particulate-­
bound pollutants in comparison with soluble particulates. Various processes incor-
porated within VFS are able to be removed by the system as compared with soluble
particulates, which can only be removed by sedimentation. The type of soil is
important because of the various processes that occur within soil. Sandy-loam soils
with a depth between 3 and 13 ft or clay soil (26–145 ft) are ideal for VFS. Vegetation
should be dense and rough and must be able to reduce the surface velocity so that
collected solids are kept within the system. Flow entering into the VFS system
should be overland sheet flow as compared to concentrated flow. Overland sheet
flow prevents sediments from leaving the VFS system, lowering the velocity of the
wastewater within the system [93]. Sheet flow is also uniform throughout the sys-
tem and is shallow [97]. Channelized or channel flow differs from overland flow
because runoff flows through a narrow channel such as a gated terrace or a water-
way. This presents a problem because water flows a velocity that is higher than one
in channelized flow. Channelized flow also requires more land because the strip will
need to be longer to accommodate the channel [96]. Loading into the VFS system is
also inconsistent. As a result, channelized flow includes a reduction in treatment and
an increase in erosion [97].
There are two types of VFS systems—vegetated infiltration basin (VIB) and veg-
etative treatment area (VTA). A VTA system plants vegetation downslope from
crops or livestock housing. On the other hand, VIB is similar to a VTA with the
exception of a berm for runoff collection. Included within the treatment system is
the presence of aerobic bacteria to treat nitrogen by means of nitrification. When
wastewater enters into the VIB system, nutrients are absorbed into the soil and are
used by plants. Runoff is collected through tiles in the system where it is transferred
to other wastewater treatment systems [97].
A VFS system is most effective if it has a depth less than 1.5 ft. In this scenario,
uptake of pollutants by plants is more feasible. Pollutant removal efficiency is also
affected by the length of the VFS—the longer the VFS, the more efficient the treat-
ment [93]. A recommended length for a VFS system is 100 ft or 1 ft/animal unit,
whichever value is greater. However, the ground slope will affect the length of the
system chosen. A 0–2% slope can have a minimum length of 100 ft, while a 6%
slope a minimum of 300 ft [94]. Other recommendations for design include 200 ft
length for a 1-year, 2-h storm, 300 ft for a 0.5% slope to 860 ft for a 4% slope. VFS
treatment system should include a pretreatment step to settle solids from the run-
off [97].
There are many types of vegetation that can be used with a VFS system. The
University of Kentucky Extension states the type of vegetation planted within the
system is contingent on the season. Five plants are suggested—tall fescue, orchard-
grass, timothy, Bermuda grass, and gamagrass. Tall fescue is an option because it is
capable of using nutrients when planted. However, it cannot be used for grazing.
Orchardgrass not only removes nutrients but is capable of being used for grazing,
1 Management and Treatment of Livestock Wastes 53

albeit only up to 4 inches, unlike tall fescue. Timothy grass is a viable option for
horses and cattle to graze provided grazing is limited to 4 inches. Bermuda grass is
a quality choice because it is capable of reducing nutrients and also drought resis-
tant. Bermuda grass can grow up to 8 inches, while grazing is limited to 3–4 inches.
Planting gamagrass will absorb nutrients deep from within the treatment sys-
tem [95].

1.6.3 Summary

In summary, this section presents several feedlot runoff control systems that are
available to divert runoff coming from a feedlot. Feedlot operators must consider
the characteristics of each control system and consult the state legislation in order to
understand what are the design requirements and limitations to use the treatment
method chosen by the feedlot.

1.7 Odors and Gases

1.7.1 Odors: Origin and Nature

Dispersed odors in the air travel and can cause great discomfort for those that live in
close proximity to livestock operations. There are three major causes for odor com-
pounds in livestock operations—“the livestock themselves, animal housing facili-
ties, feedlots, and feed storage facilities; manure storage structures; and application
of livestock manure to agricultural land” [98]. Particular examples include anaero-
bic degradation of organics in manure, feed, and silage. Odors caused by anaerobic
digestion increase in intensity when temperatures are warm. Also if manure becomes
wet, it can also be a major cause for odors [99]. In feedlot operations, incomplete
fermentation of nutrients by bacteria in manure produces odors [100].
Odors can spread in the air as a gas. Dust particles can also be agents to carry
odors. When particles that cause odors come into contact with dust particles, they
are absorbed and carried along. The effectiveness of odors spreading is contingent
on the weather. Very humid days maintain the odors in the area, while dry and
windy days will disperse them [99]. Rainfall can also increase the emission of
odors. If rainwater remains on the ground surface, anaerobic conditions can occur
on the manure [100].
54 D. H. Vanderholm et al.

1.7.2 Sources of Odors

The major sources of odors are gases, anaerobic decomposition of manure, and
other various compounds. The compounds that can provide the biggest issue include
volatile fatty acids, mercaptans, esters, carbonyls, aldehydes, alcohols, ammonia,
and amines [101]. A major proponent of odors is the formation of volatile fatty acids.
The reason why volatile fatty acids cause so many odors is because of the volatile
organic compounds that are present within the manure. Volatile fatty acid presence
within manure varies between animal types. For example, the majority of com-
pounds found in pig manure include acetic, propionic, n-butyric, iso-butyric,
n-­valeric, iso-valeric, n-caproic, and iso-caproic acids. These organic compounds
vary with the amount of carbon atoms present within the system, where butyric,
valeric, and caproic being the highest amount of odor. Other potential dangers for
volatile fatty acids increase toxic pathogens within soil base [102].
One can state that the majority of VFAs have carbon numbers between 2 and 9.
In addition, the presence of Eubacteria, Peptostreptococcus, Bacteroides,
Streptococcus, Escherichia, Megasphaera, Propionibacterium, Lactobacilli, and
Clostridium are also noted for contributing to the major problems associated with
volatile fatty acids [103]. Volatile fatty acids are generated during the process of
fermentation, when carbohydrates are broken down from sugars into pyruvate,
which is then fermented into volatile fatty acids in anaerobic conditions. Therefore,
the lack of aerobic conditions such as incomplete microbial decomposition or other
anaerobic treatment methods are the major cause of this potential issue [98].
Aromatic compounds are a major concern within animal manure due to the pres-
ence of indole, skatole, p-cresol, phenols, and 4-ethylphenol. Under anaerobic con-
ditions, bacteria such as Bifidobacterium, Clostridium, Escherichia, Eubacteria,
and Propionibacterium use aromatic amino acids such as tyrosine, phenylalanine,
and tryptophan [98].
Sulfate-reducing bacteria typically cause the presence of hydrogen sulfide due to
the reduction of amino acids cysteine and methionine. Sulfur-reducing bacteria
typically use sulfate as a terminal electron acceptor transforming sulfate compounds
into hydrogen sulfate. The most common bacteria heavily involved in this process
are Desulfovibrio desulfuricans, Veillonella, Megasphaera, and the enterobacte-
ria [98].
Ammonia emissions causing odor are commonly attributed to ammonia volatil-
ization. The reason behind such a problem can be traced back to the animal species,
diet, and age. For example, urea, the nitrogen compound within urine, typically
forms ammonium and bicarbonate ions by means of urease enzymes. Nitrogen
found in feces is broken down by bacteria, where it transfers from proteins to amino
acids and eventually into ammonium. The time in which this occurs depends on the
temperature, concentration, and pH [104].
One of the more common entities that is emitted through livestock waste is the
presence of hydrogen sulfide. Hydrogen sulfate odor emissions commonly occur
from the anaerobic decomposition of sulfur [105]. One of the most common
1 Management and Treatment of Livestock Wastes 55

methods of forming hydrogen sulfate is due to the efforts of sulfate-reducing bacte-

ria [106].

1.7.3 Odor Prevention

There are various methods to prevent the spreading of odors. These can include
animal nutrition management, manure treatment and handling, waste treatment
methods, and better livestock operation management. Tables 1.17 and 1.18 provide
various methods to mitigate odors.

1.7.3.1 Animal Nutrition Management

One of the best ways to reduce odors is to alter animal nutrition. If livestock feed
contains more crude protein concentration or blood meal, it will lead to the produc-
tion of odors. Studies have shown that feeding livestock crystalline amino acids or
peppermint as compared to a diet heavy with crude protein can reduce odorous
manure. Barley-based diets can also reduce odors by 25% as compared with a diet
dominated by sorghum [50, 107]. Fecal starches, proteins, and lipids should be
eliminated as much as possible. This will prevent incomplete fermentation, which is
the main cause of odors [100].
In addition to changing the diet of the animals, the operator should consider a
change in feeding schedule. An appropriate feeding schedule could be feeding the
animals at sunrise, noon, and sunset. This can not only eliminate the presence of
odors but also control the emission of dust in the atmosphere from cattle that move

Table 1.17 Odor emission strategies for livestock housing [50]

Method Description
Filtration and  1. Filtration traps 45% 5–10 μm particles; 40–70% particles greater
biofiltration than 10 μm
Biofilters  1. Biofilters trap and biologically degrade particles; remove odorous
emissions
 2. Biofilters can remove 90% odors, including 90% hydrogen sulfide
and 74% ammonia
Impermeable barriers  1. Dust particles retain odors preventing movement
 2. Impermeable barriers such as windbreak walls or dams are very
effective
Oil sprinkling  1. Application of vegetable oil can control dust movement
 2. Study applying oil reduced hydrogen sulfide concentrations by
40–60%
Landscaping  1. Application of trees and shrubs
 2. Landscaping reduces particulate movement and inserts dilute the
concentration of emissions
56 D. H. Vanderholm et al.

Table 1.18 Examples of odor emission strategies for manure storage [50]
Method Description
Solid separation  1. Removal of large materials, typically the size of a screen opening
 2. Removal of large material reduces the loading rates, thereby producing
less odors during decomposition of remaining material
 3. Solid separation uses processes such as sedimentation, screening,
filtration, or centrifugation
Anaerobic  1. Under anaerobic conditions, odors are biologically reduced from
digestion manure
 2. Anaerobic digestion encapsulates manure maintaining odors
Additives  3. Application of additional enzymes or chemicals to dilute manure under
anaerobic conditions
Impermeable  1. Coverage of a manure storage area will control odors from gases
cover  2. Impermeable covers can control wind and radiation
Permeable  1. Coverage of a manure storage area to control the contact between
covers manure and radiation and wind velocity
 2. Emission rates are reduced
 3. Permeable covers create an aerobic zone, encouraging aerobic
microorganism growth
Aeration  1. Application of oxygen by mechanical means to maintain aerobic
conditions
 2. Aeration can cause an increase in ammonia emissions
Composting  1. Composting provides an aerobic environment reducing the creation of
odors
 2. A more viable option for those that handle solid manure because of
high maintenance required to maintain suitable decomposition
conditions

their hooves on the ground. As a reminder, dust can be used as an agent to transfer
odors [100].

1.7.3.2 Manure Treatment and Handling

Another method for reducing odors is to consider the treatment and handling of
manure. First, operators can incorporate additives to manure. Additives can be
chemical or biological. Additives can be applied to overpower the presence of an
odor, reduce the ability for odors to be smelt, absorb constituents in manure that
cause odors, or slow microbial degradation to reduce odors [101]. Choices for addi-
tives are based on the product and the rate and frequency of application [50]. Manure
can also be chemically treated. The University of Arkansas Extension recommends
several options for chemical treatment. These include sodium bisulfate (PLT), ferric
sulfate granular (Ferric-3), alum, and zeolite [107].
Next, solid separation can be used to better hand manure. Solid separation pro-
cesses include sedimentation, screening, filtration, and centrifugation. This process
attempts to remove constituents that cannot pass through a specified screen size.
The removal of these materials decreases biological degradation and thereby reduces
1 Management and Treatment of Livestock Wastes 57

odors [50]. Solid separation also reduces odors by reducing the organic loading.
Usually solids are separated before entering a treatment basin such as a lagoon.
Some of the materials removed include cattle waste fiber and grit. There are several
machines employed for solid separation. These include vibrating screens, sloping
stationary screens, or pressure-rolling mechanical separator. Solid separation can
occur within a gravity settling basin, earthen settling basin, rectangular metallic, or
a concrete settling tank [49].
Finally, operators can make strategic choices in how they apply manure to land
for the sole purpose of preventing the spread of odors. Spreading manure can be
done in the morning or when the sun is present and on days when the direction of
the wind is away from the neighbors [101]. Manure can also be applied during the
early evening for better wind dispersion [50]. It is best for the livestock operators to
choose the weekdays as opposed to weekends when neighbors will most likely not
be at home [107]. When manure is applied, it should be applied quickly, in large
quantities, and based solely on the needs of the crop [50]. Operations should employ
a liquid waste management schedule [107].
If liquid manure is applied by irrigation equipment, operators can make choices
on nozzle size of the sprayers. An alternative would be using a low-rise, low-­
pressure, trickling system. Application of liquid manure should be done in close
range to avoid the spread of odors [50]. Instead of the land application of manure by
irrigation, operators can also make the decision to inject manure directly into soils
as compared to choosing surface application [107].
When solid manure is not directly applied, operators can select to cover the
manure before use. There are two types of covers—impermeable and permeable.
Impermeable covers prevent manure storage facilities from the emission of odors
into the atmosphere. The covers can also reduce the effects of wind and radiation.
Impermeable covers can reduce odors by 90%. Cover efficiency is contingent on the
presence of wind and snow [50].
On the other hand, permeable covers (biocovers) are used to cover places for
anaerobic digesters or manure storage facilities [50, 107]. Biocovers can consist of
straw, cornstalks, peat moss, foam geotextile fabric, or Leka rock [50]. Biocovers
can also include use closed-cell polyurethane foam with or without zeolite.
Biocovers remove radiation from the surface of the manure storage facility and also
reduce the impact of the wind blowing [107]. Biocovers contain an aerobic zone
where aerobic microorganisms thrive on the presence of chemical constituents
within the manure. These microorganisms reduced the odors. The reduction of
odors is contingent upon the material used. Covers that are primarily made of straw
reduce odors by 50%, while 85% of odors are reduced when the cover consists of a
floating mat or corrugated materials [50].
As an alternative to biocovers, manure storage facilities can be aerated to supply
molecular oxygen. This will assist in reducing odors. Nevertheless, aeration can be
dangerous because nitrogen is volatilized into the atmosphere as ammonia.
Therefore, great care should be taken to prevent this from occurring [50].
58 D. H. Vanderholm et al.

1.7.3.3 Waste Treatment Methods

There are many waste treatment methods that can reduce the potential of creating
odors. First, operators can install filters to separate odor-causing particles within the
air. There are two potential filters available—mechanical and biofilters. Mechanical
filtration devices are capable of removing odors from particles. There are indica-
tions that 45% of odors are caused by particles with a size between 5 and 10 μm,
while 80% are caused by particles greater than 10 μm. Mechanical filtration has
been proven to reduce odors between 40 and 70% [50].
Biofilters capture particles where aerobic bacteria degrade them to create prod-
ucts that do not cause odors [50]. Biofilters are supplied air by natural ventilation.
The presence of air and adequate environmental conditions allows for the bacteria
to grow within the system [99]. Bacteria grow on media consisting of wood chips or
compost [107]. For these reasons, biofilters are inexpensive as compared to mechan-
ical filtration. Efficiency of a biofilter is contingent on oxygen concentration, tem-
perature, residence time, and moisture content [50]. The design of biofilters is
contingent on the volume of air needed to be treated [107]. Biofilters have been
successful in removing 40% of hydrogen sulfide [50]. It has also been reported that
biofilters remove 90% of odors [107]. Biofilters are also capable of filtering odor-­
causing liquids from manure storage [99].
By means of Rockwool packing material, Yasuda et al. (2010) was able to pro-
duce 8.2–12.2 mg N/100 g sample of nitrification and 1.42–4.69 mg N/100 g of
denitrification [108]. Ro et al. (2008) found that a polyvinyl alcohol (PVA)-powered
activated carbon biofilter removed 80% ammonia-nitrogen with hydrogen sulfide
removal at 97% [109]. Kastner et al. (2004) found that ammonia-nitrogen concen-
tration ranging between 25 and 95% was removed in waste from swine production,
where the major factors that depended on the treatment efficiency were residence
inlet time and ammonia concentration [110].
Second, anaerobic digestion is a feasible treatment method to reduce odors. The
biological degradation of constituents under anaerobic conditions can reduce the
odors significantly in organic material. The products from anaerobic digestion can
be safely placed in a liquid storage facility [99]. A study using anaerobic digestion
for degradation of dairy waste reported a 50% reduction in odors provided the waste
remained in the digester for 20 days. While anaerobic digestion is an expensive
method, it can be viable for some operators [50]. Anaerobic digestion can profitable
as it produces biogas [99]. More information about anaerobic digestion is presented
in Sect. 1.2 of this chapter.
Various enzymes such as peroxidase, specifically horseradish peroxidase (HRP)
and trosinate [111], are used to control odors. Horseradish peroxidase (HRP) has
become a new method in research for deodorization because of the quantity of per-
oxidase within the plant, which is capable of transforming aromatic compounds into
free radicals or quinones, which ultimately form non-odor compounds [112].
Govere et al. (2007) experimented with pilot-scale reactors with volumes
between 20 and 120 L using
1 Management and Treatment of Livestock Wastes 59

minced horseradish comparing effectiveness between the addition of either cal-

cium peroxide or hydrogen peroxide to deodorize swine wastewater. From the
results, it was determined that the addition of horseradish was capable of com-
pletely removing odors [112].
The management of lagoons serves as a way of reducing odors. A healthy lagoon
will degrade organic materials into constituents that do not produce odors. Odors
can be reduced in a lagoon if manure contains a dilution of 1–2%. Lagoons should
also be refrained from having a high solid concentration. When high solids are pres-
ent, a lagoon is overloaded. Overloaded lagoons change the conditions from aerobic
to anaerobic, thereby creating odors [47, 99].

1.7.3.4 Livestock Operations Management

Livestock operators can mitigate the spread of odors by providing better manage-
ment of the buildings and facilities. This can include disposing unused or even
moldy feed, fix leaks and if necessary replace or repair pipes, and designate a loca-
tion to dispose dead animals. Another alternative is to increase ventilation within
these areas. Ventilation can be supplied by mechanical or natural means. Mechanical
methods of ventilation include fans and fresh air inlets. If cost is a barrier, an alter-
native is to use natural methods. Openings, change in roof slope, and rearranging
the orientation of the building are ways that a livestock operator can generate natu-
ral ventilation within a building or facility. Despite the fact that it saves energy,
natural ventilation may be inhibited by environmental circumstances, so the opera-
tor should make a wise decision on which method should be chosen [101].
In addition to ventilation, livestock operators can introduce landscape onto the
premises to contain odors. Landscaping provides an opportunity to prevent the con-
stituents that cause odors from further leaving the operation. These constituents are
either dispersed or diluted. Landscaping also gives an aesthetic appeal to the area.
Trees and shrubs are the two most common entities planted [50].
The design and maintenance of feedlot pens should be reviewed to better prevent
odor mitigation. Feedlot pens should maintain a dry surface to prevent the formation
of anaerobic conditions on the surface. This means that each pen should be designed
to have proper drainage. Having pens maintain a slope between 4 and 6% will pro-
vide adequate drainage and prevent pens from accumulating standing water. Also,
pen scraping should occur once every 3–4 months [100].

1.7.3.5 Summary

With many people leaving municipalities and inner-ring suburbs for rural and farm-
land communities, the discussion on odor mitigation will continue to increase.
Therefore, it is important for livestock operators to develop good relationships with
the residents living in close proximity to livestock operation facilities. Regardless of
60 D. H. Vanderholm et al.

the method(s) chosen, the ultimate goal should be to provide neighbors the ability
to feel as liberated as possible from the presence of odors.

1.7.4 Greenhouse Gas Emissions

Recent developments have discussed the relationship between greenhouse gas emis-
sions and livestock. This chapter will discuss some of the current issues related to
the relationship between greenhouse gas and livestock waste. The purpose of dis-
cussion is not to take sides but rather present what is currently found in literature.
Greenhouse gases consist of carbon dioxide (CO2), methane (CH4), and nitrous
oxide (N2O). Carbon dioxide is considered a primary greenhouse gas because in
general only 9% of greenhouse gas emissions are caused by CH4 and N2O [113].
However, in the livestock sector, CH4 production is 21 times the carbon dioxide,
while N2O 310 times the CO2 emissions. This is because animals produce methane
during the process of enteric fermentation, while nitrous oxides are formed during
the degradation of manure when nitrification and denitrification occurs. In general,
greenhouse gases maintain the temperature of the Earth to 15 °C. The current debate
with greenhouse gases involves global warming and climate change. This debate
has been whether or not greenhouse gases cause a change in climate [114]. It was
reported that from 2001 to 2010, greenhouse gas emissions from crop and livestock
operations increased by 14% [113]. In 2012, it was estimated that the agriculture
industry released 526 million metric tons of carbon dioxide equivalent (MMT of
CO2e) plus 62 MMT of CO2e related to operating electric products [114].
According to the USEPA, greenhouse gases have caused 9% of the total green-
house gas emissions in the United States, while the United Nations (UN) have stated
18% of global emissions have been caused by greenhouse gases. There are many
sources of greenhouse gases reported. The United Nations mentions that green-
house gas emissions are caused by livestock feeding, manure management, live-
stock processing, and transportation of livestock products. On the contrary, the
USEPA states that greenhouse gases have affected crop and livestock production.
Other sources have stated that deforestation (34%) and ruminant digestion (25%)
are additional factors that must be considered [114].
According to the University of Missouri Extension’s paper titled “Agriculture
and Greenhouse Gas emissions,” there are four major areas that have been major
contributors to greenhouse gases in the agriculture sector—crop and soil manage-
ment, livestock manure management, enteric fermentation, and agricultural carbon
sequestration. These values are contingent on the US production of greenhouse
gases in 2012, data produced by the USEPA [114].
1. Crop and soil management. Agricultural crop and soil management produced
307 MMT of CO2e or 48% of the total greenhouse gas emissions within the agri-
cultural sector. Ninety-eight percent of all emissions from greenhouse gas were
because of N2O. This has been attributed to the fact that cropland has produced
1 Management and Treatment of Livestock Wastes 61

more N2O than lands that are grasslands. In addition, fertilization, manure appli-
cation, crop residue collection, nitrogen-fixed crops and forage, and soils with
organic materials are major practices that lead to N2O emissions. N2O emissions
occur in the Corn Belt, cropped land in California and the Mississippi Valley, rice
production, and burnt fields.
2. Livestock manure management. Manure management accounted for 71 MMT
of CO2e in greenhouse gas emissions. Most of the greenhouse gases produced in
livestock manure are CH4. The major causes of greenhouse gases include anaero-
bic decomposition of liquids and slurry. N2O in manure management is caused
by manure, urine, and aerobic and anaerobic degradation. The dairy cattle indus-
try produced 47% of CO2 emissions, while the beef cattle industry was respon-
sible for 71% of CH4.
3. Enteric fermentation. As previously stated, enteric fermentation causes the
majority of CH4 emissions. Enteric fermentation produced a greenhouse gas
total of 141 MMT of CO2e. Varying factors determine the production of enteric
fermentation. These include the number of livestock and the type of feed.
4. Agricultural Carbon Sequestration. Land use and forestry was responsible for
979 MMT of CO2e or 15% of overall greenhouse gas emissions. A relationship
between land use and carbon sequestration was made. This relationship analyzed
the carbon sequestration of land in 2012 and its state 20 years before. Land that
remained grassland was capable of sequestering carbon where loses only
occurred because of drought. This has also been the case when land was con-
verted into grasslands. On the contrary, land that remained cropland or converted
into cropland carbon was not sequestered. However, land that remained cropland
was able to sequester carbon provided the organic content remained between
1 and 6%.
A more recent study was completed by Caro et al. to assess the global greenhouse
gas emissions between 1961 and 2010. Analysis compared the livestock greenhouse
gas emissions between developing and developed countries. The results from the
study concluded that global greenhouse gas emissions increased by 51%, where the
primary source of greenhouse gas emissions was caused by enteric fermentation. In
general, the generation of greenhouse gases decreased overall. However, there was
a difference in the trends for developing and developed countries. Greenhouse gas
emissions in developed countries increased in the 1970s and then gradually
decreased by 23%. On the contrary, greenhouse gas emissions increased in develop-
ing countries by 117%. The authors attributed increase to changes in economic and
ideological changes. The signature year for these changes occurred in 1989. These
countries transitioned from being focusing heavy on importing to exporting. With
regard to the various livestock industries, the beef cattle industry was accountable
for 54% of greenhouse gases, while only 17% was due to the dairy industry [113].
The development of numbers has created an interesting stir within the scientific
community. Various authors have published papers that attempt to support the val-
ues generated by entities recognizing global climate change (e.g. USEPA, UN, and
the International Panel on Climate Change (IPCC)). However, authors such as
62 D. H. Vanderholm et al.

Herrero et al. request for a reduction in ambiguity and more consistency in method-
ologies used to quantify greenhouse gas emissions within livestock. The areas of
concern includes the exclusion of CO2 production by livestock, quantifying emis-
sions due to land use and land change, global warming potential of methane, and the
overall allocation of processes to livestock. With a more accurate picture, the authors
state that the discussion of greenhouse gas emissions in livestock can improve
[115]. Regardless of an individual’s stance on greenhouse gas emissions and global
warning, the discussion of the livestock industry’s role in greenhouse gas emission
will continue.

1.8 Pathogens in Livestock Industries

Pathogens are an issue within the livestock industry. The impact from pathogenic
outbreak cause a loss in productivity for the livestock operation by becoming detri-
mental to the animals, the business, and employees. Pathogens can also be harmful
to the public and the environment. Survival of pathogens is predicated on the tem-
peratures, the pH, the amount of microbial activity, the routes of transfer, and the
applicable host. The routes of transfer for pathogens include fecal-to-oral, food-­
borne, aerosol, or human-to-animal contact. The applicable hosts can range from
humans, farm animals, and other carriers such as flies. There are four major catego-
ries of pathogens—viruses, bacteria, mycotic agents, and parasites [62].
For example, contact with viruses for a period of time can lead to illness or death
and can limit the product from livestock. Viruses are classified as enveloped and
non-enveloped viruses. Enveloped viruses persist within animal manure and can
stay for a long period of time without treatment and storage, while non-enveloped
viruses are incapable of being destroyed with any treatment method. On the other
hand, mycotic agents are not a major concern within the livestock industry and are
usually dangerous in soils or self-contained with the body of an animal or human
[62]. Examples of each pathogen category are listed in Table 1.19.
Livestock operators can know the quantity of pathogens within its waste by using
organisms known as fecal indicator organisms. Fecal indicator organisms are sur-
rogate organisms used in the laboratory as a method for quantifying pathogenic
presence. Typically, E. coli has been used as a fecal indicator organism, but recent
studies have used other organisms such as coliphages and C. perfringens spores. An
adequate choice for a fecal indicator organism must fulfill a series of criteria. Fecal
indicator organisms must:
1. Exist in the same conditions as pathogen.
2. Have a life span similar to pathogens.
3. Withstand disinfectants and unfavorable conditions.
4. Be easily detectable.
5. Be distributed randomly.
6. Portray similar risks in humans as pathogens.
1 Management and Treatment of Livestock Wastes 63

Table 1.19 Examples of each type of pathogen [122]

Pathogen Example
Viruses Animal enteroviruses, rotaviruses, hepatitis E
Bacteria Aeromonas hydrophila
Aerobacter
Bacillus anthracis
Chlamydia
E. coli
Salmonella
Mycotic agent Histoplasmosis capsulatum
Pneumocystis carinii
Parasites Protozoa
Ascaris and Ascariasis
Cryptosporidium parvum
Giardia
Toxoplasmosis

Table 1.20 Pathogen treatment methods [122]

Pathogen Method
Dry techniques Composting
Physical treatment Sand filtration or dry beds
Sedimentation and screening
Biological treatment Lagoon
Anaerobic digestion
Sequencing batch reactor
Constructed wetlands
Overland flow
Disinfection Chlorine
Ozone
Chlorine dioxide
Lime stabilization
Pasteurization

As an alternative, testing for microorganisms can include culture-specific microor-

ganisms, antibiotic resistance patterns, molecular fingerprinting, genotype, and
chemical indicators [62].
There are various treatment methods that can be used to reduce the pathogens
within livestock waste. The treatment of livestock waste can use dry techniques,
physical treatment, biological treatment, and chemical treatment. Examples of treat-
ment techniques found within each category are shown in Table 1.20. Many of these
methods have been discussed in grave detail in the previous sections [62].
The presence of pathogens can have a major impact on livestock operations.
While this section is not extensive, it does attempt to provide a summary of major
pathogen categories, their associated impacts, and the potential treatment methods.
64 D. H. Vanderholm et al.

1.9 Livestock Waste Management Computer Software

Within the recent century transition, there has been the presence of computer mod-
eling tools that are capable of being used to predict livestock wastes. For example,
the Animal Waste Management Software Tool (AWM) is a computer program
designed to determine parameters such as waste storage facilities, waste treatment
lagoons, and utilization [2]. Other options include the collaboration between the
University of South Carolina’s Earth Science and Resource Institute and the Natural
Resources Conservation Service (NCRS) in South Carolina to develop a suite of
products that include the geospatial tools [ArcGIS] and a nutrient management
planning software AFOPro© [116].
Ideas on the use of software for livestock waste management have not been lim-
ited to just the United States. A program known as Integrated Swine Manure
Management (ISMM) is an integrated decision support system (DSSs) used by
Canadian province decision-makers to control manure, considering various criteria
such as environmental, agronomic, social and health, greenhouse gas emission, and
economic factors [117]. The introduction of computer software for livestock man-
agement can be very significant for those that are planning to provide a consistent
method of managing livestock. Nevertheless, it is still important to remember that
computer software is a “tool” but does not replace proper education and understand-
ing of what is needed for proper livestock waste within the given area.

1.10 Recent Advances in Livestock Waste Treatment

and Management

1.10.1 Latest Technology Development, Market-Driven

Strategies, and US Policy Changes

In the United States, the major hurdles to reducing the impact of livestock waste
pollution on the nation’s watersheds are outdated American wastewater treatment
policies. Under the prevailing US legislation, the 1972 Clean Water Act (CWA), the
majority of wastewater treatment efforts have targeted “point sources of water pol-
lution” with a measurable wastewater discharge. The CWA defines point sources as
discharge pipes from industrial plants, utilities, or municipal sewage treatment facil-
ities. Many new environmental process technologies, such as improved chemical
coagulation/precipitation, clarification (dissolved air flotation and improved set-
tlers), filtration, membrane bioreactor, advanced oxidation processes, etc., have
been developed for water pollution control [118–130], but have not been seriously
considered for agricultural waste treatment.
Agricultural wastes, such as livestock manure, farm’s storm runoff water, etc.,
are considered the non-point sources of water pollution, and are not subject to CWA
regulations. In the nearly one half of a century since the passage of the CWA, the
1 Management and Treatment of Livestock Wastes 65

American agricultural industry has grown considerably. More than 70% of today’s
livestock production takes place not on small-scale family farms but on large-scale
Concentrated Animal Feeding Operations (CAFO) facilities. However, CAFOs still
use small-farm strategies for disposing of animal waste, and about half the crops in
the United States are fertilized this way. An ineffective waste strategy, coupled with
little meaningful regulation, poses a major hurdle for the rehabilitation of US water-
sheds. The agricultural water pollution problem must be dealt with its original
source. It is the opinion of Director Craig Scott of Bion Environmental Technologies
that spending billions of dollars to upgrade downstream wastewater treatment plants
and to construct large-scale stormwater projects that recollect and treat the nutrients
after they have been released to contaminate the environment is not an acceptable
solution from either a cost or a common sense perspective [127–130]. The new
market-driven strategies are treating the CAFO wastes with the best available tech-
nologies (BAT) and still considering both technical and economical feasibilities.
There are clear signs that the US Federal Government will provide funding for
nutrient control and climate control strategies and private sector solutions. In
December 2018, the USEPA and the US Department of Agriculture (USDA) noti-
fied state and tribal regulators that they are committed to working with all stake-
holders to adopt market-based approaches in the fight to clean up America’s
watersheds and prevent livestock waste from further contributing to the crisis. The
agencies said this commitment could include technical and financial support for
water quality credit trading programs and public-private partnerships [127–130].
In January 2019, former US President Donald Trump signed bipartisan legisla-
tion for federal funding to combat toxic algae blooms in the country’s water
resources. In February 2019, the USEPA issued a memorandum updating its water
quality trading policy and supporting market-based approaches to reduce nutrient
pollution in the nation’s waterways. The announcement stated “USEPA efforts seek
to modernize the agency’s water quality trading policies to leverage emerging tech-
nologies and facilitate broader adoption of market-based programs.”
There is further proof that under the leadership of US President Joseph R. Biden,
the US federal policymakers are serious about building the nation’s infrastructure
(including water and waste treatment). Controlling global warming, climate change,
and greenhouse gases are all on the horizon.

1.10.2 Livestock Water Recycling (LWR) System

Livestock Water Recycling (LWR) is one of the world’s leading providers of manure
treatment technology aiming at reducing greenhouse gas emissions, concentrating
and segregating nutrients for strategic fertilizer application, and recycling clean,
reusable water.
The LWR system is a proven and fully operating technology that reduces the
overall volume of manure, concentrates nutrients, and delivers a renewable, high-­
quality water source. According to the manufacturer, the company’s vision has
66 D. H. Vanderholm et al.

always been to help livestock farmers increase farm efficiencies while becoming
even more environmentally sustainable, and its LWR system provides a minimum
of 20–30% return on investment [118].
The LWR company is focused on developing scalable solutions that can be
applied quickly and commercialized for maximum return on investment.
LWR system is a patented process technology that uses both mechanical and
chemical treatments to remove manure contaminants and segregate valuable fertil-
izer nutrients at large livestock operations. Figures 1.23 and 1.24 show the LWR
system’s process flow diagram and process equipment, respectively [118]. As the
livestock manure effluent flows through the LWR process, solids are sequentially
removed by chemical precipitation, clarification, conventional filtration, and mem-
brane filtration. The result is valuable segregated fertilizer nutrients and clean water
that can be reused around the barns.
LWR system uses both mechanical and chemical treatments to remove manure
contaminants and segregate valuable fertilizer nutrients at large livestock operations.
Figures 1.23 and 1.24 show the LWR system’s process flow diagram and process
equipment, respectively. As the livestock manure effluent flows through the LWR
process, solids are sequentially removed by chemical coagulation/precipitation,
clarification, conventional filtration, and membrane filtration. The result is valuable
segregated fertilizer nutrients and clean water that can be reused around the barns.
The detailed process, descriptions, principles, design criteria, operational proce-
dures, terminologies, etc. of each unit process (chemical precipitation, clarification,
conventional filtration, membrane filtration, etc.) can be found in the literature
[119–126]. Either sedimentation or flotation can be used for clarification [119, 122].
The nutrient and water recovery capacity of the LWR system is so far the highest
on the market. LWR system extracts up to 75% of the water from livestock manure
while concentrating dry solids (8%) and segregating nutrients for recycling (17%).
By concentrating and segregating, the farm plant managers are given more control
over their nutrient application, which minimizes their farm’s field work. The result
is clean, potable water, dry solids that are rich in both phosphorus and organic nitro-
gen and a concentrated stable ammonium and potassium liquid. At present, LWR
system has the highest nutrient and water recovery capacity on the market, lowest

Fig. 1.23 Flow diagram of a Livestock Water Recycling (LWR) system [118]
1 Management and Treatment of Livestock Wastes 67

Fig. 1.24 Process equipment of a Livestock Water Recycling (LWR) system [118]

electrical consumption on the market, and highest number of installations in the

water and nutrient recovery market, which are all incredible.

1.10.3 BET Advanced Technologies To Benefit

From Policy Changes

There are a few commercial-ready technologies available today that can address the
problem of excess nutrient runoff from large-scale agricultural operations. Section
1.10.2 has introduced the Livestock Water Recycling (LWR) system, which is now
commercially available for livestock waste treatment.
Another advanced technology in the sector is Bion Environmental Technologies’
comprehensive environmental management system, which is also designed for the
largest CAFO livestock facilities and focused on maximizing resource recovery.
Bion Environmental Technologies’ patented 2G (second-generation) technology
has been commercially proven to substantially reduce pathogens from livestock
waste while eliminating up to 90% of greenhouse gases and ammonia emissions and
95% of nitrogen and phosphorus. The waste management system harnesses the
power of naturally occurring bacteria to convert nitrogen and phosphorus into solid
forms that are removable by other processes [127–130]. Figure 1.25 shows the flow
diagram of Bion Environmental Technologies’ comprehensive environmental man-
agement system.
Livestock waste treatment technology not only provides clean water solutions
but also creates new sources of revenue, including the production of value-added
products such as fertilizers. Bion’s patented 3G technology recovers stable concen-
trated ammonium bicarbonate, a quick-release nitrogen fertilizer, from livestock
waste without the use of chemicals. This product is well suited for a wide range of
applications in the organic markets. According to Markets and Markets researchers,
the market for global organic fertilizers is expected to grow from US$6.3 billion in
2017 to US$11.15 billion by 2022.
In 2019, Bion plans to apply to the USDA’s Organic Materials Review Institute
for use of its ammonium bicarbonate product in organic food production. The com-
pany has already applied for a Patent Cooperation Treaty for international recogni-
tion of its ammonium bicarbonate production process.
68 D. H. Vanderholm et al.

Fig. 1.25 Flow diagram of Bion’s livestock waste treatment technology

1.11 Conclusion

This chapter provides a plethora of information concerning livestock waste manage-

ment from treatment, handling, and storage. While this not an all-encompassing
manual for all given conditions, it can be used as a catalyst for research and explora-
tion in how to properly maintain and manage livestock waste for a given industry.
The readers are referred to the literature [131–135] for additional technical informa-
tion on treating the livestock’s biosolids, concentrated liquid waste stream, or
diluted liquid waste stream.

Glossary of Livestock Waste Management

Anaerobic digestion is the fermentation of organic waste by hydrolytic micro-

organisms into fatty acid chains, carbon dioxide (CO2), and hydrogen (H2).
Short fatty acids are then converted into acetic acid (CH3COOH), H2, CO2, and
microorganisms.
Biogas is a product from anaerobic digestion containing gases such as methane
(CH4), CO2, and trace elements. Biogas can be used as a source of energy.
Chemical oxygen demand (COD) is a wastewater quality index that determines
the amount of oxygen consumed by wastes.
Concentration animal feeding operations (CAFO) raises livestock within a
restricted space. It is also known as feedlot.
1 Management and Treatment of Livestock Wastes 69

Constructed wetland is a treatment method that uses plants (most commonly

water hyacinth and duckweed) to degrade organic material.
Denitrification converts nitrate into atmospheric nitrogen using microorganisms
known as dentrifiers.
Eutrophication is the condition of a water body (particularly a lake) where molec-
ular oxygen levels have been depleted. Eutrophication most commonly occurs
when nutrient levels are high within the water body, forming the presence of
algal blooms. When eutrophication occurs, all organisms rely on molecular oxy-
gen to survive.
Five-day biochemical oxygen demand (BOD5) is a wastewater quality index
that determines the amount of oxygen required for microorganisms to degrade a
given substance within a 5-day period.
Lagoon is a basin that treats wastewater and stores waste. There are three major
types of lagoons—anaerobic, aerobic, and facultative.
Liquid manure contains dry matter less than 5%.
Mesophilic is a state in an anaerobic digester or composting when the temperature
remains between 35 and 40 °C.
National Pollution Discharge Elimination System (NDPES) regulates the quan-
tity of waste entering navigable waters and point sources. It was first introduced
by the USEPA in the Clean Water Act of 1977. Livestock waste operations are
required to have NPDES permits to discharge. State legislation defines the opera-
tions that require NPDES permit.
Nitrification is the process of converting ammonium nitrogen (NH4+) into nitrate
(NO32−) with an intermediate step of producing nitrite (NO2−). Nitrification is
converted by nitrogen-fixing bacteria (nitrifiers).
Psychrophilic is a state in an anaerobic digester or composting when the tempera-
ture remains below 20 °C.
Semisolid manure contains 5–10% dry matter.
Solid manure contains dry matter greater than 15%.
Thermophilic is a state in an anaerobic digester or composting when the tempera-
ture remains between 51 and 57 °C.
Volatilization is a phase change process that converts constituents into gaseous
form. The most common volatilization experienced is ammonia volatilization or
the conversion of ammonium-nitrogen to ammonia-nitrogen. This is problematic
for livestock operations because plants’ nitrogen is lost for plant uptake.

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culture. Biotech Investing News. Retrieved from https://investingnews.com/innspired/
livestock-­waste-­treatment-­technology-­agriculture/.
128. Bion. (2021). Environmentally sustainable livestock production. Bion Environmental
Technologies, Inc., 9 East Park Court, Old Bethpage, NY. [email protected].
129. Bion. (2021, June 28). Bion files international patent applications on third generation live-
stock waste treatment technology. Bion Environmental Technologies, Inc., Old Bethpage,
NY. [email protected].
130. Bion. (2021, April 30). Bion announces letter of intent for commercial-scale third generation
project. Bion Environmental Technologies, Inc., Old Bethpage, NY. [email protected].
131. Wang, L. K., Wang, M. H. S., Cardenas, R. R., Sabiani, N. H. M., Yusoff, M. S., Hassan,
S. H., Kamaruddin, M. A., Fadugba, O. G., & Hung, Y. T. (2021). Composting processes
for disposal of municipal and agricultural solid wastes. In L. K. Wang, M. H. S. Wang, &
Y. T. Hung (Eds.), H. A. Aziz (Consul. Ed.), Solid waste engineering and management (Vol.
1, pp. 399–524). Springer Nature Switzerland.
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to solid waste management. In L. K. Wang, M. H. S. Wang, & Y. T. Hung (Eds.), H. A. Aziz
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landfill gas management. In L. K. Wang, M. H. S. Wang, & Y. T. Hung (Eds.),, H. A. Aziz
(Consul. Ed.), Solid waste engineering and management (Vol. 3, pp. 583–614), Springer
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134. Wang, L. K., Wang, M. H. S., & Shammas, N. K. (2022). Agricultural waste treatment by
water hyacinth aquaculture, wetland aquaculture, evapotranspiration, rapid rate land treat-
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& Y. T. Hung (Eds.), Waste treatment in the biotechnology, agricultural and food industries
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Chapter 2
Waste Treatment in the Pharmaceutical
Biotechnology Industry Using Green
Environmental Technologies

Lawrence K. Wang, Mu-Hao Sung Wang, Nazih K. Shammas,

and Ping Wang

Nomenclature

k Maximum substrate utilization rate

Ki The partitioning coefficient, also called the vapor-liquid equilibrium
constant
Ks Half saturation constant
P Total pressure
Pi Vapor pressure of the pure substance at the operating temperature
ri Activity coefficient of organic compound i in the wastewater at a certain
temperature
Vi Mole fraction of organic compound i in the vapor phase
Wi Mole fraction of organic compound i in the wastewater phase

L. K. Wang (*)
Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
e-mail: [email protected]
M.-H. S. Wang · N. K. Shammas
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]
P. Wang
Civil and Environmental Engineering Department, Rensselaer Polytechnic Institute, Troy,
NY, USA
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 79

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_2
80 L. K. Wang et al.

2.1 Introduction to Biotechnology

2.1.1 Pharmaceutical Industry
and Biotechnology Terminologies

Pharmaceutical industry is an industry responsible for manufacturing of drugs, vac-

cines, antibiotics, etc. using chemical reactors, biological systems, or organisms.
The chemical synthesis-based pharmaceutical industry is a part of chemical indus-
try using chemical technology and chemical reactors, while the fermentation pro-
cess-based pharmaceutical industry is a part of biotechnology industry using
biological systems or organisms in biochemical reactors.
Biotechnology is an engineering science field involving the use of biological
systems found in organisms or the use of the living organisms themselves to make
scientific advances and adapt those knowledge to various application branches, such
as medical biotechnology, agricultural biotechnology, industrial biotechnology,
environmental biotechnology, computational biotechnology, and military
biotechnology.
Medical biotechnology (including pharmaceutical biotechnology) involves the
use of living cells and other cell materials to find cures for preventing diseases and
bettering the health of humans; development of vaccines and antibiotics is a typical
example. Specific pharmaceutical biotechnology related to medicine and veterinary
products (vaccines, antibiotics, molecular diagnostics techniques, genetic engineer-
ing techniques, etc.) is also termed red biotechnology.
Agricultural biotechnology focuses on developing genetically modified plants to
increase crop yields or introduce characteristics to those plants that provide them with
an advantage growing in regions that place some kind of stress factor on the plant,
namely, weather and pests. Development of pest-resistant crops and improvement of
plant and animal breeding are typical examples. Green biotechnology refers to spe-
cific agricultural biotechnology that creates new plant varieties of agricultural interest,
biopesticides, biofertilizers, etc. This area of agricultural biotechnology is based on
transgenics (genetic modification), i.e., an extra gene or genes inserted into their
DNA. The additional gene may come from the same species or a different species.
Industrial biotechnology (including industrial fermentation biotechnology)
involves the utilization of cells, such as microorganisms, or components of cells,
like enzymes, to generate products in sectors that are industrially useful, such as
food and feed, chemicals, detergents, paper and pulp, textiles, biofuels, and biogas,
or to create genetically modified organisms (GMOs) that enhance the diversity of
applications and the economic viability of industrial biotechnology. Development
of biocatalysts (such as enzymes, to synthesize chemicals), improvement of fermen-
tation process, and production of new plastics/textiles, biofuels, etc. are typical
examples. Specific industrial biotechnology related to production of wine, cheese,
and beer by fermentation is also termed yellow biotechnology. Designing more
energy-efficient, less polluting, and low resource-consuming processes and prod-
ucts that can beat traditional ones is also termed white biotechnology.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 81

Environmental biotechnology is an interdisciplinary branch of biotechnology

using biological systems and/or organisms for conservation of environment,
resources, and energy and for protection of humans, animals, and plants on Earth
and beyond. It can be of green biotechnology, gray biotechnology, blue biotechnol-
ogy, gold biotechnology, or white biotechnology, depending on the applications or
achievement goals. Modern green environmental biotechnology has a symbol of
“green cross” that involves the construction of resource recovery facilities (RRF),
bioreactor landfills, in-vessel or in-bin composting reactors, bioremediation sites,
wildlife sanctuary areas, environmental protection parks, global warming control
facilities, salmon ladders, etc. using the best available technologies (BAT) for recla-
mation of water, air, land, nutrients, methane gas, animals, plants, etc. and produc-
tion of biofuels, bioplastics, waste-converted animal foods, etc. in turn, achieving
environmental conservation, resource sustainability, biodiversity, climate control,
ozone layer protection, etc. Gray biotechnology refers to an old traditional environ-
mental biotechnology applications to maintain biodiversity and the partial removal
of certain pollutants or contaminants using microorganisms and plants to isolate and
dispose of many kinds of substances such as heavy metals and hydrocarbons, but
without sustainability of natural resources. Typical examples are the old biological
secondary wastewater treatment plants (WWTP) and old sanitary landfills. Modern
environmental biotechnology is considered to be a green biotechnology. Blue bio-
technology is based on the use of marine resources to produce goods, generate
energy, or reduce pollution.
Computational biotechnology can be defined as “conceptualizing biotechnol-
ogy” to address biotechnology problems using computational techniques and makes
the rapid organization as well as analysis of biotechnological data possible. It can
also be termed gold biotechnology or bioinformatics.
Military biotechnology is also termed dark biotechnology because it is associ-
ated with bioterrorism or biological weapons and bio-warfare using microorgan-
isms and toxins to cause diseases and death in humans, domestic animals, and crops.
Biotechnology itself is an academic field of engineering science, while any other
academic field dealing with the law and ethical and philosophical issues around the
engineering science biotechnology is liberal art biotechnology or violet biotechnol-
ogy. This publication emphasizes environmental biotechnology to be applied to
environmental control of medical (pharmaceutical) and industrial biotechnology
industry.

2.1.2 Historical Development of Biotechnology Industry

The biotechnology industry is still young, especially compared with the automotive,
chemical, and steel industries. Despite its comparative youth, it is becoming an
important influence on many other industry segments, as well as developing an
impressive presence of its own. Its technology base continues to grow dynamically
and is melding medical science with information technology in new and exciting
82 L. K. Wang et al.

ways. While its relationship with capital markets has sometimes been stormy, that
relationship now appears to be settling into maturity as its medically oriented com-
panies bring growing numbers of new products to market.
The growth of the biotechnology industry is a unique story, and yet it rests on
foundations common to other segments of industry. Years of research, both
government-­funded and privately funded, continue to provide an ever-expanding
knowledge base. The capital market provides the ability to transform this knowl-
edge into unique products and processes for markets around the world. While there
is inevitable tension between the industry’s desire to bring new products to market
and the concerns of the industry’s regulators, both sides have found new and innova-
tive ways to work together.
Perhaps unique among industries, biotechnology is not defined by its products
but by the technologies used to make those products [1]. Biotechnology refers to a
set of enabling technologies used by a broad array of companies in their research,
development, and manufacturing activities. To date, these technologies have been
used primarily by the pharmaceutical industry, but they are being used increasingly
by a variety of other industries, such as agriculture, mining, and waste treatment.
Various US government publications have defined biotechnology as a set of tech-
niques that use organisms or their cellular, subcellular, or molecular components to
make products or modify plants, animals, and microorganisms to carry desired traits
[1]. This broad definition includes methods of treating disease developed from
recent research in molecular biology and other fields, as well as the century-old
practices of animal and plant breeding and the use of microorganisms to make leav-
ened bread and fermented beverages.
Advances in molecular biology over the past 25 years have led to the develop-
ment of genetic engineering, monoclonal antibody technologies, DNA amplifica-
tion, protein engineering, tissue engineering, and other methodologies with
applications in the medical arena. These new techniques have enabled researchers
to modify the genetic and biochemical makeup of organisms with far greater preci-
sion and speed.
In the roughly 25 years since the development of recombinant DNA technologies
in research laboratories, more than 2000 firms have been founded in the USA alone
to explore and to take advantage of these new technologies [2]. Approximately 30
new products have reached the medical market, and several hundred more are in
human clinical trials. The market for such products has grown dramatically from
$7.6 billion in 1996 to $24 billion in 2005. Similarly, the market for agricultural
biotech products has increased from $295 million to $1.74 billion in the same
period. Applications of the products will lead to enhanced pest resistance in food
crops, improved methods of food preservation, and other advances. Table 2.1 shows
the distribution of research activities and biotechnology firms in the USA.
It is clear that California and Massachusetts are the top leading biotechnology
states followed by New Jersey, North Carolina, and Maryland [3, 4].
The biotechnology industry serves both medical and nonmedical markets. The
medical market includes human therapeutics and human diagnostics as well as
applications in veterinary medicine. Nonmedical markets encompass both
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 83

Table 2.1 Leading biotechnology states in the USA [3]

Rank State Number of companies

1 California 267
2 Massachusetts 130
3 New Jersey 80
4 North Carolina 71
5 Maryland 70
6 Pennsylvania 58
7 Wisconsin 56
8 New York 55
9 Texas 50
10 Washington 40

Table 2.2 Participation of biotechnology companies by primary focus [3]

Percentage of all
Market area Number of companies companies
Therapeutics 315 29.4
Diagnostics 187 17.4
Reagents 84 7.8
Plant agriculture 68 6.3
Specially chemicals 54 5.0
Immunological products 36 3.4
Environmental testing/treatment 35 3.3
Testing/analytical services 32 3.0
Animal agriculture 29 2.7
Biotechnology equipment 26 2.4
Veterinary 26 2.4
Drug delivery systems 24 2.2
Vaccines 24 2.2

agriculture and industrial applications. Agricultural applications include making

plants and crops pest resistant, providing improved seed quality, modulating growth
and ripening times, enhancing nutrient content of foods, and providing simple and
inexpensive diagnostics for use in field testing for contaminants and toxic materials.
Industrial uses of biotechnology involve many different sectors and include indus-
trial enzymes, waste management, bioremediation, energy biomass, cosmetic for-
mulations, and diagnostics for toxicity determinations. Tables 2.2 and 2.3 show the
distribution of biotechnology firms among the various medical and nonmedical
markets by primary focus and in all areas, respectively [3, 4]. It is obvious that the
pharmaceutical industry is by far the predominant and largest area of biotechnology
[5–108].
84 L. K. Wang et al.

Table 2.3 Participation of biotechnology companies in all areas [3]

Market area Number of companies Percentage of all Companies

Therapeutics 448 41.8
Diagnostics 346 32.3
Reagents 224 20.9
Specialty chemicals 159 14.8
Immunological products 146 13.6
Cell culture products 133 12.4
Fermentation/production 116 10.8
Plant agriculture 106 9.9
Vaccines 105 9.8
Drug delivery systems 94 8.8
Environmental treatment/testing 93 8.7

2.1.3 Core Technologies

The core technique of biotechnology is elegant in its simplicity. The cell is a minia-
ture factory, containing a genetic material—DNA—that acts as a blueprint for its
structure and function. Biotechnology allows researchers to isolate, copy, and rear-
range this genetic blueprint at the molecular level to manipulate the quantity, struc-
ture, and function of the biomolecules that control cellular processes. As a result,
researchers are expanding their abilities to identify, isolate, and modify those
molecular agents.
Discoveries concerning the molecular bases of cellular processes will have a
wide range of applications. For example, in the area of health, these mechanisms
may lead to therapies that fight disease by regulating specific cellular processes.
With the help of molecular biology, biochemistry, and biophysics, the search for
molecular information is yielding an increasingly detailed guide to cell behavior
and its disruption. This knowledge allows biotechnologists to develop new prod-
ucts, processes, and therapies of commercial interest.

2.1.4 Biotechnology Materials

The raw materials of biotechnology are cells and their constituent biomolecules.
These materials may be used for a variety of purposes, including drug synthesis,
food production, and the bioremediation of hazardous waste. Examples of biotech-
nology materials include the following [1]:
1. Cytokines. Hormone-like proteins that stimulate the growth or regulate the func-
tion of various cell types. They include such agents as erythropoietin, which
stimulates the production of red blood cells and can be used to treat severe ane-
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 85

mia associated with renal disease, and granulocyte colony-stimulating factor,

which stimulates the production of white blood cells and is used to counter the
loss of such cells in patients who have received anticancer therapy, which help
regulate and target the body’s immune response and can be used to treat certain
cancers and selected viral infections.
2. Antibodies. Large protein molecules produced by the immune system that can
bind specifically to discrete antigens; foreign substances are recognized and then
attacked by the immune system.
3. Enzymes. Protein catalysts that facilitate specific chemical or metabolic reac-
tions necessary for cell growth and function. Enzymes can be used in such activi-
ties as food processing, the bioremediation of hazardous waste, and the synthesis
of certain drugs, vitamins, and fine chemicals.
4. Restriction enzymes. Enzymes that break DNA in specific locations, creating
gaps into which new genes can be inserted. These enzymes play a vital role in
genetic engineering.
5. Viral vectors. Modified, nonpathogenic viruses that deliver useful genetic infor-
mation to host cells in gene therapy and genetic engineering. In gene therapy
applications, such viruses are encoded with a specific gene, which, when incor-
porated into a host cell, confers a clinical benefit to the patient.
6. Antisense oligonucleotides. Strands of DNA that bind to targeted messenger
RNA molecules (which tell cells what proteins to make) and block the synthesis
of specific proteins. In therapeutic applications, the synthesis of disease-related
proteins is inhibited. These compounds are used in drug development and in
agricultural biotechnology.

2.1.5 Drug Development

The acceleration of the drug discovery process resulting from biotechnology

research is contributing to US competitiveness in biotechnology. Many companies
emerged in the past decade to become involved in this new approach to drug com-
mercialization. Important areas of drug-related research include the following [1]:
1. Rational drug design. Scientists are using a combination of chemistry, biology,
biophysics, and computer modeling to determine the structure of target proteins
in molecular detail and to then design specific small-molecule drugs for those
target proteins. Companies involved in rational drug design include Agouron,
Arris, BioCryst, Chiron, Procept, and Vertex.
2. Natural product screening. New methods of screening materials extracted from
animals and plants offer a rich source of potentially therapeutic compounds.
NPS Pharmaceuticals, Magainin, Shaman, and Xenova are among the biotech
firms that literally search the air, land, and sea for new drugs.
3. Combinatorial chemistry. This technology allows chemists to synthesize large,
diverse collections of molecules quickly and efficiently and to then identify the
86 L. K. Wang et al.

most active compound for a given application. Because combinatorial chemistry

can identify promising compounds in a fraction of the time required by tradi-
tional methods of drug discovery, it can significantly reduce the cost of commer-
cializing new drugs. Companies using such technology include Gilead Sciences,
Isis, and Pharmacopeia.

2.1.6 Gene Sequencing and Bioinformatics

Mutations are alterations in DNA sequence that may be associated with disease-­
causing genes. Such modified genes, and the proteins for which they encode, repre-
sent targets for drug therapy. Genes are sequenced by cutting pieces of DNA into
small segments and cloning and copying those segments millions of times over. The
order of the nucleotides (subunits of DNA) contained in those segments is then
determined. A computer program is used to analyze and correlate the nucleotide
sequences of the individual segments to create a map of the entire gene. The genes
identified by this computer analysis are then scrutinized as possible drug targets.
Rapid advances in the speed and accuracy of sequencing will revolutionize the dis-
covery of innovative drugs and diagnostics. Companies in the business of gene
sequencing include Darwin Molecular, Human Genome Sciences, Mercator
Genetics, and Sequana.

2.1.7 Applications of Biotechnology Information to Medicine

Biotechnology produces information that is used to alter and improve cell behavior.
Many biotech companies specialize in finding ways to deliver and apply biotechnol-
ogy information to cells to aid in identifying, preventing, and treating disease.
Representative applications include the following [1]:
1. Diagnostics. Tests that use biotechnology materials to detect the presence or risk
of disease or pollution of a cell or material.
2. Vaccines. Preparations of whole or significant structural portions of viruses,
microbes, plants, or other entities that are intended for active immunological
prophylaxis. Companies working in this area may specialize in the route of
administration as well as in the disease that the vaccine targets.
3. Gene therapy. The process of replacing defective genes with healthy genes,
either in vivo or ex vivo, to regulate cell replication or the production of proteins.
Alternatively, gene function may be modulated by designing and delivering mol-
ecules to cells to inhibit or promote gene action.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 87

2.1.8 Applications of Biotechnology Information

to Nonmedical Markets

Biotechnology also offers significant applications in agriculture and industry.

Industrial applications include specialty and fine chemicals and bioremediation.
Biotechnology materials, specialized software packages, and equipment used in
drug development and production are also important adjuncts to the core biotech-
nology markets.
In nonmedical areas, there are a number of potentially important developments
under way. Genetic modification of food crops, increasing protein content or salt
resistance, may help to reduce world hunger. In addition, biotechnology has the
potential to shift the world’s fish supply from an uncertain and threatened wild food
source to an agricultural analog cultivated through mariculture and freshwater aqua-
culture. The exploration, study, and harvesting of marine genetic resources through
biotechnology are expected to produce important commercial applications, includ-
ing improved diagnostics and pharmaceuticals, increased production of ocean
foods, novel energy sources, and the engineering of microorganisms to control and
eliminate environmental contaminants.

2.1.9 The Regulatory Environment

Regulation has been and will continue to be a major factor influencing the develop-
ment of the biotechnology industry and its international competitiveness, especially
for products made from recombinant DNA technology. Health, safety, and environ-
mental regulations are of critical importance, affecting the cost and time needed to
get biotech products to market and the profits thereafter. At the same time, other
federal regulations, such as those relating to the cleanup of waste sites and to air and
water quality generally, can play an important role in the development of the mar-
kets served by the bioremediation portion of the biotech industry.
The US Environmental Protection Agency’s (USEPA’s) effect on the domestic
industry is complex. On one hand, it has regulatory authorities that it intends to use
to regulate aspects of the industry’s activities and that industry fears may result in
new regulatory burdens. On the other hand, the USEPA’s responsibilities for over-
seeing the cleanup of polluted sites give it the power to create important new mar-
kets for the industry.
The USEPA’s broad responsibilities for the cleanup of hazardous waste sites
under the Comprehensive Environmental Response, Compensation, and Liability
Act (CERCLA) and the Resource Conservation and Recovery Act (RCRA) give rise
to important market opportunities for companies offering bioremediation technolo-
gies and services, but industry has pointed to several aspects of these activities that
may discourage the use of bioremediation technologies. The USEPA has initiated
proceedings to reexamine its approaches to its cleanup responsibilities, and many
88 L. K. Wang et al.

within the biotechnology industry hope this will create more opportunities for bio-
remediation technologies in both the RCRA and Superfund programs.

2.2 General Industrial Description and Classification

2.2.1 Industrial Classification of Biotechnology Industry’s

Pharmaceutical Manufacturing

The pharmaceutical industry is the biggest and most important biotech industry.
This industry produces substances that are of value for humans and other living
beings. According to the census by the US Department of Commerce (US DC), the
industry employed about 170,000 people and produced goods which were valued at
over 39 billion US dollars in 1987 [5].
The Standard Industrial Classification (SIC) has been developed and revised
since the first major version in 1972, with the purpose of promoting the comparabil-
ity of established data describing various facets of the US economy, such as man-
agement, budget, and data on production, sales, and cost for various industries.
While the pharmaceutical industry requires ultrapure water for their manufactur-
ing processes [6], their process effluents contain highly toxic pollutants which must
be properly treated before being discharged to a receiving water.
According to the Standard Industrial Classification Manual [7], the products of
the pharmaceutical industry are segregated into four categories:
1. Medical chemicals and botanical products
2. Pharmaceutical preparations
3. In vitro and in vivo diagnostic substances
4. Biological products, except diagnostic substances
The pharmaceutical industry has steadily grown because of the need to market,
develop, and discover a variety of drugs required throughout the world. This growth
of the industry has also increased the amount of waste generation and in turn dis-
posal problems. To control effluent discharge and to reduce the impact of waste
from the pharmaceutical industry, the USEPA categorized pharmaceutical manufac-
turing processes according to the SIC standard and has developed effluent discharge
limitation guidelines based on the production activities and wastes from this indus-
try [8–15].
It should be noted that the pharmaceutical SIC in the USEPA’s effluent discharge
limitation guidelines [8, 9, 11, 13–15] was based on the older versions rather than
the 1987 SIC codes cited above, although the 1987 SIC codes were used for the
recent guidelines to pollution prevention in the pharmaceutical industry [15, 16]. To
follow the effluent discharge limitation guidelines established by the USEPA, the
following sections present those SIC codes for the pharmaceutical manufacturing
quoted by the USEPA [11–15].
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 89

2.2.2 Biotechnology Industry’s Pharmaceutical SIC

Subcategory Under the USEPA’s Guidelines

According to the USEPA’s effluent discharge guidelines [11–15], pharmaceutical

manufacturing includes those plants producing or utilizing the following products,
processes, and activities:
1. Biological products
2. Medicinal chemicals and botanical products
3. Pharmaceutical products
4. All fermentation, biological and natural extraction, chemical synthesis, and for-
mulation products which are considered as pharmaceutically active ingredients
by the US Food and Drug Administration, but which are not covered by other
categories
5. Cosmetic preparations which function as a skin treatment
6. The portion of a product with multiple end uses which is attributable to pharma-
ceutical manufacturing either as a final pharmaceutical product, component of a
pharmaceutical formulation, or pharmaceutical intermediate
7. Pharmaceutical research which includes biological, microbiological, and chemi-
cal research, product development, and clinical and pilot plant activities
The pharmaceutical manufacturing under this categorization does not include all the
activities producing the substances used in medical purposes, such as some medical
instruments. Moreover, not all products containing pharmaceutical ingredients
belong to pharmaceuticals, such as milk containing vitamin D. To clarify the confu-
sion in the nature of pharmaceutical manufacturing, it is helpful to review the manu-
facturing which is similar to, but not included in, pharmaceutical manufacturing.
The following lists the production or activities specifically excluded from the phar-
maceutical manufacturing category [11]:
1.
Surgical and medical instrument and apparatus
2.
Orthopedic, prosthetic, and surgical appliances and supplies
3.
Dental equipment and supplies
4.
Medical laboratory
5.
Dental laboratory
6.
Outpatient care facilities
7.
Health and allied sources, not elsewhere classified
8.
Diagnostic devices not covered under other categories
9.
Animal feeds which include pharmaceutically active ingredients such as vita-
mins and antibiotics
10. Foods and beverages which are fortified with vitamins or other pharmaceuti-
cally active ingredients
Note, again, that these SIC codes are cited according to the earlier versions of the
Standard Industrial Classification Manual rather than the 1987 version [11, 13].
90 L. K. Wang et al.

Because each of the pharmaceutical subcategories is involved in one or more

particular processes, it is difficult to make any generalization regarding various
effluents discharged from the pharmaceutical industry. The problem is even more
complicated by the fact that pharmaceutical manufacturing uses both inorganic and
organic raw materials. To better minimize and treat pharmaceutical wastes, the man-
ufacturing processes must be first fully understood. This chapter will initially dis-
cuss the pharmaceutical manufacturing processes and waste generation, then discuss
the waste characteristics and their environmental impact, and finally discuss waste
minimization and treatment [15–108].

2.3 Manufacturing Processes and Waste Generation

While the preceding section itemizes the pharmaceutical manufacturing under the
SIC subcategorization, it is better to generalize the pharmaceutical manufacturing
with its main processes and the waste generation, so as to better understand how to
control and treat the manufacturing wastes. The five common processes used in the
manufacture of pharmaceutical products are as follows:
1. Fermentation (subcategory A)
2. Natural product extraction (subcategory B)
3. Chemical synthesis (subcategory C)
4. Formulation/mixing/compounding (subcategory D)
5. Research and development activities (subcategory E)
These five processes have been the basic pharmaceutical manufacturing processes,
although the SIC subcategory codes for the pharmaceutical industry can be revised
as stated in the preceding sections. The USEPA’s guidelines to the point source cat-
egory for pharmaceutical manufacturing (40 CFR Part 439) are established based on
these five processes and their related wastes [11, 12, 14, 15]. These five processes
are identified by the USEPA as the subcategories of pharmaceutical manufacturing
and will be used throughout this chapter, instead of using the SIC subcategories.
The USEPA [13] has reported that subcategory D (formulation/mixing/com-
pounding) is the most prevalent pharmaceutical manufacturing process, and about
80% of the plants in the industry are engaged in this activity. Furthermore, 58% of
these plants conduct subcategory D operations only.
Pharmaceutical manufacturing plants generate a variety of wastes during manu-
facturing, maintenance, and housekeeping operations. While maintenance and
housekeeping activities are similar from one plant to the next, actual processes used
in pharmaceutical manufacturing vary widely. With this diversity of processes
comes a similarly diverse set of waste streams. Typical waste streams include spent
fermentation broths, process liquors, solvents, equipment washwaters, spilled mate-
rials, off-spec products, and used processing aids [16].
The following subsections discuss those five main manufacturing processes and
their associated wastes.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 91

2.3.1 Fermentation

Although only about 6% of pharmaceutical products and their wastes are generated
by fermentation processes, fermentation is considered an important production pro-
cess for the industry [14, 16]. Most antibiotics (penicillin, streptomycin), steroids
(such as cortisone), and vitamin B12 are produced using fermentation processes.
Fermentation processes consist of three major steps:
1. Inoculum and seed preparation
2. Fermentation
3. Product recovery and purification
Figure 2.1 shows a flow diagram for a fermentation process [16]. Sterile inoculum
preparation begins with a carefully maintained population of a microbial strain. A
few cells from this culture are matured into a dense suspension through a series of
test tubes, agar slants, and shaker flasks. The cells are then transferred to a seed tank
for further propagation into a culture of sufficient quantity to function as a seed.
While tailored to a specific fermentation, the volume of the final seed tank occupies
from 1 to 20% of the volume used in full-scale production.
In the fermentation step, the material from the seed tank, along with selected raw
materials, is introduced, through a series of sterilized lines and valves, into a steril-
ized fermentor (batch vessel). Once these sterilized nutrient materials are added to
the vessel, fermentation commences. Dissolved oxygen content, pH, temperature,
and several other parameters are carefully monitored throughout the fermenta-
tion cycle.

Fig. 2.1 Fermentation process diagram [16]

92 L. K. Wang et al.

Following cell maturation, the fermentor broth from the batch vessel is often
filtered to remove the solid residues resulting from the fermentation process; the
filtrate is then processed to recover the desired product.
There are three commonly used schemes for product recovery, i.e., solvent
extraction, direct precipitation or solvent evaporation, and ion exchange or adsorp-
tion [17].
In the solvent extraction process [18], an organic solvent is used to separate a
pharmaceutical product from an aqueous filtrate and to form a more concentrated
solution. With subsequent extractions, the product is purified, especially from con-
taminants. Finally, the product is further recovered, specifically removed from the
solvent, by precipitation or crystallization or solvent evaporation.
Normally, solvents used for product recovery are recovered and reused. However,
small portions left in the aqueous phase during the solvent extraction can appear in
the plant’s wastewater stream. Typical processing solvents used in fermentation
operations are methylene chloride, benzene, chloroform, butyl acetate,
1,1-­dichloroethylene, and 1,2-transdichloroethylene [11, 12, 15, 16].
In precipitation or evaporation processes, product is recovered directly from a
treated broth. In an ion-exchange process, a product is removed from a treated broth
using ion-exchange resin and then proceeded for an additional purification and a
final isolation.
The waste characteristics of fermentation processes may vary depending on the
production. For example, the antibiotic wastes can generally be divided into four
groups [19]:
1. Group A: spent fermentation mash
2. Group B: wastes containing acids, bases, and solvents (used in the purification of
the product)
3. Group C: condensate from barometric condensers in evaporation and drying
4. Group D: washing water (used for cleaning equipment and floors)
The waste of Group A has a 5-day biological oxygen demand (5-day BOD or BOD5)
of 4000–13,000 mg/L [20] if the end product is totally absent from the effluent. For
example, in the production of streptomycin, the average 5-day BOD or the spent
mash is approximately 2500 mg/L, and for aureomycin, it is in the range of
4000–7000 mg/L. When the fermentation does not proceed satisfactorily, a batch of
the mash has to be discharged to waste together with the mycelium, which results in
the 5-day BOD of the waste rising to 20,000 mg/L or even 30,000 mg/L, while the
permanganate value increases to more than 15,000 mg/L. If the mycelium is very
carefully separated from the mash, the waste liquors are fairly clear, and the com-
bined content of organic and inorganic suspended solids in a filtered penicillin mash
is about 400 mg/L. However, the waste is commonly milky-yellow in color and
cannot be clarified easily. The waste directly from the fermentation tanks has a pH
of 2–3 units. The pH may rise to 7.5–8.0 units when it is mixed with the effluents
from Group D.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 93

Group B waste consists of the tailings from distillation apparatus used for the
recovery of organic solvents. The concentration of these components depends on
their solubility in water.
Group C waste consists of condensates from barometric condensers which are
only slightly polluted. Those wastes from the manufacturer of aureomycin, how-
ever, have a 5-day BOD of 60–120 mg/L.
Group D wastewater from washing of floor and equipment is similar to that of the
waste in Group A, with 5-day BOD from 500 to 1500 mg/L. But in penicillin pro-
duction, the washing wastewater contains alkaline, due to the use of basic sub-
stances for removing unwanted matter from equipment tanks and fermentors.
The fermentation process generates a large volume of waste such as the spent
aqueous fermentation medium and solid cell, debris. The aqueous medium is very
impure, containing unconsumed raw materials such as corn steep liquor, fish meal,
and molasses. Filtration processes result in large quantities of solids in the form of
spent filter cake including solid remains of the cells, filter aid, and some residual
product. After product recovery, spent filtrate is discharged as wastewater (known as
the “spent beers”), which contributes the most significant waste load in the fermen-
tation process. That is, this filtrate still contains a large amount of organic material,
protein, and other nutrients. Some wastewater may also come from the use of wash-
water and gas and dust scrubbers. While solvent extraction contributes relatively
small amounts of organic solvents, direct precipitation results in increased metallic
ion (particularly copper and zinc) concentration.
In general, the wastewaters from fermentation operations typically have high
5-day BOD, COD (chemical oxygen demand), and TSS (total suspended solids)
levels with a pH value in the range of 4–8 units [11, 12].
Sometimes a fermentation batch can be infested with a phage, a virus that attacks
microorganism [13]. In such a case, very large wastewater discharges may be neces-
sary in a short period of time, which causes a higher nutrient and 5-day BOD con-
centration than that of the spent broth during normal production. Some fermentation
plants use heavy-metalbearing chemicals as biocides (such as organomercury)
which will introduce heavy-metal contamination.
Volatile solvents used in product recovery operations may release vapors to the
air. Some factories may generate acid and solvent vapors such as methanol and butyl
acetate, causing air emission problems.

2.3.2 Biological Product Extraction

Biological product extraction is the production of pharmaceuticals from natural bio-

logical material sources such as roots, leaves, animal glands, and fungi. Such phar-
maceutical, which typically exhibit unique pharmacological properties, includes
allergy relief medicines, insulin, morphine, alkaloids, and papaverine [16]. Despite
their diversity, all extractive pharmaceuticals have a common characteristic: they
are too complex to synthesize commercially.
94 L. K. Wang et al.

The extraction process requires very large volumes of specialized plant or animal
matter to produce very small quantities of products. In other words, these extraction
techniques basically consist of methods to concentrate particular compounds from
either plant or animal tissue [21].
The extraction process consists of a series of subsequent extraction operations.
In almost every step, the volume of material can greatly diminish. To that end, the
volume on the final product may be less than one-thousandth of the initial volume.
Therefore, another characteristic of natural product extraction is that the amount of
finished drug product is small compared with the amount of source material used.
Because of these volume reductions, conventional batch method and continuous
processing method are not suitable for biological product extraction operations [11,
13]. Therefore, a unique assembly-line, small-scale batch processing method has
been developed. The material is transported in portable containers through the plant
in batches of 75–100 gallons (283.9–378.SL). In this method, a continuous line of
these containers is sent past a series of operating stations where technicians perform
specific tasks on each batch in turn.
An extraction plant may make one product for a few weeks and then may convert
to produce a different product after changing and redefining the tasks to be con-
ducted at each station.
Due to the nature of the extraction process, the waste material generated is prac-
tically equal to the amount of raw material processed, and most of the waste appears
in the solid or semisolid form. Wastes from biological product extraction include
spent raw materials such as leaves and roots, water-soluble solvents, solvent vapors,
and wastewaters. The wastewater is mainly from the aqueous part of the spent natu-
ral materials and from the product recovery and purification processes. The waste-
water also comprises organic solvents, heavy metals, and ammonia.
Organic solvents are used in product recovery to dissolve fats and oils which
would contaminate the product; solvents are also used to extract the product itself.
While ketones and alcohols are common extraction agents, other organic solvents,
such as benzene, chloroform, and 1,2-dichloroethane, may be used to extract the
alkali-treated plant alkaloids.
Common heavy metals are lead and zinc, which are used as precipitating agents.
Ammonia (in solution or anhydrous forms) is often used for pH control, as are the
hydroxides of various cations and also, more importantly, as a common extraction
solvent.
In general, the extraction wastewater is characterized by small flows and low
pollutant concentrations. The wastewaters typically have low BOD5, COD, and TSS
levels and a pH in the range of 6–8 [13].
Similar to the fermentation process, volatile solvents used in product recovery
operations may release vapors to the air.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 95

2.3.3 Chemical Synthesis

Most drugs are produced by chemical synthesis. In a typical manufacturing plant,

batch processing is a standard method of operation for chemical synthesis facilities,
including a series of reaction, separation, and purification steps to make a desired
product.
Chemicals used in chemical synthesis operations range widely and include
organic and inorganic reactants and catalysts. In addition, manufacturers use a wide
variety of solvents for product recovery, purification, or process reaction, which are
listed as priority pollutants [13, 15]. A large number of toxic substances are used in
chemical synthesis plants, and a correspondingly high incidence of toxic pollutants
in the plant’s wastewater has been observed.
Figure 2.2 is a process flow diagram of chemical synthesis for an anti-convulsive
drug plant [16, 22]. Raw materials, potassium permanganate, and water are mixed
in a 3000-gallon (11,355-L) reactor. A manganese dioxide precipitate is formed and
is removed from solution by a rotary drum filter coated with Celite. The wet filter
cake (manganese dioxide precipitate and Celite) is deposited into trash bins for
disposal at a municipal landfill. The filtrate is neutralized with sulfuric acid and sent
to a climbing film evaporator. Overhead water is collected and discharged into the
sewer. The enriched product solution is then sent to an 800-gallon (3028-L) Pfaudler
vessel where a final pH adjustment is made with sulfuric acid. As the mixture is
agitated and cooled, potassium sulfate is crystallized. The potassium sulfate crystals
are removed from the reaction mixture by centrifugation dissolved in water and then
discharged to the sewer. Butyl acetate is added to the concentrate, and the mixture
is azeotropically dehydrated.

Fig. 2.2 Process flow diagram of chemical synthesis for an anti-convulsive drug plant [16]
96 L. K. Wang et al.

In a continuous process, the overhead azeotropic mixture is condensed and sent

to a decanter where the lower water layer is discharged to the sewer and butyl ace-
tate is taken off the top and returned to the product mixture. This process procedure
is continued until all the water (which contains some butyl acetate) is removed. The
butyl acetate product mixture is then filtered to remove any remaining salt. The fil-
tered solution is then cooled, allowing product to crystallize and be separated by
centrifugation. Butyl acetate is recovered and stored for reuse. The product is sent
to a tumble dryer prior to packaging. Butyl acetate vapor is vented from the dryer,
condensed, and recovered for reuse [16].
Solvents serve several functions in a chemical synthesis process [11, 13]. They
dissolve gaseous, solid, or viscous reactants to bring all reactants into close molecu-
lar proximity. They also serve to transmit heat to or from the reacting molecules.
Benzene and toluene are widely used organic solvents since they are stable com-
pounds that do not easily take part in chemical reactions.
Waste streams from chemical synthesis operations are complex due to the vari-
ous operations and reactions employed. Virtually every step of an organic synthesis
generates liquor that contains unconverted reactants, reaction byproducts, and resid-
ual products in the organic solvent base. Acids, bases, cyanides, and metals may
also be generated. Typically, the spent solvents are recovered on-site by distillation
or extraction [23], which also generate solvent recovery wastes such as still bot-
tom tars.
Aqueous waste streams from synthesis processes may result from miscible sol-
vents, filtrates, concentrates, equipment cleaning, wet scrubbers, and spills.
Wastewaters typically have high 5-day BOD, COD, and TSS levels and have a pH
value in the range of 1–11 units. Solid wastes may result from filter cakes. The use
of volatile solvents can also result in air emissions.

2.3.4 Formulation, Mixing, and Compounding

Pharmaceutical formulation is a process for preparation of dosage forms such as

tablets, capsules, liquids, parenterals, and creams and ointments for consumer use.
Tablets account for over 90% of all medications taken orally [24] and are pro-
duced in three varieties: plain compressed, coated, and molded. The form of tablet
depends on the desired characteristics of active ingredient, which can be slow, fast,
or sustained, for example, spraying or tumbling the tablets with a coating material
is one of the ways controlling the release characteristics. Tablets are produced by
blending the active ingredient with fillers, such as starch or sugar, followed by com-
pressing using either wet granulation, or direct compression, or slugging.
Capsules prepared in hard or soft form are the next most widely used oral dosage
form for solid drugs. Hard capsules consist of two separate pieces which are formed
by dipping pins into a solution of gelatin maintained at a specified temperature.
When removed, a gelatin film is deposited on the pins. Unlike hard capsules, soft
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 97

capsules are prepared by placing two continuous gelatin films between rotary die
plates and then injecting in the drug.
The third type of pharmaceutical formulation is a liquid dosage form prepared
for injection or oral use, which includes solutions, syrups, elixirs, suspensions, and
tinctures, all of which are usually prepared by mixing the solutes with a selected
solvent in a glass-lined or stainless steel vessel. Suspensions and emulsions are
frequently prepared using colloid mills and homogenizers.
Parenteral dosage forms are injected into the body either intramuscularly, intra-
venously, or subcutaneously. Parenterals are prepared as solutions, as dry solids
which are dissolved immediately before injection, as suspensions, as dry insoluble
solids which are suspended before injection, and as emulsions.
Ointments and creams are semisolid dosage forms prepared for topical use.
Ointments are usually prepared by melting a base, which is typically the petroleum
derivative petrolatum. This base is then blended with the drug, and the cooled mix-
ture is passed through a colloid or roller pill. Creams are oil-in-water or water-in-oil
emulsions, rather than being petrolatum based, and are manufactured in a similar
manner [16].
Most water used in the formulation process is as cooling water, which generates
no contact wastewater. Wastewater is generally originated from cleanup, spills, and
breakage of packaged products. Some wastewaters may come from the dust scrub-
bers, which are sometimes used to control dust from tablet and capsule production.
Most wastes are nontoxic, have relatively small flows, and have low 5-day BOD,
COD, and TSS concentrations, with near neutral pH (6.0–8.0).
Air emissions may result from the use of volatile solvents in the formulation
processes.

2.3.5 Research and Development

Research and development (R & D) processes in the pharmaceutical industry

involve chemical research, microbiological research, and pharmacological research
to provide information for pharmaceutical production related in the above. The
development of a new drug with less environmental pollution requires cooperative
efforts in several fields, such as medicinal, chemical engineering, biomedical engi-
neering, environmental engineering, biology, biochemistry, pharmacology, and
toxicology.
An example is the R & D section [16] in a plant producing a wide range of der-
matological products (such as shampoos, creams, and itch soothing preparations)
and ophthalmic products (such as contact lens cleaners, eye drops, and disinfecting
solutions). These pharmaceutical compounds are formulated in the production sec-
tion after having been thoroughly researched by the R & D section. The R & D
section involved two major groups: the synthetic chemistry division and the product
development division. Halogenated and nonhalogenated solvents, such as chloro-
form, methylene chloride, acetone, methanol, acetonitrile, acetone, ethyl ether,
98 L. K. Wang et al.

xylene, and hexane, are commonly used for extraction and analyses. Acetonitrile
and methanol are extensively used as carrier liquid in high-performance liquid chro-
matography (HPLC). The plant consumed 400 gal (1514 L) of acetonitrile and
990 gal (3747 L) of methanol annually. Other chemical wastes, including photo-
graphic chemicals, radionuclides, bases, and oxidizers, can be produced from some
pharmaceutical research and development sections. Sulfuric acid is the most widely
used acid at an annual consumption of 450 gal (1703 L). In addition, a large quantity
of sulfuric acid is used in glassware washing at an annual acid consumption of
approximately 1080 gal (4088 L).
The wastes from the research and development processes can be similar to those
wastes generated from one or more or all of the above four processes, chemical
synthesis, fermentation, biological product extraction, and formulation, and can be
even more complicated, because various attempts should be made to develop a new
drug or a new pharmaceutical instrument. Radioactive wastes may also be generated.
As a result of the diverse nature of pharmaceutical research and development, a
wide range of chemical and biological laboratory wastes are produced. However,
the quantity, quality, and time schedule of discharging research and development
wastes are usually erratic, and the problem cannot be measured entirely. The quanti-
ties of materials discharged by research and development operations are in general
[25] relatively small as compared with the volumes generated by production
facilities.
Pharmaceutical production can be batch, continuous, and semi-continuous oper-
ations. Batch-type production is the most common type of manufacturing technique
for each of the subcategories. Table 2.1 summarizes the typical wastes and the asso-
ciated process origins in pharmaceutical industry. Note that most of the process
origins in the table can exist in all the five main processes but with varied qualities
(i.e., having various kinds of materials and wastes) and quantities of wastes.

2.4 Waste Characterization and Options for Waste Disposal

2.4.1 Waste Characteristics

The preceding discussions show that numerous process wastes are generated by the
pharmaceutical industry. The pharmaceutical wastes vary greatly depending upon
the manufacturing processes. The very nature of the pharmaceutical industry deter-
mines the composition of each plant effluent, which varies considerably from plant
to plant.
There are pharmaceutical plants which discharge only solid wastes, and no waste
liquors in the sense of production process. However, these plants still have to deal
with certain amounts of wastewater from washing of equipment and floors, etc.
A distinguishing feature of pharmaceutical fermentation and the biological prod-
uct extraction manufacturing is that a large proportion of the material input to the
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 99

manufacturing process ends up as process wastes. The wastes from such a low
product-­yield process may be in either solid or liquid form.
Many plants generate wastewaters with COD concentration ranging from 500 to
1500 mg/L, whereas the wastewaters from fermentation and chemical synthesis
products may have COD concentrations reaching 10,000 mg/L or even higher [26].
Generally, fermentation processes and chemical synthesis processes produce
large flows and have high levels of 5-day BOD and COD, with high TSS for the
fermentation processes, although they vary greatly from factory to factory, while the
biological product extraction, formulation, and research and development tend to
produce low flows with low levels of 5-day BOD, COD, and TSS [13]. Table 2.2
lists average waste flow and traditional pollutants from four manufacturing pro-
cesses: chemical synthesis, fermentation, biological product extraction, and formu-
lation/manufacturing.
Toxic pollutants can exist in the wastewaters. Especially, the waste from the
chemical synthesis plant usually contains significant levels of a large number of
toxic pollutants. Table 2.3 lists toxic organic pollutants associated with pharmaceu-
tical industry according to the list of organic priority pollutants by the 1977 amend-
ment to the US Clean Water Act.
Besides cyanide, many inorganic priority pollutants are commonly found in the
waste streams from pharmaceutical industry, such as arsenic, beryllium, cadmium,
chromium, copper, lead, mercury, nickel, selenium, silver, thallium, and zinc. Only
a few of these priority pollutants are widespread in their occurrence or high in con-
centration. The significance of these facts affecting the regulation of these pollutants
will be discussed later.

2.4.2 Options for Waste Disposal

There are three options of wastewater discharge for pharmaceutical manufacturing:

direct discharge after treatment, indirect discharge (i.e., discharging to publicly
owned treatment works, or POTW), and zero discharge. Many pharmaceutical man-
ufacturers treat their wastes and directly discharge their treated wastewaters to the
navigable waters. Some of pharmaceutical plants are so located that POTW are
adequate to solve their, at least a part of, waste disposal problem. Some industrial
plants generate basically no wastewater, or trade out waste, or limit the treated
wastewater on-site, resulting in zero discharge. The numbers of the three types of
wastewaters discharge by pharmaceutical industrial plants in the USA are listed in
Table 2.4.
Deep well injection [27] generates no discharge to waterways. However, most of
the deep well injections that were permitted in the early times, and at least some of
them, may not be allowed for such operation sooner or later especially if the injected
material has a great potential threat to the environment.
Datta Gupta et al. [28] described disposal of effluent by irrigation and applica-
tion of dry waste biosolids as fertilizer [29], which may generate no wastewater
100 L. K. Wang et al.

Table 2.4 Pharmaceutical process wastes [16]

Waste description Process origin Composition
Process liquors Organic syntheses Contaminated solvents
Spent fermentation broth Fermentation processes Contaminated water
Spent natural product raw Natural product extraction Leaves, tissues
materials processes
Spent aqueous solutions Solvent extraction processes Contaminated water
Leftover raw material Unloading of materials into Bags, drums (liber, plastic,
containers process equipment metal), plastic bottles
Scrubber water from pollution Dust or hazardous vapor Contaminated water
control equipment generating processes
Volatile organic compounds Chemical storage tanks, Solvents
drums
Off-spec or out-dated products Manufacturing operations Miscellaneous products
Spills Manufacturing and lab Miscellaneous chemicals
operations
Waste water Equipment cleaning, Contaminated water
extraction residues
Spent solvents Solvent extraction or wash Contaminated solvents
practices
Used production materials Manufacturing operations Miters, tubing, diatomaceous
earth
Used chemical reagents R & D operations Miscellaneous chemicals
Natural gas combustion Steam boilers Carbon compounds, oxides of
products nitrogen and sulfur

discharge. Lane [25] described an alternative treatment and disposal of spent beer
by spray irrigation. The spent beer frequently contains high amounts of nitrogen,
phosphate, and other plant growth factors. However, it is also likely to contain salts,
like sodium chloride and sodium sulfate, as a result of the extraction process. The
presence of such salts depending on their concentration can cancel out the value of
the spent beer as a fertilizer. Spray irrigation is mainly used for the purpose of dis-
posal of the spent beer, rather than just for its value as a fertilizer. This disposal
technique has a number of limitations: (a) large land areas are needed in the order
of 125 acres (505,875 m2) for 100,000 gal (378,500 L) of spent beer sprayed per day
and (b) the land should be reasonably flat so that runoff from the spraying does not
result in erosion or “puddling” in low spots [29]. The “puddling” will result in odors
that will most likely render the entire operation a public nuisance.

2.5 Environmental Regulations on Pharmaceutical

Wastewater Discharges

Wastes generated from pharmaceutical manufacturing could exert various impacts

on the environment, such as the following:
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 101

1. Color and odor problems due to the spent solvent, their raw materials, and spent
chemicals
2. The growth of bacteria in the biosolids from fermentation and natural extraction
processes
3. Oxygen depletion due to the relatively high oxygen demand load
4. Toxic materials such as heavy metal, cyanide, and toxic organic compounds
5. Air pollution due to volatilization of volatile organic solvents
The total pollution load of wastewaters generated by the pharmaceutical manufac-
turing industry in the USA was reported by the USEPA [13] as shown in Table 2.5.

2.5.1 Regulations for Direct Discharge

To ease the impact of waste discharge to the environment, the Clean Water Act
requires a permit for any discharge into the nation’s waterways. Direct discharge
into surface water must have a National Pollutant Discharge Elimination System
(NPDES) permit and/or a State Pollutant Discharge Elimination System (SPDES)
permit. The NPDES permit or the SPDES permit is granted on a case-by-case basis.
The USEPA [11, 12, 15] regulation applies to facilities organized into five sub-
categories for this pharmaceutical industry (40 CFR Part 439): (a) subcategory A
(fermentation products), (b) subcategory B (extraction products), (c) subcategory C
(chemical synthesis products), (d) subcategory D (mixing/compounding and formu-
lation), and (e) subcategory E (research).
The USEPA has regulated what is known as the Best Practicable Control
Technology Currently Available (BPT). The direct discharge limitations are pre-
sented in Table 2.6.
The regulation for cyanide is the same in the Best Available Control Technology
Economically Achievable (BAT) and the New Source Performance Standards
(NSPS). The regulations have been delineated mainly for the four subcategories:
fermentation, biological extraction, chemical synthesis, and formulation. The
USEPA tends to deregulate the effluent discharge from R & D, because only an

Table 2.5 Characteristics of major pharmaceutical wastewater streams [13]

Waste flow BOD5 COD TSS Priority
Process MGD mg/L mg/L mg/L pH pollutant
Fermentation 0.622 1668 3452 1023 4–8 Cu, Zn
Natural extraction 0.197 42 132 93 6–8 Pb, Zn.
solvents
Chemical 0.477 2.385 4.243 414 1–11 Variety
synthesis
Formulation 0.296 339 846 308 6–8
Note: MGD million gallon per day (1 MGD = 3.784 m /day)
3
102 L. K. Wang et al.

Table 2.6 Organic priority pollutants from pharmaceutical manufacturing

Concentration (μg/L)
Organic compounds Average Range
1. PAH (polynuclear aromatic
hydrocarbons)
 Acemaphtherie 12 0–100
 Naphthalene 2.8 0–14
 Anthracene 1.8 0–7
 Fluorine 3.5 0–41
 Phenanthrene 1.8 0–7
2. Nitrogen compounds
 1,2-Diphenylhydrazine 2 0–17
 N-Nitrosodiphenylamine 12 0–1400
3. Aromatic compounds
 Benzene 220 0–2100
 Chlorobenzene 67 0–600
 2.4-Dinitrotoluene 12 0–49
 Ethylbenzene 16 0–86
 Toluene 2400 0–17,000
4. Halogenatcd hydrocarbons
 Carbon tetrachloride 460 0–6000
 1,2-Dichloroethane 8.7 0–74
 1,1,1-Trichloroethane 10 0–130
 1,1,2-Trichloroethane 95 0–1300
 1,1,2,2-Tetrachloroethane 2 0–10
 Chloroform 300 0–1600
 1,1-dichloroethylene 8.9 0–95
 Methylene chloride 2600 0–20,000
 Methyl chloride 300 0–1500
 Methyl bromide 3 0–15
 Tetrachloroethylene 3.5 0–36
 Trichloroethylene 8 0–62
5. Ethers
 Bis(2-chloroethyl) ether 19 0–170
6. Phenolic compounds
 2-Chlorophenol 2.4 0–22
 2.4-Dichlorophenol 1 0–5
 4-Nitrophenol 400 0–3500
 Pentachlorophenol 4.4 0–62
7. Phthalates
 Bis (2-ethylhexyl) 37 0–170
 Butyl benzyl phthalate 33 0–360
 Di-n-butyl phthalate 10 0–90
 Diethyl phthalate 8 0–31
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 103

insignificant amount of wastes is discharged and the wastes have similarity in qual-
ity to those from the other four sections.
Note that many of the priority pollutants which may be found from pharmaceuti-
cal discharges are excluded from direct discharge regulation because either they are
present at low level or they are infrequent for occurrence, or their presence amount
is too small to be effectively reduced by the current technology.

2.5.1.1 Best Practicable Control Technology Currently Available (BPT)

The USEPA is revising the BPT effluent limitation guidelines for chemical oxygen
demand (COD) for subcategories A, B, C, and D. Appendix 1 presents these final
limitations, which are based on the application of advanced biological treatment.
The existing BPT effluent limitation guidelines for pH, BOD5, and TSS are being
maintained for all subcategories. The existing BPT effluent limitation guidelines for
cyanide are being refined; the compliance monitoring requirements for these limita-
tions have been clarified. Limitations on cyanide for B and D subcategories are
being withdrawn.

2.5.1.2 Best Available Control Technology Economically

Achievable (BAT)

The EPA is revising the BAT effluent limitation guidelines for subcategories A and
C. For subcategories A and C, the EPA is adding BAT effluent limitations for ammo-
nia as nitrogen (N), COD, and 30 priority and nonconventional organic pollutants.
For subcategories B and D, the EPA is setting a BAT effluent limitation for COD
that is equivalent to the BPT limitation. No additional BAT effluent limitations are
being set for subcategories B and D. However, EPA is withdrawing the current BAT
effluent limitations for cyanide for subcategories B and D. Appendixes 2 and 3 pres-
ent these final effluent limitation guidelines, which are based on the following: end-­
of-­pipe advanced biological treatment with nitrification for subcategories A and C
and end-of-pipe advanced biological treatment for subcategories B and D.

2.5.1.3 New Source Performance Standards (NSPS)

The USEPA is setting NSPS for priority and nonconventional pollutants for subcat-
egories A and C. The NSPS for subcategories A and C include ammonia (as N) and
30 priority and nonconventional organic pollutants, based on advanced biological
treatment with nitrification.
The USEPA is also revising the NSPS controlling discharges of BOD5, COD,
and TSS for subcategories A, B, C, and D based on advanced biological treatment.
The USEPA is withdrawing cyanide standards for subcategories B and D. Final
NSPS for subcategories A and C are presented in Appendix 4. Final NSPS for sub-
categories B and D are presented in Appendix 5.
104 L. K. Wang et al.

2.5.2 Regulations for Indirect Discharge

As mentioned earlier, an alternative way to discharge wastewaters from pharmaceu-

tical plants is discharging their wastewaters to the publicly owned treatment works
(POTW) for further treatment. However, the wastes and wash water from pharma-
ceutical plants, especially from chemical synthesis manufacturing, are not always
compatible with biological waste treatment plants. The waste and wash water may
be too concentrated or too toxic (such as heavy metal and cyanides) that will harm
the POTW biological treatment systems. Moreover, high-acid waste can seriously
destroy the material used to seal the sewer joints and can retard biological treat-
ment; flammable solvents may cause fire or explosion and then cause damage and
interruption of sewer systems.
To assist control authorities and approval authorities for industrial discharge to
POTWs, the USEPA has developed the National Categorical Pretreatment Standards
for point sources. These categorical pretreatment standards are designed to prevent
the discharge of pollutants which pass through, interfere with, or are otherwise
incompatible with the operation of POTWs. Specifically, the Pretreatment Standards
for Existing and New Sources (PSES and PSNS) were established for the indirect
dischargers to prevent the pollutants which are incompatible with or not susceptible
to treatment in a POTW [15]. The priority pollutants considered for pretreatment
standards are listed in Table 2.7.
The PSES and PSNS regulate an indirect discharge limitation for cyanide.
The waste to be discharged to the POTW must meet the influent requirements,
and the factory must pay attention to the municipal sewer system. Pretreatment is
usually required before discharging to the POTW.

2.5.2.1 Pretreatment Standards for Existing Sources (PSES)

The USEPA is revising PSES for priority and nonconventional pollutants for sub-
categories A, B, C, and D. For subcategories A and C, the USEPA is setting PSES
for ammonia (as N) and 23 priority and nonconventional organic pollutants based
on steam stripping. For subcategories B and D, the USEPA is setting PSES for five
priority and nonconventional organic pollutants based on steam stripping. Revised
PSES for subcategories A, B, C, and D are presented in Appendixes 6 and 7.

Table 2.7 Statistical data for the three types of wastewater discharges
Type of discharge Number of plants Wastewater flow MGD
Direct discharger 52 24.9 11%
Indirect discharger 285 39.9 62%
Zero discharger 127 0 27%
Total plant 464 64.8 100%
Note: MGD million gallon per day (1 MGD = 3.784 m3/day)
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 105

2.5.2.2 Pretreatment Standards for New Sources (PSNS)

The USEPA is revising PSNS for priority and nonconventional pollutants for sub-
categories A, B, C, and D equal to PSES. Revised PSNS for subcategories A, B, C,
and D are presented in Appendixes 8 and 9.

2.5.3 Historical View on Regulations

To protect the environment, the USEPA has regulated the BPT, which is basically
identical to those shown in Table 2.6. As mentioned earlier, the wastewaters from
fermentation and chemical synthesis of products may have COD ranging between
10,000 and 20,000 mg/L. According to the BPT, which is defined as a COD removal
of 74%, the fermentation and chemical plants may be able to discharge their treated
wastewater with COD concentration from 2600 to 5200 mg/L to meet 1976 BPT
[26]. In November 1982, the USEPA proposed the BAT and the NSPS to control the
discharge of nonconventional pollutant, COD, as well as other pollutants from phar-
maceutical manufacturing facilities [9, 10, 15]. However, the industry commented
that the proposed regulations could not be met based on the USEPA-proposed tech-
nology. In 1983 and modified in 1998, the USEPA promulgated final Pharmaceutical
Manufacturing Point Source Effluent Limitation Guidelines, Pretreatment
Standards, and NSPS [11, 12, 15].
The Agency decided to return to the 1976 BPT subcategorization discharge. The
1982-proposed COD regulations are no longer valid. Therefore, the BPT limitations
listed in Table 2.6 are basically the 1976 version and finalized in 1983. However, the
USEPA reserved a final decision on appropriate BAT limitations and NSPS for
COD which is postponed until additional information could be obtained on appli-
cable COD removal technologies and their achievable concentrations.
On December 16, 1986, the USEPA promulgated the BCT limitations for the
existing pharmaceutical manufacturing facilities. The existing pharmaceutical man-
ufacturers that are subcategorized A–D productions are covered by this regulation,
which set equal to the BPT limitations in 1983. All these guidelines have been reis-
sued in 1998 [15].
It should be pointed out that the US pharmaceutical industry is largely an inter-
national industry in which many companies have manufacturing facilities and sales
and distribution operations in countries other than the USA. In addition to US fed-
eral statutes and regulations, there are international laws, regulations, treaties, con-
ventions, and initiatives which are drivers of the environmental programs of
pharmaceutical companies. The Basel Convention, the ISO 14000 standards, the
environmental requirements of NAFTA, and the evolving European Union Directives
and Regulations are a few examples of important international environmental stan-
dards and programs which affect this industry [14].
106 L. K. Wang et al.

2.5.4 Regulations for Managing Pharmaceutical Wastes

Managing Pharmaceutical Waste A 10-Step Blueprint for Healthcare Facilities In

the United States [108] was published by the USEPA in 2008, and therefore, does
not cover the most recent federal and state regulations for hazardous waste pharma-
ceuticals. On February 22, 2019, the USEPA published Management Standards for
Hazardous Waste Pharmaceuticals and Amendment to the P075 Listing for Nicotine
(referred to as the “Final Rule”) [109]. The Final Rule became effective in the US
federally managed states and territories on August 21, 2019. All states in the USA
are expected to adopt the Final Rule by the deadline of July 1, 2022. One aspect of
this rule, prohibiting the disposal of hazardous waste pharmaceuticals into sewers,
took effect in all states and territories on August 21, 2019, under the Hazardous and
Solid Waste Amendments (HSWA) regulations.
In 2021, the USEPA work was initiated to update the 10-Step Blueprint to reflect
the 2019 regulatory changes. It is anticipated that this work will be completed by
March 2022 and the new document will be available on the Healthcare Environmental
Resource Center (HERC). Although the 2008 document is not current with regard
to federal regulation of hazardous waste pharmaceuticals, it does contain valuable
non-regulatory waste management information that remains valid today.
A “10-Step Blueprint for Managing Pharmaceutical Waste of Healthcare
Facilities In the United States” [108, 109] is introduced in this section. The steps in
this blueprint do not necessarily have to be taken consecutively. Some steps will
occur in parallel, and other steps will probably be referenced throughout the devel-
opment of your pharmaceutical waste management program. The following is a
summary of the ten steps and how each can be used to develop and implement your
pharmaceutical waste management program:
1. Step l begins with some action items that you can begin immediately.
2. Step 2 is an overview of how the federal Resource Conservation and Recovery
Act (RCRA) regulations apply to pharmaceutical waste management.
3. Step 3 begins where the regulations leave off providing guidance on how to
manage non-regulated hazardous pharmaceutical waste.
4. Step 4 walks you through the steps necessary to perform a drug inventory
review. This step can be very tedious and time consuming.
5. Step 5 alerts you to waste minimization opportunities. It will be helpful to
become familiar with the waste minimization opportunities before assessing
your current practices based on the guidance provided in Step 6. Review these
opportunities again upon completion of the department reviews.
6. Step 6 discusses performing department reviews and determining your genera-
tor status.
7. Step 7, taking on the communication/labeling challenge, is one of the most
critical aspects of implementing a pharmaceutical waste management program
and possibly the most challenging. How you decide to communicate pharma-
ceutical disposition information to the people handling the waste will depend
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 107

and be dependent upon which of the management options presented in Step 8

you select.
8. Step 8, considering the management options, introduces you to five implemen-
tation models that have worked for other hospitals. You may choose one model
or a hybrid.
9. Step 9, getting ready for implementation, assists you with vendor selection,
satellite and storage accumulation, and pilot program development.
10. Step 10, launching the program, is the culmination of the first nine steps, plus
the actual rollout to the entire facility.
After the program is launched, the next steps will be (a) providing additional phar-
maceutical waste management assistance to hospitals; (b) clarifying, reconsidering,
and expanding RCRA hazardous waste regulations, (c) eliminating drain disposal;
(d) making the hazardous waste determination, a communications challenge; (e)
broadening national knowledge base of pharmaceutical waste generation; (f) man-
aging waste minimization; and (g) managing routinely wasted drugs.

2.6 Waste Management

2.6.1 Strategy of Waste Management

The main objectives of pharmaceutical waste management are to reduce waste gen-
eration through improved manufacturing process and enhanced solvent recovery; to
remove suspended matter, odor, BOD matter, and hazardous and toxic materials;
and to prevent air pollution.
This section discusses three main tasks of waste management in pharmaceutical
industry:
1. In-plant control
2. In-plant treatment
3. End-of-pipe treatment
The load on the end-of-pipe treatment process depends on how well the in-plant
control is practiced. The in-plant control usually analogs to waste minimization.
However, waste minimization is defined by the USEPA as source reduction and
recycling, which covers a somewhat different practice from the traditional in-plant
control, including the interplanetary efforts to minimize wastes such as waste
exchange. In general, in-plant control is a means of waste management, and an
interplanetary waste exchange program in waste minimization cannot be practiced
without a well-oriented in-plant management. The waste exchange will be presented
in the section of in-plant control.
Since wastewater treatment and pollutant removal costs are highly influenced by
the pollutants and volume of water to be treated, the costs for treating a segregated
stream are considerably less than that would be in treating combined wastewater.
108 L. K. Wang et al.

Also, chemicals other than those being treated are less likely to interfere with the
treatment technology if treatment occurs before mixing [11, 13]. The importance of
waste separation has been recognized, which is reflected by the fact that in-plant
treatment deals with a segregated particular pollutant. The in-plant control is mainly
a source control to reduce generation of waste, while the end-of-pipe treatment
mainly deals with overall waste in the plant. From the view point of treatment,
inplant treatment can be visualized as end-of-pipe treatment or a pretreatment for a
particular production process, while from another point of view, it is an in-plant
process to reduce waste before being discharged to an overall waste stream.

2.6.2 In-Plant Control

In-plant control includes water conservation, raw material substitution, chemical

substitution, material recovery, extensive recycling of wastewater, and modification
and improvement of processes, so that the amount of wastewater can be reduced and
pollution can be minimized. The following are some examples of in-plant controls
that have been demonstrated effectively in reducing pollution loads.

2.6.2.1 Material Substitution

Material substitution is a replacement of one or more of the raw materials used in

production to reduce the toxicity or volume of wastes generated.
Material substitution has been demonstrated to be successful in pharmaceutical
tablet coating operations to reduce hazardous waste generation. Wayman and Miller
[30] reported a successful material substitution in tablet coating which reduced the
usage of methylene chloride from 60 to 8 ton/year by converting the conventional
film coating to aqueous film coating. The other example, a water-based solvent and
new spray equipment for a tablet coating developed in a manufacturing plant, elimi-
nated expensive (US $180,000) air pollution control equipment, resulting in a sav-
ings of US $15,000 per year in solvent makeup cost [31]. Other material substitutions
that may be suitable for pharmaceutical manufacturing include the use of aqueous-­
based cleaning solutions instead of solvent-based solutions and the replacement of
chlorinated solvents with non-chlorinated solvents [13]. Moreover, using nontoxic
or less toxic biocides to substitute the heavy-metal-containing biocides in the fer-
mentation processes can avoid the correlated heavy-metal contamination.
For the pharmaceutical industry, however, product reformulation seems to be
very difficult, because the reformulation must have the same therapeutic effect, sta-
bility, and purity profile as the original formulation. Moreover, it takes a consider-
able amount of time for the US Food and Drug Administration (USFDA) to approve
of the reformulated drug. Another problem that a reformulation may encounter is
the possibility of customer rejection of the product due to changes of the product’s
aesthetic qualities such as taste, color, dosage, or form. Because of the difficulties in
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 109

reformulation, waste minimization should be introduced at the research and devel-

opment phase [16].
Another sort of material substitution is to substitute the toxic materials used in
the waste recovery and cycling processes, such as using nontoxic chemicals to sub-
stitute for zinc and lead containing agents in a precipitation process.

2.6.2.2 Process Modification

Modification or modernization of the existing processes is another opportunity to

reduce waste generation.
The modification can be accomplished through, for example, controlling a suit-
able feed rate, a proper agitating and mixing, optimizing operating temperatures,
and automation control. In most cases, the product/process yield determines the
product/waste ratio. Inadequate feeding rate, mixing, or temperature control in
pharmaceutical manufacturing can cause a high byproduct yield. Reactor efficiency
can be improved, and byproduct formation can be reduced by controlling reaction
parameters.
Increased automation can reduce operation errors. For example, introducing
automation in material handling and transfer processes can reduce spillage.
Another process modification option is to redesign chemical transfer system to
reduce physical material losses [13]. For example, replacing gas pressurization with
a pumped transfer eliminates the tank pressurizing step and its associated material
losses [32].
Other design considerations for waste minimization include modifying tank and
vessel dimensions to improve drainage, installing internal recycle systems for cool-
ing wasters and solvents, selecting new or improved catalysts, switching from batch
to continuous processes for solvent recovery, and optimizing process parameters to
increase operating efficiency. Manufacturing processes have demonstrated that
excessive solvent emissions from the purging of autoclaves used for the manufac-
ture of synthetic steroids can be considerably reduced by installing rotameters with
integral needle valves to control nitrogen flow into the reactor; nitrogen flow and
resulting solvent vapor pickup can be reduced by a factor of six compared with the
baseline situation where nitrogen flow is not controlled and operated in an on-off
fashion without throttling [16].
The major obstacles of process modification to the waste minimization are new
processes must be tested and validated to ensure that the resulting product is accept-
able; a considerable amount of time may be needed for the US FDA approval, if
applicable, before instituting any change; extension process changes can be expen-
sive; and downtime will occur when production is stopped for new equipment
installation.
The routine cleanup in the pharmaceutical plant can be carried out most effec-
tively by vacuum cleaning. Wash water may be a water pollutant. Special attention
should be given to prevent such material from entering the sewer system. Lane [25]
has shown that a central wash area with portable equipment can be usable. The
110 L. K. Wang et al.

portable (even large) equipment can be moved to a central wash-up area, providing
better prevention of dumping of hazardous pollutants to the sewer system.

2.6.2.3 Recycling Wastewater and Recovering Materials

Recovering and recycling include directly reusing waste material, recovering used
materials for a separate use, and removing impurities from waste to obtain relatively
pure substances. The goal is to recover materials for reuse in the process or for reuse
in a different application. The restricted quality control requirements of the pharma-
ceutical industry often restrict reuse opportunities. After a high degree of purifica-
tion, materials recovered from manufacturing processes may be reused. Recycling
can be performed either on-site or off-site. On-site can be either integral to an opera-
tion or in a separate operating area. The value of a waste depends on the type, mar-
ket, purity, quantity and frequency of generation, and distance between the generator
and the recycling operation.
One of the important recycling programs in the pharmaceutical industry is the
recycling of solvent. Solvents are used for reaction media, extraction media, equip-
ment cleaning, and coating media. Processes for solvent recovery from concen-
trated waste streams include distillation, nebulization, evaporation, liquid-liquid
extraction, filtration, decantation, centrifugation, flotation, and sedimentation. The
commonly used and recycled solvents are acetone, cyclohexane, methylene chlo-
ride, ethyl acetate, butyl acetate, methanol, ethanol, isopropanol, butanol, pyridine,
methyl ethyl ketone, methyl isobutyl ketone, and tetrahydrofuran [33]. Solvent
waste recyclability can be improved through special arrangement of recycling pro-
cedure: for example, minimizing solid concentration in solvent wastes, segregating
chlorinated solvent wastes from non-chlorinated solvent wastes, segregating ali-
phatic from aromatic solvent wastes, segregating chlorofluorocarbons from methy-
lene chloride, and segregating water wastes from flammables.

2.6.2.4 Water Conservation and Reuse

It is more cost-effective to treat the waste with smaller volume but higher concentra-
tion than a waste with greater volume but lower concentration. Recycling and reus-
ing renovated wastewater is recommended. It has been estimated that about 1–100
tons (0.9072–90.72 metric tons) of water are used per ton of product. By modifying
processing procedures or auxiliary equipment, water usage and wastewater genera-
tion may be significantly reduced [21]. Examples are the use of surface rather than
barometric condensers, reuse of noncontact water, concentration of reaction mix-
tures to limit waste volume, and combining several processes.
King [34] has described an oil-dehydration evaporator/pyrolysis system for
energy recovery from pharmaceutical wastewater. Gas produced in the pyrolysis
unit is burned to provide steam required by the evaporator for oil dehydration.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 111

2.6.2.5 Segregation and Concentration of Wastes

Concentrating waste may reduce treatment cost. Concentration of wastewater may

also minimize the impact of intermittent hydraulic surges, specifically in fermenta-
tion operations. Segregation of waste streams, which allows concentrating the indi-
vidual waste for individual treatment, often allows more efficient removal of
particular pollutants. Segregation of wastes also allows using an individual treat-
ment method for the individual waste, such as using various evaporation or dewater-
ing methods to treat the separated waste streams for the fermentation wastes in an
in-plant treatment program. For example, cyanide destruction, metal removal, and
steam stripping to remove ammonia and organic solvents are utilized in the pharma-
ceutical industry for in-plant treatment. They need to be separated individually.
Individual process units are now commonly designed with allowance for waste
stream segregation.
For a similar reason, separation and treatment for storm runoff and sewer system
may eliminate the discharge of contaminated runoff and reduce treatment cost,
because the storm water from certain manufacturing areas can contain high levels of
toxic pollutants, while the storm runoff from some other areas and the sewer may
not. For the factories practicing in-plant treatment and direct discharge, the domes-
tic wastewater should be separated from polluted storm runoff. The latter should be
discharged directly to POTW or treated in-plant separately, while the non-polluted
storm runoff can be separated from polluted streams and discharged directly to
a river.
Sewers and pumps must be designed for peak flows to avoid flooding the mill or
bypassing the treatment plant. Also a good pipe and storage system are needed for
collecting the spills and the wastewater from various stages and storing wastewater
and biosolids.

2.6.2.6 Good Operating Practices

Good operating practices, which can help reduce waste generation, material losses,
and production cost, include closer supervision, production scheduling, material
tracking, inventory control, spill prevention, material handling and storage proce-
dures, documentation for process procedure, maintenance programs, employee
training, and management incentives. As these practices all apply to the general
waste minimization in all industries.

2.6.2.7 Reduction of Air and Dust Problems

Air pollution control in the pharmaceutical industry is mainly practiced by in-plant

control. Air and dust control technologies are fully described in Air Pollution
Control Engineering [35] and Advanced Air and Noise Pollution Control [36].
112 L. K. Wang et al.

There are three main sources of air pollution: fermentation process gas, dust, and
volatile solvents.
Most of the fermentations carried out in the pharmaceutical industry are aerobic
[25]. Air must be supplied to the fermentation organism. Compressed air is injected,
or sparged, into the lower end of the fermentor, which is simply a large, vertical,
circular tank. Supplying fresh air to the fermentation vessel on a constant basis
makes it necessary to vent or discharge an equal volume of what is termed “used”
air from the top of the fermentation vessel. The used air, or vent gas, has scrubbed a
number of materials, including carbon dioxide and many other more complex
organic materials from the fermentation as it moves up through the fermenting
mass. The organic materials generate odor. These odors vary with the material being
fermented and vary somewhat between different fermentors of the same material.
This “used” air, or vent gas, from the fermentor is the principal air pollutant. Wet
scrubbing of the vent gases may be practiced, though it may not be particularly suc-
cessful in many cases.
On large fermentors, the volume of gases is so great that the water needed to do
a scrubbing job (if water is used alone to do the job) is so large that, consequently,
generates even larger dimensions of polluted water to eliminate or even partially
reduce air pollution. Activated carbon can be used to adsorb the odor of the vent gas.
This method, however, may be practical only for large fermentors, because the
method requires a larger amount of carbon to accomplish a satisfactory end point.
Incinerating vent gas is a satisfactory solution. However, sometimes fuel is
needed to raise the vent gas temperature from fermentation temperature (generally
well below 40 °C) to an incineration level. At this point, this method may be uneco-
nomical. A possible more economical method may be piping the vent gas from the
fermentor to a boiler house and using it for combustion air in the boiler. This method
was used in large-scale operations such as in the fermentation plant at Abbott
Laboratories in North Chicago, IL, and at Eli Lilly and Company in Lafayette, IN,
both in the USA.
Air emission of volatile organic solvent can be a big air pollution problem, which
may be reduced by employing scrubbers or condensers to reclaim the solvent
vapors. Some factories may generate acid and solvent vapors such as methanol and
butyl acetate, which are sent to a house vacuum system for disposal. The waste
mycelium, or filter cake, which results from the initial separation of solids from the
fermented beer, is a frequent source of odor. The living cell biomass is quite perish-
able. If housekeeping standards are not maintained at a high level, this part of the
evaporation is also likely to contribute to the odor problem. Thus, good housekeep-
ing throughout the entire plant will do much to improve an odor situation.
Dust is a secondary pollution source. Dust inside a plant may cause “cross con-
tamination,” i.e., contamination of one drug by another. Penicillin is one of the
materials that are capable of causing extremely toxic reactions even when present in
trace quantities [25]. For example, aspirin tablet can cause a reaction of very serious
proportions (might result in death) in the presence of minute amount of penicillin.
Thus, penicillin dust should be absolutely isolated from the areas where other phar-
maceuticals are manufactured. Besides the isolation of penicillin production in a
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 113

separate area, the intake air to the areas producing other pharmaceuticals should be
carefully filtered, because the intake air may contain the air out of the penicillin
manufacturing area.
There are many methods used to remove dusts. A scrubber or RotoClone can be
used for removing many pollutants. However, the use of water with a scrubber or
RotoClone may result in water pollution problems. In such a case, a dry filter sys-
tem may be recommended. McNeil Laboratories used an extremely large Pangborn
baghouse-type dust collector to exhaust all the air from most manufacturing opera-
tions. It was 33 ft (10 m) long by 17 ft (5.2 m) wide by 20 ft (6 m) high. The inlet
duct was 44 in. (112 cm) in diameter. This single unit had a capacity of 36,000 scfm
(1019 m3/min). On this point, the pharmaceutical manufacturing areas in McNeil
Laboratories were supplied with 100% outside air [25], thus preventing secondary
pollutant from dust.

2.6.2.8 Waste Exchanges

Waste exchange is an alternative to recycling. It involves the transfer of waste to

another company for use “as is” or for reuse after treatment. Waste exchanges are
private or government-subsidized organizations that help identify the supply and
demand of various wastes. Waste exchanges have been established in some areas of
the USA to put waste generators in contact with potential users of the waste. The
USEPA [16] listed 48 state programs which offer technical and/or financial assis-
tance for waste minimization and treatment in the USA and 24 exchange operating
offices in the USA and Canada.
There are three types of waste exchanges: information exchanges, material
exchanges, and waste brokers. Metals and solvents are the most frequently recycled
materials via waste exchange, because of their high recovery value. Other wastes
commonly recycled through waste exchanges include acids, alkali salts and other
inorganic chemicals, organic chemicals, metal sludge, and solid residue from fer-
mentation and natural product extraction processes. The biosolids from the treat-
ment plant can also be beneficially reused off-site, which will be detailed in the
section of end-of-pipe treatment.

2.6.3 In-Plant Treatment

In-plant treatment in the pharmaceutical industry is mainly for treating priority pol-
lutants, such as solvents, metals, and cyanide, before combining the factory overall
waste stream. Although all three pollutants may be removed by the end-of-pipe
treatment, they can be removed more effectively by the in-plant treatment when
they are concentrated in the segregated stream. Therefore, the in-plant treatment can
also be regarded as a pretreatment to biological waste treatment.
114 L. K. Wang et al.

2.6.3.1 Cyanide Destruction Technologies

Chemical oxidation and high pressure and temperature hydrolysis are two treatment
processes which are effective in treating cyanide-bearing waste streams in the phar-
maceutical industry.
Chemical oxidation is a reaction in which one or more electrons are transferred
from the chemical being oxidized, here the cyanide waste, to the chemical initiating
the transfer, the oxidizing agent [37–39].

2.6.3.1.1 Chlorination

Cyanide can be destructed by oxidation either with chlorine gas under alkaline con-
ditions or with sodium hypochlorite. The oxidation of cyanide by chlorine under
alkaline condition can be described by the following two-step reactions:

Cl 2  NaCN  2 NaOH  NaOCN  2 NaC1  H 2 O (2.1)

3C12  6 NaOH  2 NaOCN  2 NaHCO3  N 2  6 NaC1  2H 2 O (2.2)
Cyanide is oxidized to cyanate at a pH of about 9.5–10.0. Usually 30 min are
required to complete the reaction, which markedly reduces the volatility and toxic-
ity (thousand fold reduction) of the waste. Figure 2.3 sketches a chlorination pro-
cess for a cyanide destruction system.
Since cyanate may revert to cyanide under some conditions, additional chlorine
is provided to oxidize cyanate to carbon dioxide and bicarbonate. The complete
oxidation of cyanate requires several hours at pH about 9.5–10.0 but only 1 h at a
pH between 8.0 and 8.5. Also, excess chlorine must be provided to break down
cyanogen chloride, a highly toxic intermediate compound formed during the

Fig. 2.3 Chlorination process for a cyanide destruction system [13]

2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 115

oxidation of cyanate. Although stoichiometric oxidation of a part of cyanide to cya-

nate requires only 2.73 parts of chlorine and complete oxidation of a part of cyanide
to carbon dioxide and nitrogen gas requires 6.82 parts of chlorine, nearly 3–4 parts
of chlorine are needed for oxidizing 1 part cyanide to cyanate, and 8 parts of chlo-
rine are needed for oxidizing 1 part of cyanide to gases in practice.
Iron interferes seriously with the alkaline chlorination of cyanide wastes.
However, it has been reported that ferrocyanides are treatable by alkaline chlorina-
tion at a temperature of 71 °C and at a pH of about 12.0.
Ammonia also interferes with the chlorine oxidation process by the formation of
chloramines, resulting in an increase of chlorine demand.
Cyanide levels around 0.040 mg/L are achievable by in-plant chlorination pro-
cesses in electroplating industry, if reaction interferences are not present [13]. It was
reported that in inorganic chemical industries, the free cyanide level after chemical
oxidation treatment is generally below 0.1 mg/L.
Chlorination process is a relatively low-cost system and does not require compli-
cated equipment and has received widespread application in the chemical industry.
It also fits well into the flow scheme of a wastewater treatment facility.
There are limitations and disadvantages for the chlorination process. For exam-
ple, toxic, volatile intermediate-reaction products can be formed. Thus, it is essen-
tial to control properly the pH to ensure that all reactions are carried to their end
point. Also, for waste streams containing other oxidizable matter, chlorine may be
consumed in oxidizing these materials, and this may interfere with the treatment of
the cyanide. A potential hazardous situation may exist in storage and handling when
gaseous chlorine is used.

2.6.3.1.2 Ozonation

Ozonation is an alternative oxidation treatment for cyanide destruction [13]. In fact,

ozone oxidizes many cyanide complexes (e.g., iron and nickel complexes) that are
not broken down by chlorine.
The oxidation of cyanide by ozone to cyanate occurs in about 15 min at a pH of
9.0–10.0, but the reaction is almost instantaneous in the presence of traces of copper
or manganese as catalysts. The pH of the cyanide waste is often raised to 12.0 to
obtain complete oxidation.
Oxidation of cyanate to the final end products, nitrogen and bicarbonate, is a
much slower and more difficult process unless catalysts are present. Since ozona-
tion will not readily affect further oxidation of cyanate, it is often coupled with such
independent processes as dialysis or biological oxidation.
The disadvantages of ozonation include the following:
1. Higher capital and operating costs than chlorination.
2. Toxicity problems similar to chlorination.
3. Ozone demand is increased when other oxidizable matter is present in the
waste stream.
4. The cyanide is not effectively oxidized beyond the cyanate level in most cases.
116 L. K. Wang et al.

2.6.3.1.3 Alkaline Hydrolysis

Alkaline hydrolysis is a process based on the application of heat and pressure [13].
In this process, a caustic solution is added to the cyanide-bearing wastewaters to
raise the pH to between 9.0 and 12.0. Then, the wastewater is transferred to a con-
tinuous flow reactor at temperatures in the range of 165–185 °C and pressures of
90–ll0 psi (625–763 kPa). The breakdown of cyanide in the reactor is generally
accomplished within a residence time of about 1.5 h.
It has been reported [13] that an average effluent level of 5.25 mg/L is achievable
for cyanide destruction. Alkaline hydrolysis is an economic process and has much
less storage and handling problems than chlorination. It is more likely suitable for
wastewaters with high concentrations of cyanide.

2.6.3.2 Metal Removal

Although the USEPA does not promulgate effluent guideline limitations for metals
in the pharmaceutical industry, it is useful to improve metal removal to release the
impact of heavy metals on the environment. In fact, some factories are practicing
removal of heavy metals in the waste stream [13]. The methods usually used for
metal removal are precipitation through adjustment to the optimum pH, sulfide pre-
cipitation, and chemical reduction.

2.6.3.2.1 Alkaline Precipitation

The solubility of metal hydroxides, in most cases, is a function of pH. Therefore,

adjustment to the optimal pH for precipitation of the metal hydroxide will result in
an effective removal of the metal. The alkaline precipitation for metal removal sys-
tem is schematically shown in Fig. 2.4. It should be noted that the solid contact
clarifier shown in Fig. 2.4 can be either a settling or a dissolved air flotation (DAF)
clarifier [40].
The solid metal hydroxides are coagulated (using coagulating agents) in clarifier
and deposited as sludge.
Lime is the commonly used chemical. In wastewaters containing substantial sul-
fate compounds, insoluble calcium sulfate precipitates will form when using lime.
In such instances, sodium hydroxide may be used.
The alkaline precipitation method is a well-demonstrated wastewater treatment
technology. It is easy to operate and has lower cost than other methods. Its limita-
tions and disadvantages are that (a) alkaline precipitation is subject to interference
when mixed wastes are treated and (b) relatively high quantities of residue can be
generated.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 117

Fig. 2.4 Alkaline precipitation for a metal removal system [13]

2.6.3.2.2 Sulfide Precipitation

For many heavy metals (such as copper, nickel, and zinc), their sulfides have much
lower Ksp than their hydroxides (see Table 2.8). Hence, the sulfide precipitation
method is applicable to the removal of all heavy metals by precipitating them as
metal sulfides. In the process, sulfide is supplied by the addition of a slightly soluble
metal sulfide that has solubility somewhat greater than that of the sulfide of the
metal to be removed. Normally ferrous sulfide is used [40].
Heavy metal sulfide sludges are less subject to leaching than hydroxide sludges.
However, sulfide precipitation produces sludge in greater volumes than does alka-
line precipitation. Separation of heavy metal sulfides by dissolved air flotation is
also a viable alternative [41].

2.6.3.2.3 Chemical Reduction

Some heavy metals (e.g., chromium which is a common metal contaminant in phar-
maceutical wastewater) have higher solubility in their higher valency (e.g., hexava-
lent chromium) than those in their lower valency (e.g., trivalent chromium). The
general procedure is first to reduce the valency of chromium from +6 to +3 adnd
then second to precipitate the product, chromium sulfate, at a suitable pH range by
either alkaline precipitation or sulfide precipitation, forming insoluble chromium
precipitates (either chromium hydroxide or chromium sulfide depending on the pro-
cess method used). Sulfur dioxide, sodium bisulfite, sodium metabisulfite, and fer-
rous sulfate are strong reducing agents in aqueous solution and are used for
chromium reduction. The chromium precipitates can be removed by filtration, sedi-
mentation clarification, or dissolved air flotation clarification [41, 42].
Some heavy metals are bonded in organic compounds, making their removal
more complicated. A typical example is from Merck, one of the largest
118 L. K. Wang et al.

Table 2.8 Annual mass loadings from direct and indirect pharmaceutical wastewater discharges
Mass loadings for direct Mass loadings for indirect dischargers
dischargers (1000 lb/year (1000 lb/year)+
Subcategories A, Subcategories A, B,
B, and C Subcategory D and C Subcategory D
Raw Raw Raw Raw-­
waste Final waste Final waste Discharge waste Discharge
Pollutants water effluent water effluent water to POTW water to POTW
Conventional
pollutants
 BOD5 83,000 5900 4100 300 169,000 169,000 5600 5600
 TSS 45,000 4600 1200 290 64,500 64,500 3000 3000
Priority
pollutants
 Volatile
organics 2000 77 240 6 2400 2000 18 18
 Semivolatile 120 2 17 0.2 390 330 16 16
organics
 Pesticides – – – – 0.02 0.02 – –
 Metals 60 22 1.2 0.7 51 45 2 2
 Cyanide 22 7 0.3 0.2 4.3 4.1 0.3 0.3
Nonconventional
pollutants
 COD 192,000 44,000 7500 800 411,000 411,000 24,000 24,000
 Volatile 5100 – 1000 – 7700 – 2200 –
organics
 Semivolatile 59 – 10 – 87 – 25 –
organics
 Pesticides/ 63 – II – 92 – 26 –
Herbicides
Industry
characteristics
 Number of 30 21 130 155
facilities
 Wastewater 21.38 3.54 31.1 8.8
flow, MGD
− Negligible

pharmaceutical companies. The company used an organomercury compound (thi-

merosal, RSHgEt) as a slow killing biocide in the fermentation process [43].
They developed an at-source treatment technology to remove and recover mer-
cury from the spent fermentation wastewater. The removal and reclamation of mer-
cury from wastewater is accomplished by the following four steps:
1. Using aluminum (at pH = 11.5) to reduce the sulfur-hydrogen of thimerosal to
release mercury at cationic state in water with the reaction:
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 119

Al  A13  3e  (2.3)

and one of the following reactions:

2e   RSHgEt  2H 2 O  RH  HSHgEt  2OH  (2.4)

2e   RSHgEt  H 2 O  RH  HOHHgEt  S2 
(2.5)
4e   RSHgEt  2H 2 O  RH   HHgEt   S2   2OH 
(2.6)
(Note: Since most of the biocides are associated with cell mass, caustic hydroly-
sis is used to release organomercury compound from cell paste before treatment.)
2. Using sodium borohydride to reduce mercury ions to the element state:

4Hg 2  NaBH 4  8OH   4Hg  NaBO2  6H 2 O

(2.7)
This process is at the ambient temperature and at pH = 10; the pH should be
maintained at 10 for about 10 min to complete the reaction. It should be noted that
at low pH borohydride is unstable. For example, at pH = 7, the following reaction
will occur:

BH 4  H 2 O  B  OH 3  OH   4H 2
(2.8)
3. Applying ultrafiltration: the treated water is stirred for 1 h and the colloid mer-
cury is separated by ultrafiltration; 99.7% removal can be reached (the Hg con-
centration in the effluent will be 110 ppb from an initial Hg concentration of
56 ppm).
4. Using granular activated carbon adsorption, the mercury concentration can be
reduced from the 110 to 10 ppb. The overall mercury removal can be reduced by
as much as 99.99% with the GAC filtration/polishing process (from an initial Hg
concentration of 56 ppm to l0 ppb in the effluent). Mercury can be reclaimed
from the filter cake of the ultrafiltration process.

2.6.3.3 Solvent Recovery and Removal

Solvents are used extensively in pharmaceutical manufacturing. Because solvents

are expensive, most factories try to recover and purify them for reuse whenever pos-
sible. Solvent recovery and recycling is one of the in-plant source control operations
and is also an in-plant treatment process. Typical techniques used for solvent recov-
ery are decantation, evaporation, distillation, extraction [13], and nebulization [44].
Stripping has also been proved to be an effective method to recover solvents from
pharmaceutical manufacturing processes.
120 L. K. Wang et al.

2.6.3.3.1 Steam Stripping

Steam stripping transfers the volatile constituents of a wastewater to a vapor phase

when steam is passed through preheated wastewater. The basic theory of steam
stripping is associated with the partitioning of the organic compound in the vapor
phase and in the wastewater phase. The partitioning coefficient (Ki), also called the
vapor-liquid equilibrium constant, of compound i is expressed as follows:

K i = Vi / Wi (2.9)
where Ki is the partitioning coefficient, also called the vapor-liquid equilibrium con-
stant, Vi is the mole fraction of organic compound i in the vapor phase, and Wi is the
mole fraction of organic compound i in the wastewater phase. Ki can be calculated,
for low pressures, from

K i  ri  Pi / P 
(2.10)
where ri is the activity coefficient of organic compound i in the wastewater at a cer-
tain temperature, Pi is the vapor pressure of the pure substance at the operating
temperature, and P is the total pressure.
Equations (2.9) and (2.10) show that the extent of separation is a function of the
physical properties of the volatile compounds and the temperature and pressure in
the stripper. The separation is also governed by the arrangement and type of
equipment.
The process is performed in a steam stripper which has various types, such as
packed tower, tray column, and steam flash tank. Flash tanks, which provide essen-
tially one stage of liquid-vapor contact, are used to strip extremely volatile com-
pounds. For the more difficult separations, columns filled with packing materials,
which provide large surface areas for liquid-vapor contact, can be used.
Figure 2.5 shows the processes and flow directions in a typical column stripper.
The solvent-containing wastewater is preheated, allowing the components of the
wastewater to separate by partial vaporization, then is introduced at the top or near
the middle of the column, and flows by gravity through the stripper. Steam is
injected through a sparger and rises countercurrent to the flow of the water. When
contacted with steam, the volatile organic compounds in a wastewater are driven
into the vapor phase.
Solvent-containing wastewater and condensed overhead vapors from the stripper
are allowed to accumulate in a gravity-phase separation tank. Because the conden-
sate mixes with fed wastewater accumulated in the tank, the solvent concentration
increases to the point at which it is saturated with solvent, when a two-phase mix-
ture is formed. The difference between the specific gravities of water and solvents
creates two immiscible liquid layers. One layer contains the immiscible solvents;
the other layer is an aqueous solution which is saturated with solvents.
The solvent layer is pumped to storage. The solvent can be recovered by decant-
ing the immiscible liquid layers or by recycling the condensed vapors directly to the
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 121

Fig. 2.5 Equipment for stream stripping solvents from wastewater [13]

gravity-phase separation tank, while the aqueous phase from the gravity-phase sep-
aration tank is pumped through a preheater where the temperature is raised by heat
exchange with the stripper effluent. After preheating, the solvent-saturated water is
introduced with the feed wastewater at the top or near the middle of the column and
flows by gravity through the stripper.
The hot effluent, which is discharged at the bottom of the stripper, is used as a
heating medium in the feed preheater. The temperatures of the feed, overhead, and
bottom are controlled at about boiling point. For example, the temperatures for a
methylene chloride removal in packed column steam stripper are at about 85–100 °C,
with the highest for the bottom temperature and the lowest for the feed temperature
(Table 2.9). The table indicates a poorer removal occurred under an upset condition
when the overhead temperature is too low (<85 °C). The pressure is usually under
atmospheric pressure.
122 L. K. Wang et al.

Table 2.9 Summary of BPT Regulation [11, 12]

Parameter Maximum 30-day average Daily maximum
BOD5 (mg/L) Reduction 90% from raw waste
COD (mg/L) Reduction 74% from raw waste
pH (unit) 6.0–9.0
TSS (mg/L) 1.7 times BOD concentration limitation –
Cyanide (mg/L)
Alternative Aa 9.4 33.5
Alternative Bb 9.4 (0.35) R 33.5 (0.18) R
a
Alternative A: Measure at diluent from cyanide destruction unit. Applies only when all cyanide-­
bearing wastes are diverted to a cyanide destruction unit and subsequently arc discharged to a
biological treatment system
b
Alternative B: Measure at final effluent discharge point. R: equals the dilution ratio of the cyanide
contaminated waste streams to the total process wastewater discharge flow

This practice is particularly advantageous in cases where the wastewater to be

stripped contains low concentration of the recovering solvents. The most economi-
cal operation of a wastewater steam stripper occurs when the feed is saturated with
the solvent to be recovered. The composition of the recovered solvent and economic
factors determines whether the solvent is reused within the plant, disposed of, used
as incinerator fuel, sold to solvent reclamation facility, or sold for other users.
Solvents recovered by steam stripping are normally not used directly in pharmaceu-
tical synthesis because of the US FDA purity requirements.
If the feed contains high concentrations of suspended solids, a filter may be
installed prior to the preheater to prevent fouling in the preheater and the column.
Steam stripping usually is a pretreatment method. It can effectively remove sol-
vent from wastewater. Steam stripping has been successfully used to remove methy-
lene chloride, toluene, chloroform, and benzene.
Many factories have reported that steam stripping enables the plants to meet a
POTW requirement that the concentration of explosive vapors in the plant sewer
pipes not exceed 40% of the lower explosion limit (LEL). Moreover, it has been
reported [13] that greater than 99% removal and an effluent with less than 10 mg/L
concentration have been achieved for a toluene wastewater. The stripped wastewater
is combined with other wastewater processes in another pretreatment system for
further end-of-pipe treatment, or further combined with sanitary wastewater and
then discharged to the POTW.

2.6.3.3.2 Air Stripping

Air stripping is also used to recover volatile organic compounds, such as benzene,
chloroform, 1,1,-I-trichloroethane, 1,2-dichloroethane, ethylbenzene, methyl chlo-
ride tetrachloroethylene, trichloroethylene, and toluene in pharmaceutical plants.
The air stripping process is similar to steam stripping. The basic theory of air
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 123

stripping is associated with the partitioning of the organic compound between air
and wastewater.

2.6.3.3.3 Advanced Physocochemical Treatment Processes

Carbon adsorption can also be used to remove organic solvents from a segregated
waste stream, especially in small quantities. Carbon adsorption method is widely
used in tertiary treatment.
The feasibility and extent of recovery and purification are governed largely by
the quantities involved and by the complexity of the solvent mixtures to be sepa-
rated. If recovery is not economically practicable, the used solvents may have to be
disposed of by means of incineration, landfilling, or contract disposal. It is expected
that some solvents can still be present in the wastewater even after an effort for
recovery. Further removal of solvents can be accomplished in the end-of-pipe treat-
ment in the combined overall waste stream.
Advanced physicochemical treatment processes available for treating the phar-
maceutical wastewater include coagulation and clarification, dissolved air flotation
(DAF), flotation-filtration (DAFF; filtration can be either sand filtration or GAC
filtration), granular activated carbon (GAC) adsorption, powdered activated carbon
(PAC) adsorption, wet air oxidation (WAO), supercritical water oxidation (SCWO),
Fenton oxidation, UV photocatalytic oxidation, ultrasound oxidation, air stripping,
distillation, electrochemical oxidation, ozonation, membrane filtration (MF, UF,
RO, ED, MBR), or other advanced oxidation processes (AOP), combined oxidation-­
reduction process, etc. Evaluation of these processes is presented in Sects. 2.7.7.1
and 2.7.7.2. Of these advanced treatment processes, DAF, DAFF, GAC, air strip-
ping, distillation, and membrane processes are suitable for recycling and reusing of
chemical compounds and/or water. In view of the pollution load reduction and
chemical cost saving, it is necessary to recover chemical compounds or raw materi-
als as much as possible. In view of the scarcity of water resources, it is necessary to
understand and develop methodologies for the treatment of pharmaceutical waste-
water as part of water management. While most of the advanced treatment pro-
cesses are technically feasible for treating the pharmaceutical wastewater, their
economical feasibility needs to be carefully evaluated before any implementation.

2.6.4 End-of-Pipe Treatment Technologies

End-of-pipe treatment is mainly designed to treat a number of pollutants in a plant’s

overall waste stream before it is discharged directly to a body of surface water,
although it is sometimes used for pretreating the waste stream when a wastewater is
designed for indirect discharge, i.e., discharging to the POTW for further treatment.
The pretreatment for pharmaceutical waste is mainly for reducing the toxicity of the
wastewater in order not to be harmful for the biological treatment system.
124 L. K. Wang et al.

Pretreatment is mainly accomplished by the so-called in-plant treatment as stated

previously. This section discusses the end-of-pipe treatment for direct discharge.
Generally, a secondary treatment facility is needed for an end-of-pipe treatment
for pharmaceutical wastes [13]. The treatment schemes involve primary treatment
(screening, equalization, neutralization) followed by either a secondary biological
treatment or a secondary physicochemical treatment. Additional tertiary treatments
may also be needed.

2.6.4.1 Primary Treatment

The common primary treatment methods in the pharmaceutical industry are (a)
coarse solid removal by screening; (b) primary sedimentation, applying gravity
separation to remove grit and settleable solids and using a skimmer to remove float-
ing oil and grease; (c) primary chemical flocculation/clarification; and (d) dissolved
air flotation.

2.6.4.1.1 Equalization and Neutralization

Flows are usually required to be equalized, especially if the waste from the produc-
tion plant is not equally distributed (either in flow rate or in waste characteristics)
around the clock. In this case, an equalization tank is needed to minimize or control
fluctuations in wastewater characteristics to provide optimum conditions for the
subsequent treatment processes. The main benefits of equalization are as follows:
1. Providing continuous feed to biological systems over periods when the manufac-
turing plant is not operating
2. Providing adequate dampening of organic: fluctuations to prevent shock loading
to biological systems
3. Preventing high concentrations of toxic materials from entering the biologi-
cal systems
4. Minimizing chemical requirements necessary for neutralization
Also, neutralization and nutrients addition can be accomplished in the equalization
step. A pH between 6.5 and 8.5 should be maintained in a biological system to
ensure optimum biological activities. Neutralization is important for chemical syn-
thesis plants as shown in Table 2.2.
Neutralization is performed by adding basic or acidic substances depending on
the pH of the waste stream. An economical option is by adding a proportional com-
bination of acid and basic wastewater streams.
The raw materials used in fermentation and biological product extraction manu-
facturing are mainly from natural plants and animals. Nutrients (such as nitrogen
and phosphorous) may not be needed. However, for some other wastes, nutrient
addition may be necessary prior to biological waste treatment. Mixing is usually
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 125

provided to ensure adequate equalization and to prevent settleable solids from

depositing in the basin [45].

2.6.4.1.2 Screening and Clarification

All waste flows should be passed through screens to remove large suspended matter
and through clarification (sedimentation or flotation) tanks to remove suspended
solids. Rectangular gravity clarifiers are usually used for primary sedimentation,
although circular gravity tanks or dissolved air flotation tanks are equally efficient.
Chemical coagulation and flocculation can also be combined with primary treat-
ment to increase TSS removals. Primary treatment is an important pretreatment for
the subsequent secondary biological waste treatment, which may remove 20–50%
of 5-day BOD.

2.6.4.1.3 Primary Flotation Clarification and Secondary Flotation Clarification

When conventional sedimentation cannot effectively remove suspended solids or oil

and grease, primary flotation may be used instead of primary sedimentation before
secondary biological waste treatment [46, 47].
In dissolved air flotation (DAF), wastewater is pressurized to 50–90 psi
(347–624 kPa) in the presence of sufficient air to approach saturation [40, 45, 48,
49]. When the pressure in the air-liquid mixture is released to atmospheric pressure
in the flotation unit, micro air bubbles are released from solution. The suspended
solids or oil globules are floated by these micro air bubbles, rising to the surface
where they are skimmed off.
DAF can also be used as a secondary clarifier.

2.6.4.2 Secondary Biological Treatment

2.6.4.2.1 Activated Sludge

Activated sludge is the most widely used secondary biological process for treating
pharmaceutical wastewater [50–56]. It is mainly used for medium and large waste-
water flows.
A typical activated sludge treatment system consists of an aeration tank for aero-
bic biological treatment, a secondary clarifier for solid separation, and an activated
sludge return system for sludge recycle [57]. The aeration tanks are loaded with the
equalized, neutralized, and pretreated wastewater. In the aerobic biological degrada-
tion, the soluble biodegradable wastes are transferred to insoluble microbial
biomass.
The secondary sedimentation clarifiers settle the biosolids from the biologically
treated wastewater, resulting in a clear effluent which meets the standards (mainly
126 L. K. Wang et al.

the BOD and TSS) for direct discharge. The major part of the settled biosolids is
further treated before disposal or reuse. A part of the settled biomass is returned to
the aeration tank as the return activated sludge.
The return activated sludge is fed to the aeration tank to ensure a sufficient
amount of microbial population for the degradation of the organic waste is present.
The biomass is measured by the mixed liquor volatile suspended solids (MLVSS).
Complete mixing and adequate aeration are essential in the aeration tanks.
Sufficient oxygen should be furnished to maintain dissolved oxygen throughout the
aeration volume.
There are various types of modes for operating the activated sludge system, such
as conventional, extended aeration, step aeration, contact stabilization, and com-
pletely mixed. Figure 2.6 shows the flow diagrams of a few selected activated sludge

Fig. 2.6 Flow diagrams and applications of major activated sludge processes
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 127

processes. The treatment mode is selected according to the characterization of the

wastes and the goal of treatment [58–60].
Once maximum and normal raw waste loads and flows have been determined,
the design criteria for the biological treatment plant can be established. In addition
to the removal of 5-day BOD and suspended solids, some toxic organic matters are
slightly reduced during the process. Activated sludge treatment systems can be
designed for the purpose of nitrogen removal by operating the system to accomplish
nitrification and denitrification [61, 62].
Some activated sludge treatment systems experience severe filamentous micro-
organisms buildup accompanied with very poor settling. A pilot-scale experiment
was conducted to improve sludge settling for a nitrifying activated sludge system,
treating 1.2 MGD (4.54 MLD), equivalent to 10,000–15,000 kg 5-day BOD per day,
of pharmaceutical wastewater from both synthetic and fermentation processes. The
concentration of filamentous organisms and the mixed liquor sludge volume index
(SVI) can be reduced by changing the aeration pattern from three aeration basins in
parallel flow to three completely mixed compartments in series. Such process
change results in reducing the filamentous population and improving settling
characteristics.
Alternatively, a secondary flotation clarifier can be adopted to replace a second-
ary sedimentation clarifier to solve the problems of sludge bulking and rising [40,
57, 63].
According to Mayabhate et al. [64], an oxidation ditch activated sludge system
was capable of providing acceptable treatment for pharmaceutical wastes.
Datta Gupta et al. [28] described a complete treatment system for antibiotic pro-
duction wastewater including lime neutralization, clarification, activated sludge
treatment, postaeration, and chlorination. The effluent was disposed of by irrigation,
while the biosolids were dried and utilized as fertilizer.
Schumann [65] described a treatment system for high-strength pharmaceutical
wastewater, which included neutralization and aerobic activated sludge treatment
with aerobic sludge stabilization [29].

2.6.4.2.2 Aerated Lagoon

Aerated lagoons are usually rectangular in shape, with a length-to-width ratio of

2:1. The depth of lagoons is usually about 8–12 ft (2.44–3.66 m). The lagoon bot-
tom and sides are lined and have a freeboard of at least 3 ft About 1–2 months of
retention time are required for treatment by an aerated lagoon. The detention time
and waste loading determine the required lagoon volume, which in turn determines
the surface area of the lagoon [66].
Complete mixing and adequate aeration are essential. Sufficient oxygen should
be furnished to maintain dissolved oxygen throughout the entire 8–12-ft depth
(Fig. 2.7). Aerators should be spaced to provide uniform blending for dispersion of
dissolved oxygen and suspension of microbial mass. The oxygen provided for aer-
ated lagoons is commonly provided by mechanical aeration, diffused aeration, or
128 L. K. Wang et al.

Fig. 2.7 Aerated lagoon system

induced surface aeration. The mechanical aeration units can be either floating or
platform-mounted.
The aerated lagoon is the second widely used biological treatment method for
treating pharmaceutical wastewater. It is mainly used for relatively small plants and
can achieve 85–95% reduction of 5-day BOD.

2.6.4.2.3 Trickling Filter

Trickling filters are fixed film reactors using a biological process for wastewater
treatment [67]. It is widely used in pharmaceutical waste treatment for plants
medium to large in size. The filter medium consists of a bed of coarse material such
as broken stones, plastic rings, corrugated plastic sheets, or plastic tubes over which
wastewater is distributed. The plastic media are predominant for high-rate filters
such as for strong industrial wastewaters with high loading rates. Nitrification-­
denitrification can be accomplished by using low loading rates and multistage trick-
ling filtration.
Wastewater is applied to trickling filters by a rotary distributing system. The
wastewater then trickles downward through the media, on which a zoogleal slime
layer is formed (Fig. 2.8). Dissolved organic material in the wastewater is trans-
ported into the slime layer where biological oxidation takes place. The effluent liq-
uid is then collected by an underdrain system. Organic removal occurs by adsorption
and assimilation of the soluble and suspended waste materials by microorganisms
attached to the media. Oxygen for the process is supplied from air circulating
through the interstices between the filter media, which increases dissolved oxygen
in wastewater.
The quantity of biological slime produced is controlled by available food. Growth
will increase as the organic load increases until a maximum effective thickness is
reached. This maximum growth is controlled by physical factors including hydrau-
lic dosage rate, type of media, type of organic matter, amount of essential nutrients
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 129

Fig. 2.8 Trickling filter

130 L. K. Wang et al.

present, temperature, and the nature of the particular biological growth. During
trickling filter operations, biological slime is sloughed off, either periodically or
continuously. The sloughed biomass is removed in the subsequent clarification pro-
cess. Recirculation of trickling filter effluent is practiced in high-rate trickling filters
which improve the filter efficiency.
The overall performance of trickling filters is related to the hydraulic and organic
loading. The performance can be correlated to either hydraulic loading or organic
loading when the BOD concentration in wastewater and the depth of the filter
remain constant [67–69]. Other factors that affect the performance of trickling filter
plants include the specific surface area of media, flow distribution and dosing fre-
quencies, wastewater temperature, recirculation rate, underdrain and ventilation
system, filter staging, and secondary clarification [67, 70, 71].
It is important to note that either sedimentation clarifiers or dissolved air flotation
clarifiers can be used as the secondary clarification units for separating the biomass
from the effluent of trickling filters [63].

2.6.4.2.4 Anaerobic Treatment

Anaerobic treatment involves the breakdown of organic wastes to gas (mainly meth-
ane and carbon dioxide) in the absence of oxygen. This process involves two steps:
the breakdown of organics by facultative and anaerobic organisms to organic acids
and the subsequent breakdown of these acids to methane and carbon dioxide
[51, 72].
Since the anaerobic process has less cell synthesis than that in the aerobic sys-
tem, the nutrient requirements are correspondingly less. The conversion of organic
acids to methane gas yields little energy. The rate of growth is slow, and the yield of
organisms by synthesis is low. Therefore, the kinetic rate of removal and the sludge
yield are considerably less than those in the activated sludge process or the trickling
filter process. Figure 2.9 illustrates several anaerobic processes that have been used
in the treatment of pharmaceutical wastewater [73–76].
The conventional anaerobic treatment process provides a continuous or intermit-
tent feeding without solid separation. The detention time is usually 10–30 days and
the minimum time is 3–5 days.
An anaerobic-contact process provides for separation and recirculation of seed
organisms, therefore allowing process operation at detention periods of 6–12 h. A
90% removal of COD was reported for wastewater at a loading of 2.5 kg COD/m3/
day [77].
In an anaerobic filter, the growth of the anaerobic microorganisms occurs on the
surface of packed media. The filter is operated either in the upflow or downflow
mode, and part of the effluent is recirculated. The packed filter media also provide
for the separation of solids and the gas generated in the anaerobic process. Jennet
and Dennis [78] treated pharmaceutical wastewater and achieved a 97% removal of
COD at a loading of 3.5 kg COD/m3/day at 37 °C. Sachs et al. [79] used an anaero-
bic filter to treat biological or chemically synthesized pharmaceutical wastewater.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 131

Fig. 2.9 Anaerobic wastewater treatment processes

With a loading of 0.56 kg COD/m3/day at 35 °C and 36 h hydraulic retention time,

they achieved 80% COD removal.
In a fluidized bed reactor, the wastewater is pumped upward through a sand bed.
Part of the effluent is recycled. Stronach et al. [80] utilized anaerobic fluidized beds
to treat two types of wastes. The first waste, a propanol-containing waste, was
132 L. K. Wang et al.

nutrient limited and caused inhibition of methanogenesis, whereas the second waste,
a methylformamidecontaining waste, appeared to contain a non-biodegradable and
toxic fraction, which did not inhibit methanogenesis but caused a reduction in COD
removal and erratic volatile acid production. The feed flow had a COD concentra-
tion of 2500 mg/L, which was applied at an organic loading rate of 4.5 kg COD/m3/
day and with a hydraulic retention time of 0.53 day. Final COD removal was 54 and
45% for the first and second wastes, respectively.
In an upflow anaerobic sludge blanket process reactor, wastewater is directed to
the bottom of the reactor where it is distributed uniformly. Methane and carbon
dioxide rise upward and are captured in a gas dome. The flow passes into the settling
portion of the reactor where solid-liquid separation takes place.
An anaerobic degradation of pharmaceutical antibiotic fermentation wastewater
was studied at a pilot scale [81] and then was applied to a full-scale treatment plant.
The waste contained a high proportion of suspended solids representing about 40%
of the COD as well as residual amounts of antibiotics, extraction solvents, grain
flours, sugars, protein, and nutrients. Four treatment configurations were piloted: a
downflow anaerobic filter, a downflow/upflow anaerobic filter, an upflow anaerobic
sludge blanket, and a low- rate anaerobic reactor. The high-rate systems were ulti-
mately incapable of assimilating the feed pollutants, resulting in excessive loss of
biomass and, therefore, low soluble COD removals. The low-rate system adequately
hydrolyzed the feed pollutants and yielded 70% COD and 80–90% TSS removals.
The presence of antibiotic residuals did not affect the system.
Shafai and Oleszkiewicz [82] investigated the anaerobic ammonification of
wastewater from an estrogen-extracting pharmaceutical plant. Both flow-through
and batch anaerobic reactors were used to treat a waste with high loading of total
dissolved solids (TDS), TKN nitrogen, and total organic carbon (TOC). It was
found TDS concentrations over 17 g/L in the flow-through reactors and in excess of
10 g/L in the batch reactors to be inhibitory to both ammonification and
methanogenesis.
Anaerobic treatment has also been used as an additional treatment to supplement
the main treatment system. One example is at the Abbott Laboratories in North
Chicago, Illinois. The healthcare product manufacturer operates a large fermenta-
tion and chemical synthesis plant. The total wastewater flow from the factory is 0.92
MGD (3.48 MLD); the COD, BOD, and TSS loads are 25,000, 11,500, and 3500 lb/
day, respectively (11,340, 5216, and 1588 kg/day, respectively). About 70–85% of
the waste is from the fermentation process. The wastewater flow was treated in an
extended aeration activated sludge plant. To accommodate the growth and expand-
ing load from the fermentation process, a low-rate anaerobic reactor was added as a
pretreatment step for the high-strength fermentation wastewater prior to aerobic
treatment. The anaerobic reactor was also used for the digestion of the raw waste
solids from fermentation and for the wasted sludge from the aerobic system. The
flow diagram of the treatment plant is shown in Fig. 2.10. The low-rate anaerobic
reactor performance operating at a temperature of 28.5–32.5 °C and with a hydrau-
lic retention time of 9.5–10.0 days was as follows: 79% removal of COD, 86%
removal of 5-day BOD, and 83% removal of TSS.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 133

Fig. 2.10 Aerobic-anaerobic treatment of chemical synthesis and fermentation wastewater

effluents

Fig. 2.11 Schematic diagram of rotating biological contactors

2.6.4.2.5 Advanced Biological Treatment Methods

Other biological treatment methods utilized in pharmaceutical wastewater are waste

stabilization ponds [66], rotating biological contactors (RBC) [83–85] (see
Fig. 2.11), polishing ponds, sequencing batch reactors (SBR) [86], membrane bio-
reactor (MBR), and sequencing batch biofilters [87]. For detailed description of
these processes, the readers are referred to the books Biological Treatment Processes
[51], Advanced Biological Processes [72], Membrane and Desalination Technologies
[123], and Environmental Flotation Engineering [124]. Additional biological treat-
ment processes are introduced in other chapters of this book [175–176].
134 L. K. Wang et al.

2.6.4.3 Tertiary Treatment

Tertiary treatment using physicochemical processes is usually applied for further

improving the quality of the secondary effluent following biological treatment.
Examples of these additional treatment methods are the polishing pond, coagula-
tion/flocculation/clarification, secondary neutralization, chlorination, ion exchange,
and filtration (multimedia, sand, and granular activated carbon) [40, 88].

2.6.4.3.1 Filtration and Carbon Adsorption

Filtration is widely used for polishing wastewater. The most common filter type is a
multimedia of activated carbon and sand. The filter needs a periodical backwash and
is used mainly for removal of relatively coarse particles. Granular activated carbon
is more versatile in dealing with various kinds of small suspended solid particles,
colloidal, and dissolved pollutants [164, 169].
Carbon adsorption uses activated carbon which has a great specific surface area
(surface area per unit volume) to effectively adsorb pollutants [40, 88]. Granular
activated carbon is an effective and economical adsorbent because besides its higher
specific surface area, it has a high hardness, which lends itself to reactivation and
repeated use.
The granular activated carbon adsorption process is usually preceded by prelimi-
nary filtration or clarification to remove insoluble particles. Once the carbon is
depleted, it can be reactivated by heating to a temperature between 1600 and 1800 °F
(871–982 °C) to volatilize and oxidize the adsorbed contaminates. Oxygen in the
furnace is normally controlled at less than 1% to avoid loss of carbon by combus-
tion [13].
The application of carbon adsorption in pharmaceutical industry is limited. Most
of the priority pollutants (heavy metals, volatile organics, and cyanide) are gener-
ally reduced more effectively and with less cost by other technologies. This method
is particularly applicable in situations where pollutants in low concentrations not
amenable to treatment by other technologies must be removed from waste streams.
Holler and Schinner [89] arrived at the same conclusion and stated that for eco-
nomic reasons carbon adsorption should be mainly used as a tertiary treatment for
final polishing of secondary effluents. Bauer et al. [90] used activated carbon filtra-
tion in an activated sludge system to remove toxic compounds. More details on the
removal of organics and toxic material from pharmaceutical wastewater effluents
can be found in [91–99].
Besides the usage of granular activated carbon as a filtration media, powdered
activated carbon (PAC) has been used as an additive in an activated sludge system
[26]. One of the experiments showed that the MLSS concentration increased from
5850 to 8830 mg/L as the PAC dosage to the influent was increased from 208 to
1520 mg/L. The 0.7 mg/mg PAC dosage resulted in 50% additional removal of COD.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 135

2.6.4.3.2 Coagulation, Flocculation, and Clarification

Coagulation is a process used for the removal of colloidal and fine suspended par-
ticles [100, 101]. Kharlamova et al. [102] used alum, lime, and bentonite clay as
coagulants to treat pharmaceutical waste effluents. The treated effluents had lighter
coloring and increased transparency. The reduction in BOD and COD, however,
was limited. On the other hand, the researchers were successful in destroying syn-
thetic surfactants used in the production of antibiotics using hydrogen peroxide as
an oxidant and iron and aluminum ions as catalysts. However, flocculation and
coagulation may not be effective or cost-efficient for pharmaceutical wastewater
treatment, although it is able to reduce COD concentrations [64].
PAC can also be applied to a coagulation/flocculation/clarification system for
removal of toxic substances [63]. Clarification can be either a sedimentation clarifi-
cation or a flotation clarification.

2.6.4.3.3 Chlorination

Chlorination as a means of disinfection is needed before the discharge of effluent

after biological treatment. For example, post-aeration and chlorination are used in
addition to activated sludge treatment for wastewater treatment at a penicillin pro-
duction facility [28].
Table 2.10 shows a summary of end-of-pipe treatment methods used for waste-
water treatment in the pharmaceutical industry. It is estimated [13] that the activated
sludge process is the most widely used biological treatment method, at about 60%
of the biological treatment plants. Physicochemical treatment methods have been
used in only 20% of the plants, out of which thermal oxidation is the most
widely used.

2.6.4.4 Residue Treatment and Waste Disposal

A large proportion of the material input to the manufacturing process ends up as

process waste. Fermentation and biological extraction, as well as the formulation
processes, are typical examples. Besides excess sludges generated during produc-
tion processes, sludge can also be generated in the processes of pretreatment, pri-
mary treatment, secondary treatment, and tertiary treatment.
Fat and oil may also occur during biological extraction manufacturing proce-
dures, which are skimmed-off in flotation or settling tanks. The sludges generated in
the pretreatment stages usually contain contaminants such as traces of solvents and
heavy metals. Organic contaminants in the sludge are either (a) traces of solvents
used in the fermentation, chemical synthesis, and biological extraction manufactur-
ing steps or (b) reactants or byproducts of the chemical synthesis steps. Biological
sludges, also known as biosolids, need to be thickened, dewatered, conditioned, and
stabilized before disposal. Disposal methods of sludge include incineration, landfill,
136 L. K. Wang et al.

Table 2.10 Pretreatment pollutants standards [13]

No. of occurrences in Max. wastewater concentration level
Pollutant wastewaters (μg/L)
Cyanide 5 590
Acrolein 2 100
Acrylonitrile 1 100
Benzene 6 580
Carbon tetrachloride 1 300
Chlorobenzene 2 11
1,2-dichloroethane 2 290
1,1,1-trichloroethane 4 360.000
1,1-dichloroethane 3 27
Chloroform 6 1350
1,1-dichloroethylene 2 10
1,2-trans-­ 1 550
dichloroethylene
Ethylbenzene 3 21
Methylene chloride 9 890.000
Bromoform 1 12
Tetrachloroelhylene 1 2
Toluene 6 1050
Trichloroethylene 1 7

and reuse. In the latter two cases, sludge stabilization and disinfection will be
needed [29, 65].
Recovered solvents may be used as fuel for incineration or other kinds of benefi-
cial uses. Fats and oil may be incinerated or landfilled along with sludge or may also
be transferred to other industry such as soap manufacturing to be used as raw mate-
rials. Such a beneficial usage of residue is one of the waste exchange programs that
should be encouraged.
Sludge may be spread on land for agricultural purposes [103] or sold as an ani-
mal feed supplement. However, the wasted biological sludges are generally con-
taminated with varying degrees of potentially toxic materials, which may exclude
the above two types of beneficial usage.
Wickramanyake [104, 105] discussed the treatment of sludge generated at a
DNA processing facility. The sludge consisted mainly of biological solids (i.e., bio-
solids), such as cells and cell debris. The solid levels in the sludge samples can vary
depending on the process used to concentrate solid materials. The solid content and
physical properties of biosolids significantly affect decontamination processes
including incineration, thermal (dry heat and steam) treatment, gamma and electron
radiation, microwave radiation, and chemical decontamination [29]. Each of these
microbial inactivation techniques can be effective in the treatment of the DNA bio-
solids. Since verification of the extent of decontamination is difficult with biosolids,
high safety factors should be incorporated into the design of treatment units, and
good maintenance and operating procedures should be employed.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 137

Incineration may not be legally practiced in some areas, such as New York City.
The New York City Department of Environmental Protection has developed com-
prehensive plans to handle sludge problems [106]. The plan includes heat drying,
composting, chemical stabilizing of dewatered biosolids, landfilling (mainly for
toxic-containing biosolids), and, more importantly, beneficial usage. The beneficial
applications include the spreading of biosolids on or just below the surface of land
to benefit soil and plants and as a substitute for soils imported by the city for daily
cover at active landfills or as capping material for closed landfills.

2.7 Case Study

This section uses a factory producing antibiotics by fermentation as an example of

waste generation and end-of-pipe treatment in the fermentation pharmaceutical
industry.

2.7.1 Factory Profiles

Ansa, a plant at Izmit, Turkey, produces antibiotic pharmaceutical products by fer-

mentation. It has the capacity to produce 120 metric ton/year of tetracycline and
oxytetracycline derivatives and 1.5–2.0 metric ton/year of gentamicin sulfate. The
following description covers the period when the production rate of the factory was
50–60% of full capacity. The production was carried out year round, 7 days a week
and 24 h a day with three shifts. The maximum daily production capacity was
400 kg/day for tetracycline and oxytetracycline and 20 kg per 3 days (intermittent
production) for gentamicin [107].

2.7.2 Raw Materials and Production Process

The production used different raw materials from agricultural sources and used
various chemicals (Table 2.11).
Figure 2.12 shows the production mode. A bacterial-based mycelium was first
produced in the microbiology laboratory.
The fermentation involved two phases: solubilization of antibiotics by acidifica-
tion and filtration. The whole process was carried out on a batch basis.
The processes following the filtration of fermentation product were slightly dif-
ferent between tetracycline and oxytetracycline production and gentamicin produc-
tion. For tetracycline and oxytetracycline production, the fermentor filtrates were
treated by extraction, pH adjustment, filtration, precipitation, centrifugation, com-
plex formation and crystallization, and purification, before yielding the final
138 L. K. Wang et al.

Table 2. 11 Solubility Products (Ksp) for Insoluble Metal Salts [13]

Compound Ksp Metal Ion Conc. (μg/L)
CuS 6 × 10−36 1 × 10−10
NiS 2 × 10−25 8 × 10−6
ZnS 1.6 × 10−25 2 × 10−5
Cu(OH)2 3.5 × 10−19 25
Ni(OH)2 1.5 × 10−15 400
Zn(OH)2 1.8 × 10−14 1 × 10−3

Fig. 2.12 Antibiotic production process system [107]

product. For gentamicin production, the filtrates were treated by extraction, chro-
matographic resin adsorption, evaporation, filtration, crystallization, or spray drying
to yield the final product.

2.7.3 Waste Generation and Characteristics

The production generated 33 sources of wastewater discharges. They can be grouped

into seven main processes:
1. Wastewaters from fermentation processes (strong)
2. Wastewaters from extraction and purification processes (strong)
3. Wastewaters from recovery process (strong)
4. Floor and equipment washings (dilute)
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 139

5. Laboratory wastes, miscellaneous wastes (varied)

6. Sanitary wastes
7. Waste cooling water (uncontaminated)
These waste streams can be further grouped into three groups: the strong process
wastes, the diluted wastes, and the cooling water. The strong process wastes were
from fermentation process, extraction and purification processes, and recovery pro-
cess. The diluted wastes were from the floor and equipment washings, laboratory
wastes, and miscellaneous wastes (varied). The cooling water was confined, without
contacting with processing water, which, in fact, was uncontaminated and generated
no waste.
The flow rates for the three main streams were as follows:
1. Strong process wastes: Q = 120 m3/day
2. Diluted wastes: Q = 160 m3/day
3. Cooling water: Q = 1000 m3/day
Table 2.12 lists the flow and concentrations of some major traditional wastes for
the above first two major types of wastewater. The process wastes were very strong
in organic content, having a 5-day BOD of 13,500 mg/L, a COD of 34,000 mg/L,
and a BOD/COD ratio of 1:2. The total loads were 1680 kg/day of 5-day BOD and
4180 kg/day of COD. The diluted wastes had 400 mg/L of 5-day BOD and
600 mg/L of COD.
In fact, full segregation of the strong and dilute waste streams was not possible
due to the complexity of existing piping system. The process wastes and dilute
wastes were actually diluted with the wasted cooling water down to a 5-day BOD of
8400 and 50 mg/L, respectively, and the flow rates at 200 and 800 m3/day,

Table 2.12 Methylene chloride removal in packed column steam stripper [13]
Feed Methylene chloride
Sample temp. Overhead Bottoms Feed rate Steam (mg/L)
number (°C) temp. (°C) temp. (°C) (gpm) rate (L/h) Influent Effluent
1 87 97 104 9.6 160 NAa 0.926
2 86 98 102 8.9 160 NA 5.10
3 86 94 101 9.0 150 NA 4.94
4 86 89 102 9.0 150 NA 3.00
5 85 89 102 9.0 150 NA 1.99
6 85 86 102 9.0 150 NA 5.70
7 85 84 102 9.0 155 NA 22.80b
8 84 84 101 9.0 155 NA 38.05b
Composite of Influent samples 260 NA
Average of all effluent datum points 10.31
Average of effluent datum points obtained under normal operating 3.61
conditions
a
NA means not analyzed. 1 gpm = 3.785 LPM = 3.785 L/min
b
Efffluent concentrations under upset conditions, overhead temperature < 85°
140 L. K. Wang et al.

respectively, as shown in Table 2.12. Combining the waste streams yielded a total
flow of 1000 m3/day and 5-day BOD of 1720 mg/L.
The strong process waste didn’t maintain a uniform composition, which was
drastically affected when tetracycline and oxytetracycline were alternately pro-
duced together with gentamicin. Moreover, the strong waste had strong sulfate level
and frequent changes in the products and wastewater properties. An adequate dilu-
tion of process waste could avoid the toxicity and BOD shock load when otherwise
treating a smaller flow and stronger waste, where a high concentration of sulfate and
more variable discharge were encountered. These factors all affected the treatability
properties of the wastes.

2.7.4 End-of-Pipe Treatment Case Histories and Green

Environmental Technologies

2.7.4.1 Case Histories of Current Technologies

Table 2.13 presents a summary of all end-of-pipe treatment processes [11, 12].
However, aerobic treatment scheme was selected for end-of-pipe waste treatment as
an engineering project. Anaerobic treatment was not chosen because (a) a total of
360,000 m3/day of air, with oxygen content, was regularly discharged from the
plant, favoring an aerobic process as an economic treatment system, and (b) the
inhibition problems were possibly due to high sulfate levels, frequent changes in
products, and fluctuation in wastewater characteristics.
An activated sludge treatment system shown in Fig. 2.13 was selected and
designed for the pharmaceutical plant [107]. Tables 2.14 and 2.15 introduce the raw
material consumption and the wastewater characteristics, respectively, of the antibi-
otic production plant [107]. It basically involved a separate equalization of waste
streams, pH adjustment, aeration, activated sludge system, secondary clarification,
and biosolid treatment.
The strong and diluted wastes (flow rates of 200 and 800 m3/day and with 5-day
BOD at 8400 and 50 mg/L, respectively) were equalized in separate tanks, because
they had quite different waste discharge rates and continuous variation in waste
characters around the clock. The two equalized waste streams were then combined
for the next treatment step: pH adjustment. The combined waste had a 5-day BOD
of 1720 mg/L and a flow rate of 1000 m3/day.
The waste stream was then sent to a single-stage activated sludge unit. The aera-
tion tank had four aeration compartments in series and was designed for a hydraulic
detention time of 24 h.
The two alternating process wastes (i.e., tetracycline and oxytetracycline were
alternately produced together with gentamicin) showed substantially different prop-
erties affecting the mode of treatment. The yield value was much lower for oxytet-
racycline waste. Oxytetracycline had also a very high maximum substrate utilization
rate (k), but it took a significantly large range of substrate concentration to reach this
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 141

Table 2.13 Summary of end-of-pipe treatment processes [11, 12]

End-of-pipe technology Number of plants

Equalization 62
Neutralization 80
Primary treatment 61
Coarse settleable solids removal 41
Primary sedimentation 37
Primary chemical flocculation/clarification 12
Dissolved air dotation 3
Biological treatment 76
Activated sludge 52
Pure oxygen 1
Powdered activated carbon 2
Trickling filter 9
Aerated lagoon 23
Waste stabilization pond 9
Rotating biological contactor 1
Other biological treatment 2
Physical/chemical treatment 17
Thermal oxidation 3
Evaporation 6
Additional treatment 40
Polishing ponds 10
Filtration 17
Multimedia 7
Activated carbon 4
Sand 5
Other polishing 17
Secondary chemical flocculation/clarification 5
Secondary neutralization 5
Chlorination 11

level as attested by a high half saturation constant (Ks). The tetracycline waste
appeared to be biodegradable at a much slower rate (k = 0.5/day), but it had an
inherent instability as far as substrate removal rates to be employed in the treatment,
since its half saturation constant was comparatively too low. The operation showed
that, under the hydraulic detention time of 1 day, the activated sludge system could
yield an effluent 5-day BOD of 120 mg/L with a substrate removal rate of 0.31/day
and an MLVSS concentration of 4200 mg/L. The designed treatment plant was
capable to achieve 90% removal for 5-day BOD and 80% removal for COD.
142 L. K. Wang et al.

Fig. 2.13 Wastewater treatment system: a case study [107]

Table 2.14 Raw materials consumption for antibiotic production—case study [107]
Usage (tone
Raw materials per year)
Carbohydrate sources: Starch, dextrin, sugars, vegetable oils 1500
Protein sources: Soy meal, soy flour, com. Steep liquor gluten 300–100
Minerals: ammonium sulfate, ferrous sulfate, manganese sulfate, cobalt chloride, 25
calcium chloride, sodium ferrocyanide, sodium hydrogen sulfide. Phosphates
Ammonia. 23% 100–200
Acids, Bases: NaOH, HCl, H2SO4, oxalic acid 600–700
Quarternary ammonium salts 100–125
Antifoams 30
Solvents (all regenerated): acetone, methanol, oxitol. n-butanol 500
Urea 150–200
Note: 1 ton/year = 907.2 kg/year

2.7.4.2 Green Environmental Technologies Developed by the Lenox

Institute of Water Technology (LIWT)

The Lenox Institute of Water Technology (LIWT) has developed many modern dis-
solved air flotation (DAF) clarifiers, dissolved air flotation-filtration (DAFF) pack-
age plants, and DAF-DAFF package plants. The filtration portion can be either sand
filtration or GAC filtration. Through Krofta Engineering Corporation (KEC) and its
partners, over 3000 DAF, DAFF, and combined DAF-DAFF water and wastewater
treatment plants have been installed around the world. The LIWT has also devel-
oped several multistage hybrid green environmental technologies which are both
technically feasible and economically feasible for treating the pharmaceutical and
other industrial wastewater depending on the characteristics of original industrial
effluent: (a) DAF-DAFF-UV photocatalytic oxidation (one of AOP) for pretreat-
ment, (b) DAF-aerobic biological-DAFF-UV photocatalytic oxidation for treating
low-concentration wastewater, (c) DAF-anaerobic-aerobic biological-DAFF-UV
photocatalytic oxidation for treating medium- to high-concentration wastewater,
and (d) DAF-anaerobic-aerobic biological-DAFF-UF-UV photocatalytic oxidation
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 143

Table 2.15 Characteristics of wastewater streams—case study [107]

Parameters Process wastes Other diluted wastes

Flow, m3/day 120 160
pH 6.5–8.5 7.0–8.0
Alkalinity, mg/L 2000 –
BOD5. mg/L 13,500 400
COD, mg/L 34,000 600
SS, mg/L 1500 300
TKN-N, mg/L 1500 40
Total P, mg/L 70 10
Sulfates, mg/L 3000
Temperature, °C Ambient Ambient

for treating extremely high-concentration wastewater. The researchers and PhD stu-
dents are invited to study the LIWT systems further. Adopting a hybrid green envi-
ronmental technology consisting of both biological system (aerobic alone or
anaerobic-aerobic depending on the organic concentration of the wastewater) and
UV-photocatalytic oxidation will be a feasible solution to treating the pharmaceuti-
cal wastewater or similar.

2.7.5 Pharmaceutical Waste Minimization Case Study

of Hennepin County Medical Center

2.7.5.1 Company Overview [108]

Hennepin County Medical Center (HCMC), a public teaching hospital in

Minneapolis, Minnesota, is a nationally recognized level one trauma center and the
third largest hospital in the Twin Cities. HCMC has over 356,000 patient visits
annually.

2.7.5.2 Waste Reduction Project [108]

In 2006, HCMC returned over 900 different outdated pharmaceuticals, most in mul-
tiple quantities, through the reverse distribution process. The total cost to purchase
was $146,411. Of this amount, only 202 items were credited for a total of $75,657.
Therefore, a waste reduction project was conducted at HCMC that focused on
reducing pharmaceutical waste from the reverse distribution process at the inpatient
pharmacy. Waste reduction resulted in over $80,000 in cost savings and 378 lbs of
pharmaceutical waste.
144 L. K. Wang et al.

2.7.5.2.1 Crash Boxes

Crash boxes, similar to crash carts, were found to be a significant source of waste.
These boxes contain emergency medicine needed to revive someone in the event of
a cardiac event. Waste occurs when boxes contain drugs that are not used by their
expiration date. When this occurred in the past, the pharmacy exchanged the box
and updated all the drugs so they are good for about 1 year. Outdated and nearly
outdated drugs were sent for reverse distribution.
In investigating the crash boxes, it was determined that many of the drugs found
in the boxes are regularly used in other locations in the hospital. It was recom-
mended the pharmacy bring back the crash boxes 3 months prior to expiration and
move the drugs to locations where they are used more frequently, potentially using
them prior to expiration.
Other recommendations for the crash box drugs included the following:
(a) Replacing the specialty epinephrine intracardiac syringe that was rarely used. It
expired and was returned 98% of the time. The use of a more commonly used
epinephrine syringe and an 18-gauge, 3-inch needle banded to the epinephrine
box was recommended.
(b) Changing the dosage of glutose gel from the 45 g dosage type, much of which
was wasted. In most cases, a 30 g dose of glucose is used. A recommendation
was made to lower the dosage carried from 45 g to 15 g for the drug to be used
in more applications.
(c) Lowering the size of the nitroglycerin bottle from 100 count to 25 count and
switching to a generic form from a brand name.

2.7.5.2.2 Other Reverse Distribution Drugs

Review of reverse distribution manifests helped identify the most common and
costly drugs returned. HCMC also found they were returning 4% of their inventory,
which is 2% above the average as determined by the American Society of Health-­
System Pharmacists. The top ten of these were (a) crash box epinephrine, (b) epi-
nephrine, (c) glucagon, (d) glutose gel, (e) Nitrostat, (f) hydralazine, (g) lidocaine,
(h) amiodarone, (i) adenosine, and (j) naloxone.
It was recommended that HCMC review par usage reports for the top ten returns
and adjust inventory quantities accordingly. Doing so would save at least $80,000
and eliminate 210 pounds of pharmaceutical waste.

2.7.5.3 Results

HCMC implemented all the recommendations. It is estimated they are saving

$80,000 annually and have eliminated 378 pounds of drug waste.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 145

2.7.6 Pharmaceutical Waste Minimization Case Study of Falls

Memorial Hospital

2.7.6.1 Company Overview [108]

Falls Memorial Hospital (FMH), a 25-bed facility, is a charitable, not-for-profit

Critical Access Hospital located in International Falls, Minnesota. It has planned to
reduce its drug inventory and pharmaceutical waste.

2.7.6.2 Inventory Reduction Project [108]

Prior to undergoing the inventory reduction project, FMH was checking for out-
dated drugs every other month, stock was not rotated regularly, and par usage reports
were not available. In 2006, a staff pharmacist noticed many drugs on site were
outdated and the facility was stocking too many extra medications. Because par
usage reports had not been used previously, FMH, at that point, did not know how
many drugs were required for the facility. Due to these factors, the facility began to
look at ways to reduce inventory, save money, and decrease pharmaceutical waste.

2.7.6.2.1 Chemotherapy Drugs [108, 109]

Looking closely into quantities ordered and costs, FMH realized that chemotherapy
drugs were the largest expense for the facility. They were being ordered monthly,
and in December 2007, the facility spent over $90,000. Because of the long holding
time for some of the chemotherapeutics, they were outdating on the shelf. FMH also
realized, through facility-wide research, that some chemotherapy drugs were
extremely expensive and came in multiple strengths. FMH changed their ordering
for chemotherapeutic drugs from once per month to once a week.

2.7.6.2.2 Routine Stock on Floors

FMH utilizes AcuDose, an automated dispensing machine, to supply most of their

stock of drugs. AcuDose machines were stationed in the emergency room, medical/
surgical area, operating room, and intensive care unit. As part of the inventory
reduction project, the pharmacist noted that the AcuDose machine in the intensive
care unit was rarely used because most of the pharmaceuticals were special order
for the patients and resulted in numerous expired drugs and the inventory not being
rotated frequently enough. Therefore, the pharmacist recommended placing the
medications only where they are needed and rotating the stock on a more regu-
lar basis.
146 L. K. Wang et al.

2.7.6.2.3 Therapeutic Substitution

In order to reduce the amount of drugs at the hospital, the pharmacist recommended
using therapeutic substitution lists. For example, there are five medications in a
class of drugs called proton pump inhibitors, or PPI. Instead of having all five medi-
cations on the formulary, FMH chose to carry just two of them. This would ensure
that the hospital was not carrying multiple medications in the same category and
make it easier to rotate stock. If a patient comes into the hospital on a PPI not on the
formulary, they will be automatically switched to an equivalent dose of a PPI that is
on the formulary.
Multiple dosage types were also noted. The number of dosage forms has been
reduced to those used most often and multiples of those to achieve the strengths for
esoteric doses. The pharmacy now also searches out and purchases only from those
vendors that have the least packaging.

2.7.6.3 Pollution Prevention Impacts

Due to FMH’s inventory reduction project, the facility is ordering and stocking
fewer drugs, reducing packaging waste and shipping costs. This project reduced
FMH’s monthly overhead from $210,000 in January 2006 to $87,000 in October
2007 and dramatically reduced the amount of waste from expiring medications and
excess stock.

2.7.7 Recent Investigations of Pharmaceutical Wastewater

Treatment Technologies

A detailed review of available technologies for wastewater treatment and water

reuse in pharmaceutical industry has been conducted by Gadipelly et al. [110]. In
their review, the various sources of wastewaters in the pharmaceutical industry are
identified, and the best available technologies (BAT) for removing pollutants from
them are critically evaluated. Effluents arising from different sectors of active phar-
maceutical ingredients (API), bulk drugs, and related pharmaceutics, which use
large quantities of water, are analyzed, and strategies are proposed to recover valu-
able compounds, and finally the treatment of very dilute but detrimental wastewa-
ters is discussed [110]. It appears that no single technology can completely remove
pharmaceuticals and other pollutants from pharmaceutical wastewaters. The use of
conventional biological treatment methods along with innovative membrane reac-
tors and advanced posttreatment methods resulting in a combined hybrid wastewa-
ter treatment system appears to be the best [110–136]. Appendix I and Appendix II
document many researchers’ investigations reviewed by Gadipelly et al. [110]. The
authors of this publication list the original research sources of many useful
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 147

investigations for reference by the readers. The environmental technologies used for
treating various pharmaceutical wastes and their process terminologies have been
introduced in the previous sections of this book chapter, and they can also be found
from the literature [1–136] for further research.

2.7.7.1 Chemical Synthesis-Based Pharmaceutical Wastewater

Treatment Technologies

The chemical synthesis-based pharmaceutical wastewater treatment technologies,

which have been investigated with various degrees of success, are as follows:
1. Sulfate anion radical oxidation (Fe and Co sulfate salts used with hydrogen
peroxide and ozone) [132]
2. Dissolved air precipitation with solvent sublation simulated water: mineral oil
layer with organic solvents (toluene, methylene chloride, benzene, chloroben-
zene, hexane, butyl acetate) [133]
3. Electrocoagulation (EC) followed by heterogeneous photocatalysis (TiO2; iron
electrodes were used as cathode and anode) [130]
4. Upflow anaerobic sludge blanket (UASB) + micro-aerobic hydrolysis acidifica-
tion reactor (NHAR) + two-stage aerobic process, cyclic activated sludge sys-
tem (CASS), and biological contact oxidation tank (BCOT) [143]
5. Two-phase anaerobic digestion (TPAD) system and a subsequential membrane
bioreactor (MBR). TPAD system comprised of a continuous stirred tank reactor
(CSTR) and an upflow anaerobic sludge blanket-anaerobic filter (UASBAF),
working as the acidogenic and methanogenic phases [131]
6. Adsorption: granular activated carbon (a series of columns of GAC were
used) [164]
7. Electrochemical treatment (boron doped diamond BDD anode for corrosion
stability) [168]
8. Continuous heterogeneous catalytic wet peroxide oxidation (CWPO) process
using a Fe2O3/SBA-15 nanocomposite catalyst [134]
9. Acidogenic reactor (USAB sludge from an alcohol industry was used with high
glucose as initial feed and then varying pharmaceutical wastewater) [135]
10. Hybrid upflow anaerobic sludge blanket reactor [149]
11. Conventional treatment: activated sludge reactor using sequencing batch reac-
tor (SBR) [142]
12. Hybrid upflow anaerobic sludge blanket reactor (ASBR) [153]
13. Catalytic wet air oxidation [136]
14. Membrane bioreactor (MBR) [143]
15. Photo/Fenton followed by lime or sodium hydroxide precipitation/coagula-
tion [160]
148 L. K. Wang et al.

2.7.7.2 Fermentation Process-Based Pharmaceutical Wastewater

Treatment Technology

The fermentation process-based pharmaceutical wastewater treatment technologies,

which have been investigated with various degrees of success, are as follows:
1. Photocatalysis (TiO2) + H2O2; a single baffled reactor for the process [159].
2. Biodegradation using bacterial strains (Pseudomonas aeruginosa and
Pseudomonas pseudomallei) [165].
3. Photocatalysis (Fenton + photo-Fenton + ozonation) [155].
4. Ozonation (pretreatment) + biological activated sludge reactor combination in
series [157].
5. Fenton-biological process: first Fenton coagulation and then biological treat-
ment by activated sludge [161].
6. Chemical oxidation ozonation and ozonation coupled with treatment with
hydrogen peroxide [156].
7. Membrane bioreactor technology (hollow fiber membrane) [144].
8. Upflow anaerobic stage reactor (UASR) [148, 153].
9. Upflow anaerobic stage reactor (UASR) [152].
10. Ozonation (pretreatment) + biological activated sludge treatment by synthetic
biomass with 30% COD [158].
11. Activated sludge reactor in batch and continuous flow [146].
12. Anaerobic biological treatment using activated sludge reactor [151].
13. Hybrid treatment technology (aerobic biological pretreatment + ozonation +
MBR), the biological treatment for reducing the ozone demands. Ozonation
reduces almost all of the organic compounds [145, 157].
14. Anaerobic granulation batch/column reactor [150].
15. Catalytic wet air oxidation coupled with anaerobic biological oxidation [154].
16. Aerobic biological treatment with variable temperature study [51].
17. Biological treatment by activated sludge: in seven stages, a pilot plant study [55].
18. Suspended growth photo-bioreactor: non-sulfur photosynthetic bacterium iso-
lated from the soil and fluorescent light reactor [137].
19. Membrane bioreactor (GE ZeeWeed membrane bioreactor technology) [138].
20. Semiconductor photocatalysis Ti/TiO2: RuO2-IrO2 as anode, graphite as cath-
ode, and chloride as electrolyte [163].
21. Penraporation through water-selective membranes [139].
22. Sequencing batch reactor (SBR): an activated sludge reactor [140].
23. Solar photo-Fenton and biological treatment [162].
24. Anaerobic multichamber bed reactor (AMCBR) + AMCBR with continuous
stirred tank reactor (CSTR) [166].
25. ANAMMOX (anaerobic ammonium oxidation) process with sequential bio-
catalyst (ANAM-MOX granules) addition (SBA-ANAMMOX process) [167].
26. Fenton oxidation (pretreatment) by oxidation and coagulation stage followed
by aerobic biological degradation in sequencing batch reactor [147].
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 149

27. Catalytic wet air oxidation (CWAO) mixtures of waste streams used in auto-
clave to form polyoxometalates (POMs) as a cocatalyst system [141].

2.8 Summary and Conclusions

1. Toxic or hazardous pharmaceutical pollutants are typically produced in batch

pharmaceutical manufacturing processes leading to the presence of a wide vari-
ety of undesirable pharmaceuticals in wastewaters, air, and soil. Common use
of pharmaceutical compounds by human consumption and farming operations
is also an input source of undesirable pharmaceuticals in the environment. It is
concluded that the presence of pharmaceutical compounds in drinking water,
livestock, and human body comes from both of the above two sources: (a) pro-
duction processes of the pharmaceutical industry and (b) common use of phar-
maceutical compounds resulting in their presence in urban and farm wastewaters.
2. The pharmaceutical wastewaters generated in different processes in the manu-
facture of pharmaceuticals and drugs contain a wide variety of chemical com-
pounds. Some pharmaceutical pollutants are biodegradable; some are not
biodegradable or toxic to microorganisms. Conventional cost-effective biologi-
cal waste treatment technologies (i.e., gray environmental technologies), such
as activated sludge, trickling filters, lagoons, sequencing batch reactor, mem-
brane bioreactor, composting, sanitary landfill, etc., alone cannot properly treat
the liquid and solid wastes. An integrated approach must be taken to manage all
wastes within a pharmaceutical manufacturing plant. A “10-Step Blueprint for
Managing Pharmaceutical Waste of Healthcare Facilities In the United States”
has been developed by the US Environmental Protection Agency (USEPA), and
this blueprint must be examined and followed in order to reduce the hazardous
pharmaceuticals; in turn, a sustainable green biotechnology, bioreactor landfill,
can be used to treat the nonhazardous pharmaceutical solid waste, generate
methane gas as biofuel, and protect groundwater.
3. Pharmaceutical industry manufactures drugs, vaccines, antibiotics, products
with therapeutic value, etc. using chemical reactors, biological systems or
organisms, and many different raw materials. Pharmaceutical products are pro-
duced by chemical synthesis, fermentation, extraction from naturally occurring
plant or animal substances, or by refining a technical grade product. The USEPA
regulation applies to pharmaceutical industrial facilities which are organized
into five subcategories: (a) subcategory A (fermentation products), (b) subcat-
egory B (extraction products), (c) subcategory C (chemical synthesis products),
(d) subcategory D (mixing, compounding, and formulation), and (e) subcate-
gory E (research organizations).
4. Fermentation process of pharmaceutical plants produces most antibiotics and
steroids using three basic steps: inoculum and seed preparation, fermentation,
and product recovery. Fermentation is conventionally a large-scale batch pro-
cess. The fermentation step begins with a wash water and steam sterilization of
150 L. K. Wang et al.

the fermenter vessel. Sterilized nutrient raw materials in water are then charged
to the fermenter. The process wastewater from fermentation plants is character-
ized by high BOD, COD, and TSS concentrations, relatively large flows, and a
pH range of approximately 4.0–8.0.
5. Biological and natural extraction operations of pharmaceutical plants use many
materials as pharmaceuticals are derived from such natural sources as the roots
and leaves of plants, animal glands, and parasitic fungi. These products have
numerous and diverse pharmaceutical applications, ranging from tranquilizers
and allergy-relief medications to insulin, morphine, plasma, and its derivatives.
The extraction process consists of a series of operating steps beginning with the
processing of a large quantity of natural or biological material containing the
desired active ingredient. Residual wastes from an extraction plant essentially
will be equal to the weight of raw material. Solid wastes are the greatest source
of the pollutant load; however, solvents used in the processing steps can cause
both air and water pollution. The principal sources of wastewater from biologi-
cal/natural extraction operations are (a) spent raw materials, (b) floor and
equipment wash water, (c) chemical wastes (e.g., spent solvents), and (d)
cleanup of spills. Wastewater from extraction plants is generally characterized
by low BOD, COD, and TSS concentrations, small flows, and pH values of
approximately 6.0–8.0.
6. Chemical synthesis operations of pharmaceutical plants manufacture most of
the active ingredients marketed and sold as drugs using organic and inorganic
chemical reactions. The conventional batch reaction vessel is the major piece of
equipment used on the process line. The reaction vessel is one of the most stan-
dardized equipment designs in the industry. Chemical synthesis effluent gener-
ally has a high BOD and COD waste load. The pollutants in chemical synthesis
wastewater vary with respect to toxicity and biodegradability. Chemical synthe-
sis wastewater may be incompatible with biological treatment systems because
it is too concentrated or too toxic for the biomass in the treatment system. Thus,
it may be necessary to equalize and/or chemically pretreat some chemical syn-
thesis wastewater prior to biological treatment. Primary sources of wastewater
from chemical synthesis operations are (a) process wastes such as spent sol-
vents, filtrates, and concentrates; (b) floor and equipment wash water, (c) pump
seal water, (d) wet scrubber wastewater, and (e) spills. Wastewater from chemi-
cal synthesis plants can be characterized as having high BOD, COD, and TSS
concentrations, large flows, and extremely variable pH values, ranging from 1.0
to 11.0.
7. Mixing, compounding, or formulating operations of pharmaceutical plants pro-
duce pharmaceutically active ingredients batch processes in bulk form and con-
vert them to dosage form such as tablets, capsules, liquids, and ointments, for
consumer use. In addition, active ingredients can also be incorporated into
patches and time release capsules. Wastewater sources from mixing, com-
pounding, or formulating operations are (a) floor and equipment wash water,
(b) wet scrubbers, and (c) spills. The use of water to clean out mixing tanks can
periodically flush dilute wastewaters of unusual composition into the plant
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 151

sewer system. In general, this wastewater is readily treatable by biological

treatment systems. The wastewater from mixing, compounding, or formulating
plants normally has low BOD5, COD, and TSS concentrations, relatively small
flows, and pH values of 6.0–8.0.
8. The USEPA pharmaceutical industry effluent limitations (pretreatment or end-­
of-­the pipe treatment) are provided in this publication for the US readers. The
readers in other countries must contact their own country for the effluent limita-
tion details. Knowing both the government effluent limitations and the waste-
water characteristics will help select an integrated waste management plan and
a feasible wastewater treatment system.
9. Advanced treatment processes available for treating the pharmaceutical waste-
water include coagulation and clarification, dissolved air flotation (DAF),
flotation-­filtration (DAFF; filtration can be either sand filtration or GAC filtra-
tion), granular activated carbon (GAC) adsorption, powdered activated carbon
(PAC) adsorption, wet air oxidation (WAO), supercritical water oxidation
(SCWO), Fenton oxidation, UV photocatalytic oxidation, ultrasound oxidation,
air stripping, distillation, electrochemical oxidation, ozonation, membrane fil-
tration (MF, UF, RO, ED, MBR), or other advanced oxidation processes (AOP),
combined oxidation-reduction process, etc. Of these advanced treatment pro-
cesses, DAF, DAFF, GAC, air stripping, distillation, and membrane processes
are suitable for recycling and reusing chemical compounds, and/or water. In
view of the pollution load reduction and chemical cost saving, it is necessary to
recover chemical compounds or raw materials as much as possible. In view of
the scarcity of water resources, it is necessary to understand and develop meth-
odologies for treatment of pharmaceutical wastewater as part of water manage-
ment. While most of the advanced treatment processes are technically feasible
for treating the pharmaceutical wastewater, their economical feasibility needs
to be carefully evaluated before any implementation.
10. The Lenox Institute of Water Technology (LIWT), a nonprofit educational
institute, has developed several multistage hybrid green environmental tech-
nologies which are both technically feasible and economically feasible for
treating the pharmaceutical and other industrial wastewater depending on the
characteristics of original industrial effluent: (a) DAF-DAFF-UV photocata-
lytic oxidation (one of AOP) for pretreatment, (b) DAF-aerobic biological-­
DAFF-­UV photocatalytic oxidation for treating low-concentration wastewater,
(c) DAF-anaerobic-aerobic biological-DAFF-UV photocatalytic oxidation for
treating medium- to high-concentration wastewater, and (d) DAF-anaerobic-­
aerobic biological-DAFF-UF-UV photocatalytic oxidation for treating
extremely high-concentration wastewater. The researchers are invited to study
the LIWT systems further. Adopting a hybrid green environmental technology
consisting of both biological system (aerobic alone or anaerobic-aerobic
depending on the organic concentration of the wastewater) and UV photocata-
lytic oxidation will be a feasible solution to treating the pharmaceutical waste-
water or similar.
152 L. K. Wang et al.

Glossary of Biotechnology and Pharmaceutical Industry

Agricultural Biotechnology (a) It focuses on developing genetically modified

plants to increase crop yields or introduce characteristics to those plants that
provide them with an advantage growing in regions that place some kind of stress
factor on the plant, namely, weather and pests; (b) development of pest-resistant
crops and improvement of plant and animal breeding are typical examples; (c)
green biotechnology refers to specific agricultural biotechnology that creates
new plant varieties of agricultural interest, biopesticides, biofertilizers, etc. This
area of agricultural biotechnology is based on transgenics (genetic modification),
i.e., an extra gene or genes inserted into their DNA. The additional gene may
come from the same species or a different species.
Biological and Natural Extraction (Pharmaceutical) Many materials used as
pharmaceuticals are derived from such natural sources as the roots and leaves
of plants, animal glands, and parasitic fungi. These products have numerous
and diverse pharmaceutical applications, ranging from tranquilizers and allergy-­
relief medications to insulin and morphine. Also included in this group is blood
fractionation, which involves the production of plasma and its derivatives.
The extraction process consists of a series of operating steps beginning with
the processing of a large quantity of natural or biological material containing
the desired active ingredient. After almost every step, the volume of material
being handled is reduced significantly. Neither continuous processing methods
nor conventional batch methods are suitable for extraction processing. Residual
wastes from an extraction plant essentially will be equal to the weight of raw
material, since the active ingredients extracted are generally present in the raw
materials at very low levels. Solid wastes are the greatest source of the pollutant
load; however, solvents used in the processing steps can cause both air and water
pollution. Detergents and disinfectants used in equipment cleaning operations
are normally found in the wastewater. Priority pollutants, including methylene
chloride, toluene, chloroform, 1,2-dichloroethane, and phenol, were identified
as being used in the manufacturing of extractive pharmaceuticals. The principal
sources of wastewater from biological/natural extraction operations are (a) spent
raw materials (e.g., waste plasma fractions, spent media broth, plant residues),
(b) floor and equipment wash water, (c) chemical wastes (e.g., spent solvents),
and (d) cleanup of spills. Wastewater from extraction plants is generally charac-
terized by low BOD, COD, and TSS concentrations, small flows, and pH values
of approximately 6.0–8.0.
Biotechnology It is an engineering science field involving the use of biological sys-
tems found in organisms or the use of the living organisms themselves to make
scientific advances and adapt those knowledge to various application branches,
such as (a) medical biotechnology (including pharmaceutical biotechnology), (b)
agricultural biotechnology, (c) industrial biotechnology (including industrial fer-
mentation biotechnology), (d) environmental biotechnology, (e) computational
biotechnology, and (f), military biotechnology:
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 153

Blue Biotechnology It is a specific environmental biotechnology which is based

on the use of marine resources to create products, energy, or pollution control.
Chemical Synthesis (Pharmaceutical) Most of the active ingredients marketed
and sold as drugs are manufactured by chemical synthesis. Chemical synthesis
is the process of manufacturing pharmaceuticals using organic and inorganic
chemical reactions. The conventional batch reaction vessel is the major piece
of equipment used on the process line. The reaction vessel is one of the most
standardized equipment designs in the industry. By using heating or refrigera-
tion devices, the chemicals may be boiled or chilled in them, according to pro-
cess needs. By adding reflux condensation equipment, the vessel may perform
complete reflux operations (i.e., recycling of condensed vapors). The vessels can
also become evaporators if vacuum is applied. The reactors may also be used to
perform solvent extraction operations, and, by operating the agitator at a slow
speed, the vessels can serve as crystallizers. Synthetic pharmaceutical manufac-
ture consists of using one or more of these reactor vessels to perform, in a step-­
by-­step fashion, the various operations necessary to make the product. Chemical
synthesis effluent generally has a high BOD and COD waste load. The pollutants
in chemical synthesis wastewater vary with respect to toxicity and biodegrad-
ability. The production steps may generate acids, bases, cyanides, metals, and
other pollutants, while the waste process solutions and vessel wash water may
contain residual organic solvents. Occasionally, chemical synthesis wastewater
is incompatible with biological treatment systems because it is too concentrated
or too toxic for the biomass in the treatment system. Thus, it may be necessary
to equalize and/or chemically pretreat some chemical synthesis wastewater prior
to biological treatment. Primary sources of wastewater from chemical synthesis
operations are (a) process wastes such as spent solvents, filtrates, and concen-
trates, (b) floor and equipment wash water, (c) pump seal water, (d) wet scrub-
ber wastewater, and 5) spills. Wastewater from chemical synthesis plants can be
characterized as having high BOD, COD, and TSS concentrations, large flows,
and extremely variable pH values, ranging from 1.0 to 11.0.
Computational Biotechnology (a) It can be defined as “conceptualizing biotech-
nology” to address biotechnology problems using computational techniques and
makes the rapid organization as well as analysis of biotechnological data pos-
sible; (b) it can also be termed gold biotechnology or bioinformatics.
Dark Biotechnology It means the military biotechnology that is associated with
bioterrorism or biological weapons and bio-warfare using microorganisms and
toxins to cause diseases and death in humans, domestic animals, and crops.
Environmental Biotechnology (a) It is an interdisciplinary branch of biotech-
nology using biological systems and/or organisms for conservation of environ-
ment, resources, and energy and for protection of humans, animals, and plants
on Earth and beyond; it can be of green biotechnology, gray biotechnology, blue
biotechnology, gold biotechnology, or white biotechnology; (b) modern green
environmental biotechnology has a symbol of “green cross” that involves the
construction of resource recovery facilities (RRF), bioreactor landfills, in-vessel
or in-bin composting reactors, bioremediation sites, wildlife sanctuary areas,
154 L. K. Wang et al.

environmental protection parks, global warming control facilities, salmon lad-

ders, etc. using the best available technologies (BAT) for reclamation of water,
air, land, nutrients, methane gas, animals, plants, etc. and production of biofuels,
bio-plastics, waste-converted animal foods, etc., in turn achieving environmental
conservation, resource sustainability, biodiversity, climate control, ozone layer
protection, etc. (c) Gray biotechnology refers to an old traditional environmental
biotechnology applications to maintain biodiversity and the partial removal of
certain pollutants or contaminants using microorganisms and plants to isolate
and dispose of many kinds of substances such as heavy metals and hydrocar-
bons, but without sustainability of natural resources. Typical examples are the
old biological secondary wastewater treatment plants (WWTP) and old sanitary
landfills. Modern environmental biotechnology is considered to be a green bio-
technology. (d) Blue biotechnology is based on the use of marine resources to
create products, energy, or pollution control.
Fermentation (Pharmaceutical) Most antibiotics and steroids are produced by
the fermentation process, which involves three basic steps: inoculum and seed
preparation, fermentation, and product recovery. Fermentation is conventionally
a large-scale batch process. The fermentation step begins with a wash water and
steam sterilization of the fermenter vessel. Sterilized nutrient raw materials in
water are then charged to the fermenter. Microorganisms grown from seed to aid
in the fermentation process are transferred to the fermenter from the seed tank
and fermentation begins. During fermentation, air is sparged into the batch, and
temperature is carefully controlled. After a period that may last from 12 h to
1 week, the fermenter batch whole broth is ready for filtration. Filtration removes
mycelia (i.e., remains of the microorganisms), leaving the filtered aqueous broth
containing product and residual nutrients that are ready to enter the product
recovery phase. There are three common methods of product recovery: solvent
extraction, direct precipitation, and ion exchange or adsorption. Fermentation
broth contributes pollutants to wastewater from the food materials contained
in the broth, such as sugars, starches, protein, nitrogen, phosphate, and other
nutrients. Fermentation wastes are very amenable to biological treatment. The
spent broth can be satisfactorily handled by biological treatment systems in a
concentrated form. Equalizing the broth prior to treatment helps avoid system
upsets that may occur if the biota receive too high feed concentrations at one
time. The process wastewater from fermentation plants is characterized by high
BOD, COD, and TSS concentrations, relatively large flows, and a pH range of
approximately 4.0–8.0.
Gold Biotechnology It is equivalent to bioinformatics, or computational biotech-
nology, that addresses biotechnology problems using computational techniques
and makes the rapid organization as well as analysis of biotechnological data
possible.
Gray Biotechnology It refers to an old traditional environmental biotechnology
applications to maintain biodiversity and the partial removal of certain pollut-
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 155

ants or contaminants using microorganisms and plants to isolate and dispose of

many kinds of substances such as heavy metals and hydrocarbons, but without
sustainability of natural resources. Typical examples are the old biological sec-
ondary wastewater treatment plants (WWTP) and old sanitary landfills. Modern
environmental biotechnology is considered to be a green biotechnology.
Green Biotechnology (a) It is modern environmental biotechnology that achieves
environmental conservation and resource sustainability, or a specific agricultural
biotechnology that creates new plant varieties of agricultural interest, biopes-
ticides, biofertilizers, etc. (b) Modern green environmental biotechnology has
a symbol of “green cross” that involves the construction of resource recovery
facilities (RRF), bioreactor landfills, in-vessel or in-bin composting reactors,
bioremediation sites, wildlife sanctuary areas, environmental protection parks,
global warming control facilities, salmon ladders, etc. using the best available
technologies (BAT) for reclamation of water, air, land, nutrients, methane gas,
animals, plants, etc., in turn achieving environmental conservation, resource sus-
tainability, biodiversity, climate control, ozone layer protection, etc.; (c) the area
of green agricultural biotechnology is based on transgenics (genetic modifica-
tion), i.e., an extra gene or genes inserted into their DNA. The additional gene
may come from the same species or a different species.
Industrial Biotechnology (including industrial fermentation biotechnol-
ogy) (a) It is the utilization of cells, such as microorganisms, or components of
cells like enzymes, to generate products in sectors that are industrially useful,
such as food and feed, chemicals, detergents, paper and pulp, textiles, biofuels,
and biogas, or to create genetically modified organisms (GMOs) that enhance
the diversity of applications and the economic viability of industrial biotechnol-
ogy; (b) development of biocatalysts (such as enzymes, to synthesize chemicals),
improvement of fermentation process, and production of new plastics/textiles,
biofuels, etc. are typical examples; (c) a specific industrial biotechnology related
to production of wine, cheese, and beer by fermentation is also termed yellow
biotechnology; (d) designing more energy-efficient, less polluting, and low
resource-­consuming processes and products that can beat traditional ones is also
termed white biotechnology.
Medical biotechnology (including pharmaceutical biotechnology) (a) It has a
symbol of “red cross” and involves the use of living cells and other cell materi-
als to find cures for preventing diseases and bettering the health of humans; (b)
development of vaccines and antibiotics is a typical example; (c) a specific phar-
maceutical biotechnology related to medicine and veterinary products (vaccines,
antibiotics, molecular diagnostics techniques, genetic engineering techniques,
etc.) is also termed red biotechnology.
Military Biotechnology It is also termed dark biotechnology because it is associ-
ated with bioterrorism or biological weapons and bio-warfare using microor-
ganisms and toxins to cause diseases and death in humans, domestic animals,
and crops.
156 L. K. Wang et al.

Mixing, Compounding, or Formulating (Pharmaceutical) Pharmaceutically

active ingredients are generally produced by batch processes in bulk form and
must be converted to dosage form for consumer use. Common dosage forms for
the consumer market are tablets, capsules, liquids, and ointments. In addition,
active ingredients can also be incorporated into patches and time release cap-
sules. Wastewater sources from mixing, compounding, or formulating operations
are (a) floor and equipment wash water, (b) wet scrubbers, and (c) spills. The use
of water to clean out mixing tanks can periodically flush dilute wastewaters of
unusual composition into the plant sewer system. In general, this wastewater is
readily treatable by biological treatment systems. The wastewater from mixing,
compounding, or formulating plants normally has low BOD5, COD, and TSS
concentrations, relatively small flows, and pH values of 6.0–8.0.
Pharmaceutical Biotechnology It is a part of medical biotechnology (or a part of
red biotechnology) related to manufacturing of drugs, vaccines, antibiotics, etc.
using biological systems or organisms.
Pharmaceutical Industry Pharmaceutical industry manufactures drugs, vaccines,
antibiotics, products with therapeutic value, etc. using chemical reactors, bio-
logical systems or organisms, and many different raw materials Pharmaceutical
products are produced by chemical synthesis, fermentation, extraction from
naturally occurring plant or animal substances, or refining a technical grade
product. The USEPA regulation applies to pharmaceutical industrial facilities
are organized into five subcategories: (a) subcategory A (fermentation products),
(b) subcategory B (extraction products), (c) subcategory C (chemical synthesis
products), (d) subcategory D (mixing, compounding, and formulation), and (e)
subcategory E (research organizations).
Red Biotechnology It is a specific medical (including pharmaceutical) biotechnol-
ogy related to medicine and veterinary products (vaccines, antibiotics, molecular
diagnostics techniques, genetic engineering techniques, etc.).
White Biotechnology It is a specific industrial biotechnology involving white bio-
technology designing more energy-efficient, less polluting, and low resource-­
consuming processes and products that can beat traditional ones.
Yellow Biotechnology It is a specific industrial biotechnology related to produc-
tion of wine, cheese, and beer by fermentation.
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 157

 ppendix 1: BPT effluent limitations for subcategory

A
A (fermentation operations), subcategory B (biological
and natural extraction operations), subcategory C (chemical
synthesis operations), and subcategory D (mixing,
compounding, or formulating operations)

BPT effluent limitation for end-of-pipe

monitoring
points
Pollutant or Maximum for any Monthly
Subcategory pollutant property one day (mg/L) average (mg/L)
A: Fermentation operations COD 1675 856
B: Biological and natural COD 228 86
extraction operations
C: Chemical synthesis COD 1675 856
operations
D: Mixing, compounding, or COD 228 86
formulating operations

 ppendix 2: BAT effluent limitations for subcategory

A
A (fermentation operations) and subcategory C (chemical
synthesis operations)

BAT effluent limitations for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Acetone 0.5 0.2
Acetonitrile 25.0 10.2
Ammonia as N 84.1 29.4
n-Amyl acetate 1.3 0.5
Amyl alcohol 10.0 4.1
Benzene 0.05 0.02
n-Butyl acetate 1.3 0.5
Chemical oxygen demand (COD) 1675 856
Chlorobenzene 0.15 0.06
Chloroform 0.02 0.01
o-Dichlorobenzene 0.15 0.06
1,2-Dichloroethane 0.4 0.1
Diethylamine 250.0 102.0
Dimethyl sulfoxide 91.5 37.5
158 L. K. Wang et al.

BAT effluent limitations for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Ethanol 10.0 4.1
Ethyl acetate 1.3 0.5
n-Heptane 0.05 0.02
n-Hexane 0.03 0.02
Isobutyraldehyde 1.2 0.5
Isopropanol 3.9 1.6
Isopropyl acetate 1.3 0.5
Isopropyl ether 8.4 2.6
Methanol 10.0 4.1
Methyl cellosolve 100.0 40.6
Methylene chloride 0.9 0.3
Methyl formate 1.3 0.5
MIBK 0.5 0.2
Phenol 0.05 0.02
Tetrahydrofuran 8.4 2.6
Toluene 0.06 0.02
Triethylamine 250.0 102.0
Xylenes 0.03 0.01

 ppendix 3: BAT effluent limitations for subcategory B

A
(biological and natural extraction operations) and subcategory
D (mixing, compounding, or formulating operations)

BAT effluent limitations for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Chemical oxygen demand (COD) 228 86

 ppendix 4: NSPS for subcategory A (fermentation

A
operations) and subcategory C (chemical synthesis operations)

NSPS for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Acetone 0.5 0.2
Acetonitrile 25.0 10.2
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 159

NSPS for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Ammonia as N 84.1 29.4
n-Amyl acetate 1.3 0.5
Amyl alcohol 10.0 4.1
Benzene 0.05 0.02
n-Butyl acetate 1.3 0.5
Chlorobenzene 0.15 0.06
Chloroform 0.02 0.01
o-Dichlorobenzene 0.15 0.06
1,2-Dichloroethane 0.4 0.1
Diethylamine 250.0 102.0
Dimethyl sulfoxide 91.5 37.5
Ethanol 10.0 4.1
Ethyl acetate 1.3 0.5
n-Heptane 0.05 0.02
n-Hexane 0.03 0.02
Isobutyraldehyde 1.2 0.5
Isopropanol 3.9 1.6
Isopropyl acetate 1.3 0.5
Isopropyl ether 8.4 2.6
Methanol 10.0 4.1
Methyl cellosolve 100.0 40.6
Methylene chloride 0.9 0.3
Methyl formate 1.3 0.5
MIBK 0.5 0.2
Phenol 0.05 0.02
Tetrahydrofuran 8.4 2.6
Toluene 0.06 0.02
Triethylamine 250.0 102.0
Xylenes 0.03 0.01
bod5 267 111
COD 1675 856
TSS 472 166
160 L. K. Wang et al.

 ppendix 5: NSPS for subcategory B (biological and natural

A
extraction operations) and subcategory D (mixing,
compounding, or formulating operations)

NSPS for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
bod5 35 18
COD 228 86
TSS 58 31

 ppendix 6: PSES for subcategory A (fermentation

A
operations) and subcategory C (chemical synthesis operations)

PSES for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Acetone 20.7 8.2
Ammonia as N 84.1 29.4
n-Amyl acetate 20.7 8.2
Benzene 3.0 0.6
n-Butyl acetate 20.7 8.2
Chlorobenzene 3.0 0.7
Chloroform 0.1 0.03
o-Dichlorobenzene 20.7 8.2
1,2-Dichloroethane 20.7 8.2
Diethylamine 255.0 100.0
Ethyl acetate 20.7 8.2
n-Heptane 3.0 0.7
n-Hexane 3.0 0.7
Isobutyraldehyde 20.7 8.2
Isopropyl acetate 20.7 8.2
Isopropyl ether 20.7 8.2
Methyl cellosolve 275.0 59.7
Methylene chloride 3.0 0.7
Methyl formate 20.7 8.2
MIBK 20.7 8.2
Tetrahydrofuran 9.2 3.4
Toluene 0.3 0.1
Triethylamine 255.0 100.0
Xylenes 3.0 0.7
2 Waste Treatment in the Pharmaceutical Biotechnology Industry Using Green… 161

 ppendix 7: PSES for subcategory B (biological and natural

A
extraction operations) and subcategory D (mixing,
compounding, or formulating operations)

PSES for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Acetone 20.7 8.2
n-Amyl acetate 20.7 8.2
Ethyl acetate 20.7 8.2
Isopropyl acetate 20.7 8.2
Methylene chloride 3.0 0.7

 ppendix 8: PSNS for subcategory A (fermentation

A
operations) and subcategory C (chemical synthesis operations)

PSNS for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Acetone 20.7 8.2
Ammonia as N 84.1 29.4
n-Amyl Acetate 20.7 8.2
Benzene 3.0 0.6
n-Butyl acetate 20.7 8.2
Chlorobenzene 3.0 0.7
Chloroform 0.1 0.03
o-Dichlorobenzene 20.7 8.2
1,2-Dichloroethane 20.7 8.2
Diethylamine 255.0 100.0
Ethyl acetate 20.7 8.2
n-Heptane 3.0 0.7
n-Hexane 3.0 0.7
Isobutyraldehyde 20.7 8.2
Isopropyl acetate 20.7 8.2
Isopropyl ether 20.7 8.2
Methyl cellosolve 275.0 59.7
Methylene chloride 3.0 0.7
Methyl formate 20.7 8.2
MIBK 20.7 8.2
Tetrahydrofuran 9.2 3.4
Toluene 0.3 0.1
162 L. K. Wang et al.

PSNS for end-of-pipe monitoring points

Pollutant or pollutant property Maximum for any 1 day mg/L Monthly average mg/L
Triethylamine 255.0 100.0
Xylenes 3.0 0.7

 ppendix 9: PSNS for subcategory B (biological and natural

A
extraction operations) and subcategory D (mixing,
compounding, or formulating operations)

PSNS for end-of-pipe monitoring points

Pollutant or pollutant psroperty Maximum for any 1 day mg/L Monthly average mg/L
Acetone 20.7 8.2
n-Amyl Acetate 20.7 8.2
Ethyl acetate 20.7 8.2
Isopropyl acetate 20.7 8.2
Methylene chloride 3.0 0.7

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Chapter 3
Vermicomposting Process for Treating
Agricultural and Food Wastes

Lawrence K. Wang, Mu-Hao Sung Wang, Yung-Tse Hung, Kathleen Hung Li,
Hamidi Abdul Aziz, Mohd Suffian Yusoff, and Puganeshwary Palaniandy

Acronyms and Nomenclature

B Biochar
C Carbon
C:N Carbon-to-nitrogen ratio
CEC Cation exchange capacity
CPMV Cowpea mosaic virus

L. K. Wang (*)
Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
e-mail: [email protected]
M.-H. S. Wang
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]
Y.-T. Hung
Department of Civil and Environmental Engineering, Cleveland State University, Cleveland,
OH, USA
e-mail: [email protected], [email protected], [email protected]
K. H. Li
NEC Business Network Solutions, Inc., Irving, TX, USA
e-mail: [email protected]
H. A. Aziz
School of Civil Engineering and Solid Waste Management Cluster, Universiti Sains Malaysia,
Pulau Pinang, Malaysia
e-mail: [email protected], [email protected]
M. S. Yusoff · P. Palaniandy
School of Civil Engineering, Universiti Sains Malaysia, Pulau Pinang, Malaysia
e-mail: [email protected]; [email protected]

© Springer Nature Switzerland AG 2022 173

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_3
174 L. K. Wang et al.

DEH Dehydrogenase
EC Electric conductivity
ELISA Enzyme-linked immunosorbent assay
FTIR Fourier transform infrared
GC-MS Gas chromatography-mass spectrometry
GW Green waste
HA Humic acid
K Potassium
MSW Municipal solid waste
N Nitrogen
P Phosphorus
PFRP Process to Further Reduce Pathogens
SC Skin coffee
TCLP Toxicity characteristic leaching procedure
TMV Tobacco mosaic virus
TOC Total organic carbon
US EPA US Environmental Protection Agency
WSC Water-soluble carbon
WSP Water-soluble phosphorus

3.1 Introduction

3.1.1 Summary

Vermicomposting is a novel municipal/agricultural sludge and solid waste treatment

process that uses earthworms (oligochaete annelids) for the biodegradation of the
sludge and/or organic solid wastes, such as agricultural and food wastes. This novel
biological system is alternately called earthworm conversion, vermicomposting,
vermistabilization, worm composting, or annelidic consumption. The worms main-
tain aerobic conditions in the organic substances while accelerating and enhancing
the biological decomposition of the organic substances. The main product of the
vermicomposting (earthworm conversion) process is the worm’s castings. In some
process arrangements, there may be a net earthworm production. The excess earth-
worms may then be sold for fish bait or animal protein supplement. Earthworm
marketing is a complex problem; for municipal sludge applications, surplus earth-
worms may be considered a by-product, while the principal product is the castings,
which can be a resource, called vermicompost, compost, soil conditioner, or com-
post fertilizer.
This chapter presents the following: (a) an introduction and review of the vermi-
composting process; (b) technology development, technical problems, legal prob-
lems, and technology breakthrough of the process; (c) current status and resources;
(d) vermicomposting process design considerations; (e) process applications with
special emphasis on agricultural and food waste treatment; and (f) future
3 Vermicomposting Process for Treating Agricultural and Food Wastes 175

development and directions of the process. Recent advances in vermicomposting

process research and new process applications are reported.

3.1.2 Process Description

Vermicomposting differs from the conventional composting of wastewater treat-

ment plant sludge. In the vermicomposting process, worms are used to develop an
optimum environment for consuming or metabolizing the sludge and produce feces
or castings. These castings may be used as a soil conditioner [1–38, 41–63]. In the
conventional composting process, microorganisms are used for the degradation of
sludge and other putrescible organic solid materials under an aerobic metabolism
environment. Conventional composting is also suitable for converting undigested
primary sludge, secondary sludge, and certain solid wastes into an end product ame-
nable to resource recovery with a minimum capital investment and relatively small
operating commitment [39–40, 45].
Figure 3.1 (Source: US EPA) shows a basic simple vermicomposting process
[60, 62] that requires worm beds and an ample supply of worms. Generally, digested
and dewatered sludge is put into the beds, although experiments are underway,
where raw liquid sludge is placed in beds. If anaerobic digestion is used prior to
earthworm conversion, additional pretreatment may be needed. A bulking agent
such as wood chips may be useful in some cases for keeping the bed porous and

Fig. 3.1 Diagram of an earthworm conversion process

176 L. K. Wang et al.

aerobic, especially if moisture is high. Sludge is, however, generally applied with-
out any bulking agent. A worm bed may take the form of a simple tray. Windrows
similar to those for composting may also be used. After the worms have consumed
the sludge, they must be separated from the castings. This may be done with an
earthworm harvester, a drum screen that rotates on a nearly horizontal axis. Castings
fall through the screen openings, while worms tumble through the length of the
drum. Section 3.6 contains some critical operational parameters for the earthworm
conversion process.

3.2 Technology Development

Conversion of sludges (or biosolids) into topsoil by earthworms was initially

attempted by Mitchell et al. of the State University of New York at Syracuse, College
of Environmental Science and Forestry in 1977 [1]. Later, Mitchell et al. investi-
gated the potential role of the earthworm, Eisenia foetida, on the decomposition of
sewage sludge in drying beds and reported the results in 1980 [2]. Specifically,
Mitchell et al. sought to determine the decomposition rates of biosolids in drying
beds as indexed by consumption of oxygen and evolution of carbon dioxide and
methane, to ascertain whether E. foetida can alter the form and rate of decomposi-
tion, and to ascertain the relationship among specific biotic and abiotic components
in decomposition. At two facilities tested, the aerobic and anaerobic bacteria were
abundant, and the dominant bacteria were not enteric. A computer simulation model
regarding the role of macroinvertebrates in decomposition was used to analyze the
effects of the earthworm.
In August 1980, Camp, Dresser and McKee, Inc., of Boston, MA, USA, com-
pleted a technical report [3] which assessed the technical and economic feasibility
of vermicomposting or vermistabilization process based on several pilot-scale stud-
ies conducted by private entrepreneurs. The assessment was based on examining
facilities and costs for a municipal operation serving (a) a community of 50,000
persons and (b) a community of about 500,000 persons. Vermicomposting was com-
pared to three other methods of solid waste management: sanitary landfill, windrow
composting, and combustion. In 1980, vermicomposting was estimated to cost
about $24–32 per ton of waste processed (note: 1 ton = 2000 pounds; 1
pound = 0.454 kg).
In 1981, Hornor and Mitchell [4] studied the effect of the earthworm, Eisenia
foetida, on fluxes of volatile carbon and sulfur compounds from sewage sludges.
Hartenstein [5] suggested the potential use of earthworms as a solution to sludge
management. In Hartenstein’s study at the State University of New York at Syracuse
[5], the feasibility of using earthworms in the management of municipal sludges
was examined in detail. Results of tests performed by Hartenstein on two earth-
worm species—E. eugeniae and E. foetida—were reported. The following observa-
tions were made: (a) the toxicity of worm casts to the earthworms signifies the need
to retain E. foetida in its source of food (biosolids) as long as, or slightly longer
3 Vermicomposting Process for Treating Agricultural and Food Wastes 177

than, the time required to convert the sludge into castings; (b) knowledge of the
quantity of material passing through the earthworm gut per unit of time, for a par-
ticular ingestible sludge, permits prediction of sludge quantity manageable per unit
time; and (c) E. foetida fails to gain weight rapidly, if at all, on unlimited supplies
of certain organic materials.
Also in 1981, Collier and Livingstone [6] completed research sponsored by the
National Science Foundation. They used earthworms of the redworm (E. foetida)
species to accomplish vermicomposting or vermistabilization of biosolids from the
San Jose and Santa Clara Wastewater Treatment Plants in California, USA. Ninety
tons of earthworm manure were produced from the sludge over a 5-year period.
Different size windrows were populated with different densities of earthworms, and
castings were harvested by passing windrow contents through a rotating screen
which separated the worms from the castings for reuse. Plants in castings outgrew
plants in topsoil by a factor of 4 to 1. Their 1981 cost analysis showed the system to
be cost effective at a cost of $29.45 per dry ton in a 10 ton per day facility and to
return a profit of $3.34 per dry ton if castings were produced at the rate of 50 tons
per day.
In 1982, Hartenstein [8] reported (a) the metabolic parameters of the earthworm
Eisenia foetida in relation to temperature and (b) the potential use for manure man-
agement and as a source of protein biomass. In 1983, Chosson and Dupuy [9] dem-
onstrated their improvement of the cellulolytic activity of a natural population of
aerobic bacteria (enrichment culture) and presented their isolation and characteriza-
tion of worm gut and compost cellulolytic strains. In 1984, Hartenstein et al. [10]
attempted to use earthworms in trickling filters for wastewater treatment.
In March 1984, Loehr et al. [11] presented the results of an investigation of the
vermistabilization process using stabilized and unstabilized wastewater treatment
sludges. Four earthworm species were evaluated: E. foetida, E. eugeniae, P. haway-
ana, and P. excavatus. E. foetida was found to have the greatest overall reproductive
capacity. The best growth of E. foetida in terms of total biomass weight gain
occurred in media that had a total solids content, wet basis, of between 9 and 17%.
The best growth and cocoon production for this earthworm species was shown to
occur at temperatures of 20–25 °C. With both dewatered and liquid sludges, ver-
mistabilization units functioned successfully for long periods of time—up to 1 year
for dewatered sludge and at least 6 months for the liquid sludges. Cost estimates
indicated that the capital and annual costs of liquid vermistabilization were com-
petitive with those for other sludge management systems.
In 1985, Loehr et al. of Cornell University [12] evaluated several fundamental
factors that affect the performance of the vermistabilization process such as tem-
perature, moisture content of the waste material, and the combined use of several
earthworm species (polyculture). The earthworms Dendrobaena veneta, Eisenia
foetida, Eudrilus eugeniae, Perionyx excavatus, and Pheretima hawayana were
used in one or more of the studies. The best growth and reproduction of these spe-
cies occurred at temperatures of 20–25 °C. The growth of all five species was
reduced at 30 °C and death occurred at 35 °C. Of the five species, Eisenia foetida
produced the largest number of young in a 20-week study. The growth of Eisenia
178 L. K. Wang et al.

foetida occurred optimally in media with a total solids content, wet basis, of between
9 and 16%. Polyculture did not exhibit any obvious advantages over monoculture.
Stabilization of liquid sludge, or biosolids, by the vermistabilization process was
also reported by Loehr et al. of the University of Texas at Austin, TX, USA [13].
The investigators conducted basic studies to identify fundamental factors that affect
the performance of the vermistabilization process and applied studies to determine
design and management relationships. As earthworms are a key component of the
liquid sludge vermistabilization (LSVS) process, control reactors that did not con-
tain worms failed in a much shorter period of time than did the reactors with the
worms. LSVS reactors that were not overloaded functioned successfully for
140–198 days and were stopped only because the project ended. Oxidized nitrogen
(nitrates) in the drainage from the LSVS reactors indicated that aerobic conditions
were being maintained. Liquid primary sludge and liquid waste activated sludge
(biosolids) can be stabilized by the LSVS process.
LSVS reactors were not adversely affected by short-term, large variations in
loading rates. Liquid primary sludge was stabilized to about the same degree as
liquid aerobically digested sludge in the LSVS process. Moisture balances indicated
an overall moisture loss of 4–20%. Loading rates of about 21,000 g/m2/week vola-
tile solids or less resulted in satisfactory operation of LSVS reactors stabilizing
liquid primary and liquid waste activated sludge. Loading rates greater than 1200 g/
m2/week volatile solids could be used for LSVS reactors stabilizing liquid aerobi-
cally digested sludge. With LSVS reactors, the disposal of residual stabilized solids
occurs at long intervals. The total solids content of the stabilized residual solids in
the LSVS reactors was from 14 to 24%, a considerable increase from the 0.6 to
1.3% that was added. LSVS proved to be a successful process for both dewatering
and stabilization. The stabilized residual solids had approximately the same charac-
teristics regardless of the type of liquid sludge added to the reactors. Size and cost
estimates indicated that LSVS might be an economically feasible sludge manage-
ment process.
Reviews of the literature on sludge characteristics, solids concentration and con-
ditioning, stabilization and inactivation, incineration, and ultimate disposal and uti-
lization were conducted by Hasit of Weston, Inc., West Chester, PA, USA, in 1985
[14] and 1986 [15]. Vermistabilization was one of the sludge management technolo-
gies reviewed and assessed.
In 1986, Stafford and Edwards [16] of Rothamsted Experimental Station,
Harpenden, England, used earthworms in the field to indicate levels of soil pollution
and in the laboratory for the ecotoxicological testing of industrial chemicals. An
earthworm bioassay procedure developed at the Waterways Experiment Station in
Vicksburg, Mississippi, USA, was modified and evaluated as a method of providing
information on heavy metal bioavailability in contaminated soils and sediments
from Europe. Eight soils/sediments containing elevated levels of at least one of the
elements Zn, Cu, Cd, and Pb were selected, as well as a control and a reference soil.
Six earthworm species, including the WES bioassay earthworm E. foetida, and five
field species were grown in the soil for periods of 15, 28, or 56 days. Concentrations
of the elements Zn, Cu, Cd, Ni, Cr, and Pb present in the earthworm samples
3 Vermicomposting Process for Treating Agricultural and Food Wastes 179

(corrected for the presence of soil-derived metals within the earthworm gut) were
compared between earthworm species from the same soil and for each earthworm
species from a range of metal-contaminated soils/sediments.
A US Patent No. 4971616, entitled “Process for Preparing Organic Compost
from Municipal Refuse,” was awarded to Mark E. Glogowski on November 20,
1990 [17]. The patent involved the use of earthworms for treatment and disposal of
shredded cellulose refuse.
The earthworm Eisenia foetida is known to contain bactericidal enzymes. In
1990, Amaravadi et al. tested the earthworm for virucidal activity using cowpea
mosaic virus (CPMV) and tobacco mosaic virus (TMV) as model agents [18].
Earthworms were fed cellulose saturated with a virus suspension, and their excreted
castings were analyzed for structurally intact virus protein using enzyme-linked
immunosorbent assay (ELISA) and virus infectivity by local lesion assays.
Observations of the feeding experiments indicated a considerable reduction in the
infectivity of both viruses. Virucidal activity was also observed when virus suspen-
sions were incubated with the earthworm enzyme extract and analyzed by local
lesion assay. The observed reductions in the infectivity of both viruses suggested
that E. foetida might possess a virucidal enzyme system and, accordingly, might
contribute to the inactivation of pathogenic viruses potentially associated with land
application of sewage sludges and livestock manure.
Another US Patent No. 5055402, entitled “Removal of Metal Ions with
Immobilized Metal Ion-Binding Microorganisms,” was awarded to Greene et al. on
October 8, 1991 [19]. The inventors cited the use of earthworms.

3.3 Problems and Technology Breakthrough

3.3.1 Introduction

While vermicomposting has demonstrated its benefits, the process faces obstacles in
meeting US regulatory requirements. This section presents the problems and prog-
ress made in vermicomposting, i.e., new technologies that have been developed to
overcome the technical and legal problems.

3.3.2 Problems

Scientific interest in earthworms is on the rise worldwide [20–26]. At the Fifth

International Symposium on Earthworm Ecology in 1994, 183 presentations were
given at the 1994 International Symposium that were divided into two general cat-
egories: using earthworms directly in horticulture and agriculture to enhance crop
growth and using earthworms to turn various residuals into beneficial composts for
180 L. K. Wang et al.

reuse. Despite the increasing number of studies, however, financial support for ver-
micomposting research has been cut by the funding agencies in the USA since 1990.
Another problem is the process’s failure to meet regulatory requirements. The
US Environmental Protection Agency’s “Process to Further Reduce Pathogens
(PFRP) Requirements” for in-vessel or aerated static pile composting of biosolids
requires maintaining a temperature of 55 °C or higher in composting for 3 days.
Worms can survive in thermophilic composting windrows, but they tend to stick to
the edges of the pile. Temperatures above 35 °C, which is the heat generated by
thermophilic composting, are too high for earthworms and will kill them. In vermi-
composting, temperatures are generally kept below 30 °C. While organic substances
can be effectively processed by worms at low temperature range, the US EPA’s
PFRP requirements cannot be met. Progress in vermicomposting of organic sub-
stances proceeded slowly due to the above technical and legal problems.
There has been continuous debate in the State of California, USA, regarding the
classification and potential regulation of composting facilities. A draft of regula-
tions released in August 1994 by the California Integrated Waste Management
Board (CIWMB) excludes vermicomposting operations from the notification and
permitting that would be required of most larger facilities using conventional ther-
mophilic composting to process yard trimmings, manure, biosolids, and other
organic substances [24]. Under current California ruling, vermicomposting may be
considered an agricultural operation, in which vermiculture uses organics as a feed-
stock for raising worms in a worm farm. The advantage is that the owners and
operators of the vermicomposting facilities have free rein in process control and
management and are not subject to the state inspections. The disadvantage is that as
long as vermicomposting is not recognized as solid waste disposal process, the
progress for its technology development and application will be slow.
Noting the US federal requirements on PFRP, vermiculturists now precompost
the organic substances in the thermophilic temperature range for pretreatment and
disinfection. Worms are added to compost windrows for a subsequent vermiphilic
decomposition after the heat of initial thermophilic decomposition subsides. In
comparison with conventional thermophilic composting as a process, the modified
vermicomposting process has a shorter processing time. With conventional thermo-
philic composting alone, it is difficult to produce high-quality products under
6 months, while with the modified vermicomposting (i.e., thermophilic composting
pretreatment plus vermicomposting posttreatment), it is possible to create a market-
able end product in one-sixth of the operating time. Compared to the conventional
thermophilic compost end product, vermicompost contains smaller particles and
worm cocoons (meaning a free workforce for the future) and has lower odor and
enhanced microbial activity. According to commercial estimates, consumers would
be willing to pay up to three times more for the vermicompost, or worm castings,
than they would pay for most normal thermophilic compost. Many commercial-­
scale breakthroughs in vermicomposting technology have been noted and are intro-
duced below [23–25].
The Resource Conversion Corporation (7825 Fay Avenue, Suite 380, La Jolla,
CA 92037, USA) has developed a proprietary “Vermiconversion System” which
3 Vermicomposting Process for Treating Agricultural and Food Wastes 181

significantly modifies traditional vermiculture windrow methods. Variations include

sloped plastic liner beneath the windrow, reclaim water, aeration piping, and a
sprinkler to maintain proper temperature levels. In July 1994, Resource Conversion
Corporation and Sanifill, a national landfill company, together opened Canyon
Recycling outside of San Diego, which is a 6-acre (note: 1 acre = 4047 m2 = 0.4046 ha)
facility currently processing around 100 tons per day of brush, green material, and
wood from construction and demolition operations and manure from the San Diego
Zoo. After grinding and screening, some woody materials are marketed “as is.”
Leafy greens, wood fines, and manures proceed through a blending plant, then
“cured” via thermophilic composting to neutralize pathogens. After curing, the pre-
processed material is applied to the vermiculture windrows in thin layers. The rows
are carefully segregated and check for biological reactions to new feedstock. Two to
four inches of material are applied every other day continuously. The rows are com-
partmentalized to prevent possible contamination of the entire facility. The facility
adopts both the thermophilic composting pretreatment (for 3–15 days aiming at
pathogen reduction and decomposition) and the vermicomposting posttreatment
(for additional 15–30 days aiming at final curing and decomposition). Their worm
castings product is being sold for $33 per ton on the bulk market. The company is
now building a 100-acre facility to manage San Diego’s biosolids under a 20-year
contract.
The Oregon Soil Corporation (17,810 SSW Bunker Oak Road, Aloha, OR, USA)
has developed a technology to reduce the space requirements for a vermiculture
operation using a “continuous flow system.” The newly developed continuous flow
system utilizes a raised, 120-foot trough (note: 1 foot = 0.3048 meter) that is 2.5 ft
deep and 8 ft wide, with a mesh floor. An adapted manure spreader makes a daily
pass over the trough, laying down about 3 inches (note: 1 inch = 2.54 cm) of pre-
pared organic materials, or roughly 6 tons per day (note: 1 ton = 2000 pounds; 1
pound = 0.454 kg). As the worms eat up through it, the worm castings sink down
and are mechanically scraped off the bottom of the screen and collected. Under the
protection of a greenhouse-like structure, the worm reactor can handle about 2500
tons of organic residuals a year. Currently, the Oregon Soil Corporation accepts year
trimmings deliveries from local landscapers and picks up food scraps and paper
from 15 Fred Meyers grocery stores around Portland. They process around 5 or 6
tons of food scraps, over 2 tons of supplemental yard trimmings or compost, and
around half a ton of paper per day. It takes only 21 days to make earthworm castings
using the continuous flow system.
The Worm Concern (note: it is The Worm Connection now in California, USA)
had grown to a 22-acre spread during its 18 years in business. Around 100 tons per
day of brush, leaves, tree limbs, grass clippings, and horse manure are delivered to
the site for processing. Incoming material first passes through a grinder and a tram-
mel before being placed in windrows by a front-end loader. The facility adopts both
anaerobic windrow preprocessing (in which the piles are not turned at all until mate-
rial is moved to the worm rows) and vermicomposting posttreatment using worms.
At harvest time, worm rows are scooped up with a front-end loader and placed in
screen. Castings come out one end and the worms come out the other, unharmed.
182 L. K. Wang et al.

Their vermicastings are sold in bulk, blended on site with mulch or other landscape
products, and bagged for retail sale.
Finally, the Environmental Earthworm Projects, Inc. (8114 Port Said Street,
Orlando, FL 32813, USA) currently operates two sites handling a combined total of
30 tons per month of composted yard trimmings from the Orange County landfill
and 20 tons per month of shredded cardboard. They also have conducted earthworm
trials with RDF fines from Palm Beach County and other organics.

3.3.3 Progress in Vermicomposting Outside the USA

Engineers and scientists in the countries other than the USA have shown their inter-
est in the theories, principles, and applications of the vermistabilization process
since 1992. Practical applications of the vermicomposting process in disposal of
biosolids and organic solid wastes have been attempted by many entrepreneurs
around the world. The progress in vermicomposting process development and appli-
cations outside of the USA is discussed below [20–26].
In November 1992, Concheri et al. of Italy reported humification of organic
waste materials during earthworm composting [20]. In March 1993, Anton et al. of
Spanish Council for Scientific Research, Madrid, Spain, reported carbofuran acute
toxicity to Eisenia foetida earthworms [21].
In 1993, Van-Gestel and Ma of the National Institute of Public Health and
Environmental Protection, Bilthoven, Netherlands, reported their results on the
development of QSARs in soil ecotoxicology [22]. The earthworm toxicity and its
soil sorption of chlorophenols, chlorobenzenes, and chloroanilines were docu-
mented by the investigators of Netherlands.
Also in 1993, Original Vermitech Systems, Ltd. (2328 Queen Street East,
Toronto, Ontario M4E1G9, Canada; Tel. No. 416-693-1027) installed a composting
unit with a capacity of up to 600 pounds of organics per day at the Brockville
Psychiatric Hospital in Ontario, Canada. It is the largest composter in Canada right
now [23]. The system is equipped with panels and temperature sensors for maintain-
ing a tolerable environment for the worms.
At the Fifth International Symposium on Earthworm Ecology, held at Ohio State
University in 1994, scientists from the University of Agricultural Sciences in
Dharwad, India, told conference attendees that in their experiments, earthworms
could turn crop and weed residuals into vermicompost at the rate of 8–10 tons per
year from a bed area of 100 m2 [24, 25]. At the same symposium, scientists from the
Biosystems Research Group at the Open University, Milton Keynes, in England,
reported on their experiments of the modified vermicomposting process [24, 25].
The English scientists added earthworms to compost windrows after the heat of
initial decomposition subsided. Their worms worked well in this situation and
shortened the time of curing and stabilization of the compost.
Changes in heavy metal extractability and organic matter fractions after vermi-
composting of sludges from a paper mill industry and wastewater treatment plant
3 Vermicomposting Process for Treating Agricultural and Food Wastes 183

were reported by Elvira et al. of the University of Vigo, Spain, in 1995 [24–26].
According to the researchers from the Department of Natural Resources, University
of Vigo, vermicomposting of paper mill sludge has been proven to be viable in their
country.

3.4 Pioneers, Current Status, and Resources

The pioneers of the vermistabilization process, as well as its current status and
resources, are introduced in this section in detail.

3.4.1 Pioneers and Current Status

Many pioneers of the vermicomposting process deserved to be recognized. Jack

E. Collier and Diane Livingstone were the principal investigators of a milestone
research project sponsored by the National Science Foundation entitled “Conversion
of Municipal Wastewater Treatment Plant Residual Sludges into Earthworm
Castings for Use as Topsoil” [7]. Collier and his wife still operate an earthworm
farm in California, USA, which provides high-quality earthworms for all types of
earthworm research including vermistabilization. The Colliers often serve as con-
sultants on their vermistabilization technology to individuals or organizations. Dr.
Mark Buchannon, a soil scientist of the University California at Santa Cruz, USA,
recently collaborated with the Colliers to complete his PhD research in a simi-
lar field.
Dr. Raymond C. Loehr of the University of Texas at Austin, Department of Civil
Engineering, Austin, TX, USA, is another legend in vermistabilization technology
development [11–13]. Dr. Loehr, too, consults on vermistabilization research and
applications, if requested.
Dr. Clive Edwards, Professor of Entomology at Ohio State University, has also
been instrumental as the founder of the International Symposium on Earthworm
Ecology and has conducted several key vermicomposting projects leading to com-
mercialization of the process.
Practicing vermicomposting technologists who can provide assistance in vermi-
composting facility installation and process operation include Frank Stevenson of
the Environmental Earthworm Projects, Inc.; Dan Holcombe of Oregon Soil
Corporation; Albert Eggen of Original Vermitech Systems, Ltd.; Joseph Roberts of
Resource Conversion Corporation; Tim Morhar of The Worm Connection; and
Sandra Kandracs of Enviro-Ganics.
Writers/reporters Gene Logsdon, David Riggle, and Hannah Holmes discussed
the progress of vermicomposting technology in two articles for BioCycle [24, 25], a
trade journal that documents and reports the scientific knowledge and commercial
news involving worms.
184 L. K. Wang et al.

Steven Zorba Frankel and Stephen White of the Edible City Resource Center
have published a 32-page quarterly newspaper, Worm Digest [27–39], which pro-
motes vermicomposting technology as well as other technologies involving the use
of earthworms. Today, Worm Digest reports on the subjects of worms and worm
composting for organic waste conversion and soil enrichment. The newspaper gen-
erally features a wide variety of interesting and practical information to help pro-
mote awareness of vermiculture eco-technology on all levels. Columns such as the
following appear intermittently in each issue [27–29]: Worm Shorts x New Products
x International Worm News x The Industrious Worm (large-scale projects) x
Hands-On x Worm Workers x Kids’ Corner/Page x Questions & Answers x Eco-­
Logic x Worm Stories x Cyber-Worm x Advertisements & Resource Listings x
Calendar of Events.
At the request of environmental engineers in Ukraine, the authors conducted an
investigation on the current status and future direction of the vermistabilization pro-
cess. It was discovered that the vermistabilization (vermicomposting) operations/
research in sites such as Syracuse, NY; Ithaca, NY; West Chester, PA; San Jose, CA;
and Austin, TX, in the USA was terminated due to minor technical and legal prob-
lems and a lack of financial and public support. It is encouraging to learn, however,
that several companies in the USA and Canada have seriously conducted their
research for modification and optimization of the vermicomposting (or vermistabi-
lization) process despite the lack of proper funding. Now the process has been
improved and commercialized, and many large-scale vermicomposting or vermicul-
ture projects in Florida, California, Oregon, and Ontario are in progress.
Earthworm research is still being widely conducted by soil scientists and envi-
ronmental scientists around the world. Earthworms are tested as the organisms for
organic waste disposal, the toxicity indicators of the ecological system, or as the
topsoil producers. As mentioned, there is even an annual International Symposium
on Earthworm Ecology.
Interest in the vermistabilization process for sludge management has quickly
spread from the USA to European and Asian countries [20–105], indicating that
there will always be ample room for additional research on process improvement.
To explore or establish any international cooperative programs in the field of
environmental engineering, readers are encouraged to contact the authors and the
experts listed in Sect. 3.4.2 for technical or managerial assistance.

3.4.2 Resources

Important resources of the vermicomposting process around the world are intro-
duced in this section. It should be noted that the first letter of each resource defines
its nature in accordance with the following key: Associations (A), Publications (P),
Retail Businesses (R), Consultants (C), Distributors (D).
3 Vermicomposting Process for Treating Agricultural and Food Wastes 185

P 1. Edible City Resource Center, Worm Digest, PO Box 544, Eugene, OR 97440,
U.S. Tel. No./Fax No. (call first) 541-485-0456.
R 2. The Worm Factory, RR # 3, Perth, Ontario, Canada K7H 3C5. Tel. No.
613-267-5540.
A 3. The Composting Council of Canada, Canada. Tel. No. 416-535-0240; Fax No.
416-536-9892. e-mail address: [email protected].
A 4. Association of Oregon Recyclers, PO Box 15279, Portland, OR 97210,
U.S. Tel. No. 503-661-4475.
P 5. BioCycle, Journal of Composting & Recycling (monthly), 419 State Avenue,
Emmaus, PA 18049, U.S. Tel. No. 800-661-4905; 610-967-4135.
W 6. Lake County Worm Farm, PO Box 1332, Kelseyville, CA 95451, U.S. Tel. No.
800-399-9464; Fax No. 707-279-8031.
P 7. Australian Worm Growers Association, PO Box 318, Ferntree Gully, VIC 3156,
Australia.
R 8. Arlan & Sons (bookseller), 11881 Arroyo, Santa Ana, CA 92705, U.S. Tel. No.
714-838-8539; Fax No. 714-838-4950. e-mail address: [email protected].
R 9. Avant Garden Vermicomposting Systems (worm bins), PO Box 1047, Point
Reyes Station, CA 94956, U.S. Tel. No. 415-663-1975; Fax No. 415-663-1975.
C 10. Vermitechnology Unlimited, Inc., PO Box 130, Orange Lake, FL 32681, U.S.
A 11. International Worm Growers Association, PO Box 887, Littlerock, CA 93543
U.S. Tel. No. 805-9442994, Fax No. 805-944-3965.
R 12. WormWide Books, 20 Forest Avenue, Kingston Park, South Australia 5049,
Australia. Tel. No. 610412-112285; Fax No. 61-08-377-2668.
W 13. Rainbow Worm Farm, 24700 County Road, No. 95, Davis, CA 95616,
U.S. Tel. No. 916-758-9906; Fax No. 916-756-0414.
C 14. Oregon Soil Corporation, 1324 Beaver Lane, Oregon City, OR 97045, U.S. Tel
Nos. 503-557-9742, 503-629-5933.
R 15. Flowerfield Enterprises, 10332 Shaver Road, Kalamazoo, MI 49002, U.S. Tel.
No. 616-327-0108.
C 16. Roberta Trombley, 3030 Marshall, Cincinnati, OH 45220, U.S. Tel. No.
513-683-2340.
D 17. Viscor Distribution Inc. (Worm Bins), 12165 Cherrywood Drive, Maple
Ridge, BC, Canada V2X OB7. Tel. No. 800-609-1223; Fax No. 604-467-9661.
R 18. Worms & Worm Boxes, 968 Valencia Street, San Francisco, CA 94110,
U.S. Tel. No. 415-282- WORM.
W 19. Willingham Worm Farm, Rt. # 1, Box 241, Butler, GA 31006, U.S. Tel. No.
912-862-5545.
W 20. Manchester Worm Farm, 1131-0 Tolland Turnpike, Manchester, CT 06040,
U.S. Tel. No. 203-647-8067.
C 21. Environmental Recycling Systems, PO Box 904, Alpine, CA 91903, U.S. Tel.
No. 619-445-1873; Fax No. 619-445-6057.
C 22. Vermiculture Services International, U.S. Tel. No. 800-399-9464; Fax No.
707-279-8031.
D 23. Recycle-It Corporation, U.S. (distributor of worm bins, curbside recycling
bins, and backyard composting bins) Tel. No. 800-769-1044.
186 L. K. Wang et al.

W 24. Olympic Worm Casting Farm, McCleary, WA, U.S. Tel. No. 206-495-3762.
C 25. Casting a New Future, Portland, OR, U.S. Tel. No. 503-246-7382.
D 26. RPM, 2829 152nd Ave. NE, Redmond WA 98052, U.S. Tel. No. 800-867-3201.
27. Sound Resource Management Group, Inc., 119 Pine Street, Seattle, WA 98101,
U.S. Tel. No. 206-622-9454; Fax No. 206-622-9569.
R 28. Worm World, 26 Ihnat Lane, Avella, PA 15312, U.S. Tel. No. 412-356-2397.
C 29. Resource Conversion Corporation, 7825 Fay Avenue, Suite 380, La Jolla, CA
92037, U.S. Tel. No. 619551-4800.
C 30. Environmental Earthworm Projects, Inc., 8114 Port Said Street, Orlando, FL
32813, U.S. Tel. No. 407678-6454.
C 31. Original Vermitech Systems, Ltd., 2328 Queen Street East, Toronto, Ontario,
M4E1G9, Canada. Tel. No. 416-693-1027.
C 32. The Worm Connection, 581 Camino Manzanas, Thousand Oaks, CA 91360,
U.S. Tel. No. 805-496-2872, Tel/Fax No. 805-376-9918.

3.5 Process Design Considerations

3.5.1 Process Adoption and Advantages

Earthworm castings are essentially odorless when dry; when damp, they have a mild
odor like a good quality topsoil. Also, castings have a favorable appearance. When
sifted and dry, they are granular, about 0.02–0.1 inches (0.5–3 mm) in maximum
dimension (with some fines); color is brownish gray. In a study where municipal
sludge was applied to a wheat crop, it was found that when earthworms were added
to the sludge, the germination rate of the wheat improved [52]. The odor, appear-
ance, and soil supplementation advantages of the earthworm conversion process
may help in the acceptance of sludge by farmers and householders.
Earthworm conversion affects several other sludge characteristics. The oxygen
uptake rate increases [48]; the acid-extractable fraction of various nutrients increases
[52]. The volatile content of the solids drops slightly, and humic acid concentrations
fluctuate [48]. While these effects may be beneficial, there are no data to show how
the results affect design or operation of earthworm conversion installations.
The earthworm conversion process would appear to be low in cost, although this
cannot be said with certainty, since no cost data are available for full-scale opera-
tions on sludge. The process does not require chemicals, high temperatures, or large
amounts of electricity. Only a small amount of low-speed mechanical equipment is
needed. Significant expenditures may be required to offset the potential operating
difficulties discussed below.
3 Vermicomposting Process for Treating Agricultural and Food Wastes 187

3.5.2 Process Operation and Troubleshooting

A number of potential operating difficulties and their solutions exist in the earth-
worm conversion process. None of these difficulties are insurmountable, however.
Probably the most difficult problem is to economically pretreat anaerobically
digested sludge so that it is nontoxic to the worms [61, 63]. Other problems that
must be considered include:
• Worm drowning: Worms must be protected from flooding.
• Worm loss due to migration from the process: Caused by flooding, toxic sludge,
unpalatable sludge, adjoining areas attractive to worms, lack of artificial lighting
on rainy nights.
• Toxicity of sludge to worms: Significant for anaerobically digested sludge.
However, toxicity is eliminated by exposing the sludge to air for 2 months [48]
or wetting sun-dried sludge daily for 14 days [52]. Stabilization by lime or chlo-
rine is not recommended for sludge that will be fed to earthworms. Toxicants
such as copper salts might also cause problems. Aerobic digestion is best suited
for sludge to be converted by earthworms.
• Toxicity or unpalatable nature of dewatering chemicals: Avoided at Hagerstown,
Md., by use of food-grade polymer [50]. Drying beds may be used; drying beds
do not usually require chemicals.
• Worm shortage in the process, so that worm additions are required: Worms
reproduce via egg capsules, which may be lost from the process in the castings.
Also, toxic conditions, drowning, and other problems will cause worm popula-
tions to drop. At Hagerstown, Md., a worm-raising operation has been proposed
to supply the necessary makeup worms to the sludge conversion process [50].
• Shortage of worms for initial inventory or restart: To begin operation, a large
worm inventory may be needed, but local worm suppliers may be unable to meet
this demand. Gradual start-up is therefore desirable, especially for large plants.
Also, earthworm exchanges may become available nationwide so that sludge
operations can draw on larger numbers of earthworm suppliers.
• Temperature extremes: Worm feed most rapidly at 15–20 °C; about 5 °C, feeding
is quite slow [48]. Freezing will kill worms. High temperatures can also cause
problems. It may be necessary to stockpile sludge during the winter or provide a
heated building for the conversion process.
• Shortage of enzymes: Not a problem, despite claims by marketers of enzyme
preparations that these preparations are valuable to the process [54].
• Exposure to light: Worms avoid bright light. Some sort of cover or shade should
be provided so that worms will convert the top layer of the sludge.
• Dehydration: There is a minimum moisture content for the worm bed [54].
• Salinity in castings: Under some conditions, castings may have sufficient dis-
solved salts to inhibit plant growth. This problem may be eliminated by leaching
or by mixing the castings with other materials with lower dissolved salts [55, 56].
188 L. K. Wang et al.

• Contamination of castings by heavy metals, motor oil, rags, and similar materi-
als: Source control may be used where feasible, as for other processes aimed at
reuse of sludge as a soil conditioner.
• Odors: The most likely source is raw or aerobically digested sludge, which has
been stockpiled to await earthworm conversion.

3.5.3 Process Limitations

Limitations of the earthworm conversion process include, but are not limited to, the
following [60, 62]:
• Earthworm conversion decreases the total nitrogen values in the sludge, as
ammonia nitrogen will be lost to the atmosphere.
• Costs are unpredictable.
• Two common ions in municipal wastewater sludge, ammonium and copper, may
be toxic to worms. Studies have found that these ions were lethal at additions
equivalent to 180 mg NH4-N and 2500 mg Cu per kg of wet substrate [57, 58].
Safe limits for these elements are not known.
• Cadmium accumulates in the worm Eisenia foetida. Zinc apparently does not
accumulate in Eisenia foetida but does accumulate in other species [58, 59]. If
the worms are to be used as animal feed, the system must be operated such that
cadmium and zinc concentrations in the worms do not exceed recommended
levels for animal consumption.
• Space requirements may rule out earthworm conversion at some treatment plants.
• The earthworm business has been afflicted with unsound investments and exces-
sive claims. For example, it has been claimed that earthworm processing is able
to reduce concentrations of heavy metals [60]. Any such reduction could only be
caused by simple dilution with uncontaminated waste or by concentration of the
contaminants in the earthworms.
• If a particular sludge is suitable for earthworm conversion, that sludge should
also be suitable for reuse as a soil conditioner without being processed by earth-
worms. However, earthworm conversion reduces odor, improves texture, and
may increase germination rate.
These limitations may seem significant but are not overwhelming. Considerable
research and development is underway, and it appears that earthworm conversion
may soon have a role in municipal wastewater treatment plant sludge processing.

3.5.4 Process Design Criteria

Design criteria have been generated by the operators and researchers in the field
[48–51, 61, 63, 69–90, 102] for the vermicomposting process.
3 Vermicomposting Process for Treating Agricultural and Food Wastes 189

Species of worm being tested were Eisenia foetida (redworm, hybrid redworm,
tiger worm, dung worm) [48, 51], Lumbricus rubellus (red manure worm, red wig-
gler worm) [49], and Lumbricus terrestris (night crawler) [48]. The following are
the compiled design criteria:
Detention time of sludge in worm beds = 2–32 days [49, 50].
Worm reproductive cycle = 1–2 months.
Rate of worm feeding (15 °C) = 0.17–1.7 g dry sludge per gram dry worm weight
per day [48].
Optimum temperature = 15–20 °C.
Dry matter content of worms = 20–25% (Eisenia foetida) [51].
Minimum solids content of the worm bed mixture = 20%: Actual minimum sol-
ids content depends on such factors as porosity, type of sludge, and ability to keep
aerobic. Experiments are being conducted to better define these parameters.

3.6 Process Application Examples

The Wright-Patterson Air Force Base in Dayton, Ohio, USA [45], launched a ver-
micomposting program in July 2002, using earthworms to consume a daily average
of 500 pounds of solid waste. The worms digest vegetable matter and old newspa-
pers, saving the base about $25 per day on transporting and disposing of waste. As
the number of worms grows, so does the amount of waste they consume. The base
acquired 250,000 worms and their climate-controlled home (at a constant 70 degree
F) for the environmental project. At the base, which produces fruit and vegetable
waste from its commissary, the earthworms have flourished, now numbering more
than 300,000. Their numbers eventually could top one million. The worm casings
replace chemical fertilizer at the base’s golf course, which saves additional money.
More successful stories can be found in the literature [42–61].
Vermicomposting has gained popularity in schools and municipalities, according
to Stuckey and Hudak [62]. In Boston, Massachusetts, USA, Josiah Quincy
Elementary School received a grant to build a rooftop organic garden. The students
maintain garbage-eating red wiggler worms to break down fruits and vegetables.
Once processed in the bin, the compost is applied to the garden. In Orange County,
Florida, USA, a revolutionary worm-use concept has been promoted where worms
stabilize biosolids to a “Class A pathogen standard” substance.
Mudrunka et al. [92] attempted to eliminate microbial pollution of domestic ani-
mal excrements using vermicomposting. Their investigations were carried out in a
three-chamber domestic wooden vermicomposter, in which aerobic degradation of
three types of animal excrements (cow, pig, dog) using the earthworm Eisenia
andrei. Before laying the individual excrements to the compost batch, the appropri-
ate input samples were taken for the microbiological examination of the biopatho-
gens. After 6 months, final samples of the final substrate were taken to determine
whether proper compost sanitization took place during the vermicomposting
190 L. K. Wang et al.

process. According to valid legislation, the bacteria Escherichia coli, Enterococcus

sp., and Salmonella sp. were identified as indicator microorganisms. After the eval-
uation of the performed laboratory analyses, it was proved that the use of earthworm
bioactivity resulted in elimination or at least significant reduction of the concentra-
tions of these bacterial strains in the final vermicompost samples [92].
Agricultural wastes include mainly the organic wastes generated from various
operations in the agricultural and forestry industries, such as crop residues, weeds,
leaf litter, sawdust, forest wastes, livestock waste, biosolids, fruit pomace, animal
dung, cardboard compost, rice straw, paper mill sludge and fibers, vegetable plant
debris, fruit plant debris, etc. Theoretically, all organic agricultural and food wastes
can be properly treated by the vermicomposting process for solid waste volume
reduction, pathogen reduction, and vermicompost recycle.
Among the various agricultural wastes, livestock waste is always a preferred
choice for researchers as feedstock for earthworms and as bulking substrate for
vermicomposting [90, 105]. Livestock waste is considered as the suitable organic
amendment to enhance the process of vermicomposting because of its low cost,
easy availability, sufficient nutrient content, and ideal C/N ratio [71]. The chemical
composition of livestock waste depends on the type of feed given to the animal,
bedding material, and fresh or dried including the manner how excreta is collected,
stored, and handled prior to vermicomposting [74]. Hence, differences in physico-
chemical characteristics of livestock waste have effects on the life cycle of earth-
worm species [71].
Many researchers [69–89] have conducted vermicomposting studies on the use
of various agricultural wastes as feedstock. Sharma and Garg [90] have compiled
their research data together as shown in Table 3.1.
An extensive vermicomposting research involving the use of a skin coffee (SC)
amended with green waste (GW) and biochar (B) has been conducted by Zulhipri
et al. [102]. Skin coffee is a food manufacturing waste from Cibulao coffee farm,
Bogor, Indonesia. Green wastes are mainly the branch cuttings and leaves from the
garden of the Universitas Negeri Jakarta. Biochar is a carbon-rich product made
from the rice husk pyrolysis process at 450 °C. Biochar was mixed by the research-
ers [102] with SC and GW in different proportions, i.e., 6%, 8%, and 10%, along
with control and allowed to pass through earthworm guts for 2 months’ processing.
The 8% biochar addition rate achieved maturity of vermicomposting and resulted in
the highest-quality vermicompost based on parameters such as organic C, C:N ratio,
total N, P, K content, and pH in comparison with a control vermicompost. They
further used the produced vermicompost for cultivating the growth of coffee plant.
Physiological parameters and morphology of coffee plant growth such as number of
leaves, height, plant diameter, and shoot dried weight were recorded. Their impor-
tant research data are presented in Tables 3.2, 3.3, and 3.4 [102].
The analytical results on the characteristics of earthworm media are shown in
Table 3.2 [102]. The high nutrient content in biochar is expected to increase the
nutrient content in vermicompost. Table 3.3 shows that the earthworm media sig-
nificantly affected the chemical content and macro- and microelements in vermi-
compost. There is no significant increase in nitrogen content with an increase in
3 Vermicomposting Process for Treating Agricultural and Food Wastes 191

Table 3.1 Vermicomposting of various agricultural wastes. Credit: K Sharma and VK Garg [90]
Type of waste
(bulking
No. material) Earthworm Duration Results References
1 Buffalo waste, Eisenia 90 days Maximum earthworm growth Sharma and
sheep waste, foetida rate was achieved in the various Garg [71]
goat waste, cow combinations of buffalo dung and
waste minimum growth rate in sheep
waste. TOC content and C/N
ratio decreased during
vermicomposting, whereas total
nutrient content increased
2 Rice straw + E. foetida 105 days Paper waste and rice straw Sharma and
paper waste + effectively convert into nutrient-­ Garg [72]
cow dung rich vermicompost.
Vermicompost is more
fragmented than parent
feedstocks. Use of rice straw in
higher ratio was not
recommended
3 Salvinia E. foetida 45 days Chemical compounds responsible Hussain et al.
molesta for weed allelopathic effects [79]
destroyed completely. The C/N
ratio of Salvinia was reduced
sharply from 53.9 to 9.35
4 Sewage sludge E. foetida 80 days Vermicomposting modifies the Lv et al. [74]
(cattle dung) structure of bacterial community
in the waste and reduces the
pathogenic human bacteria
population
5 Pig manure and E. foetida 40 days Vermicompost has higher pH, P, Zhu et al.
rice straw K, Zn, and CEC but lower [82]
available N and Cu than the
parent substrate. Increment in
aromatic compounds indicated
high humification during
vermicomposting. Earthworm
tissues accumulated 13C
6 Crop/tree Eudrilus Earthworm growth and Thomas et al.
residues sp. conversion efficiency vary with [69]
waste. In all the crop residues,
pH, EC, and N and P levels
increased, whereas C/N and C/P
ratios decreased
(continued)
192 L. K. Wang et al.

Table 3.1 (continued)

Type of waste
(bulking
No. material) Earthworm Duration Results References
7 Horse manure, Eisenia 240 days Study evaluated vermicompost Sanchez et al.
apple pomace, andrei characteristics based on [70]
grape pomace, 120-day-old layer and 240-day-­
and digestate old layer in vermireactor.
(manure slurry, Maximum biomass of
corn silage, earthworms was in 120-day-old
haylage) layer. After 240 days, microbial
biomass activity decreased due to
a decrease in the earthworm
activity, indicating a high degree
of stabilization. Enzyme
activities differ according to the
age of the layers and the type of
waste. Germination index
increased after vermicomposting
and was higher with apple
pomace and digestate than that
with horse manure and grape
pomace
8 Cow manure E. foetida 60 days Urease activity is a suitable Sudkolai and
and wheat indicator of vermicompost Nourbakhsh
residues maturity and waste stabilization [83]
during the process of
vermicomposting. Urease activity
was highly correlated with the
time of vermicomposting
resulting in r = 0.97 for cattle
manure and r = 0.99 for wheat
waste. Urease activity showed
significant correlations with the
C/N ratio
9 Wheat straw, E. foetida 90 days Highest worm production and Vodounnou
pig dung, growth rate were obtained with et al. [75]
poultry dung, cow dung followed by pig dung;
rabbit dung, however, earthworm growth
cattle dung, decreased in vegetable compost.
sheep dung, Maximum earthworm growth
and vegetal rate was found on the 90th day.
compost Growth and worm production
depend on the biochemical
quality of the feedstocks
(continued)
3 Vermicomposting Process for Treating Agricultural and Food Wastes 193

Table 3.1 (continued)

Type of waste
(bulking
No. material) Earthworm Duration Results References
10 Sawdust, E. foetida 100 days MSW and carbonaceous Alidadi et al.
boxwood materials in different proportions, [73]
leaves, and viz., 50:50, 70:30, 85:15, and
cardboard 100:0, were vermicomposted for
compost 100 days. Vermicomposting for
(MSW) 75 days is sufficient for
vermicompost maturity in terms
of EC, WSC, DEH, and C/N
ratio. Phosphorus, nitrogen, and
pH levels were higher in the
vermicompost
11 Salvinia natans E. foetida 45 days Total concentration of heavy Singh and
(cattle manure metals (Zn, Cu, Mn, Fe, Cr, Pb, Kalamdhad
and sawdust) Cd, Ni) increased; however, [84]
concentration of water-soluble
and plant-available heavy metals
was reduced in the final
vermicompost. TCLP tests
confirmed the suitability of
vermicompost for agriculture
12 Leaf litter Perionyx 60 days Cashew leaf litter mixed with Parthasarathi
(horse dung, excavatus cow dung at 2:2 ratio was found et al. [85]
sheep dung) to best in terms of vermicompost
properties. The vermicompost
produced had lower pH, organic
carbon content, C/N ratio, C/P
ratio, and lignin, cellulose,
hemicellulose, and phenol
content but higher NPK, DEH,
and HA content than the waste
and compost. Reduction in the
lignocellulose and phenol content
is due to the combined action of
the gut lignocellulolytic
microflora and earthworms
during the vermicomposting
process
13 Ipomoea E. foetida 30 days Total carbon contents decreased Hussain et al.
from 527.3 to 282.8 g/kg and [76]
total nitrogen contents increased
from 20.2 to 28.5 g/kg. C/N ratio
of Ipomoea vermicompost was
9.9. Spectroscopic analysis
revealed transformation of weed
into potent organic fertilizer
(continued)
194 L. K. Wang et al.

Table 3.1 (continued)

Type of waste
(bulking
No. material) Earthworm Duration Results References
14 Coconut husk Eudrilus 21 days Highest recovery of relative N Swarnam
poultry manure, eugeniae (1.6) and K (1.3) was in 20% et al. [86]
pig slurry feedstock substitution by pig
slurry, and highest P recovery
(2.4) was with poultry manure
substitution. Vermicompost
contains higher pH, microbial
biomass carbon, and macro- and
micronutrients than the initial
waste
15 Cow dung, E. foetida After vermicomposting, pH, TOC Lv et al. [74]
poultry manure content, and C/N ratio were
reduced but EC and HA were
increased. Heavy metals
stabilized
16 Decanter cake E. 2 weeks Four treatments with different Lim et al.
+ rice straw eugeniae ratios of decanter cake and rice [87]
straw (2:1, 1:1, 1:2, 1:3) were
prepared. Two parts decanter
cake and one part rice straw
(w/w) was found to best among
all the treatments
17 Crop residue E. 90 days Highest earthworm weight and Aynehband
(rice, wheat, eugeniae vermicomposted matter were et al. [85]
corn, achieved in wheat and lowest
sugarcane) with corn residue
18 Lantana E. foetida – C/N ratio reduced from 22.7 to Hussain et al.
8.1; humification index from 8.38 [78]
to 2.03. FTIR spectra revealed
complete degradation of phenols
and sesquiterpene lactones and
formation of simple compounds.
GC-MS analysis revealed
transformation of 24–86
constituents
19 Parthenium E. foetida – Chemicals responsible for the Hussain et al.
allelopathic effect of parthenium [77]
weed are destroyed. Scanning
electron microscopy shows
marked disaggregation of the
material in the vermicompost as
compared with the well-formed
matrix of Salvinia leaves
(continued)
3 Vermicomposting Process for Treating Agricultural and Food Wastes 195

Table 3.1 (continued)

Type of waste
(bulking
No. material) Earthworm Duration Results References
20 Tomato plant E. foetida 6 months Characterize HA isolated from Fernandez
debris + paper different waste mixtures before Gomez et al.
mill sludge and after vermicomposting. HA [81]
content increased by 15.9–16.2%.
Vermicompost produced from
tomato debris/paper mill sludge
(2:1) recorded higher C content
and C/N ratio. HA from tomato
debris/paper mill sludge (1:1)
vermicompost showed a higher O
content and O/C ratio
21 Filter cake E. foetida 30 days Positive correlation of Busato et al.
(cattle manure) phosphatase activities with TOC [89]
content, pH, and WSP but
negative correlation with HA
content. Nanopore volume found
to be negatively correlated with
phosphatase activities for filter
cake but not for cattle manure.
HA content of filter cake
vermicompost was higher than
that of cattle manure
vermicompost
Note: CEC cation exchange capacity; DEH dehydrogenase; EC electric conductivity; FTIR Fourier
transform infrared; GC-MS gas chromatography-mass spectrometry; HA humic acid; MSW munic-
ipal solid waste; TCLP toxicity characteristic leaching procedure; TOC total organic carbon; WSC
water-soluble carbon; WSP water-soluble phosphorus

Table 3.2 Chemical content of the worming media. Credit: Zulhipri et al. [102]
No Parameter Skin coffee Cow dung Biochar
1 pH 6.21 7.74 7.82
2 C organic (%) 34.48 48
3 N (%) 1.27 1.05 2.26
4 P (ppm) 29 84.62 97.80
5 K (ppm) 2.46 8.08 12.04
6 C/N ratio 32.84 21.23

%B. The pH and organic C, however, increase with an increase in biochar addition.
By increasing C, the C/N ratio also increases. After organic waste becomes vermi-
compost, the mineral content of vermicompost increases with increasing biochar
content. This is because biochar contains porous organic carbon with high surface
area which is able to adsorb metallic metals from earthworm media. It was observed
in the experiment [102] that the media containing 8% biochar produced more ver-
micompost than media that contained 4% biochar or did not contain biochar. The
196 L. K. Wang et al.

Table 3.3 Vermicompost characteristic of various media. Credit: Zulhipri et al. [102]
No Chemicals Compost +0%B Compost +6%B Compost +8%B
1 pH 5.36 5.48 5.60
2 Organic 53.20 61.65 65.80
3 Carbon organic 26.30 34.50 36.40
4 Nitrogen 2.70 2.50 2.76
5 Sodium 1.73 1.29 1.32
6 Potassium 3.48 3.62 3.84
7 Calcium 51.46 52.70 56.00
8 Magnesium 6.22 6.24 6.85
9 Phosphorus 28.42 28.90 32.74
10 Zinc 2245 2264 2676
11 Manganese 5.24 5.18 5.42

Table 3.4 Effect of vermicompost on coffee plant. Credit: Vanderholm et al. [105]
No Sample Composition Height Leaf
1 EC Soil + compost 12.00 4
2 EV5 Soil + 5% vermicompost 13.14 7
3 EV10 Soil + 10% vermicompost 15.24 11
4 EV15 Soil + 15% vermicompost 17.22 13
5 EV20 Soil + 20% vermicompost 18.86 15

composting time also has a significant effect on the weight of the vermicompost
produced. The composting time of 2 weeks gives the highest vermicompost results,
namely, 469 g. This is due to the fact that in the second week the number of young
worms increases, and with increasing time, the mother worms decrease, so that less
vermicompost is produced. Table 3.4 shows the effect of vermicompost on the
growth of coffee plant. The highest height of coffee seedlings was in the EV20, but
it was not significantly different from the EV15 treatment. The lowest plant height
was in EC treatment and significantly different from other treatments as shown in
Table 3.4. EV20 treatment, soil containing 20% vermicompost, gave the best results
against all variables observed. This is because in this composition there is more soil
containing vermicompost fertilizer. Vermicompost can fertilize the soil through its
influence on soil physical, chemical, and biological properties. Physically, vermi-
compost can (a) affect soil texture, (b) improve soil structure, (c) improve soil con-
sistency, (d) improve soil drainage, (e) improve soil pores, (f) increase soil maturity,
(g) increase plant growth power, and (h) loose the soil so that the space for the roots
will increase. Chemically, vermicompost fertilizer will (a) contribute macro- and
micronutrients, (b) increase soil reaction (soil pH), (c) improve soil colloids (min-
eral matter), (d) increase ion exchange capacity, and (e) gain base saturation so that
the availability of nutrients is getting better. Biologically, vermicompost can increase
the population of soil microorganisms so that the soil becomes more fertile.
3 Vermicomposting Process for Treating Agricultural and Food Wastes 197

Vermicompost also functions as a biological control tool in suppressing plant dis-

eases, namely, by inhibiting disease growth through natural processes by increasing
competitive activity and antibiotics in the inoculum. The best media composition is
P5 treatment (one part of subsoil mixed with four parts of vermicompost from cof-
fee husk waste). The use of vermicompost fertilizer from coffee husk waste can
substitute NPK inorganic fertilizer for coffee nurseries in the main nursery.

3.7 Future Development and Direction

Vermicomposting (or vermistabilization) should be encouraged by governments in

the field of environmental engineering as promising processes for disposal of bio-
solids and other organic solid wastes. Special efforts should be made in the near
future to obtain recognition for the process, and funding sources should be explored
at all levels for economical analysis and optimization of the process. At the global
level, international agencies should encourage and fund the transfer of vermicom-
posting technology between the USA and all other countries.

Glossary

Agricultural wastes They include mainly the organic wastes generated from vari-
ous operations in the agricultural and forestry industries, such as crop residues,
weeds, leaf litter, sawdust, forest wastes, livestock waste, biosolids, fruit pom-
ace, animal dung, cardboard compost, rice straw, paper mill sludge and fibers,
vegetable plant debris, fruit plant debris, etc.
Biochar It is a carbon-rich product made from the rice husk pyrolysis process
at 450 °C.
Green waste They are mainly the branch cuttings and leaves from the gardens.
Vermicompost The end product of a vermicomposting process which is an effi-
cient growth promoter for plants, as it contains plant-available nutrients, rich
microbial population, humic substances, growth hormones, and enzymes. It is an
organic fertilizer that improves crop growth and yield.
Vermicomposting It is a bio-oxidative natural decomposition process that occurs
under mesophilic conditions further aided by the biochemical action of micro-
organisms, such as different earthworm species. The mutual action of worms
and microbes converts organic waste, such as agricultural wastes, food wastes,
biosolids, etc., into fine, homogenized, odor-free, nutrient-rich, and humus-rich
manure that is called vermicompost. Vermicomposting helps achieve a circular
bioeconomy by converting waste into useful products that are necessary for the
overall sustainable development of a country.
198 L. K. Wang et al.

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Chapter 4
The Impacts of Climate Change
on Agricultural, Food, and Public Utility
Industries

Josephine O. Wong, Erick Butler, Nai-Yi Wang, Mu-Hao Sung Wang,

and Lawrence K. Wang

Acronyms and Nomenclature

BC Black carbon
CCS Carbon capture and sequestration
CFC Chlorofluorocarbon
CH4 Methane
CO2 Carbon dioxide
ENSO El Niño-Southern Oscillation
ERF Effective radiative forcing
GHG Greenhouse gas

J. O. Wong (*)
City of San Francisco, San Francisco, CA, USA
e-mail: [email protected]
E. Butler
School of Engineering and Computer Science, West Texas A&M University,
Canyon, TX, USA
e-mail: [email protected]
N.-Y. Wang
University of Buffalo, Buffalo, NY, USA
e-mail: [email protected]
M.-H. S. Wang
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]
L. K. Wang
Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 205

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_4
206 J. O. Wong et al.

GWP Global warming potential

HBFC Hydrobromofluorocarbon
HC Hydrocarbon
HCFCs Halogenated fluorocarbons
HFCs Hydrofluorocarbons
IPCC Intergovernmental Panel on Climate Change
LFG Landfill gas
MCL Maximum contaminant level
N2O Nitrous oxide
NPDWR National Primary Drinking Water Regulations
ODS Ozone-depleting substance
O3 Ozone
PFCs Perfluorinated carbons
PM Particulate matter
RF Radiative forcing
USEPA US Environmental Protection Agency
USGS US Geological Survey
WMO World Meteorological Organization
W m−2 Watts per square meter

4.1 Introduction

Global climate change, also referred to as global warming, is a serious threat to our
environment. This chapter will present a summary of scientific facts about climate
change and discuss its impacts in particular on water resources and planning. The
underlying science of climate change is undisputable. Climate change has become
a contentious political issue, which is unfortunate if only because it distracts society
and our policymakers from necessary discussions and decisions about how to
respond to the impacts of climate change on our communities.

4.1.1 Weather, Climate, and Climate Change

The climate of a region can be thought of as the “average” of that region’s weather.
Climate is predictable, or at least enough so that we rely on it for planning. We
speak of the sunny, temperate climate of the Mediterranean, or the harsh, cold cli-
mate of Siberia (even though there are both cold winter storms in Italy and warm
summer days on the taiga), and vacations and population growth projections are
adjusted accordingly. In simple modeling terms, if an input that was previously
thought to be constant—climate—is found to be variable, or to have become vari-
able, then the model output will also necessarily change from what was previously
expected. If the weather we are used to expecting is no longer what can be expected
in the future, then what should current and future infrastructure planning be based
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 207

on? Land use planners and water and wastewater utility operators and regulators
need to understand what the impacts of climate change will be on water resources
and how to prepare for these changes.
It is well known that the Earth has gone through multiple ice ages in periods of
dramatic cooling and dramatic warming. Among the many ice ages the Earth has
undergone, we are currently going through a warming trend. Questions we would
ask are: “Why is the Earth warming?” “How does the source warm the Earth?”
“What are the implications on our water resources, animals, and agriculture?” There
are more questions than answers [1–38]. But before we discuss how the Earth’s
climate is changing, there are some technical terminologies presented below as well
as in the “Glossary” section of this chapter [26–30].
Stratospheric ozone plays a decisive role in the stratospheric radiative balance.
Since ozone absorbs a band of ultraviolet radiation called UVB that is particularly
harmful to living organisms, the ozone layer prevents most UVB from reaching the
ground. Depletion of stratospheric ozone, due to chemical reactions that may be
enhanced by climate change, results in an increased ground-level flux of ultraviolet
(UV) B radiation.
Ozone “Hole” is a large area of the stratosphere with extremely low amounts
of ozone.
Ozone-depleting substance (ODS) is a family of man-made compounds that
includes, but is not limited to, chlorofluorocarbons (CFCs), bromofluorocarbons
(halons), methyl chloroform, carbon tetrachloride, methyl bromide, and hydrochlo-
rofluorocarbons (HCFCs). These compounds have been shown to deplete strato-
spheric ozone and therefore are typically referred to as ODSs.
Ozone layer depletion means chemical destruction of ozone molecules in the
ozone layer. Depletion of this ozone layer by ozone-depleting substances will lead
to higher UVB levels (a band of ultraviolet radiation), which in turn will cause
increased skin cancers and cataracts and potential damage to some marine organ-
isms, plants, and plastics.
Particulate matter (PM) are very small pieces of solid or liquid matter such as
particles of soot, dust, fumes, mists, or aerosols. The physical characteristics of
particles, and how they combine with other particles, are part of the feedback mech-
anisms of the atmosphere.
Photosynthesis is a process by which plants take CO2 from the air (or bicarbonate
in water) to build carbohydrates, releasing O2 in the process. There are several path-
ways of photosynthesis with different responses to atmospheric CO2
concentrations.
Phytoplankton are microscopic plants that live in salt and freshwater
environments.
Precession means the wobble over thousands of years of the tilt of the Earth’s
axis with respect to the plane of the solar system.
Precipitation includes rain, hail, mist, sleet, snow, or any other moisture that falls
to the Earth.
Radiation is a type of energy transfer in the form of electromagnetic waves or
particles that release energy when absorbed by an object.
208 J. O. Wong et al.

Radiative forcing means (a) a change in the balance between incoming solar
radiation and outgoing infrared radiation and (b) a measure of the influence of a
particular factor (e.g., greenhouse gas (GHG), aerosol, or land use change) on the
net change in the Earth’s energy balance.
Soil is a complex mixture of inorganic minerals (i.e., mostly clay, silt, and sand),
decaying organic matter, water, air, and living organisms.
Soil carbon is a major component of the terrestrial biosphere pool in the carbon
cycle. The amount of carbon in the soil is a function of the historical vegetative
cover and productivity, which in turn is dependent in part upon climatic variables.
Solar energy is also called solar radiation, energy from the Sun, and also referred
to as shortwave radiation. Of importance to the climate system, solar radiation
includes ultraviolet radiation, visible radiation, and infrared radiation. It also
includes indirect forms of energy such as wind falling or flowing water’s hydro-
power, ocean thermal gradients, and biomass, which are produced when direct solar
energy interacts with the Earth.
Solar radiation is a radiation emitted by the Sun. It is also referred to as short-
wave radiation. Solar radiation has a distinctive range of wavelengths (spectrum)
determined by the temperature of the Sun.
Source means any process or activity that releases a greenhouse gas, an aerosol,
or a precursor of greenhouse gas into the atmosphere.
Stratosphere is the region of the atmosphere above the troposphere and between
the troposphere and the mesosphere. The stratosphere extends from about 8–50 km
(6–31 miles) in altitude. Specifically, it has a lower boundary of approximately
8 km at the poles to 15 km at the equator and an upper boundary of approximately
50 km. Depending upon latitude and season, the temperature in the lower strato-
sphere can increase, be isothermal, or even decrease with altitude, but the tempera-
ture in the upper stratosphere generally increases with height due to absorption of
solar radiation by ozone. So the stratosphere gets warmer at higher altitudes. In fact,
this warming is caused by ozone absorbing ultraviolet radiation. Warm air remains
in the upper stratosphere, and cool air remains lower, so there is much less vertical
mixing in this region than in the troposphere. Commercial airlines fly in the lower
stratosphere.
Terrestrial radiation means the total infrared radiation emitted by the Earth and
its atmosphere in the temperature range of approximately 200–300 Kelvin.
Terrestrial radiation provides a major part of the potential energy changes necessary
to drive the atmospheric wind system and is responsible for maintaining the surface
air temperature within limits of livability.
Troposphere is (a) the region of the atmosphere closest to the Earth. The tropo-
sphere extends from the surface up to about 10 km (6 miles) in altitude, although
this height varies with latitude. Almost all weather takes place in the troposphere.
Mt. Everest, the highest mountain on Earth, is only 8.8 km (5.5 miles) high.
Temperatures decrease with altitude in the troposphere. As warm air rises, it cools,
falling back to Earth. This process, known as convection, means there are huge air
movements that mix the troposphere very efficiently; or (b) the lowest part of the
atmosphere from the surface to about 10 km in altitude in midlatitudes (ranging
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 209

from 9 km in high latitudes to 16 km in the tropics on average) where clouds and

“weather” phenomena occur. In the troposphere, temperatures generally decrease
with height. All weather processes take place in the troposphere. Ozone that is
formed in the troposphere plays a significant role in both the greenhouse gas effect
and urban smog. The troposphere contains about 95% of the mass of air in the
Earth’s atmosphere.
Weather is the specific condition of the atmosphere at a particular place and time.
It is measured in terms of such things as wind, temperature, humidity, atmospheric
pressure, cloudiness, and precipitation. In most places, weather can change from
hour to hour, day to day, and season to season. Climate in a narrow sense is usually
defined as the “average weather” or, more rigorously, as the statistical description in
terms of the mean and variability of relevant quantities over a period of time ranging
from months to thousands or millions of years. The classical period is 30 years, as
defined by the World Meteorological Organization (WMO). These quantities are
most often surface variables such as temperature, precipitation, and wind. Climate
in a wider sense is the state, including a statistical description, of the climate system.
A simple way of remembering the difference is that climate is what you expect (e.g.,
cold winters) and “weather” is what you get (e.g., a blizzard) [30].

4.1.2 Greenhouse Gases, Greenhouse Effect, Global Warming,

Global Warming Potential

A greenhouse gas (GHG) is any gas that absorbs infrared radiation in the atmo-
sphere. Greenhouse gases include water vapor, carbon dioxide (CO2), methane
(CH4), nitrous oxide (N2O), halogenated fluorocarbons (HCFCs), ozone (O3), per-
fluorinated carbons (PFCs), hydrofluorocarbons (HFCs), and sulfur hexafluoride.
Gases absorb heat in the atmosphere near the Earth’s surface, preventing it from
escaping into the space. If the atmospheric concentrations of these gases rise, the
average temperature of the lower atmosphere will gradually increase, a phenome-
non known as the greenhouse effect.
Specifically, greenhouse effect is produced as greenhouse gases allow incoming
solar radiation to pass through the Earth’s atmosphere, but prevent most of the out-
going infrared radiation from the surface and lower atmosphere from escaping into
the outer space. This process occurs naturally and has kept the Earth’s temperature
about 60 °F warmer than it would otherwise be. Current life on Earth could not be
sustained without the natural greenhouse effect. The greenhouse effect is trapping
heat and build-up of heat in the atmosphere (troposphere) near the Earth’s surface.
Some of the heat flowing back toward the space from the Earth’s surface is absorbed
by water vapor, carbon dioxide, ozone, and several other gases in the atmosphere
and then reradiated back toward the Earth’s surface [2, 30].
Global warming is known due to the recent and ongoing global average increase
in temperature near the Earth’s surface. It is the observed increase in average tem-
perature near the Earth’s surface and in the lowest layer of the atmosphere. In
210 J. O. Wong et al.

common usage, “global warming” often refers to the warming that has occurred as
a result of increased emissions of greenhouse gases from human activities. Global
warming is a type of climate change; it can also lead to other changes in climate
conditions, such as changes in precipitation patterns.
Global warming potential (GWP) is a measure of the total energy that a gas
absorbs over a particular period of time (usually 100 years), compared to carbon
dioxide. GWP is a number that refers to the amount of global warming caused by a
substance. The GWP is also the ratio of the warming caused by a substance to the
warming caused by a similar mass of carbon dioxide (CO2). Thus, the GWP of CO2
is 1.0. Chlorofluorocarbon (CFC)-12 has a GWP of 8500; CFC-11 has a GWP of
5000; hydrochlorofluorocarbons and hydrofluorocarbons have GWPs ranging from
93 to 12,100; and water has a GWP of 0 [2, 30].

4.2 Main Contributors to Greenhouse Gases

The main reason for the Earth’s warming is due to the greenhouse effect (Figs. 4.1
and 4.2). Before human activity, natural activities such as volcanic activity and natu-
ral forest fires would emit greenhouse gases in the atmosphere. As greenhouse gases
absorb heat and solar radiation, the concentrations of gases get trapped near the
Earth’s surface and sustain life [2]. A microcosm of this balance can be seen in the
relationship between humans and trees. Humans take in oxygen and release carbon

Fig. 4.1 The natural greenhouse effect before human activity ([2], Permission to use)
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 211

Fig. 4.2 The greenhouse effect after human activity ([2], Permission to use)

dioxide (CO2), while trees take in carbon dioxide and release oxygen, thereby creat-
ing a balance in nature.
However, due to increased human activity (from electricity, transportation,
industry, and population increase) [3], large amounts of CO2 and other greenhouse
gases have been released to the atmosphere. Also, as trees get cut down and defor-
ested, there are less resources using up the CO2, causing levels to rise beyond natu-
ral levels. So, what happens to the additional greenhouse gases floating in the air?
The excess CO2 and other greenhouse gases trap the extra radiation near the Earth’s
surface, causing global temperatures to rise, or global warming [2, 30].
The definitions of greenhouse gases and other technical terms have been intro-
duced in Sect. 4.1. The main contributor to the greenhouse effect is carbon dioxide.
Following carbon dioxide are methane gas (CH4) and nitrous oxide (N2O), and the
halocarbons as the leading greenhouse gases. Figure 4.3 below shows the major
climate changing agents of greenhouse gases and their radiative forcing (W m−2),
showing their emission ability to retain heat, and their great amounts on the Earth’s
surface [4]. As seen in Fig. 4.3, carbon dioxide has the greatest amount where it
absorbs heat at a radiative forcing above 1.5 W m−2 of increased and retained solar
radiation at the Earth’s surface. Although halocarbons, methane, and nitrous oxide
have a greater warming potential than carbon dioxide, the larger quantity of carbon
dioxide has a greater impact [28, 30]. Ozone depends on the location. In the tropo-
sphere where people live, ozone is a greenhouse gas where it absorbs heat; however,
ozone in the stratosphere actually absorbs UV radiation and holds back the radiation
from hitting the Earth’s surface, thereby keeping the Earth cooler. Water does not
affect the warming of the Earth too much since the concentration levels are fairly
constant. Land use goes both ways where dark forested areas or black carbon on
snow or from diesel engines in the troposphere would absorb heat, whereas planting
212 J. O. Wong et al.

Fig. 4.3 Radiative forcing estimates in 2011 relative to 1750 and aggregated uncertainties for the
main drivers of climate change [4]

lighter colored plants on arid regions where light reflects back to the space would
actually cool the Earth. Aerosols’ effects are uncertain; the concentration of aero-
sols can be monitored based on the brightness of clouds; the higher the concentra-
tion of aerosols, the brighter the clouds are. The reason is the aerosols feed the water
droplets that contribute to the clouds, and the more water droplets there are, the
more the droplets reflect light more. Aerosols in the stratosphere from volcanic
activity block the radiation to help cool the Earth. There is still much to investigate
and discover about aerosols [2, 4].
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 213

Figure 4.3 shows the total weighted average of all the climate changing agents;
there is a total of 2.29 W m−2 increase in the amount of solar energy absorbed at the
surface of the Earth [2, 4]. Because carbon dioxide shows the greatest quantity and
has the greatest impact, carbon dioxide will be the main focus and should often be
a reference point to compare to other greenhouse gases. Values in Fig. 4.3 are global
average radiative forcing (RF), partitioned according to the emitted compounds or
processes that result in a combination of drivers. The best estimates of the net radia-
tive forcing are shown as black diamonds with corresponding uncertainty intervals;
the numerical values are provided on the right of the figure, together with the confi-
dence level in the net forcing (VH, very high; H, high; M, medium; L, low; VL, very
low). Albedo forcing due to black carbon on snow and ice is included in the black
carbon aerosol bar. Small forcings due to contrails (0.05 W m−2, including contrail-­
induced cirrus) and HFCs, PFCs, and SF6 (total 0.03 W m−2) are not shown.
Concentration-based RFs for gases can be obtained by summing the like-colored
bars. Volcanic forcing is not included as its episodic nature makes it difficult to
compare to other forcing mechanisms. Total anthropogenic radiative forcing is pro-
vided for three different years relative to 1750.
The strength of drivers in Fig. 4.3 is quantified as radiative forcing (RF) in units
watts per square meter (W m−2) as in previous IPCC assessments. RF is the change
in energy flux caused by a driver and is calculated at the tropopause or at the top of
the atmosphere. In the traditional RF concept employed in previous IPCC reports,
all surface and tropospheric conditions are kept fixed. In calculations of RF for well-­
mixed greenhouse gases and aerosols in this report, physical variables, except for
the ocean and sea ice, are allowed to respond to perturbations with rapid adjust-
ments. The resulting forcing is called effective radiative forcing (ERF) in the under-
lying report. This change reflects the scientific progress from previous assessments
and results in a better indication of the eventual temperature response for these
drivers. For all drivers other than well-mixed greenhouse gases and aerosols, rapid
adjustments are less well characterized and assumed to be small, and thus the tradi-
tional RF is used [4].

4.3 Global Warming Potential and Its Limitations

In the earlier sections of this chapter, global warming potential (GWP) is a measure-
ment of how well heat is absorbed by greenhouse gases. The IPCC defines global
warming potential (GWP) as “the ratio of the time integrated radiative forcing from
a pulse emission of 1 kg of a substance, relative to that of 1 kg of carbon dioxide,
over a fixed horizon period. GWP is a relative index used to compare the climate
impact of an emitted greenhouse gas, relative to an equal amount of Carbon Dioxide”
[10]. Also, the IPCC examines the GWP for 1 g of carbon dioxide at a 20-, 100-, and
500-year time horizon in comparison to other greenhouse gases [9]. The six major
greenhouse gases are determined by the Kyoto Protocol. The Kyoto Protocol is an
international treaty that sets obligations on industrialized countries to lower the
214 J. O. Wong et al.

Table 4.1 IPCC global warming potential consensus [2, 8, 9]

Global warming potential for the
Six greenhouse gases listed designated by Lifetime given time horizon
the Kyoto Protocol (years) 20 years 100 years 500 years
Carbon dioxide, CO
2 ~150 1 1 1
Methane, CH
4 12 72 25 7.6
Nitrous oxide, N O
2 114 289 298 153
HFC-23 270 12,000 14,800 12,200
PFC-116 10,000 8630 12,200 18,200
Sulfur hexafluoride, SF
6 3200 5210 7390 11,200

emissions of greenhouse gases. The GWP values can be seen in Table 4.1; the table
shows the six major greenhouse gases from the Kyoto Protocol: carbon dioxide
(CO2), methane (CH4), nitrous oxide (N2O), HFCs, PFCs, and sulfur hexafluoride
(SF6). The Intergovernmental Panel on Climate Change (IPCC) includes many
greenhouse gases in their consensus and a list of HFCs and PFCs. The HFC and
PFC chosen in Table 4.1 are for the greatest GWP values within the consensus list.
The lifetime among the six gases ranges from 12 to 10,000 years. The value closest
to the median is sulfur hexafluoride, with a lifetime of 3200 years. At a time horizon
of 500 years, while carbon dioxide releases 1 g, sulfur hexafluoride releases 11,200 g
of carbon dioxide (11,200 times more) for the same time horizon. A stronger green-
house gas can easily leak and create a major impact.
The global warming potential has limitations where radiative properties are
uncertain and nonlinear (CO2, CH4, N2O); the actual resident life of greenhouse
gases and how long it actually stays in the atmosphere vary and some are unknown
(CO2, ozone precursors, diesel PM, and PM); if the resident lifetimes are short-lived
in the atmosphere, the GWP is not useful; there are not only direct radiative forcings
but also indirect radiative forcings with uncertainties (i.e., ozone precursors are not
only a gas, but they also form ozone) [2]. While the graphs and data interpretations
are accepted, people have challenged methodologies and how data is used and inter-
preted; however, in this case, the US Environmental Protection Agency (USEPA),
the United Nations (UN), and the Intergovernmental Panel Data Analysis reports
and technical books [3–6, 8–11, 16–17, 19–25] are widely accepted. Comparing
global warming potentials to carbon dioxide, greenhouse gases are better under-
stood when examining why carbon dioxide absorbs heat on a molecular level.

4.4 Heat Absorption by Carbon Dioxide

Carbon dioxide’s ability to absorb heat is characterized by the molecular structure,

the wavelength, and radiative properties. Visible light from the Sun is able to pass
the carbon dioxide molecules without its energy being absorbed since the frequency
of visible light does induce a dipole moment on the atmospheric CO2 molecules.
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 215

Carbon dioxide does however absorb “infrared” radiation (heat from the Earth’s
surface) and also re-emits that energy at the same wavelength as what was absorbed
(also as heat) [6]. As for its molecular structure, “Carbon dioxide doesn’t have a
molecular dipole in its ground state. However, some CO2 vibrations produce a struc-
ture with a molecular dipole. Because of this, CO2 strongly absorbs infrared radia-
tion” [7].
The energy of a molecule can change due to a change in the energy state of the
electrons of which it is composed. Thus, the molecule also has electronic energy.
The energy levels are quantized and take discrete values only. Absorption and emis-
sion of radiation take place when the atoms or molecules undergo transitions from
one energy state to another. In general, these transitions are governed by selection
rules. Atoms exhibit line spectra associated with electronic energy levels.
The dipole moment is determined by the magnitude of the charge difference and
the distance between the two centers of charge. If there is a match in frequency of
the radiation and the natural vibration of the molecule, absorption occurs and this
alters the amplitude of the molecular vibration. This also occurs when the rotation
of asymmetric molecules around their centers results in a dipole moment change,
which permits interaction with the radiation field. Dipole moment is a vector quan-
tity and depends on the orientation of the molecule and the photon electric vec-
tor [12].
In accordance with Kirchhoff’s laws, the following are noted:
1. Materials that are strong absorbers at a given wavelength are also strong emitters
at that wavelength; similarly weak absorbers are weak emitters.
2. Emission, reflection, and transmission account for all the incident radiation for
media in thermodynamic equilibrium [2, 6].

4.5 Rising Temperature Trend in the Environment

4.5.1 Atmosphere Temperature Increase

As a result of increased human activity, more greenhouse gases are warming the
Earth, resulting in an increased temperature trend around the world. The following
graphs suggest the increase in emissions has led to the increase in CO2 in the atmo-
sphere, thereby increasing the temperature of the Earth’s surface on land. Figure 4.4
shows the global per capita carbon emission estimates versus years. It appears that
the global per capita carbon emission increases significantly after the year 2000 [15].
216 J. O. Wong et al.

Fig. 4.4 Global per capita carbon emission estimates versus years [15]

4.5.2 Land and Ocean Temperature Increase

As land temperatures rise, we find ocean temperatures rise as well. “The globally
averaged combined land and ocean surface temperature data as calculated by a lin-
ear trend, show a warming of 0.85 [0.65–1.06] °C, over the period 1880–2012,
when multiple independently produced datasets exist. The total increase between
the average of the 1850–1900 period and the 2003–2012 period is 0.78 [0.72–0.85]
°C, based on the single longest dataset available. There are two methods: The first
calculates the difference using a best fit linear trend of all points between 1880 and
2012. The second calculates the difference between averages for the two periods
1850–1900 and 2003–2012 [4].
Figure 4.5 shows an increase of carbon dioxide in the atmosphere from 1958 to
2012 [4], while Fig. 4.6 shows the annual temperature anomalies from land ocean
in the period of 1880–2012 [16]. Based on the presented figures, an increase of
global carbon emissions shown in Fig. 4.4 leads to an increase of carbon dioxide in
the atmosphere shown in Fig. 4.5 and finally results in a temperature increase on
land and ocean shown in Fig. 4.6.
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 217

Fig. 4.5 Increase of carbon dioxide in the atmosphere [4]

4.5.3 Rising Temperatures of Land, Air, Sea, and Ice

Figure 4.7 summarizes the temperatures of land, air, and sea, with a dramatic
increase beginning in 1980. Similarly, 1920–1940 also experienced an upward
trend; however, from 1940 approaching 1980, the temperatures slightly decreased.
The reason the temperatures dropped is due to the industrial revolution’s emissions,
where manufacturers and factories sent a layer of soot in the atmosphere. The layer
of soot became a barrier and blocked solar radiation from hitting the Earth’s surface,
causing a cooling effect. However, when the Clean Air Act of 1970 was enforced,
the layer of soot moved out of the atmosphere and so began the true and actual
warming trend [2, 18, 19].
218 J. O. Wong et al.

Fig. 4.6 Land and ocean temperature increase: annual temperature anomalies from land and
ocean 1880–2012 [16]

4.6 Increased Temperatures on Land and Its Impacts

on Agriculture

As discussed previously, climate change results in increased temperatures in the

atmosphere, land, and sea. Weather patterns will become more extreme and forceful
where storms become cyclones such as Hurricanes Katrina, Irene, and Sandy in the
United States. Winters will reach record cooling temperatures, and summers will
reach record hotter temperatures. Dry land will become more dry, and droughts will
become so severe, agriculture and food shortages may eventually lead to famine.
Arid regions will be impacted where there once was water, will be no more water at
all, or will experience significantly lower water levels. Agriculture will become a
great challenge as soil becomes too dry or arid to harvest food [34].
Table 4.2 demonstrates the impacts of increased temperatures to our food supply.
Table 4.2 illustrates the types of impacts that could be experienced as the world
comes into equilibrium with more greenhouse gases. The top panel shows the range
of temperatures projected at stabilization levels between 400 ppm and 750 ppm CO2
at equilibrium. The solid horizontal lines indicate the 5–95% range based on climate
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 219

Fig. 4.7 Observed globally averaged combined land and ocean surface temperature anomaly
1895–2012

sensitivity estimates from the IPCC 20012 and a recent Hadley Centre ensemble
study. The vertical line indicates the mean of the 50th percentile point. The dashed
lines show the 5–95% range based on 11 recent studies. The bottom panel illustrates
the range of impacts expected at different levels of warming. The relationship
between global average temperature changes and regional climate changes is very
uncertain, especially with regard to changes in precipitation. This figure shows
potential changes based on current scientific literature [34].

4.7 Effect of Global Warming and Climate Change on Sea

Level Rise

As global average surface temperatures rise, and global average sea levels increase,
the snow cover and ice will decrease and melt. Figure 4.8 reports the changes of
temperature, sea level, and Northern Hemisphere snow cover. Specifically observed
changes in Fig. 4.8(a) show global average surface temperature versus years.
Figure 4.8(b) shows global average sea level from tide gauge (blue) and satellite
(red) data versus years. Figure 4.8(c) shows Northern Hemisphere snow cover for
220 J. O. Wong et al.

Table 4.2 Stabilization levels and probability ranges for temperature increases

March–April versus years. All differences are relative to corresponding averages for
the period 1961–2000. The smoothed curves represent decadal averaged values,
while the circles show yearly values. The shaded areas are the uncertainty intervals
estimated from a comprehensive analysis of known uncertainties shown in the
figures.
The impacts of sea level rise include increased flood risk, infrastructure invest-
ment implications around the world as seen in other countries like Italy, Netherlands,
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 221

Fig. 4.8 Changes in temperature, sea level, and Northern Hemisphere snow cover

and the South Pacific Island. Sea level rise will also contribute to increases in salin-
ity of rivers and estuaries, saltwater intrusion, as seen in the United States and all
around the world. Saltwater intrusion can affect the water supplies for drinking and
irrigation water and depletes the available freshwater habitat, as seen in the
United States.
The Asia-Pacific Region’s climate change adaptation involves the risk insurance
scheme as a social safety net through risk transfer and creates resilient societies. The
advantages of risk transfer are that it promotes risk mitigation compared to current
response-driven mechanisms, provides a cost-effective way to deal with expensive
impacts from the effects of climate change, “supports climate change adaptation by
covering the residual risks uncovered by other risk reduction mechanisms such as
222 J. O. Wong et al.

building regulations, landuse planning and disaster risk management plans,” stabi-
lizes incomes in rural areas and minimizes the gap in income fluctuation and socio-
economic development, provides partnerships between public and private sectors,
reduces government dependence after a disaster to reconstruct, helps people and
communities to recover and get back to everyday life quickly, and addresses various
risks from climatic and non-climatic origin, depending on how insurance is set up.
One of the greatest challenges to sea level rise impacts is increased salinity intru-
sion in rivers and estuaries, putting our water supply at risk.

4.8 Increased Salinity Intrusion in Rivers and Estuaries

According to the IPCC, studies have shown freshwaters to become more saline over
time and seawater has become fresher. This has affected rivers and estuaries, drink-
ing water supplies, irrigation, sea levels, and ecosystems.

4.8.1 Salinity Intrusion in Rivers and Estuaries

Saltwater intrusion (or salinity intrusion) is the movement of saline water into fresh-
water aquifers, which can become contaminated and undrinkable. Freshwater aqui-
fers can experience saline intrusion due to the hydraulic connection between
groundwater and seawater. Because saltwater is more mineral rich than freshwater,
it is denser and has higher water pressure. And so the heavier saltwater is able to
push inland beneath the freshwater.
Sources of saltwater intrusion include, but are not limited to, (a) activities like
groundwater pumping from coastal freshwater wells as seen in coastal areas; (b)
water extraction which drops the level of fresh groundwater, reducing its water pres-
sure and allowing saltwater to flow inland; and (c) water channels or agricultural
and drainage channels, carrying saltwater inland, and causing sea level rise.
Saltwater intrusion can also be worsened by extreme weather events like tropical
cyclones and hurricane storm surges. All over the world, rivers and estuaries experi-
ence salinity intrusion as a result of rising sea levels.
A case study to consider is California’s Sacramento-San Joaquin Delta. The
Sacramento-San Joaquin Delta is at the heart of most discussions about water in
California. The 1153 square mile of twist-and-turn islands and interconnected
waterways is located where the Sacramento and San Joaquin Rivers converge and
flow into San Francisco Bay through the Golden Gate Bridge. About 42% of the
state’s annual runoff flows through the Delta serving more than 23 million
Californians and irrigating millions of acres in the Central Valley. Two-thirds of
Californians get all or part of their drinking water from the Delta by government
water projects that export water to the San Francisco Bay Area and Central and
Southern California. The Delta is also the largest estuary on the West Coast with
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 223

hundreds of species of birds that travel along the Pacific Flyway and dozens of fish
species including salmon and steelhead that migrate through the Delta on their jour-
ney to and from the ocean. The Delta is strongly influenced by freshwater inflow
from tributary rivers, by tides in the SF Bay, and by salinity upstream. Since 1860,
the Delta waters have seen an increase in salinity [1, 2].
Plant pollen revealed the Delta was mainly a freshwater marsh for the past
2500 years; however, in the past 100 years, because of human activity, the Delta has
become more saline. Today, salinity intrusion is approximately 3–15 miles deeper
into the Delta than the early twentieth century. Between 1860 and 1920, human
activity modified the Delta when marshland was reclaimed, hydraulic mining
caused increased deposition and erosion sediment, and the expansion of the Delta
channel’s width, depth, and connections took place.
Before freshwater diversions increased in the 1940s, the Delta and Suisun Bay
would freshen every winter, even during extreme droughts as seen in the 1930s.
However, the Delta did not freshen during recent droughts (1976–1977, 1987–1994,
2007–2009, and 2014–2015), resulting in contaminants and toxins accumulating in
the system. The past 25 years have been relatively wet; the Delta’s autumn salinity
levels have shown to be in drought-like conditions due to human activity and water
diversions.
The historical record and published studies demonstrate that the Delta is far salt-
ier now due to human interference. Starting in 1917, local industries and residents
observed unprecedented salinity levels, causing a local sugar refinery to find a new
water supply, the Town of Antioch to file a lawsuit against upstream water users, and
the State of California to start a salinity monitoring program and investigation
(Fig. 4.9).
The colored portion on each chart represents the amount of freshwater available
within Suisun Bay downstream of the Delta boundary (approximately 18 miles
above Crockett). From 2001 to 2005, freshwater was seldom available below the
Delta boundary, indicating that the Delta did not “flush” as it used to. Without the
seasonal freshening of the Delta, contaminants and toxics can accumulate in the
system; and in this case, toxics were found to be a factor in the decline of the Delta
ecosystem. Note: While hydrological conditions were similar in the three time peri-
ods shown in the table, the sequence of wet and dry periods differs.

4.8.2 Water Quality and Water Supply Impacted by Climate

Change and Salinity Intrusion

Because of salinity intrusion, water quality impacts to the water supply are affected.
Many coastal cities in the United States have experienced saltwater intrusion
through water supply wells. Impacts of saltwater intrusion depend on how far the
intrusion extends, the plans for the water use, and how concentrations exceed the
standard of dissolved ions for its intended use.
224 J. O. Wong et al.

Fig. 4.9 Freshwater was available within Suisun Bay for a longer time period each year during the
early 1900s

For example, a coastal state such as Washington State and Southern California
reaches portions of the aquifer, affecting only certain water supply wells, whereas
Cape May, New Jersey, United States, experienced saltwater encroachment laterally
within each aquifer; it led to closing 20 or more public and industrial supply wells.
Not only in the United States, but salinity intrusion is a threat to water quality
and water supply all over the world. Since the autumn of 2003, a strong salinity
intrusion has caused a serious threat to the water supply in the Pearl River Delta, in
Macau and Zhuhai, cities in China. The salinity intrusion is caused by rapid indus-
trialization and urbanization, leading to greater water demand in the middle and
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 225

upper stream of the river basin. When the flow runs low, either by water demand or
during a dry season, salinity intrusion often occurs. Other sources of salinity intru-
sion include the rise of temperatures and sea levels, thereby affecting the water
supply and quality.
Also, in Bangladesh, drinking water from natural sources by the coast has
become contaminated by varying degrees of salinity due to saltwater intrusion from
rising sea levels, cyclone and storm surges, and upstream withdrawal of freshwater.
Not only in Asia, but also in Africa, the Ada peninsular in Ghana has suffered rapid
coastal erosion and inundation for over 50 years. As a result, it has led to loss of
property and livelihoods, economic stagnation, and saltwater intrusion. While prop-
erty loss and economic opportunities are addressed, the solutions do not benefit
increasing salinity intrusion in the Volta River. In fact, it aggravates the salinity
intrusion, increasing salt in the Volta Estuary. The government intervened to imple-
ment a sea defense project to keep seawater from intruding and is concerned about
the water supply and water quality due to saltwater intrusion.
Ada, situated at the mouth of the Volta Estuary, the Impact Assessment Report
writes there is a likely significant impact on the physicochemical water characteris-
tic of the estuary: “The most significant change will be an increased salinity in this
zone, whereby the salinity gradient will shift to the north… The intrusion of salt
water further into the estuary will probably lead to local changes in water quality. In
places where the fresh water is high in particulate organic matter content comes in
contact with salty coastal water, the organic matter starts to flocculate creating
depositions of dark material in which toxins and nutrients tend to accumulate. This
compromises the quality of water in the far southern part of the estuary, even more
so because it creates nutrient rich conditions in which many bacterial and viral
organisms, capable of causing diseases thrive. Hence, by opening up the estuary,
this zone could be drawn into the estuary, negatively affecting water quality.
However, the spatial extent of salinity shift is uncertain, and rated low to moderate
compared to some natural phases in the estuary dynamics where openings have
been created naturally that are far larger than the planned access channel.”
In the case of Australia’s dry weather challenges, droughts and extreme low
flows in the water systems reduce ecosystem capacity to absorb and process con-
taminated water. As sea levels rise, the estuaries in the Murray-Darling Basin expe-
rience an increase in saltwater intrusion, affecting major urban water supplies, as
well as freshwater ecosystem stability and productivity.
According to a case study of salinity of Israel’s Lake Kinneret, the conclusion
was made that increased salinity did not show obvious signs to effects on the lake
ecology; however, the study showed a reduced water quality; this brought attention
to implement changes in the Israeli water supply system.
All around the world, water quality impacts to water supply are affected by
increased salinity in drinking water, such as well water, aquifers, and other sources,
where the salt concentrations exceed the allowable. In the United States, a second-
ary maximum contaminant level is applied to 15 contaminants as a nonmandatory
measure.
226 J. O. Wong et al.

Water naturally accumulates a variety of dissolved solids, or salts, as it passes

through soils and rocks on its way to the sea. These salts typically include such
cations as sodium, calcium, magnesium, and potassium and anions such as chloride,
sulfate, and bicarbonate. A careful analysis of salinity would result in a list of the
concentrations of the primary cations and anions, but a simpler, more commonly
used measure of salinity is the concentration of total dissolved solids (TDS). As a
rough approximation, freshwater can be considered to be water with less than rough
approximation, freshwater can be considered to be water with less than 1500 mg/L
TDS, brackish waters may have TDS values up to 5000 mg/L, and saline waters are
those with concentrations above 5000 mg/L, whereas seawater contains
30,000–34,000 mg/L TDS.
In some parts of the country, salty water may be encountered. Since the saltwater
generally is overlain by freshwater, the lower part of the well in the saltwater zone
can be sealed off. But when this is done, the yield of the well is decreased.
Sometimes, waste saltwater resulting from the backwashing of a home ion
exchange water softener is discharged close to the well. Since saltwater is not fil-
tered out in seeping through the soil, it may find its way into the well. The best thing
to do is to discharge the wastewater as far as possible and downgrade from the well
or utilize a commercial water softener service. Saltwater is corrosive; it will damage
grass and plants and sterilize soil. Road salting or salt storage areas may also con-
tribute to well pollution.
Special desalting units (using distillation, deionization, and reverse osmosis) are
available for residential use, but they are of limited capacity and are relatively
expensive, and pretreatment of the water may be needed. Complete information,
including effectiveness with the water in question and annual cost, should be
obtained before purchase.
The US Environmental Protection Agency (USEPA) has established National
Primary Drinking Water Regulations (NPDWR) that set mandatory water quality
standards for drinking water contaminants. One enforceable standard is the mea-
surement of MCL (maximum contaminant levels), which were established to pro-
tect the public against consumption of drinking water contaminants that present a
risk to human health. An MCL is the maximum allowable amount of a contaminant
in drinking water, which is delivered to the consumer.

4.8.3 Agricultural Irrigation and Operations Impacted by

Climate Change and Salinity Intrusion

According to Hung and Forrider [37], the impacts of global warming and climate
change include degradation of natural resources, reduced agricultural production,
and human dislocation, and these impacts as a driver of future forced migration
depend on several factors:
1. Quantity of future GHG emissions
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 227

2. Rate of future population growth and distribution

3. Meteorological evolution of climate change
4. Effectiveness of local and national adaption strategies
Climate change has affected the food security either directly or indirectly caus-
ing stress on the production of food [39]. Climate change has also created pressures
on the hydrological cycle and impacts water availability, which strongly influences
agriculture. Effects on crop production are hard to predict as it depends on the fre-
quency or intensity of extreme weather events. Global aridity has increased since
the 1970s due to desertification. The areas under aridity have increased from 17 to
27% from the 1950s until now. This has notable effect on crop production and
decreasing crop yields. For example, maize yields diminish up to 1.7% under
drought conditions [39].
Higher CO2 levels can positively affect food crop growth. However, other factors
such as temperature, ozone, water availability, and nutrient constraints may counter-
act potential increases in yield. Many weeds, pests, and fungi thrive under warmer
temperatures and wetter climates. Currently, farms spend more than $11 billion per
year to fight weeds which compete with crops.
Drought may threaten livestock yield. In 2011, exposure to temperature events
caused over $1 billion in heat-related losses. Heat stress affects animals directly and
indirectly, making them vulnerable to disease and infertility. The prevalence of par-
asites increases, and the productivity of pastures is affected. The quality of the for-
age found in pastureland decreases with higher GHG; as a result, the cattle would
need to eat more to get the same nutritional value. This can lead to overgrazing and
misuse of land management [40].
Fisheries are also impacted [30]. American anglers catch or harvest five million
metric tons of seafood each year and contribute $1.5 billion to the US economy
annually. Aquatic species migrate to colder waters, and shell-building animals
decrease in number. Fishermen experience decreases in harvest, which increase the
price and availability of seafood. American fisheries are analogous to the global
fishing economy.
Salt affects plant growth in three ways: (a) osmotic effects, caused by the total
dissolved salt concentration in the soil water; (b) specific ion toxicity, caused by the
concentration of individual ions; and (c) soil particle dispersion, caused by high
sodium and low salinity. With increasing soil salinity in the root zone, plants expend
more of their available energy on adjusting the salt concentration within the tissue
(osmotic adjustment) to obtain needed water from the soil. The consequence is less
energy is available for plant growth.
In irrigated areas, salts originate from the local groundwater or from salts in the
applied water. Salts tend to concentrate in the root zone due to evapotranspiration,
and plant damage is tied closely to an increase in soil salinity. Establishing a net
downward flux of water and salt through the root zone is the only practical way to
manage a salinity problem. Under such conditions, good drainage is essential to
allow a continuous movement of water and salt below the root zone. Long-term use
228 J. O. Wong et al.

of reclaimed water for irrigation in which only the conventional constituents have
been removed is not possible without adequate drainage.
Specific ion toxicity is another factor to be studied. If the decline of crop growth
is due to excessive concentrations of specific ions, rather than osmotic effects alone,
it is referred to as “specific ion toxicity.” The ions of most concern in wastewater are
sodium, chloride, and boron. The most prevalent toxicity from the use of reclaimed
water is boron. The source of boron is usually household detergents or discharges
from industrial plants. The quantities of chloride and sodium also increase as a
result of domestic usage, especially where water softeners are used.
For sensitive crops, specific ion toxicity is difficult to correct without changing
the crop or the water source. The problem is also accentuated by hot and dry cli-
matic conditions due to high evapotranspiration rates. Regulations for maximum
trace element concentrations for irrigation water are reported. In severe cases, these
elements tend to accumulate in plants and soils, which could result in human and
animal health hazards or cause phytotoxicity in plants.
The concentration of dissolved solids is an important indicator of the usefulness
of water for various applications. Drinking water, for example, has a recommended
maximum total dissolved solids (TDS) concentration of 500 mg/L. Many people
will begin to notice water tastes salty at about 1000 mg/L of TDS, although this is
very dependent on the particular dissolved constituents. Livestock can tolerate
higher concentrations. Upper limits for stock water concentrations quoted by the US
Geological Survey (1985) include poultry at 2860 mg/L, pigs at 4290 mg/L, and
beef cattle at 10,100 mg/L. Of greater importance, however, is the salt tolerance of
crops. As the concentration of salts in irrigation water increases above 500 mg/L,
the need for careful water management to maintain crop yields becomes increas-
ingly important. With sufficient drainage to keep salts from accumulating in the soil,
up to 1500 mg/L TDS can be tolerated by most crops with little loss of yield, but at
concentrations above 2100 mg/L, water is generally unsuitable for irrigation except
for the most salt-tolerant crops. All naturally occurring water has some amount of
salt in it. In addition, many industries discharge high concentrations of salts, and
urban runoff may contain large amounts in areas where salt is used to keep ice from
forming on roads in the winter. Although such human activities may increase salin-
ity by adding salts to a given volume of water, it is more often the opposite process,
the removal of freshwater by evaporation, that causes salinity problems. When
water evaporates, the salts are left behind, and since there is less remaining freshwa-
ter to dilute them, their concentration increases.
Irrigated agriculture, especially in arid areas, is always vulnerable to an accumu-
lation of salts due to this evapotranspiration on the cropland itself. The salinity is
enhanced by the increased evaporation in reservoirs that typically accompany irri-
gation projects. In addition, irrigation drainage water may pick up additional salt as
it passes over and through soils. As a result, irrigation drainage water is always
higher in salinity than the supply water and, with every reuse, its salt concentration
increases even more. In rivers that are heavily used for irrigation, the salt concentra-
tion progressively increases downstream as the volume of water available to dilute
salts decreases due to evaporation and as the salt load increases due to salty drainage
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 229

water returning from irrigated lands. It has been estimated that roughly one-third of
the irrigated lands in the western part of the United States have a salinity problem
that is increasing with time, including regions in the Lower Colorado River Basin
and the west side of the San Joaquin Valley in California, United States. An esti-
mated 100,000 tons of salt are imported annually into Southern Arizona through its
consumption of Colorado River water via the 300 mile long, Central Arizona Project
canal. Salinity problems are also having major impacts on irrigated lands in Iraq,
Pakistan, India, Mexico, Argentina, Mali, and North Africa, among others. The col-
lapse of ancient civilizations, once known as the Fertile Crescent, and is now Iraq,
is thought to have formed by accumulating salt from irrigated agriculture. Agriculture
that depends on irrigation from affected rivers would be directly impacted by sea
level rise. Crops do not grow as well with salty water, as seen in the resulting smaller
leaves, shorter stature, and sometimes fewer leaves. The severity of salinity on crops
is based on the environment’s humidity, temperature, radiation, and air pollution.
Some of the agricultural production could be shifted to salt-tolerant crops. Irrigation
with salty water tends to accumulate salt in the soil, decreasing soil productivity.
Note that this is not the only way that climate change can disrupt food production.

4.8.4 Food Production Impacted by Climate Change

and Salinity Intrusion

Drought and other climate extremes have a direct impact on food crop, food supply,
and economics. During a dry spell, there will be excessive water loss from the
plants; thus, the process of photosynthesis is greatly reduced and it is difficult for
the plants to survive [36]. On the contrary, during a flooding event, plants will be
inundated and damaged due to depleted oxygen (approaches zero after 24 h flood-
ing event) and nitrogen levels in the flooded soils. In addition, the affected plant’s
stomata will be closed for a period of time which will subsequently reduce the res-
piration, transpiration. Loss in crop yield may lead to economic collapse (as the
price of staple crops could rapidly escalate causing major inflation) and food short-
age, where hunger will be the biggest battle and create conflicts in some countries.
Coming out of the last ice age, the climate change was maintained and steady
and the human population was small and nomadic, whereas now, large communities
of an increasing population live away from agriculture that naturally gets rain, but
instead relies on irrigation. Demand for water is greater than the renewable supply
of freshwater for a community’s demand and supply needs. Irrigation demand is of
poor water quality containing dissolved salts that collect in the soil. Irrigation and
removing native perennial vegetation have led to rising water tables—some rise into
the root zone and soak the land. Over 50% of groundwater is saline, especially in
dry and semiarid regions, so as water tables rise, the saltwater gets brought into root
zone areas. How sustainable are irrigated systems as we work through issues of
water resource availability and allocation? The changes in climate and population
230 J. O. Wong et al.

are projected to increase, and so irrigation and water supply would follow. Unless
dramatic changes are made, continued increasing salinity will be found in
agriculture.

4.8.5 Ecosystem Impacts Due to Loss of Freshwater Habitat

(Recreation, Fishing)

Salt accumulation in soils is often controlled by flushing the salts away with addi-
tional amounts of irrigation water. This increases costs; wastes water, which may
not be abundantly available in the first place; and unless adequate drainage is avail-
able, increases the likelihood that a rising water table will drown plant roots in salt-­
laden water. Providing adequate drainage can be an expensive and challenging task
involving extensive on-farm subsurface drainage systems coupled with a central
drain and disposal system. Even when salt is removed from the agricultural land by
good drainage, it can have later repercussions. In the mid-1980s, it was found that
birdlife in the natural freshwater marshes of the Kesterson National Wildlife Refuge
in Central California was being poisoned by selenium draining from the region’s
agricultural fields. Since irrigation return water contains not only salts but also fer-
tilizers and pesticides, finding an acceptable method of disposal is difficult. These
issues with salts highlight how important it is to not only deal with the immediate
impacts of pollution but to develop the remedies so that further downstream impacts
are not created.

4.9 Impacts of Solid Waste Landfill Gas on Sanitary Landfill

Utility, Ecosystem, and Human

4.9.1 Impacts on Sanitary Landfill Operations

and Surrounding Environment

Shammas, Wang, Wang, and Chen [35] have discussed the ecological impact of
sanitary landfill gas (LFG) on the landfill utility’s operation as well as LFG collec-
tion, control, and utilization. The result of a 214-year study of the time phase evolu-
tion of various gases in a landfill has shown the following [38]:
1. Hydrogen, oxygen, nitrogen, carbon dioxide, and methane constitute the
major gases.
2. Hydrogen in great quantities appears during the first 3 weeks (20% during the
first and second weeks).
3. Hydrogen sulfide appears in a trace form during the first 2 years.
4. Carbon dioxide reaches 35% after 2 weeks (40% after 2 months).
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 231

5. Methane reaches 2.7% after 2 months, 6% in 6 months, 13% after 1 year, and
20% after 2 years.
6. Composition of gas is dependent on compaction densities. Higher compaction
densities yield more gas per unit volume.
7. The most pronounced changes in the organic materials occur within the first
2 months.
8. A landfill is still far from being stabilized at the end of 2 years.
9. Dry refuse and saturated refuse produce 0.0022 and 0.0131 m3 gas/kg refuse,
respectively, on a dry basis.
10. Carbon dioxide increases the hardness and level of bicarbonates in groundwa-
ter. Depending on the pH, the water may become acidic and corrosive.
Aziz, Rosli, and Hung [36] have reported that methane is a shorter lifetime potent
gas (9–15 years) with a high global warming potential due to a strong molar absorp-
tion coefficient. As the concentrations in the atmosphere increased due to uncon-
trolled anthropogenic methane production, it has become more long-lived and
causes damages by creating an imbalance between methane emissions and remov-
als. Their publication [36] discusses about methane generation in landfills (anaero-
bic decomposition process, source of methane in landfills, and methane reduction),
methane emissions (mechanisms and factors influencing the mechanisms), methane
in the atmosphere (methane sink and removal), and the impact of landfill methane
emissions.
The migration and emission of LFG may potentially lead to negative effects in
the surroundings, for example, fire and explosion hazards, health risks, damage to
vegetation, groundwater contamination, and global climate effects. The main envi-
ronmental hazards related to methane emissions are believed to be explosion haz-
ards and global climate effects.
The potential for methane gas to explode is determined by its lower explosive
limit (LEL) and upper explosive limit (UEL), which lies between 5 and 15% in air
at ambient temperature and atmospheric pressure. Even though explosion will not
occur if the concentration is above the UEL, methane concentrations equal to or
greater than LEL will be considered hazardous as it exceeds the LEL. Thus, it is
essential to monitor and keep the methane concentration below the LEL.

4.9.2 Impacts on Human Health

Extreme climate affects human health with exposure to both extreme hot and cold
weather being associated with cardiovascular disease (CVD) and mortality [36].
The Europe episode in summer 2013 is one example of mortality effect, when the
temperature increased to 3.5 °C above normal and caused 22,000–45,000 heat-­
related deaths within 2 weeks in August 2003. In addition, changes in the rainfall
pattern in many areas affect the distribution of infectious diseases/vector-borne dis-
eases (malaria, dengue, plague, elephantiasis, and bluetongue disease) due to the
232 J. O. Wong et al.

nature of the infectious agents (bacteria, virus, and protozoa) and their vector organ-
isms (mosquitoes, snails, and other insects) that are temperature dependent, with a
warm environment boosting their rate of reproduction. This was seen during an El
Niño episode in Peru (1997–1998) when the ambient temperature increased more
than 5 °C above normal and caused the number of daily admissions for diarrhea to
increase by twofold from the previous rate.
At a low concentration in the air, methane and carbon dioxide do not affect the
health. Nevertheless, high concentrations of methane and carbon dioxide in the
atmosphere will contribute to adverse health effects, not by breathing the gases
itself but through the displacement of oxygen, which can reduce the concentration
of oxygen (below 16%) in the air [41]. As a result, there is a risk of asphyxiation,
which can lead to dizziness, fatigue, vomiting, headache, visual disturbance, faster
heartbeat, asthma, reduced lung function, unconsciousness, and even death if the
condition is prolonged [42].

4.9.3 Impacts on Vegetation

Methane does not have a direct toxicity effect on the plant or vegetation growth.
Nevertheless, a high methane concentration in ambient air will result in a lack of
oxygen in the root zone, and the displacement of oxygen by methane can cause
anaerobic soil conditions which are detrimental to plants [35].

4.10 Natural Variability

Is it possible that the natural environment could make a small contributing factor
toward climate change? When assessing climate change, one must also consider the
impacts of the natural environment. Natural contributions to climate can happen
through either internal impacts or external forces. Internal impacts are those factors
that occur directly within a climate system. These can occur within the atmosphere,
through entities within the climate system, or among phenomena that drive climate
variations on Earth. The effects of internal impacts can happen almost immediately
or incrementally over a long period. On the other hand, external forces are factors
outside of the climate system that can result in changes in the climate. For example,
ash and sulfuric aerosols from a volcanic eruption may cause temporary changes
both locally and thousands of miles away from the eruption. The consequence of
these natural particulate emissions is that these emissions create a layer of particu-
lates that keep sunlight from penetrating the atmosphere. As a result, there is an
expectation that temperatures will be cooler for a period before recovering back to
levels experienced prior to the event [31–32].
The change to climate due to nature can happen over a short or long period of
time. Changes that happen over thousands of years are known as millennial climate
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 233

cycles. These cycles can happen every 10,000–100,000 years and can cause signifi-
cant periods of warming and cooling. The cause of these changes can be attributed
to Milankovitch cycles, or changes in the Earth’s orbit around the Sun. While these
changes do not directly cause warming and cooling, they can provide a mechanism
for these phenomena to take place. For example, a change in solar reflectivity can
increase ice melt. Periods where warming and cooling can occur between 250 and
1000 years are known as century-scale climate changes. The shorter periods between
events could be attributed to the Sun or ocean circulation patterns. Finally, there are
periods where climate can change in as short of a time as year to every 10 years.
Most of the time, this is caused by interactions between the ocean and the atmo-
sphere. The most common example is the El Niño-Southern Oscillation (ENSO).
This can bring warmer weather to some areas and an increase in precipitation
in others.
It is important to quantify if climate change is directly related to the climate sys-
tem or some external factors. The primary way to determine this is by having an
understanding of the physical attributes of the climate. Data based on climatic
observations is also beneficial. Models can also be considered because they provide
a way to simulate the unpredictable effects of varying phenomena over time.
Nevertheless, models must be compared with climatic observations and the known
physical attributes of a climate to ensure that the models are put into proper context.
This is done by placing proper boundaries around scenarios and outcomes that
would not otherwise fit within what has been observed historically through data or
by prior knowledge of the climate. This would in turn minimize potential biases that
could occur in situations where data was unavailable [32].
So what have the models reported? The following is a summary of key points as
described from the IPCC [31–32]:
1. With an exception of a few locals, the models show both model and observed
data agree that warming occurs around the world. During the first half of the
century, warming was due to a combination of anthropogenic and natural events
(volcanic, solar, and internal). Anthropogenic sources caused warming during
the second half of the century. The anthropogenic forcing appears to be primar-
ily greenhouse gases (GHGs).
2. An increase in Northern Hemisphere (NH) temperatures has occurred within
the last 50 years regardless of methods of reconstruction or external factors
employed.
3. It is impossible to see a significant increase in NH temperatures without human
influence, but natural variability would play some role since the warming is not
consistent.
4. Greenhouse gas, volcanic eruptions, and solar irradiance have played some role
in temperature change over the past 1000 years. Volcanic activity during 1675
and 1715 might have led to cooling during this time.
5. Models found that data that only considered natural variability did not match
global mean surface temperature data. This data appeared to match better to
what is seen in simulations comparing with what was observed.
234 J. O. Wong et al.

6. There is uncertainty surrounding the effects from the Sun and volcanoes. This
is primarily because of the changes to methods in modeling (e.g., number of
sample sizes, scaling factors to account for unknown factors, and internal vari-
ability). Therefore, one must consider the assumptions and factors each author
makes within a model.
7. Regional climate change may be hard to predict due to internal impacts that are
unique to a particular area. These impacts will become more important at the
regional level as opposed to considering the larger area. This is also true on
shorter time scales of less than 50 years.
8. There has been documented evidence of a change in tropospheric height, ozone-­
induced stratospheric cooling, and tropospheric warming by GHGs. It appears
that natural causes alone are simply unable to explain these changes.
9. Oceans have gained 14.2 × 1022 J of energy from 1961 to 2003. The reason
behind such a gain may be attributed to GHGs and sulfur aerosols. Volcanic
eruptions can explain some cooling events within an ocean.
10. The sea level rise might be explained by anthropogenic reasons, specifically
upper ocean and glacier loss. There have been small changes to sea level pres-
sure changes due to ozone depletion. Anthropogenic impacts have affected
Asian monsoon circulation (black carbon aerosols), an increase of tropical
cyclones, atmospheric water vapor, and saturated vapor pressure. The combina-
tion of both anthropogenic and natural causes has also contributed to an increase
in land mass mean precipitation.
11. Greenhouse gases may have also caused changes in precipitation values and
glacial retreat. Warming may have altered the movement of water vapor from
the tropics to high-altitude regions may have led to changes in the precipita-
tion values.

4.11 Applications to Take Action

To avoid the consequences, solutions begin with us. We can start by [2]:
1. Carpooling.
2. Get a vehicle with better gas mileage
3. Use compact florescent lights
4. Make your home more energy efficient by replacing appliances
5. Turn off your power strip when you are done; it conserves 25%
6. Be a better consumer by buying recycled things, and recycle simple things like
the disposable coffee sleeve from your coffee shop
7. Get off junk mail
8. Stop buying bottled water and use a water filter
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 235

4.12 Summary

What is certain? It is certain that:

1. The study of climate change begun as early as the 1820s with scientists such as
Fourier, Tyndall, and Arrhenius. Therefore, this field of science is older than the
first trans-Atlantic flight and was before the invention of the atomic bomb!
2. Contemporary recognition of climate change began in the 1950s and continued
to become a staple within the scientific community during the environmental
movement of the 1960s and 1970s.
3. The increasing amount of human activity is changing the composition of the
atmosphere with overwhelming supporting data.
4. Carbon dioxide, methane, and nitrous oxide are increasing dramatically because
of human activities.
5. Greenhouse gases absorb heat and emit heat; since they get trapped in the atmo-
sphere, the heat gets trapped in the atmosphere and warms the Earth.
6. Human activity produces greenhouse gases that remain in the atmosphere
for years.
7. It is estimated that the average global temperatures have risen between 1 and 4 °F.
8. During the first half of the twentieth century, increases in temperature have been
due to natural causes. During the latter half, temperature increases have been due
to anthropogenic activities.
9. Greenhouse gases, global warming, and climate change have negative impacts
on agricultural irrigation, agricultural operations, food production, water utility,
and sanitary landfill utility.
What is uncertain? It is uncertain that:
1. Forecasting exact impacts to health, agriculture, water resources, forests, wild-
life, and coastal areas in regional basis is difficult.
2. There is also uncertainty in quantifying the exact magnitude and extent of
adverse effects, projecting the magnitude of sea level rise, and quantifying the
indirect effects of aerosol particles to the Earth’s energy balance (i.e., cloud for-
mation and its radiative properties, precipitation efficiencies).
3. The negative impacts of greenhouse gases, global warming, and climate change
on agricultural irrigation, agricultural operations, food production, water utility,
and sanitary landfill utility cannot be quantified at present.
Adaptation/mitigation for the effects of climate change is necessary because evi-
dence shows it is too late for complete prevention. The responsible thing to do is to
start preparing now.
236 J. O. Wong et al.

Glossary

Agricultural irrigation It is a large-scale agricultural process of applying con-

trolled amounts of water to land to assist in the production of crops, as well as to
grow landscape plants. Small-scale irrigation applied to lawn is called watering.
There are different types of irrigation, such as sprinkler irrigation, surface irriga-
tion, drip irrigation, subirrigation, and manual irrigation.
Climate (a) Climate in a narrow sense is usually defined as the “average weather”
or, more rigorously, as the statistical description in terms of the mean and vari-
ability of relevant quantities over a period of time ranging from months to thou-
sands of years. The classical period is three decades, as defined by the World
Meteorological Organization (WMO). These quantities are most often surface
variables such as temperature, precipitation, and wind. Climate in a wider sense
is the state, including a statistical description, of the climate system. (b) The
average weather (usually taken over a 30-year time period) for a particular region
and time period. Climate is not the same as weather, but rather, it is the average
pattern of weather for a particular region. Weather describes the short-term state
of the atmosphere. Climatic elements include precipitation; temperature; humid-
ity; sunshine; wind velocity; phenomena such as fog, frost, and hailstorms; and
other measures of the weather.
Climate change (1) Changes in average weather conditions that persist over multi-
ple decades or longer. Climate change encompasses both increases and decreases
in temperature, as well as shifts in precipitation, changing risk of certain types
of severe weather events, and changes to other features of the climate system.
(2) Climate change refers to any significant change in the measures of climate
lasting for an extended period of time. In other words, climate change includes
major changes in temperature, precipitation, or wind patterns, among others, that
occur over several decades or longer.
Enhanced greenhouse effect The concept that the natural greenhouse effect has
been enhanced by increased atmospheric concentrations of greenhouse gases
(such as CO2 and methane) emitted as a result of human activities. These added
greenhouse gases cause the Earth to warm.
Environment The complex of physical, chemical, and biotic factors (as climate,
soil, and living things) that act upon an organism (a living thing) or an ecological
community (a collection of living things) and ultimately determine its form and
survival. The circumstances, objects, and conditions that surround each of us.
Public utility (a) A public utility is an organization that maintains the infrastruc-
ture for a public service and, therefore, is subject to forms of public control and
regulation ranging from local community-based groups to statewide government
monopolies. (b) It is an organization supplying the community with electricity,
gas, water, solid waste disposal service, or sewerage management service.
Rain-fed agriculture It is an agriculture that does not use irrigation but instead
relies only on direct rainfall.
4 The Impacts of Climate Change on Agricultural, Food, and Public Utility Industries 237

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Chapter 5
Innovative PACT Activated Sludge,
CAPTOR Activated Sludge, Activated
Bio-­Filter, Vertical Loop Reactor,
and PhoStrip Processes

Lawrence K. Wang, Mu-Hao Sung Wang, and Nazih K. Shammas

Acronyms

ABF Activated bio-filters

BOD Biochemical oxygen demand
CAST CAPTOR in activated sludge treatment
CBOD Carbonaceous biochemical oxygen demand
COD Chemical oxygen demand
DAF Dissolved air flotation
F/M ratio Food-to-microorganism ratio
HRT Hydraulic retention time, d
MF Membrane filters
MG Million gallons
MGD Million gallons per day
MLSS Mixed liquor suspended solids
NH3-N Ammonia nitrogen
NO2-N Nitrite nitrogen
NO3-N Nitrate nitrogen
NSFC National Small Flows Clearinghouse
PAC Powdered activated carbon
PACE Effluent PAC concentration, mg/L
PACI Influent PAC concentration, mg/L
PACR Mixed liquor PAC concentration in the reactor, mg/L

L. K. Wang (*)
Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
e-mail: [email protected]
M.-H. S. Wang · N. K. Shammas
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 241

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_5
242 L. K. Wang et al.

PACT Powdered Activated Carbon Treatment

SRT Design solids retention time, d
TKN Total Kjeldahl nitrogen
TSS Total suspended solids
UMIST University of Manchester Institute of Science and Technology
UNIDO United Nations Industrial Development Organization
USACE US Army Corps of Engineers
USEPA US Environmental Protection Agency
VLR Vertical Loop Reactor
WRC British Water Research Centre

5.1 Powdered Activated Carbon Treatment (PACT)

5.1.1 Types of PACT Systems

The powdered activated carbon (PAC) activated sludge system is a process modifi-
cation of the activated sludge process. PAC is added to the aeration tank where it is
mixed with the biological solids (Fig. 5.1). The mixed liquor solids are settled and
separated from the treated effluent. In a gravity clarifier, polyelectrolyte will nor-
mally be added prior to the clarification step to enhance solids-liquid separation. If
phosphorus removal is necessary, alum is often added at this point also. Even with
polyelectrolyte addition, tertiary filtration is normally required to reduce the level of
effluent suspended solids. The clarifier underflow solids are continuously returned
to the aeration tank. A portion of the carbon-biomass mixture is wasted periodically
to maintain the desired solids inventory in the system.
There are six types of combined biological and physicochemical PAC process
systems [1–7]:

Fig. 5.1 Powdered activated carbon activated sludge process (PACT) [10, 14]
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 243

(a) Continuous combined biological and physicochemical PAC process systems

involving the use of sedimentation clarifiers
(b) Combined biological and physicochemical PAC sequencing batch reactor sys-
tems involving the use of sedimentation clarifiers
(c) Continuous combined biological and physicochemical PAC process systems
involving the use of dissolved air flotation (DAF) clarifiers
(d) Combined biological and physicochemical PAC sequencing batch reactor sys-
tems involving the use of DAF clarifiers
(e) Continuous combined biological and physicochemical PAC process systems
involving the use of membrane filters (MF)
(f) Combined biological and physicochemical PAC sequencing batch reactor
involving the use of membrane filters (MF)
When PAC is dosed into an activated sludge process for combined adsorption and
biochemical reactions, the combined process is also called the PACT process, in
which PAC still stands for powdered activated carbon, while ACT stands for acti-
vated sludge.

5.1.2 Applications and Performance

The addition of PAC to plug flow and complete mix suspended growth reactors is a
more common process modification for industrial wastewater treatment than for
municipal systems. Demonstrated advantages of PAC addition to suspended growth
reactors include [8]:
(a) Improved solids settling and dewatering characteristics
(b) The ability of PAC to adsorb biorefractory materials and inhibitory compounds
(c) Improving effluent quality and reducing the impact of organic shock loads
(d) Reduction in odor, foaming, and sludge bulking
(e) Improved color and 5-day BOD removal
Because PAC is wasted with excess biomass, virgin or regenerated PAC addition is
required to maintain the desired concentration in the biological reactor. This can
represent a significant cost factor for the system. When carbon addition require-
ments exceed 900–1800 kg/day (2400–4000 lb/day), wet air oxidation/regeneration
(WAR) is claimed to represent an economical approach to carbon recovery and
waste biomass destruction [9]. However, an ash separation step is needed in this
case, affecting the economics of carbon regeneration and recovery [10]. The eco-
nomic analysis is further clouded by the inability to analytically differentiate pow-
dered carbon from background refractory volatile materials, thus making it difficult
to quantify the value of the volatile suspended material recovered after WAR. Although
ash separation processes have been reported to be effective in at least two municipal
PAC activated sludge plants, the economics of complete PAC/WAR systems relative
to other activated sludge nitrification systems are unclear [7, 10, 11].
In the United States, PACT systems for nitrification generally have been applied
at municipal treatment plants where industrial sources contribute a significant
244 L. K. Wang et al.

fraction of the incoming wastewater. In all instances, PAC regeneration was included
in the flowsheet [12]. A summary of selected municipal PACT facilities is presented
in Table 5.1.
The procedure to follow in designing PACT systems for nitrification involves a
modification to those for complete mix or conventional plug flow systems in order
to account for the effects of the addition of PAC [13]. According to the major sup-
plier of the technology [12, 14], most PAC process systems are designed at MLSS
concentrations of approximately 15 g/L. The mixed liquor is composed of volatile
activated carbon, biomass, nonvolatile PAC ash, biomass decay components, and
influent inert material. The relative proportions of these materials are strongly influ-
enced by whether carbon regeneration via wet air oxidation and a return of this
material to the aerator is practiced. The intent is to maintain the PAC concentration
at approximately 1.5 times the biomass level in nitrification PAC reactors [12, 14].
The most appropriate PAC concentration will be dictated by the specific wastewater
characteristics and often cannot be specified without bench- or pilot-scale studies.
The PAC concentration to be added will depend on the design solids retention time,
the hydraulic retention time, and the required PAC concentration in the reactor.
According to the US Environmental Protection Agency [14], for practical engineer-
ing design considering the loss, the PAC concentration to be added can be calculated
from Eq. (5.1):

Table 5.1 Summary of PACT process systems using wet air oxidation for APC regeneration
[10, 14]
Permit limits
NH 4  N
Current/design PAC/WARa Reason for BOD5, TSS, ,
Facility flow, m3/s status PACa mg/L mg/L mg/L
Vemon, CT 0.18/0.28 MA C 10 20 –
Mt. Holly, NJ 0.11/0.22 MA C,S 30 30 20
E. Burlington, 0.31/0.53 MA C,N,T 12–24 30 4.0–8.0
NC
S. Burlington, 0.30/0.42 AS C,N,T 12–24 30 4.0–8.0
NC
Kalamazoo, MI 1.1/2.4 MA C,N,T 7–30 20–30 2.0–10.0
Bedford Hts., 0.15/0.15 NAC N,S 10 12 5.1
OH
Medina Co., 0.31/0.44 MA N 10 12 1.5–8.0
OH
N. Olmsted,b 0.26/0.31 AS N,S 30 30 2.3–6.9
OH
Sauget, IL 0.70/1.2 AS T 20 25 –
El Paso, TX 0.20/0.44 MA N,O SDd SD SD
a
C = Color Removal; S = Space; N = Nitrification; T = Toxics; O = Organics
b
Plan to convert to NAC without regeneration
c
MA = Modified operation and/or design for ash control. AS = Converted to conventional activated
sludge. NAC = Converted to the use of nonactivated carbon without regeneration
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 245

PACI  PACE   PACR  HRT / SRT

(5.1)
where
PACI is the influent PAC concentration, mg/L
PACR is the mixed liquor PAC concentration in the reactor, mg/L
PACE is the effluent PAC concentration, mg/L
HRT is the hydraulic retention time, day
SRT is the design solids retention time, day
The value of PACE in Eq. (5.1) can be estimated by assuming that the carbon
fraction in the effluent TSS (total suspended solids) is the same as the fraction of
PAC in the MLSS (mixed liquor suspended solids).
PACT nitrification systems are normally selected when the municipal wastewater
contains compounds originating from industrial operations, as stated previously.
Nitrifiers are susceptible to a number of organic and inorganic inhibitors found in
many industrial wastewaters [14]. Researchers have provided evidence that the
addition of PAC to nitrifying activated sludge systems receiving industrial wastewa-
ters improved nitrification rates [14–16]. More recent studies have been completed
with the goal of determining the mechanism of nitrification enhancement in PAC
activated sludge systems in the presence of adsorbable and nonadsorbable inhibitors
[17]. The results indicated that the addition of the proper amount of PAC can com-
pletely nullify the toxic effects of an adsorbable nitrification inhibitor. A minor posi-
tive effect on nitrification rates was observed when PAC was added to a nitrifying
activated sludge system receiving nonadsorbable inhibitors. The activated sludge
used in these studies was not acclimated to the inhibiting compounds. Another pos-
sible contributing factor to the enhancement of nitrification could be attributed to
the fact that the addition of PAC provides particulate matter for attachment of the
nitrifying microorganisms, thereby promoting nitrification [18].

5.1.3 Process Equipment

PAC can be fed in the dry state using volumetric or gravimetric feeders or can be fed
in slurry form. There are more than 3 major PAC producers, over 50 manufacturers
of volumetric and gravimetric feeders, and over 50 manufacturers of slurry feeders
[19–21]. There are also many manufacturers of sequencing batch reactors (SBR)
[2], dissolved air flotation (DAF) clarifiers [7], and membrane filtration (MF) reac-
tors [6].
246 L. K. Wang et al.

5.1.4 Process Limitations

The process limitations of PACT process systems are identical to that of the PAC
physicochemical process. The PACT process will increase the amount of generated
sludge. Regeneration will be necessary at higher dosages in order to maintain rea-
sonable costs. Most systems will require post-filtration to capture any residual car-
bon particles. Some sort of flocculating agent such as an organic polyelectrolyte is
usually required to maintain efficient solids capture in the clarifier.
About 1 pound of dry sludge will be generated per pound of carbon added. If
regeneration is practiced, carbon sludge is reactivated and reused with only a small
portion removed to prevent the buildup of inert material. PAC physicochemical pro-
cess systems are reasonably reliable. In fact, PAC systems can be used to improve
process reliability of existing systems.
Additional information on carbon adsorption and combined biological and phys-
icochemical PACT process systems can be found in Refs. [22–31].

5.2 Carrier-Activated Sludge Processes (CAPTOR

and CAST Systems)

There has been a substantial interest in recent years in the potential benefits of high
biomass wastewater treatment. The major obstacle for achieving this has been the
inability of biosolids separation in secondary clarifiers. For the most part, this has
been overcome by using various forms of support media or carriers that have the
ability to attach high concentrations of aerobic bacterial growth [32–34]. The
increase in immobilized biomass reduces the process dependence on secondary set-
tling basins for clarification. In such hybrid systems where attached growth coexists
with suspended growth, one gets more stable systems which possess the combined
advantages of both fixed and suspended growth reactors.

5.2.1 Advantages of Biomass Carrier Systems

The performance of carrier systems is dependent on the amount of attached bio-

mass, the characteristics of attached and suspended microorganisms, and the type of
carriers. The advantages of such hybrid systems are:
(a) Heterogeneity of the microbial population. This is brought about by the differ-
ences in the microhabitat of organisms attached to the surface of a carrier and
those in the bulk of the solution with respect to pH, ionic strength, and concen-
tration of organics [35–39].
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 247

(b) Increased persistence in reactor. This leads to an increase in biomass of organ-

isms, reduction of hydraulic retention time, and thus smaller reactor volumes
[40–42].
(c) Higher growth rate [43–45].
(d) Increased metabolic activity. This leads to an increase in respiration and sub-
strate utilization, hence higher removal rates [46–49].
(e) Better resistance to toxicity [50–53].

5.2.2 The CAPTOR Process

One interesting concept of hybrid systems is the CAPTOR process developed

jointly by the University of Manchester Institute of Science and Technology
(UMIST) and Simon-Hartley, Ltd., in the United Kingdom. This high biomass
approach uses small reticulated polyurethane pads as the bacterial growth medium
[54]. The pads are added to standard activated sludge aeration reactor, and the sys-
tem is operated without sludge recycle, essentially combining suspended growth
with a fixed film in one process. Excess growth is removed from the pads by peri-
odically passing them through specially designed pressure rollers.
The British Water Research Centre (WRC) and Severn Trent Water Authority
conducted a full-scale evaluation of the CAPTOR process for upgrading the acti-
vated sludge plant at the Freehold Sewage Treatment Works, in the West Midlands
area of England, to achieve year-round nitrification. This full-scale study was jointly
sponsored by the US Environmental Protection Agency [55, 56].

5.2.3 Development of CAPTOR Process

As mentioned earlier, the CAPTOR process originated from research work on pure
systems in the Chemical Engineering Department of UMIST. Single strands of
stainless steel wire were woven into a knitted formation and then crushed into a
sphere of about 6 mm (0.25 in.) diameter. These particles of known surface area
were used for modeling liquid-fluidized bed systems. From this work derived the
idea of using porous support pads for growing biomass at high concentrations that
could be used in wastewater treatment systems. The idea was jointly developed and
patented by UMIST and their industrial partner Simon-Hartley, Ltd. The present
form of the CAPTOR process uses 25 mm × 25 mm × 12 mm (1 in. × 1 in. × 0.5 in.)
reticulated polyether foam pads containing pores nominally of about 0.5–0.9 mm
(0.02–0.035 in.) diameter and 94% free space [57–59].
248 L. K. Wang et al.

5.2.4 Pilot-Plant Study

The conducted pilot-plant work indicated that it was possible to achieve the follow-
ing [55, 56]:
(a) Biomass concentrations of 7000–10,000 mg/L
(b) Waste sludge concentrations of 4–6% dry solids using a special pad cleaner
(c) Improved oxygen transfer efficiencies
(d) High BOD volumetric removal rates

5.2.5 Full-Scale Study of CAPTOR and CAST

The full-scale evaluation of the CAPTOR process was undertaken at the Freehold
Sewage Treatment Works near Stourbridge, West Midlands. The Freehold plant did
not achieve any nitrification in the winter and only partial nitrification in the sum-
mer. Freehold’s activated sludge system consisted of five trains equipped with
tapered fine bubble dome diffusers arranged in a grid configuration. The system was
modified as shown in Fig. 5.2 to split the wastewater flow into two equal volumes.
Half went to two trains that were modified by adding CAPTOR pads to the first
quarter of two aeration basins, and the other half went to two trains that remained
unaltered and served as a control. The CAPTOR modified trains were each equipped
with a CAPTOR pad cleaner (Fig. 5.3), and the CAPTOR pads were prevented from
escaping into the remainder of the experimental system aeration basins by screens
placed at the effluent ends of the CAPTOR zones.
The Simon-Hartley design predicted that, with a concentration of 40 pads/L, an
annual average removal of 75% of the BOD5 coming into the plant could be achieved
in the CAPTOR zones, resulting in a reduced food-to-microorganism (F/M) loading
on the follow-on activated sludge stage of 0.08 kg BOD5/day/kg MLSS. With the

Fig. 5.2 Schematic of treatment plant showing incorporation of CAPTOR [56]

5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 249

Fig. 5.3 CAPTOR pad cleaner [56]

reduced load, it was predicted that the modified system would achieve year-round
nitrification with an effluent ammonia nitrogen concentration of 5 mg/L or less [56].

5.2.5.1 Full-Scale Plant Initial Results

The Freehold modified CAPTOR activated sludge system was put in operation and
immediately encountered a major problem. The CAPTOR pads floated on the sur-
face of the tanks and would not become incorporated into the tank liquor. A solution
was found by removing three of the seven longitudinal rows of fine bubble diffusers
in the CAPTOR aeration basins. This was done to create a spiral roll in the tanks,
which leads to areas of rising and falling liquid with quite large channels down
where the pads can fall. The spiral roll modification provided the necessary falling
zone and produced complete mixing of the CAPTOR pads.
Another problem that occurred was maldistribution of the pads. The flow of
wastewater tended to push the CAPTOR pads to the outlet of their zones, resulting
in a concentration of 50–60 pads/L at the outlet and only 10–20 pads/L at the
inlet end.
One other disturbing feature was the rapid deterioration in the CAPTOR pads.
The CAPTOR pads used initially were black and were wearing at such a rate that
they would not have lasted for more than 3 years, rendering the process uneconomical.
It had also become evident by this time that with the Freehold wastewater it
would be possible to achieve the concentration of 200 mg biomass/pad predicted in
the design. However, it was found that if the biomass was allowed to grow beyond
180 mg/pad, the biomass in the center of the pad became anaerobic. The control of
pad biomass was difficult because the pad cleaners provided were not reliable and
250 L. K. Wang et al.

were situated at the CAPTOR zone inlets while most of the pads gravitated to the
outlet ends of the zones.
During this early period, while the above problems were being tackled on the
full-scale plant, there were some occasions when the effluent from the CAPTOR
units was reasonable (BOD removals of 40–50%), but BOD removal never
approached the average of 75% predicted based on the earlier pilot-plant results.
Poor BOD removals were being experienced because the suspended solids concen-
tration in the effluent was always high (>80 mg/L).
Consequently, more pilot-scale studies were used to find solutions to the operat-
ing problems described above before attempting further full-scale evaluation at
Freehold.

5.2.5.2 Pilot-Scale Studies for Project Development

It was decided to evaluate two variations of the CAPTOR process. The new varia-
tion differed from the original CAPTOR in that the pads were placed directly into
the mixed liquor of the activated sludge aeration tank rather than in a separate stage
before the activated sludge tank. WRC named this process variation CAST
(CAPTOR in activated sludge treatment). The CAST system had been applied to
upgrade several overloaded wastewater treatment plants in Germany and France and
was found to be useful in improving the treatment efficiency and plants’ perfor-
mance [60–62].
In addition, a single aeration tank filled with 40 CAPTOR pads/L was fed efflu-
ent from the above activated sludge control unit to assess the potential of CAPTOR
as a second-stage nitrification process. Neither pad cleaning nor final clarification
was necessary with this process variation because of the low sludge yields charac-
teristic of nitrifier growth.
Studies were conducted using two well-mixed CAPTOR tanks in series. A range
of loading and pad cleaning rates were used to evaluate process removal capabilities
for CAPTOR. The intermediate effluent was used as a measure of process efficiency
of the primary reactor and the final effluent for the entire system. This permitted
plotting (Fig. 5.3) of % BOD5 removal (total and soluble) vs. volumetric organic
loading rate over the range of 1–3.5 kg BOD5/day/m3 (62–218 lb/day/1000 ft3).
High and low pad cleaning rates are differentiated in Fig. 5.4 as ≥16% and <16% of
the total pad inventory/d, respectively [56].
Total BOD5 removal efficiency was less than soluble BOD5 removal efficiency
because of the oxygen demand exerted by the biomass solids lost in the process
effluent. The higher pad cleaning rates are believed to have contributed to the
improved total and soluble BOD removals shown in Fig. 5.4, although low bulk
liquid DOs may have adversely affected removals on some of the low cleaning runs.
Low cleaning rates (<16%/day) were detrimental to soluble BOD5 removal effi-
ciency because of a gradual decline in activity of the biomass remaining in the pad.
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 251

Fig. 5.4 Pilot-scale CAPTOR BOD5 removals as a function of organic loading rate [56]

Cleaning rates greater than 24%/day, however, resulted in reduced biomass levels in
the pads and a reduction in performance.
The problem of maldistribution of CAPTOR pads in the aeration tank (i.e.,
crowding of pads into the effluent end of the tank when operated in plug flow fash-
ion as at Freehold) was solved by modifying the flow pattern to transverse flow
(across the width of the tank rather than down the length). When implemented later
at Freehold, this pattern resulted in a fourfold decrease in flow velocity.
Several mixing intensities and diffuser arrangements were tried to decrease bio-
mass shedding into the process effluent. It became obvious, however, that produc-
tion of effluent biomass solids was not significantly affected by changes in mixing
intensity or diffuser arrangement. High effluent suspended solids proved to be far
more dependent on pad cleaning rate, biochemical activity of the biomass, and bio-
mass growth directly in the liquor.
Using the transverse flow scheme and a regular pad cleaning regimen, CAPTOR
process performance was similar to that experienced in the small tanks. Operating
parameters and process performance are summarized in Table 5.2 for two different
volumetric loading rates [56].
Respiration studies conducted on pads indicated that biomass held within the
pads respires at up to 40–50% less than equivalent biomass in free suspension. Any
increase in net biomass concentration achieved in a CAPTOR reactor above that in
a conventional activated sludge reactor may not produce noticeable benefits, there-
fore, due to the lower specific activity. These observations suggest that diffusion
limitations were occurring in the CAPTOR pads.
The CAST variation of CAPTOR was operated in conjunction with a final clari-
fier to settle the mixed liquor solids component of the total biomass inventory and
252 L. K. Wang et al.

Table 5.2 Pilot-scale operating conditions and process performance [56]

Period
Parameter 1 2
Volumetric loading (lb BOD5/day/1000 ft3)a 113 213
HRT (h) 2.32 1.52
Pads/L 40 40
Biomass/pad (mg) 121 126
Equivalent MLSS (mg/L) 4.840 5.040
F/M loading (kg BOD5/day/kgMLSS) 0.37 0.68
SRT (days) 3.23 1.72
DO (mg/L) 4.2 4.7
In Out In Out
Total BOD5 (mg/L) 175 93 216 129
Soluble BOD5 (mg/L) 86 24 85 33
SS (mg/L) 116 120 178 160
Total BOD5 removal (%) 47 40
Soluble BOD5 removal (%) 72 61
SS removal (%) −3 10
a
1 lb/day/1000 ft3 = 0016 kg/day/m3

return it to the aeration tank. CAPTOR pads and biomass retained therein were kept
in the reactor by screens. Operating and performance data are compared in Table 5.3
for the CAST unit and the parallel activated sludge control unit for a 25-day period
when the volumetric loadings and hydraulic residence times (HRTs) for both units
were identical.
In the nitrification experiments conducted on the CAPTOR process, the biomass
concentrations per pad ranged from 99 to 124 mg. This is within the range of
100–150 mg/L reported by other researchers [63]. With a pad concentration of 40/L,
equivalent MLSS levels varied from 3960 to 4960 mg/L. Liquor DO concentrations
were maintained between 6.4 and 8.4 mg/L, and liquor temperature ranged from
11.50 to 6.5°C.
Secondary effluent from the control activated sludge pilot unit used in the CAST
experiments was applied to the nitrification reactor over a range of loading condi-
tions. Essentially complete nitrification was achieved at TKN and ammonia nitro-
gen loadings of approximately 0.25 kg/day/m3 (15.6 lb/day/1000 ft3) and 0.20 kg/
day/m3 (12.5 lb/day/1000 ft3), respectively.

5.2.5.3 Full-Scale Plant Results After Modifications

Following the successful testing of the transverse mixing arrangement in the pilot-­
scale study, the two Freehold CAPTOR trains were modified. The modifications
involved the following [56]:
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 253

Table 5.3 Pilot-scale CAST and activated sludge operating conditions and performance [56]
System
Parameter CAST Activated Sludge
Volumetric loading (lb BOD5/day/1.000 ft3)a 148 148
HRT (h) 1.8 1.8
Pads/L 34 –
Biomass/pad (mg) 116 –
Equivalent MLSS in pads (mg/L) 3930 –
MLSS in suspension (mg/L) 3720 6030
Total MLSS (mg/L) 7650 6030
F/M loading (kg BOD5/day/kg total MLSS) 0.31 0.39
SRT, based on total MLSS (days) 3.6 3.0
DO (mg/L) 2.5 3.0
In Out In Out
Total BOD5 (mg/L) 178 12 178 20
Soluble BOD5 (mg/L) 101 5 101 4
SS (mg/L) 121 15 121 23
Total BOD5 removal (%) 93 89
Soluble BOD5 removal (%) 95 96
SS removal (%) 88 81
a
1 lb/day/1000 ft3 = 0.016 kg/day/m3

(a) Splitting each of the CAPTOR trains, C1 and C2, into two compartments, C1A
and C1B and C2A and C2B, as shown in Fig. 5.5
(b) Feeding influent flow along long weirs at the side of the trains instead of at the
narrow inlet ends
(c) Modifying the aeration pipework to place all three rows of dome diffusers
directly below the outlet screens (covering about 25% of the width of the tanks),
thereby creating a spiral roll of pads and liquid countercurrent to the flow of
wastewater entering along the weirs on the sidewalls
(d) Installing two extra pad cleaners so that each CAPTOR subunit was provided
with a cleaner
(e) Installing fine screens at the outlet from the primary clarifiers to reduce the
quantity of floating plastic material entering the CAPTOR units that created
problems with the cleaners
The objective of the first three modifications was to achieve uniform mixing of the
pads in the CAPTOR units and prevent the situation that had occurred previously
where high concentrations of pads (50–60 pads/L) collected at the outlet end and
very low concentrations (10–20 pads/L) at the inlet end. Pads were removed from
the tanks during the modifications. After the modifications were completed, the
number of pads in each compartment was equalized at about 35/L.
The changes were completely successful in obtaining uniform distribution and
complete mixing of the CAPTOR pads. A lithium chloride tracer test conducted on
254 L. K. Wang et al.

Fig. 5.5 Modifications to full-scale CAPTOR system flow pattern [56]

the modified tanks indicated that no dead zone was occurring in the “eye” of the roll.
Formation of floating pad rafts (which had occurred at the outlet end of the tank
with the original arrangement) was completely eliminated. The modifications, how-
ever, had no effect on the high level of suspended solids present in the liquor. The
modified CAPTOR system was operated at an average volumetric loading rate of
1.24 kg BOD5/day/m3 (77 lb/day/1000 ft3), an average HRT (excluding sludge recy-
cle) of 2.55 h, and an overall biomass concentration of 4830 mg/L.
The CAST variation of the CAPTOR process, which had exhibited somewhat
better performance than conventional activated sludge in the small tank experi-
ments, was also field evaluated at Freehold. The CAPTOR trains were further modi-
fied so that return sludge could be introduced to the CAPTOR zones (35 pads/L),
providing an activated sludge component throughout the entire aeration tanks, not
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 255

just in the nitrification stage. The average volumetric organic loadings and HRTs
(excluding sludge recycle) were 1.11 kg BOD5/day/m3 (69 lb/day/1000 ft3) and
3.40 h, respectively.
Performance data summarized in Tables 5.4 and 5.5 indicate that the CAST sys-
tem exhibits somewhat better performance than the CAPTOR version. In the CAST
process, the removal of soluble BOD5 is 96% compared to 90% in CAPTOR; the
removal of total BOD5 is 88% compared to 83%; and the removal of SS is about the
same at about 78%.

5.2.5.4 Overall Conclusions

The US Environmental Protection Agency (USEPA) conclusions and recommenda-

tions for the CAPTOR/CAST treatment systems are as follows [55, 56, 64]:
(a) In the initial phase when the CAPTOR process was installed at the Freehold
Sewage Treatment Works, several problems were immediately evident. There
were major problems with respect to pad mixing, suspension, and distribution,
and the process performance was adversely affected by the high level of sus-
pended solids in the CAPTOR stage effluent. The problems of pad mixing and
distribution were solved by pilot- and full-scale development work.
(b) The performance of the CAPTOR process was still adversely affected by the
high level of suspended solids in the CAPTOR stage effluent after correction of
the pad mixing, suspension, and distribution problems. This prevented the
achievement of nitrification in the follow-on activated sludge stage.
(c) The presence of CAPTOR pads in the tank liquid did not improve oxygen trans-
fer efficiency.
(d) The durability of the CAPTOR pads was solved by switching to different pads.
(e) The peak biomass concentration in the pads is unpredictable. It does not appear
to be related to the BOD concentration of the wastewater. There were indica-
tions in the various studies, however, that the frequency of pad cleaning (and,
hence, the biomass/pad concentration) was critical to the performance of the
process. Regular pad cleaning is essential to prevent anaerobic conditions from
developing in the pads.
(f) It is possible to raise the biomass concentration in a CAPTOR stage to
6000–8000 mg/L, but the respiration rate of the biomass in the pads is lower
than the respiration of the same biomass if freely suspended and less than that
of normal activated sludge. These data suggest that the geometry of the

Table 5.4 Full-scale modified CAPTOR performance results [56]

Parameter Influent, mg/L Effluent, mg/L Removal, %
Total BOD5 128 22 83
Soluble BOD5 40 4 90
SS 138 32 77
NH4-N 24 24.4 0
256 L. K. Wang et al.

Table 5.5 Full-scale modified CAST performance results [56]

Parameter Influent, mg/L Effluent, mg/L Removal, %
Total BOD5 138 16 88
Soluble BOD5 56 2 96
SS 120 27 78
NH4-N 26.7 17.2 36

CAPTOR pads results in diffusion limitations, which demands further pad

design improvement to enhance the potential for economic utilization of the
CAPTOR process in wastewater treatment.
(g) The CAST variation of the CAPTOR process performs well.
(h) CAPTOR has the potential as an add-on package for tertiary nitrification.
(i) The CAPTOR option was projected to be more cost effective than extending the
activated sludge plant for upgrading Freehold to complete year-round
nitrification.
(j) For CAPTOR and CAST to achieve their full potential, as predicted by the
pilot-scale studies, further design development and improvements are needed.

5.3 Activated Bio-filter (ABF)

5.3.1 Description

Activated bio-filters (ABF) are a recent innovation in the biological treatment field.
This process consists of the series combination of an aerobic tower (bio-cell) with
wood or other packing material, followed by an activated sludge aeration tank and
secondary clarifier. Settled sludge from the clarifier is recycled to the top of the
tower. In addition, the mixture of wastewater and recycle sludge passing through the
tower is also recycled around the tower, in a similar manner to a high-rate trickling
filter. No intermediate clarifier is utilized. Forward flow passes directly from the
tower discharge to the aeration tank (Fig. 5.6). The use of the two forms of biological
treatment combines the effects of both fixed and suspended growth processes in one
system. The microorganisms formed in the fixed growth phase are passed along to
the suspended growth unit, whereas the suspended growth microorganisms are recy-
cled to the top of the fixed media unit [65]. This combination of the two processes
results in the formation of a highly stable system that has excellent performance and
good settling biological floc when treating wastewaters that have variable loads [66].
The bio-media in the bio-cell consists of individual racks made of wooden laths
fixed to supporting rails. The wooden laths are placed in the horizontal direction,
permitting wastewater to pass downward, and air horizontally and vertically. The
horizontal surfaces reduce premature sloughing of biota. Droplet formation and
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 257

Fig. 5.6 ABF process flow diagram [65]

breakup induced by wastewater dripping from lath to lath enhances oxygen transfer.
Other types of material for the bio-media have also been reported by other research-
ers and equipment manufacturers [67–70]. The aeration basin is a short detention
unit that can be designed for either plug flow or complete mix operation. The efflu-
ent from the aeration basin passes to a secondary clarifier where the activated sludge
is collected and recycled to the top of the bio-cell tower and to waste.
ABF units can be used for the removal of either carbonaceous material or for
carbonaceous removal plus nitrification by appropriately modifying the detention
time of the aeration basin. When nitrification is desired, the bio-cell acts as a first-­
stage roughing unit and the aeration basin as a second-stage nitrification unit [71,
72]. ABF bio-cells can be either rectangular or round. Various types of aeration
equipment can be used in the aeration system, including both surface and diffused
aerators. The detention time of the aeration tank can be modified, depending on
influent quality and desired effluent quality. ABF units can be supplied with mixed
media effluent filters for enhanced treatment.

5.3.2 Applications

Activated bio-filters can be used for treating municipal wastewater and biodegrad-
able industrial wastewater. ABF systems are especially useful where [65, 66]:
(a) Both BOD5 removal and nitrification are required.
(b) Land availability is low.
(c) Raw wastewater organic loadings fluctuate greatly, due to its ability to handle
shock conditions.
(d) Existing trickling filter facilities and overloaded existing secondary plants need
to be upgraded at reduced cost.
A typical ABF application is the Burwood Beach Wastewater Treatment Works in
Australia [73]. The plant was upgraded in the 1990s using ABF at a cost of
$48 M. The facility currently serves a population of 180,000 with a flow of 43 ML
258 L. K. Wang et al.

a day and has the capacity to treat 53 ML/day for a population of 220,000 in the year
2020. The bio-filter is 30 m in diameter and has a design organic loading of 3.2 kg
BOD5/m3/day. The aeration tank is designed for 1.5 h of hydraulic detention time.
The plant has been in operation for around 10 years producing an effluent that is
consistently within the required USEPA set limits.

5.3.3 Design Criteria

The design criteria for the ABF system are reported to be as follows [65, 74, 75]:
(a) Bio-cell organic load: 100–200 lb BOD5/day/1000 ft3
(b) Return sludge rate: 25–100%
(c) Bio-cell recycle rate: 0–100%
(d) Bio-cell hydraulic load: 1–5.5 gpm/ft2
(e) Aeration basin detention time: 0.5–3.0 h for BOD5 removal only
5.8–7.5 h for two-stage nitrification
(f) System F/M: 0.25–1.5 lb BOD5/day/lb MLVSS for BOD removal
0.18 lb BOD5/day/lb MLVSS for two-stage nitrification.

5.3.4 Performance

ABF systems are quite stable and highly reliable. They can treat standard municipal,
combined municipal/industrial, or industrial wastewaters to BOD5 and suspended
solids levels of 20 mg/L or less. Test study on a package system showed at least 90%
removal of BOD5, TSS, and NH4-N [65]. The detailed results are shown in Table 5.6.
Sludge production was reported at 0.25–1.0 lb of waste VSS per lb of BOD5
removed. The mean yield over the course of the study was 0.60 lb VSS per lb of
BOD removed.

Table 5.6 Performance of BAF systems [65]

Parameter Influent, mg/L Effluent, mg/L Removal, %
BOD5 153 14 91
COD 330 58 82
TSS 222 20 91
NH4-Na 20 1 90
a
When used for nitrification
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 259

5.4 Vertical Loop Reactor (VLR)

5.4.1 Description

A Vertical Loop Reactor (VLR) is an activated sludge biological treatment process

similar to an oxidation ditch [76, 77]. The wastewater in an oxidation ditch circu-
lates in a horizontal loop; the water in a VLR circulates in a vertical loop around a
horizontal baffle, as shown in Fig. 5.7 [78]. A typical VLR consists of an 18 ft deep
concrete or steel basin with a horizontal baffle extending the entire width of the
reactor and most of its length. Operating basins are reported to have sidewall depths
which range from approximately 10–22 ft [79]. The length and width of the VLR
are determined by the required capacity, but, as a rule, the length is at least twice the
width. The baffle is generally 5–11 ft below the surface of the water. Because a VLR
is typically deeper than an oxidation ditch, the VLR requires less land area.
Aeration in a VLR is provided by coarse bubble diffusers, which are located
below the horizontal baffle, and by disc aeration mixers. The disc mixers also circu-
late the wastewater around the baffle at a velocity of 1–1.5 ft/s [80]. Because the
diffusers are positioned below the baffle, the air bubble residence time in a VLR is
as much as six times longer than the bubble residence time in a conventional aera-
tion system. This extended bubble contact time increases the process aeration effi-
ciency. Denitrification in an anoxic zone also reduces oxygen requirements.
The VLR process is usually preceded by preliminary treatment such as screen-
ing, comminution, or grit removal. Secondary settling of the VLR effluent is typi-
cally provided by a separate clarifier. An intra-channel clarifier may be used for
secondary settling in place of a separate clarifier.
Vertical loop reactors may be operated in parallel or series. When a series of
VLRs are used, the dissolved oxygen profile can be controlled to provide nitrifica-
tion, denitrification, and biological phosphorus removal at hydraulic detention times
of 10–15 h.

Fig. 5.7 Diagram of the Vertical Loop Reactor [77, 78]

260 L. K. Wang et al.

5.4.2 Applications

VLR technology is applicable in any situation where conventional or extended aera-

tion activated sludge treatment is appropriate. The technology is applicable for nitri-
fication and denitrification. Biological phosphorus removal may be incorporated in
the system design. Power costs may be lower for a VLR system than for other aer-
ated biological treatment systems, due to improved oxygen transfer efficiency.
There are currently more than ten municipal wastewater treatment facilities in the
United States with VLRs. One such example is the City of Willard, OH, United
States, wastewater treatment plant [81]. The facility is designed for an average daily
flow of 4.5 MGD and is capable of handling a peak flow of 7.2 MGD.
The following advantages have been reported for VLR systems [82]:
(a) The land area required for VLRs is about 40% less than for oxidation ditches.
(b) The VLR aeration basin cost is about 30% less than for oxidation ditches.
(c) The multiple tank basin series arrangement is an advantage for facilities with
highly variable flow.
(d) VLRs are useful for retrofitting existing basins for plant upgrade to suit
increased flows or more stringent effluent requirements.

5.4.3 Design Criteria

The design criteria for the VLR process are reported to be as follows [76]:
BOD loading: 14–22 lb BOD5/1000 ft3/day
SRT: 17–36 day
Detention Time: 12–24 h

5.4.4 Performance

The average effluent BOD5 and TSS concentrations for the five studied operating
VLR facilities are 4.2 and 7.1 mg/L, respectively. The average effluent ammonia
concentration is 0.8 mg/L. Only one of the VLRs studied was designed for biologi-
cal phosphorus removal; the average effluent phosphorus concentration for this
plant was 1.45 mg/L, and alum was added in the final clarifiers. A second VLR facil-
ity was not designed for biological phosphorus removal but was required to monitor
phosphorus. This plant had an average effluent phosphorus concentration of 2.19
without any chemical addition.
The VLR system is quite reliable. Table 5.7 indicates the percent of time the
monthly average effluent concentration of the given pollutants was less than the
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 261

Table 5.7 Reliability of the VLR treatment process [76]

Concentration, mg/L BODa NH3-Na TSSa Pa
0.2 0 30 0 2
0.5 0 63 1 10
1.0 0 83 1 24
2.0 20 88 5 63
3.0 71 95 43 93
10.0 97 96 75 100
20.0 100 100 96 100
Number of plants 5 5 5 1
a
Percentage of time the monthly average concentration of the pollutant was less than the stated
value in the first column

concentration given in the first column. No significant difference in results was

observed between winter and summer data.

5.4.5 USEPA Evaluation of VLR

The following summarizes the major findings and conclusions of USEPA evaluation
of VLRs [77]. The information is based on analysis of available information from
site visits, a detailed design of a full-scale VLR system, and information from con-
sultants and manufacturers.
(a) The VLR is a modification of the conventional activated sludge process. The
unique features of the process are circulating mixed liquor around a horizontal
baffle with a dual aeration system, bubble diffused air beneath the horizontal
baffle, and disc aerators at the surface of the aeration tank. The process operates
as a plug flow reactor with capability for varying dissolved oxygen profiles to
achieve biological phosphorus and nitrogen removal. The VLR process also
features a stormwater bypass design for treatment of high peak to average flows.
(b) There are currently over ten operating VLRs in the United States ranging in size
from 0.22 to 5.0 MGD.
(c) Performance data from operating VLRs show that this process is capable of
achieving effluent carbonaceous biochemical oxygen demand levels of less
than 10 mg/L, effluent total suspended solids levels of less than 10 mg/L, and
effluent ammonia nitrogen levels of less than 1.0 mg/L. The process is further
capable of achieving total nitrogen and phosphorus removals of 60–80%.
(d) The VLR process is applicable for flows ranging from 0.05 to over 10 MGD.
(e) The claimed advantages of this process by the manufacturer include the
following:
• Higher dissolved oxygen transfer than conventional equivalent technology
• Improved response to peak flows due to a stormwater bypass feature
262 L. K. Wang et al.

• A credit for oxygen release due to denitrification with the credit based on
80% denitrification
• Increased mixed liquor settleability and process stability
(f) The design criteria for the existing VLRs are conservative. HRTs range from
11.9 to 24 h. Volumetric loading ranged from 13.6 to 23.1 lbs CBOD/1000 ft3.
This loading is similar to that used for extended aeration systems and is about
1/3 to 1/2 of that normally used for conventional activated sludge designs.
(g) The VLR technology has been designated as Innovative Technology by the EPA
for three plants due to a 20% claimed energy savings.
(h) Based on this assessment, the 20% energy savings over competing technology
could not be verified.
(i) The VLR was compared to oxidation ditches as “Equivalent Technology.” The
results of this comparison indicated:
• The VLR technology produces comparable to slightly improved effluent lev-
els of BOD, TSS, and NH3-N than oxidation ditch plants.
• Total removal of phosphorus and total nitrogen are equivalent to oxidation
ditches designed for the same level of treatment.
• The energy requirements for aeration were found to be similar to 10% less
than for oxidation ditches.
• The land area required for VLRs was found to be approximately 40% less
than for oxidation ditches based on equivalent aeration tank loadings.
• The VLR aeration basin cost was found to be approximately 30% less than
for oxidation ditches for situations where rock excavation is not required for
the deeper VLR basin.
• A definitive comparison of total VLR plant costs to total oxidation plant
costs could not be made. Data submitted from both manufacturers indicated
a comparable cost for plants in the 0–2 MGD range. The reported VLR costs
at plants ranging from 2 to 10 MGD were significantly less than oxidation
ditch plant costs. This would be expected because of the modular design and
common wall construction of the VLR compared to oxidation ditches.
• The total operation and maintenance costs of the two technologies were
found to be similar.

5.4.6 Energy Requirements

The VLR energy requirements are shown in Fig. 5.8. The requirements are based on
the following assumptions [76]:
(a) Water quality
BOD5: influent = 200 mg/L, effluent = 20 mg/L
TKN: influent = 35 mg/L, effluent = 1 mg/L
(b) Design basis
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 263

Fig. 5.8 VLR energy requirements and construction cost [76, 77]

Oxygen transfer efficiency: 2.5 lb O2/Hp hour

Nitrification occurs
(c) Operating parameters
Oxygen requirement: 1.5 lb O2/lb BOD5 removed
4.57 lb O2/lb TKN removed
(d) Type of energy: electrical
264 L. K. Wang et al.

5.4.7 Costs

The construction costs (1991 Dollars, Utilities Index = 392.35) for VLR are shown
in Fig. 5.8. To obtain the values in terms of the present 2004 US Dollars, using the
Cost Index for Utilities (Appendix 1), multiply the costs by a factor of
506.13/392.35 = 1.29 [83]. The operation costs are similar to oxidation ditch type
treatment plant.

5.5 PhoStrip Process

5.5.1 Description

“PhoStrip” is a combined biological-chemical precipitation process based on the

use of activated sludge microorganisms to transfer phosphorus from incoming
wastewater to a small concentrated substream for precipitation. As illustrated in
Fig. 5.9, the activated sludge is subjected to anoxic conditions to induce phosphorus
release into the substream and to provide phosphorus uptake capacity when the
sludge is returned to the aeration tank. Settled wastewater is mixed with return acti-
vated sludge in the aeration tank. Under aeration, sludge microorganisms can be
induced to take up dissolved phosphorus in excess of the amount required for
growth. The mixed liquor then flows to the secondary clarifier where liquid effluent,
now largely free of phosphorus, is separated from the sludge and discharged. A por-
tion of the phosphorus-rich sludge is transferred from the bottom of the clarifier to
a thickener-type holding tank: the phosphate stripper. The settling sludge quickly
becomes anoxic and, thereupon, the organisms surrender phosphorus, which is
mixed into the supernatant. The phosphorus-rich supernatant, a low-volume, high-­
concentration substream, is removed from the stripper and treated with lime for
phosphorus precipitation. The thickened sludge, now depleted in phosphorus, is
returned to the aeration tank for a new cycle [65]. The readers are referred to the

Fig. 5.9 PhoStrip process flow diagram [65]

5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 265

literature [84–97] for additional innovative wastewater and sludge treatment pro-
cesses, such as biological sequencing batch reactor, physicochemical sequencing
batch reactor, membrane bioreactor, flotation bioreactor, membrane flotation biore-
actor, Symbio process, column bioreactor clarifier process, upflow sludge blanket
filtration, deep well injection, land application, aerobic granulation technology, ver-
tical shaft bioreactor, vertical shaft digestion, bioreactor landfill, post aeration, etc.
The PhoStrip process has demonstrated a compatibility with the conventional
activated sludge process and is compatible with its modifications. The process can
operate in various flow schemes, including full or split flow of return activated
sludge through the phosphate stripper, use of an elutriate to aid in the release of
phosphorus from the anoxic zone of the stripper, or returning lime-treated stripper
supernatant to the primary clarifier for removal of chemical sludge.
This technique is a new development in municipal wastewater treatment and has
been demonstrated in pilot-plant and full-scale studies. Notable large-scale evalua-
tions have been conducted at Seneca Falls, New York, United States, and, more
recently, Reno/Sparks, Nevada, United States. Nearly a dozen commercial installa-
tions are reported to be in the operational phase.

5.5.2 Applications

This method, which involves a modification of the activated sludge process, can be
used in removing phosphorus from municipal wastewaters to comply with most
effluent standards. Direct chemical treatment is simple and reliable, but it has the
two disadvantages of significant sludge production and high operating costs. The
PhoStrip system reduces the volume of the substream to be treated, thereby reduc-
ing the chemical dosage required, the amount of chemical sludge produced, and
associated costs. Lime is used to remove phosphorus from the stripper supernatant
at lower pH levels (8.5–9.0) than normally required. The cycling of sludge through
an anoxic phase may also assist in the control of bulking by the destruction of fila-
mentous organisms to which bulking is generally attributed [65].
On the negative side, it should be pointed out that more equipment and automa-
tion, along with a greater capital investment, are normally required than for conven-
tional chemical addition systems. Since this method relies on activated sludge
microorganisms for phosphorus removal, any biological upset that hinders uptake
ability will also affect effluent concentrations. It has been found that sludge in the
stripper tank is very sensitive to the presence of oxygen. Anoxic conditions must be
maintained for phosphorus release to occur.
266 L. K. Wang et al.

Table 5.8 Typical design criteria for the PhoStrip process [74]
Design parameter Unit Value
Food-to-microorganism ratio (F/M) lb BOD/lb MLSS/day 0.1–0.5
Solids retention time (SRT) day 10–30
Mixed liquor suspended solids (MLSS) mg/L 600–5000
Hydraulic retention time in stripper (t) h 8–12
Hydraulic retention time in aeration tank (t) h 4–10
Return activated sludge (RAS) % of influent 20–50
Internal recycle (stripper underflow) % of influent 10–20

5.5.3 Design Criteria

The fraction of the total sludge flow which must be processed through the stripper
tank is determined by the phosphorus concentration in the influent wastewater to the
treatment plant and the level required in the treated effluent. The required detention
time in the stripper tank ranges from 5 to 15 h. Typical phosphorus concentrations
produced in the stripper are in the range of 40–70 mg/L. The volume of the
phosphorus-­rich supernatant stream to be lime treated is 10–20% of the total flow
[65]. Typical design criteria for the PhoStrip process are shown in Table 5.8 [74].

5.5.4 Performance

Pilot- and full-scale studies of the process have shown it to be capable of reducing
the total phosphorus concentration of typical municipal wastewaters to 1.5 mg/L
[74] or even to 0.5 mg/L or less [75]. A plant-scale evaluation of the method treating
6 MGD of municipal wastewater at the Reno/Sparks Joint Water Pollution Control
Plant in Nevada demonstrated satisfactory performance for achieving greater than
90% phosphorus removal. Results showed that the process enhanced the overall
operation and performance of the activated sludge process, since it produced a more
stable, better settling sludge. Regular maintenance of mechanical equipment,
including pumps and mixers, is necessary to ensure proper functioning of the
entire system.

5.5.5 Cost
5.5.5.1 Construction Cost

The construction costs (1980 Dollars, Utilities Index = 277.60) for PhoStrip are
shown in Fig. 5.10. To obtain the values in terms of the present 2004 US Dollars,
using the Cost Index for Utilities (Appendix 1), multiply the costs by a factor of
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 267

CONSTRUCTION COST
100

10
Millions of Dollars

1.0

0.1
0.1 1.0 10 100
Wastewater Flow, Mgal/d

Fig. 5.10 PhoStrip construction cost [65]

506.13/277.60 = 1.82 [83]. Construction costs include stripper (10 h detention time
at 50% of return sludge), flash mixer, flocculator/clarifier, thickeners, and lime feed
and storage facilities [65].

5.5.5.2 Operation and Maintenance Cost

The electrical energy required for operation of pumps, lime mixing equipment, and
clarifiers is shown in Fig. 5.11. The operation and maintenance costs (1980 Dollars,
Utilities Index = 277.60) for PhoStrip are shown in Fig. 5.12. To obtain the values
in terms of the present 2004 US Dollars, using the Cost Index for Utilities (Appendix
1), multiply the costs by a factor of 506.13/277.60 = 1.82 [83]. Operation and main-
tenance costs include labor for operation, preventive maintenance, and minor
repairs; materials to include replacement parts and major repair work; and lime and
power cost based on the electrical energy requirement shown in Fig. 5.11 [65].

Glossary

Activated bio-filter (ABF) Activated bio-filters are a recent innovation in the

biological treatment field. This process consists of the series combination of an
aerobic tower (bio-cell) with wood or other packing material, followed by an
268 L. K. Wang et al.

107

Electrical Energy Required, kWh/yr

106

105

104
0.1 1.0 10 100
Wastewater Flow, Mgal/d

Fig. 5.11 PhoStrip electrical energy requirement [65]

OPERATION & MAINTENANCE COST

1.0 0.1

Chemicals
Annual Cost. Millions of Dollars

Total
0.01
Total-Labor-Materials

0.1
Power-Chemicals

Labor

Power

0.01 0.001
Materials

0.001 0.0001
0.1 1.0 10 100
Wastewater Flow, Mgal/d

Fig. 5.12 PhoStrip operation and maintenance cost [65]

activated sludge aeration tank and secondary clarifier. Settled sludge from the
clarifier is recycled to the top of the tower. In addition, the mixture of wastewater
and recycle sludge passing through the tower is also recycled around the tower,
in a similar manner to a high-rate trickling filter. No intermediate clarifier is
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 269

utilized. Forward flow passes directly from the tower discharge to the aeration
tank. The use of the two forms of biological treatment combines the effects of
both fixed and suspended growth processes in one system. The microorganisms
formed in the fixed growth phase are passed along to the suspended growth unit,
whereas the suspended growth microorganisms are recycled to the top of the
fixed media unit. This combination of the two processes results in the formation
of a highly stable system that has excellent performance and good settling bio-
logical floc when treating wastewaters that have variable loads.
Carrier-activated sludge processes (CAPTOR and CAST systems) There has
been a substantial interest in recent years in the potential benefits of high bio-
mass wastewater treatment. The major obstacle for achieving this has been the
inability of biosolids separation in secondary clarifiers. For the most part, this
has been overcome by using various forms of support media or carriers that have
the ability to attach high concentrations of aerobic bacterial growth. The increase
in immobilized biomass reduces the process dependence on secondary settling
basins for clarification. In such hybrid systems where attached growth coexists
with suspended growth, one gets more stable systems which possess the com-
bined advantages of both fixed and suspended growth reactors.
PACT activated sludge process The powdered activated carbon (PAC) activated
sludge system is a process modification of the activated sludge process. PAC is
added to the aeration tank where it is mixed with the biological solids. The mixed
liquor solids are settled and separated from the treated effluent. In a ­gravity
clarifier, polyelectrolyte will normally be added prior to the clarification step
to enhance solids-liquid separation. If phosphorus removal is necessary, alum is
often added at this point also. Even with polyelectrolyte addition, tertiary filtra-
tion is normally required to reduce the level of effluent suspended solids. The
clarifier underflow solids are continuously returned to the aeration tank. A por-
tion of the carbon-biomass mixture is wasted periodically to maintain the desired
solids inventory in the system.
PhoStrip process “PhoStrip” is a combined biological-chemical precipitation pro-
cess based on the use of activated sludge microorganisms to transfer phosphorus
from incoming wastewater to a small concentrated substream for precipitation.
The activated sludge is subjected to anoxic conditions to induce phosphorus
release into the substream and to provide phosphorus uptake capacity when the
sludge is returned to the aeration tank. Settled wastewater is mixed with return
activated sludge in the aeration tank. Under aeration, sludge microorganisms can
be induced to take up dissolved phosphorus in excess of the amount required
for growth. The mixed liquor then flows to the secondary clarifier where liquid
effluent, now largely free of phosphorus, is separated from the sludge and dis-
charged. A portion of the phosphorus-rich sludge is transferred from the bottom
of the clarifier to a thickener-type holding tank: the phosphate stripper. The set-
tling sludge quickly becomes anoxic and, thereupon, the organisms surrender
phosphorus, which is mixed into the supernatant. The phosphorus-rich superna-
270 L. K. Wang et al.

tant, a low-volume, high-concentration substream, is removed from the stripper

and treated with lime for phosphorus precipitation. The thickened sludge, now
depleted in phosphorus, is returned to the aeration tank for a new cycle.
Vertical Loop Reactor (VLR) A Vertical Loop Reactor (VLR) is an activated
sludge biological treatment process similar to an oxidation ditch. The wastewater
in an oxidation ditch circulates in a horizontal loop; the water in a VLR circulates
in a vertical loop around a horizontal baffle. A typical VLR consists of an 18 ft
deep concrete or steel basin with a horizontal baffle extending the entire width of
the reactor and most of its length. Operating basins are reported to have sidewall
depths which range from approximately 10–22 ft. The length and width of the
VLR are determined by the required capacity but, as a rule, the length is at least
twice the width. The baffle is generally 5–11 ft below the surface of the water.
Because a VLR is typically deeper than an oxidation ditch, the VLR requires less
land area.

Appendix 1: US Yearly Average Cost Index for Utilities [83]

Year Index Year Index

1967 100 1995 439.72
1968 104.83 1996 445.58
1969 112.17 1997 454.99
1970 119.75 1998 459.40
1971 131.73 1999 460.16
1972 141.94 2000 468.05
1973 149.36 2001 472.18
1974 170.45 2002 486.16
1975 190.49 2003 497.40
1976 202.61 2004 563.78
1977 215.84 2005 605.47
1978 235.78 2006 645.52
1979 257.20 2007 681.88
1980 277.60 2008 741.36
1981 302.25 2009 699.70
1982 320.13 2010 720.80
1983 330.82 2011 758.79
1984 341.06 2012 769.30
1985 346.12 2013 776.44
1986 347.33 2014 791.59
1987 353.35 2015 786.32
1988 369.45 2016 782.46
1989 383.14 2017 803.93
(continued)
5 Innovative PACT Activated Sludge, CAPTOR Activated Sludge, Activated… 271

Year Index Year Index

1990 386.75 2018 841.84
1991 392.35 2019 866.18
1992 399.07 2020 867.71
1993 410.63 2021 893.02a
1994 424.91 2022 918.91a
a
Projected future cost index values

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Chapter 6
Agricultural Waste Treatment by Water
Hyacinth Aquaculture, Wetland
Aquaculture, Evapotranspiration, Rapid
Rate Land Treatment, Slow Rate Land
Treatment, and Subsurface Infiltration

Lawrence K. Wang, Mu-Hao Sung Wang, and Nazih K. Shammas

Acronyms

Al2(SO4)3 Aluminum sulfate

BOD5 5-day biochemical oxygen demand
CaCO3 Calcium carbonate
COD Chemical oxygen demand
ET Evapotranspiration
FeC13 Ferric chloride
FWS Free water surface
MG Million gallons
MPN Most probable number
NEHA National Environmental Health Association
SFS Subsurface flow system
TDH Total dynamic head
TSS Total suspended solids
UNIDO United Nations Industrial Development Organization
USACE US Army Corps of Engineers
USEPA US Environmental Protection Agency
VSB Vegetated submerged bed

L. K. Wang (*)
Lenox Institute of Water Technology, Latham, NY, USA
Agricultural Engineering Department, University of Illinois, Urbana-Champaign, IL, USA
e-mail: [email protected]
M.-H. S. Wang · N. K. Shammas
Lenox Institute of Water Technology, Latham, NY, USA
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 277

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_6
278 L. K. Wang et al.

6.1 Aquaculture Treatment: Water Hyacinth System

6.1.1 Description

Aquaculture or the production of aquatic organisms (both flora and fauna) under
controlled conditions has been practiced for centuries, primarily for the generation
of food, fiber, and fertilizer. The water hyacinth (Eichhornia crassipes) appears to
be the most promising organism for wastewater treatment and has received the most
attention [1]. However, other organisms are being studied. Among them are duck-
weed, seaweed, midge larvae, alligator weeds, and a host of other organisms. Water
hyacinths are large fast-growing floating aquatic plants with broad, glossy green
leaves and light lavender flowers. A native of South America, water hyacinths are
found naturally in waterways, bayous, and other backwaters throughout the South.
Insects and disease have little effect on the hyacinth, and they thrive in raw, as well
as partially treated, wastewater. Wastewater treatment by water hyacinths is accom-
plished by passing the wastewater through a hyacinth-covered basin (Fig. 6.1),
where the plants remove nutrients, BOD5 (5-day biochemical oxygen demand), TSS
(total suspended solids), heavy metals, etc. Batch treatment and flow-through sys-
tems, using single and multiple cell units, are possible. Hyacinths harvested from
these systems have been investigated as a fertilizer/soil conditioner after compost-
ing, animal feed, and a source of methane when anaerobically digested [2].

6.1.2 Applications

Water hyacinths are generally used in combination with (following) lagoons, with
or without chemical phosphorus removal. A number of full-scale systems are in
operation, most often considered for nutrient removal and additional treatment of
secondary effluent [1–3]. Also, research is being conducted on the use of water
hyacinths for raw and primary treated wastewater or industrial wastes, but present
data favor combination systems. Very good heavy metal uptake by the hyacinth has
been reported. Hyacinth treatment may be suitable for seasonal use in treating
wastewaters from recreational facilities and those generated from processing of
agricultural products. Other organisms and methods with wider climatological
applicability are being studied. The ability of hyacinths to remove nitrogen during

Fig. 6.1 Aquaculture treatment: water hyacinth system. Source: U. S. EPA [2]
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 279

active growth periods and some phosphorus and retard algal growth provides poten-
tial applications in [2, 3]:
(a) The upgrading of lagoons
(b) Renovation of small lakes and reservoirs
(c) Pretreatment of surface waters used for domestic supply
(d) Stormwater treatment
(e) Demineralization of water
(f) Recycling fish culture water
(g) For biomonitoring purposes

6.1.3 Limitations

Climate or climate control is the major limitation. Active growth begins when the
water temperature rises above 10 °C and flourishes when the water temperature is
approximately 21 °C. Plants die rapidly when the water temperature approaches the
freezing point; therefore, greenhouse structures are necessary in northern locations.
Water hyacinths are sensitive to high salinity. Removal of phosphorus and potas-
sium is restricted to the active growth period of the plants.
Metals such as arsenic, chromium, copper, mercury, lead, nickel, and zinc can
accumulate in hyacinths and limit their suitability as a fertilizer or feed material.
The hyacinths may also create small pools of stagnant surface water which can
breed mosquitoes. Mosquito problems can generally be avoided by maintaining
mosquito fish in the system. The spread of the hyacinth plant itself must be con-
trolled by barriers since the plant can spread and grow rapidly and clog affected
waterways. Hyacinth treatment may prove impractical for large treatment plants
due to land requirements. Removal must be at regular intervals to avoid heavy inter-
twined growth conditions. Evapotranspiration can be increased by two to seven
times greater than evaporation alone.

6.1.4 Design Criteria

Ponds, channels, or basins are in use. In northern climates, covers and heat would
be required. Harvesting and processing equipment are needed. Operation is by grav-
ity flow and requires no energy. Hyacinth growth energy is supplied by sunlight. All
experimental data is from southern climates where no auxiliary heat was needed.
Data is not available on heating requirements for northern climates, but it can be
assumed proportional to northern latitude of location and to the desired growth rate
of hyacinths.
280 L. K. Wang et al.

Design data vary widely. Table 6.1 shows the design criteria for water hyacinth
systems [4]. The following ranges refer to hyacinth treatment as a tertiary process
on secondary effluent [2]:
(a) Depth should be sufficient to maximize plant rooting and plant absorption.
(b) Detention time depends on effluent requirements and flow, range 4–15 days.
(c) Phosphorus reduction, 10–75%.
(d) Nitrogen reduction, 40–75%.
(e) Land requirement is usually high, 2–15 acres/MG/day.

6.1.5 Performance

The process appears to be reliable from mechanical and process standpoints, subject
to temperature constraints. In tests on five different wastewater streams including
raw wastewater and secondary effluents, the following removals were reported [2]:
(a) BOD5, 35–97%
(b) TSS, 71–83%
(c) Nitrogen, 44–92%
(d) Total P, 11–74%
Takeda and coworkers [3] reported using aquaculture wastewater effluent for straw-
berry production in a hydroponic system which reduced the final effluent phospho-
rus concentration to as low as 0.1 mg/L which meets the stringent phosphorus
discharge regulations. There is also evidence that in aquaculture system coliform,
heavy metals and organics are also reduced, as well as pH neutralization.
Hyacinth harvesting may be continuous or intermittent. Studies indicate that
average hyacinth production (including 95% water) is on the order of 1000–10,000 lb/
day/acre. Basin cleaning at least once per year results in harvested hyacinths. For

Table 6.1 Design criteria for water hyacinth systems. Source: U. S. EPA [4]
Factor Aerobic non-aerated Aerobic non-aerated Aerobic aerated
Influent wastewater Screened or Settled Secondary Screened or settled
Influent BOD5, mg/L 130–180 30 130–180
BOD5 loading, kg/ha day 40–80 10–40 150–300
Expected effluent, mg/L
 bod5 <30 < 10 <15
 SS <30 <10 <15
 TN <15 <5 <15
Water depth, m 0.5–0.8 0.6–0.9 0.9–1.4
Detention time, days 10–36 6–18 4–8
Hydraulic loading, m3/ha day >200 <800 550–1000
Harvest schedule Annually Twice per month Monthly
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 281

further detailed information on water hyacinth systems, the reader is referred to

Refs. [5–13].

6.2 Aquaculture Treatment: Wetland System

6.2.1 Description

Aquaculture-wetland systems for wastewater treatment include natural and artificial

wetlands as well as other aquatic systems involving the production of algae and
higher plants (both submerged and emergent), invertebrates, and fish. Natural wet-
lands, both marine and freshwater, have inadvertently served as natural waste treat-
ment systems for centuries; however, in recent years, marshes, swamps, bogs, and
other wetland areas have been successfully utilized as managed natural “nutrient
sinks” for polishing partially treated effluents under relatively controlled condi-
tions. Constructed wetlands can be designed to meet specific project conditions
while providing new wetland areas that also improve available wildlife wetland
habitats and the other numerous benefits of wetland areas. Managed plantings of
reeds (e.g., Phragmites spp.) and rushes (e.g., Scirpus spp. and Schoenoplectus
spp.) as well as managed natural and constructed marshes, swamps, and bogs have
been demonstrated to reliably provide pH neutralization and reduction of nutrients,
heavy metals, organics, BOD5, COD (chemical oxygen demand), TSS, fecal coli-
forms, and pathogenic bacteria [2, 4].
Wastewater treatment by natural and constructed wetland systems is generally
accomplished by sprinkling or flood irrigating the wastewater into the wetland area
or by passing the wastewater through a system of shallow ponds, channels, basins,
or other constructed areas where the emergent aquatic vegetation has been planted
or naturally occurs and is actively growing (see Fig. 6.2). The vegetation produced
as a result of the system’s operation may or may not be removed and can be utilized
for various purposes [2]:
(a) Composted for use as a source of fertilizer/soil conditioner

Fig. 6.2 Aquaculture treatment: wetland system. Source: U. S. EPA [2]

282 L. K. Wang et al.

(b) Dried or otherwise processed for use as animal feed supplements

(c) Digested to produce methane

6.2.2 Constructed Wetlands

Constructed wetlands are classified as a function of water flow [2, 4]: surface and
subsurface which are known as free water surface (FWS) and subsurface flow sys-
tem (SFS) (also termed vegetated submerged bed (VSB)). When simply expressed,
constructed wetland treatment technology makes artificial receiving water and its
vegetation part of the treatment process. In comparison to algae, the higher forms of
plant life—floating (duckweed, water hyacinths), submerged, and emergent (cat-
tails, rushes, and reeds)—perform less efficiently per unit weight of biomass.
FWS constructed wetland treatment conceptually relies on attached growth bac-
terial performance, receiving oxygen from the evapotranspiration response of the
aquatic vegetation. Practically, the dominant bacterial action is anaerobic. The
ammonium and nitrogen removal mechanisms [14–17] are a combination of aerobic
oxidation, particulate removal, and synthesis of new plant protoplasm.
An FWS wetland is nothing more than a lagoon, except that a far greater expanse
is needed to maximize the productivity per unit area. In practice, very large systems
may achieve significant, if not complete, nitrogen oxidation, with surface reaeration
contributing to the oxygen supply. Some nitrification and denitrification undoubt-
edly occur in all systems.
If it is assumed that the wetland vegetation will not be harvested, as is the case
with natural wetland systems, its capacity for nitrogen control is finite, reflecting the
site-specific vegetation and the ability to expand in the available space. Thus, the
bigger the natural wetland that is called part of the process, the better, since there is
dilution of the wastewater to the point that it is no longer significant in comparison
to the naturally occurring background flow and water quality.
Constructed FWS wetlands yield a managed vegetative habitat that becomes an
aquaculture system. Examination of the evolution of this technology shows the
emergence of concepts that include organic load distribution or artificial aeration to
avoid aesthetic nuisances and emphasis on plants that grow the fastest. Duckweed
and water hyacinth systems (classified as aquaculture) have been reported to achieve
long-term total nitrogen residuals of less than 10 mg/L and may be manageable,
with harvesting and sensitive operation, to values of less than 3 mg/L on a seasonal,
if not sustained, basis.
Submerged-flow constructed wetlands are simply horizontal-flow gravel filters
with the added component of emergent plants within the media. They have been
classically used for BOD removal following sedimentation and/or additional BOD
and SS removal from lagoon effluents as with FWS approaches. This technology
has the potential for high-level denitrification when a nitrified wastewater is applied;
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 283

the naturally occurring environment promotes anoxic (denitrification) pathways for

oxidized nitrogen elimination.
Ultimately, the success or failure of the wetland approach for nitrogen control
may rest with the harvest of the vegetation, the need for backup (so that areas under
harvest have the backup of areas in active growth), and often natural seasonal growth
and decay cycles. If biomass production is an unacceptable goal, the designer should
think of a more tolerant mixed vegetation system that minimizes the need to harvest
the accumulated vegetation and maximizes the promotion of concurrent or staged
nitrification and denitrification in some fashion. Conceptually, the optimization has
to begin with promotion of nitrogen oxidation systems that may be shallow (better
aeration for attached and suspended bacterial growth) with vegetation that mini-
mizes light penetration and avoids as much algal growth as possible. Cyclic staging,
recycle, forced aeration, and mixing represent some of the enhancements that natu-
rally follow [17].

6.2.3 Applications

Several full-scale systems are in operation or under construction [18]. Wetlands are
useful for polishing treated effluents. They have potential as a low-cost, low-energy-­
consuming alternative or addition to conventional treatment systems, especially for
smaller flows. Wetlands have been successfully used in combination with chemical
addition and overland flow land treatment systems. Wetland systems may also be
suitable for seasonal use in treating wastewaters from recreational facilities, some
agricultural operations, or other waste-producing units where the necessary land
area is available [18]. Potential application as an alternative waste discharge tech-
nology to lengthy outfalls extended into rivers, etc. and as a method of pretreatment
of surface waters for domestic supply, stormwater treatment, recycling fish culture
water, and biomonitoring purposes.

6.2.4 Limitations

Temperature (climate) is a major limitation since effective treatment is linked to the

active growth phase of the emergent vegetation. Tie-ins with cooling water from
power plants to recover waste heat have potential for extending growing seasons in
colder climates. Enclosed and covered systems are possible for very small flows.
Herbicides and other materials toxic to the plants can affect their health and lead
to poor treatment. Duckweeds are prized as food for waterfowl and fish and can be
seriously depleted by these species. Winds may blow duckweeds to the shore if
wind screens or deep trenches are not employed. Small pools of stagnant surface
water which can allow mosquitoes to breed can develop, but problems can generally
be avoided by maintaining mosquito fish or a healthy mix of aquatic flora and fauna
284 L. K. Wang et al.

in the system. Wetland systems may prove impractical for large treatment plants due
to the large land requirements. They also may cause loss of water due to increases
in evapotranspiration.

6.2.5 Design Criteria

Natural or artificial marshes, swamps, bogs, shallow ponds, channels, or basins

could be used. Irrigation, harvesting, and processing equipment are optional.
Aquatic vegetation is usually locally acquired.
Design criteria are very site and project specific. Available data vary widely. The
values below refer to one type of constructed wetland system used as a tertiary pro-
cess on secondary effluent [2]:
(a) Detention time = 13 days.
(b) Land requirement = 8 acres/MG/day.
(c) Depth may vary with type of system, generally 1–5 ft.

6.2.6 Performance

The process appears reliable from mechanical and performance standpoints, subject
to seasonality of vegetation growth. Low operator attention is required if properly
designed.
Tables 6.2 and 6.3 illustrate the capacities of both natural and constructed wet-
lands for nutrient removal [4]. In test units and operating artificial marsh facilities
using various wastewater streams, the following removals have been reported for
secondary effluent treatment (10-day detention) [2]:
(a) BOD5, 80–95%
(b) TSS, 29–87%
(c) COD, 43–87%
(d) Nitrogen, 42–94% depending upon vegetative uptake and frequency of
harvesting
(e) Total P, 0–94% (high levels possible with warm climates and harvesting)
(f) Coliforms, 86–99%
(g) Heavy metals, highly variable depending on species
There is also evidence of reductions in wastewater concentrations of chlorinated
organics and pathogens, as well as pH neutralization without causing detectable
harm to the wetland ecosystem.
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 285

Table 6.2 Nutrient removal from natural wetlands. Source: U. S. EPA [4]
Percent reduction
Flow, m3/ Wetland NH3-­ NO3-­
Project day type TDPa N N TNb
Brillion Marsh, WI 757 Marsh 13 – 51 –
Houghton Lake, MI 379 Peatland 95 71 99c –
Wildwood, FL 946 Swamp/ 98 – – 90
Marsh
Concord, MA 2309 Marsh 47 58 20 –
Bellaire, MI 1,136d Peatland 88 – – 84
Coots Paradise, Town of Dundas, – Marsh 80 – – 60–
Ontario, Canada 70
Whitney Mobile Park, Home Park, FL ≈227 Cypress 91 – – 89
Dome
a
Total dissolved phosphorus
b
Total nitrogen
c
Nitrate and nitrite
d
May–November only

Table 6.3 Nutrient removal from constructed wetlands. Source: U. S. EPA [4]
Percent Hydraulic
Wetland BOD5, mg/L SS, mg/L reduction surface
loading
Flow, rate, m3/
Project m3/day Type Influent Effluent Influent Effluent BOD5 SS ha day
Listowel, 17 FWSa 56 10 111 8 82 93 –
Ontario [12]
Santee, CA – SFSb 118 30 57 5.5 75 90 –
[10]
Sidney, 240 SFS 33 4.6 57 4.5 86 92 –
Australia
[13]
Arcata, CA 11,350 FWS 36 13 43 31 64 28 907
Emmitsburg, 132 SFS 62 18 30 8.3 71 73 1543
MD
Gustine, CA 3785 FWS 150 24 140 19 84 86 412
a
Free water surface system
b
Subsurface flow system

Residuals are dependent upon the type of system and whether or not harvesting
is employed. Duckweed, for example, yields 50–60 lb/acre/day (dry weight) during
peak growing period to about half of this figure during colder months. For further
detailed information on wetland systems, the reader is referred to Refs. [19–23].
286 L. K. Wang et al.

6.3 Evapotranspiration System

6.3.1 Description

Evapotranspiration (ET) system is a means of on-site wastewater disposal that may

be utilized in some localities where site conditions preclude soil absorption.
Evaporation of moisture from the soil surface and/or transpiration by plants is the
mechanism of ultimate disposal. Thus, in areas where the annual evaporation rate
equals or exceeds the rate of annual added moisture from rainfall and wastewater
application, ET systems can provide a means of liquid disposal without danger of
surface or groundwater contamination.
If evaporation is to be continuous, at least three conditions must be met [2]:
(a) There must be a continuous supply of heat to meet the latent heat requirement,
approximately 590 cal/g of water evaporated at 15 °C.
(b) A vapor pressure gradient must exist between the evaporative surface and the
atmosphere to remove vapor by diffusion, convection, or both. Meteorological
factors, such as air temperature, humidity, wind velocity, and radiation, influ-
ence both energy supply and vapor removal.
(c) There must be a continuous supply of water to the evaporative surface. The soil
material must be fine textured enough to draw up the water from the saturated
zone to the surface by capillary action but not so fine as to restrict the rate of
flow to the surface.
Evapotranspiration is also influenced by vegetation on the disposal field and can
theoretically remove significant volumes of effluent in late spring, summer, and
early fall, particularly if large silhouette, good transpiring bushes and trees are
present.
A typical ET bed system (Fig. 6.3) consists of a 1½ to 3 ft depth of selected sand
over an impermeable plastic liner. A perforated plastic piping system with rock
cover is often used to distribute pretreated effluent in the bed. The bed may be
square shaped on relatively flatland or a series of trenches on slopes. The surface
area of the bed must be large enough for sufficient ET to occur to prevent the water
level in the bed from rising to the surface.
Beds are usually preceded by septic tanks or aerobic units to provide the
­necessary pretreatment. Given the proper subsurface conditions, systems can be
constructed to perform as both evapotranspiration and absorption beds. Nearly
three-fourths of all the ET beds in operation were designed to use both
disposal methods. Mechanical evaporators have been developed, but are not used at
full scale.
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 287

Fig. 6.3 Section through an evapotranspiration bed. Source: U. S. EPA [2]

6.3.2 Applications

There are estimated to be 4000–5000 year-round evapotranspiration beds in opera-

tion in the United States, particularly in the semiarid regions of the Southwest.
ET beds are used as an alternative to subsurface disposal in areas where these
methods are either undesirable due to groundwater pollution potential or not feasi-
ble due to certain geological or physical constraints of land. The ET system can also
be designed to supplement soil absorption for sites with slowly permeable soils. The
use of ET systems for summer homes extends the range of application, which is
otherwise limited by annual ET rates. Since summer evaporation rates are generally
higher and plants with high transpiration rates are in an active growing state, many
areas of the country can utilize ET beds for this seasonal application.

6.3.3 Limitations

The use of an evapotranspiration system is limited by climate and its effect on the
local ET rate. In practice, lined ET bed systems are generally limited to areas of the
country where pan evaporation exceeds annual rainfall by at least 24 in. The decrease
of ET in winter at middle and high latitudes greatly limits its use. Snow cover
reflects solar radiation, which reduces EF. In addition, when temperatures are below
freezing, more heat is required to change frozen water to vapor. When vegetation is
dormant, both transpiration and evaporation are reduced. An ET system requires a
large amount of land in most regions. Salt accumulation may eventually eliminate
vegetation and thus transpiration. Bed liner (where needed) must be kept watertight
to prevent the possibility of groundwater contamination. Therefore, proper
288 L. K. Wang et al.

construction methods should be employed to keep the liner from being punctured
during installation.

6.3.4 Design Criteria

Design of an evapotranspiration bed is based on the local annual weather cycle. The
total expected inflow based on household wastewater generation and rainfall rates is
compared with an average design evaporation value established from the annual
pattern. It is recommended to use a 10-year frequency rainfall rate to provide suffi-
cient bed surface area [2]. A mass balance is used to establish the storage require-
ments of the bed. Vegetative cover can substantially increase the ET rate during the
summer growing season, but may reduce evaporation during the non-growing sea-
son. Uniform sand in the size range of D50 of approximately 0.10 mm is capable of
raising water about 3 ft to the top of the bed. The polyethylene liner thickness is
typically greater than or equal to 10 mil. Special attention should be paid to storm-
water drainage to make sure that surface runoff is drained away from the bed prox-
imity by proper lot grading.

6.3.5 Performance

Performance is a function of climate conditions, volume of wastewater, and physi-

cal design of the system. Evapotranspiration (ET) is an effective and reliable means
of domestic wastewater disposal. An ET system that has been properly designed and
constructed is an efficient method for the disposal of pretreated wastewater and
requires a minimum of maintenance. Healthy vegetative covers are aesthetically
pleasing, and the large land requirement, although it limits the land use, does con-
serve the open space. Neither energy is required, nor is head loss of any value
incurred.

6.3.6 Costs

The following site-specific costs serve to illustrate the major components of an

evapotranspiration bed in Boulder, Colorado, United States, with an annual net ET
rate in the range of 0.04 gpd/ft2 [2]. A 200 gpd household discharge would require
a 2 ft deep bed with an area of approximately 5000 ft2. All costs have been adjusted
to 2020 US Dollars using the Cost Index for Utilities shown in Appendix 1 [24].
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 289

Construction cost (2020)

Building sewer with 1000 gal septic tank, design, and permit $2616
Excavation and hauling (375 yd3) $3767
Liner (5200 ft2) $2442
Distribution piping (625 ft) $1099
Sand (340 yd3) and gravel (38 yd3) $6557
Supervision and labor $1831
Total $18,312
Annual operation and maintenance cost (2020)
Pumping septage from septic tank (every 3–5 years) $17–73
Total $17–73

The 2020 construction cost for this particular system would be approximately
$3.66/ft2, which is consistent with a reported national range of $2.72–5.86/ft2. The
cost of an evapotranspiration bed is highly dependent upon local material and labor
costs. As shown, the cost of sand is a significant portion of the cost of the bed. The
restrictive sand size requirement makes availability and cost sensitive to location.
If an aerobic pretreatment unit is used instead of the septic tank, add $600–6000
to the 2020 construction cost and an amount of $218–750/year to the annual opera-
tion and maintenance cost.

6.4 Land Treatment: Rapid Rate System

The land-based technologies have been in use since the beginning of civilization.
Their greater value may be the use of the wastewater for beneficial return (agricul-
tural and recharge) in water-poor areas, as well as nitrogen control benefits. If nitro-
gen control benefits are desired, some key issues arise concerning the type of plant
crop with its growing and harvesting needs and/or the cycling of the water applica-
tion and restorative oxygenation resting periods. Native soils and climate add the
remaining variables.
Generally, the wastewater applications are cyclic in land-based technologies,
making some form of storage or land rotation mandatory to ensure the restorative
oxygenation derived from the resting period. Surface wastewater applications allow
additional beneficial soil aeration (plowing, tilling, and raking), which can become
mandatory for the heavily loaded systems after an elapsed season, or number of
loading cycles. Actual surface cleaning programs, to remove the plastic, rubber, and
other debris found in pretreated municipal wastewaters, also may be necessary,
although not at the frequency used for beneficial soil aeration.
In this and the following sections, detailed information on the four most common
land-based technologies will be provided. Subsurface, slow, and rapid infiltration
systems do not discharge to surface waters and conceptually may allow a more
relaxed nitrogen control standard in comparison to the overland flow system,
depending on local groundwater regulations.
290 L. K. Wang et al.

6.4.1 Description

Rapid rate infiltration was developed approximately 100 years ago and has remained
unaltered since then. It has been widely used for municipal and certain industrial
wastewaters throughout the world. Wastewater is applied to deep and permeable
deposits such as sand or sandy loam usually by distributing in basins (Fig. 6.4) or
infrequently by sprinkling and is treated as it travels through the soil matrix by fil-
tration, adsorption, ion exchange precipitation, and microbial action [25]. Most
metals are retained on the soil; many toxic organics are degraded or adsorbed. An
underdrainage system consisting of a network of drainage pipe buried below the
surface serves to recover the effluent, to control groundwater mounding, or to mini-
mize trespass of wastewater onto adjoining property by horizontal subsurface flow.
To recover renovated water for reuse or discharge, underdrains are usually inter-
cepted at one end of the field by a ditch. If groundwater is shallow, underdrains are
placed at or in the groundwater to remove the appropriate volume of water [2].
Thus, the designed soil depth, soil detention time, and underground travel distance
to achieve the desired water quality can be controlled. Effluent can also be recov-
ered by pumped wells.
Basins or beds are constructed by removing the fine-textured topsoil from which
shallow banks are constructed. The underlying sandy soil serves as the filtration
media. Underdrainage is provided by using plastic, concrete (sulfate resistant if
necessary), or clay tile lines. The distribution system applies wastewater at a rate
which constantly floods the basin throughout the application period of several hours
to a couple of weeks. The waste floods the bed and then drains uniformly away,
driving air downward through the soil and drawing fresh air from above. A cycle of
flooding and drying maintains the infiltration capacity of the soil material. Infiltration
diminishes slowly with time due to clogging. Full infiltration is readily restored by
occasional tillage of the surface layer and, when appropriate, removal of several
inches from the surface of the basin. Preapplication treatment to remove solids
improves distribution system reliability, reduces nuisance conditions, and may
reduce clogging rates. Common preapplication treatment practices include the
following:
(a) Primary treatment for isolated locations with restricted public access [26].
(b) Biological treatment for urban locations with controlled public access.
(c) Storage is sometimes provided for flow equalization and for nonoperating
periods.
Nitrogen removals are improved by [17, 27]:

Fig. 6.4 Flow diagram of land treatment using rapid rate system. Source: U. S. EPA [2]
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 291

(a) Establishing specific operating procedures to maximize denitrification

(b) Adjusting application cycles
(c) Supplying an additional carbon source
(d) Using vegetated basins (at low rates)
(e) Recycling portions of wastewater containing high nitrate concentrations
(f) Reducing application rates
Rapid rate infiltration systems require relatively permeable, sandy to loamy soils.
Vegetation is typically not used for nitrogen control purposes but may have value
for stabilization and maintenance of percolation rates. The application of algae-­
laden wastewater to rapid infiltration systems is not recommended because of clog-
ging considerations but could be considered with attendant additional tolerance for
surface maintenance, drying, and soil aeration needs.

6.4.2 Applications

Rapid infiltration is a simple wastewater treatment system that is [2]:

(a) Less land intensive than other land application systems and provides a means of
controlling groundwater levels and lateral subsurface flow.
(b) It provides a means of recovering renovated water for reuse or for discharge to
a particular surface water body.
(c) It is suitable for small plants where operator expertise is limited.
(d) It is applicable for primary and secondary effluent and for many types of indus-
trial wastes, including those from breweries, distilleries, paper mills, and wool
scouring plants [26, 28, 29].
In very cold weather, the ice layer floats atop the effluent and also protects the soil
surface from freezing. Generated residuals may require occasional removals of top
layer of soil. The collected material is disposed of on-site.

6.4.3 Limitations

The rapid infiltration process is limited by [2]:

(a) Soil type
(b) Soil depth
(c) The hydraulic capacity of the soil
(d) The underlying geology
(e) The slope of the land
Nitrate and nitrite removals are low unless special management practices are used.
292 L. K. Wang et al.

6.4.4 Design Criteria

The design criteria for the rapid rate system can be summarized as follows [2]:
(a) Field area, 3–56 acres/MG/day
(b) Application rate, 20–400 ft/year, 4–92 in./week
(c) BOD5 loading rate, 20–100 lb/acre/day
(d) Soil depth, 10–15 ft or more
(e) Soil permeability, 0.6 in./h or more
(f) Hydraulic loading cycle, 9 h to 2 weeks’ application period, 15 h to 2 weeks’
resting period
(g) Soil texture sands, sandy barns
(h) Basin size, 1–10 acres, at least 2 basins/site
(i) Height of dikes, 4 ft; underdrains, 6 or more ft deep
(j) Application techniques: flooding or sprinkling
(k) Preapplication treatment: primary or secondary
Designs can be developed that foster only nitrification or nitrification and denitrifi-
cation [17, 27]. Nitrification is promoted by low hydraulic loadings and short appli-
cation periods (1–2 days) followed by long drying periods (10–16 days).
Denitrification can vary from 0% to 80%. For significant denitrification, the applica-
tion period must be long enough to ensure depletion of the soil (and nitrate nitrogen)
oxygen. Higher denitrification values predictably track higher BOD/nitrogen ratios.
Enhancement may be promoted by recycling or by adding an external driving sub-
strate (methanol). Nitrogen elimination strategies also may reduce the drying period
by about half to yield lower overall nitrogen residuals with higher ammonium-­
nitrogen concentrations. Suggested loading cycles [25] to maximize infiltration
rates, nitrogen removal, and nitrification rates are given in Table 6.4.

6.4.5 Performance

The effluent quality is generally excellent where sufficient soil depth exists and is
not normally dependent on the quality of wastewater applied within limits. Well-­
designed systems provide for high-quality effluent that may meet or exceed primary
drinking water standards. Percent removals for typical pollution parameters are [2]:
(a) BOD5, 95–99%
(b) TSS, 95–99%
(c) Total N, 25–90%
(d) Total P, 0–90% until flooding exceeds adsorptive capacity [30]
(e) Fecal coliform, 99.9–99.99 + % [31]
The process is extremely reliable, as long as sufficient resting periods are provided.
However, it has a potential for contamination of groundwater by nitrates. Heavy
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 293

Table 6.4 Loading cycles for high rate infiltration systems. Source: U. S. EPA [25]
Applied Application period, Drying period,
Loading cycle objective wastewater Season daya day
Maximize infiltration Primary Summer 1–2 5–7
rates Winter 1–2 7–12
Secondary Summer 1–3 4–5
Winter 1–3 5–10
Maximize nitrogen Primary Summer 1–2 10–14
removal Winter 1–2 12–16
Secondary Summer 7–9 10–15
Winter 9–12 12–16
Maximize nitrification Primary Summer 1–2 5–7
Winter 1–2 7–12
Secondary Summer 1–3 4–5
Winter 1–3 5–10
a
Regardless of season or cycle objective, application periods for primary effluent should be limited
to 1–2 days to prevent excessive soil clogging

metals could be eliminated by pretreatment techniques as necessary. Monitoring for

metals and toxic organics is needed where they are not removed by pretreatment.
The process requires long-term commitment of relatively large land areas, although
small by comparison to other land treatment systems [32, 33].

6.4.6 Costs

The construction and operation and maintenance costs are shown in Figs. 6.5 and
6.6, respectively [2]. The costs are based on 1973 (Utilities Index = 149.36, USEPA
Index 194.2, ENR Index = 1850) figures. To obtain the values in terms of the present
2020 US Dollars, using the Cost Index for Utilities (Appendix 1), multiply the costs
by a factor of 5.81 [24].
Assumptions applied in preparing the costs given in Figs. 6.5 and 6.6:
(a) Application rate, 182 ft/year.
(b) Construction costs include field preparations (removal of brush and trees) for
multiple unit infiltration basins with 4 ft dike formed from native excavated
material, and storage is not assumed necessary.
(c) Drain pipes buried 6–8 ft with 400 ft spacing, interception ditch along length of
field, and weir for control of discharge; gravel service roads and 4 ft stock fence
around perimeter.
(d) O & M cost includes inspection and unclogging of drain pipes at outlets; annual
tilling of infiltration surface and major repair of dikes after 10 years; high pres-
sure jet cleaning of drain pipes every 5 years, annual cleaning of interceptor
ditch, and major repair of ditches, fences, and roads after 10 years.
294 L. K. Wang et al.

Fig. 6.5 Construction costs for rapid rate system. Source: U. S. EPA [2]

(e) Costs of pretreatment monitoring wells, land, and transmission to and from
pretreatment facility not included.

6.5 Land Treatment: Slow Rate System

6.5.1 Description

Slow rate land treatment system represents the predominant municipal land treat-
ment practice in the United States. In this process, wastewater is applied by sprin-
kling to vegetated soils that are slow to moderate in permeability (clay barns to
sandy barns) and is treated as it travels through the soil matrix by filtration, adsorp-
tion, ion exchange, precipitation, microbial action, and plant uptake (Fig. 6.7). An
underdrainage system consisting of a network of drainage pipe buried below the
surface serves to recover the effluent, to control groundwater, or to minimize tres-
pass of leachate onto adjoining property by horizontal subsurface flow. To recover
renovated water for reuse or discharge, underdrains are usually intercepted at one
end of the field by a ditch. Underdrainage for groundwater control is installed as
needed to prevent waterlogging of the application site or to recover the renovated
water for reuse. Proper crop management also depends on the drainage conditions.
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 295

Fig. 6.6 Operation and maintenance costs of rapid rate system. Source: U. S. EPA [2]

Fig. 6.7 Flow diagram of land treatment using slow rate system. Source: U. S. EPA [2]

Sprinklers can be categorized as hand moved, mechanically moved, and permanent

set, the selection of which includes the following considerations [2]:
(a) Field conditions (shape, slope, vegetation, and soil type)
(b) Climate
(c) Operating conditions
(d) Economics
Vegetation is a vital part of the process and serves to extract nutrients, reduce
erosion, and maintain soil permeability. Considerations for crop selection include:
(a) Suitability to local climate and soil conditions
(b) Consumptive water use and water tolerance
(c) Nutrient uptake and sensitivity to wastewater constituents
(d) Economic value and marketability
(e) Length of growing season
(f) Ease of management
(g) Public health regulations
296 L. K. Wang et al.

Common preapplication treatment practices include the following:

(a) Primary treatment for isolated locations with restricted public access and when
limited to crops not for direct human consumption
(b) Biological treatment plus control of coliform to 1000 MPN/100 mL for agricul-
tural irrigation, except for human food crops to be eaten raw
(c) Secondary treatment plus disinfection to 200 MPN/100 mL fecal coliform for
public access areas (parks)
Wastewaters high in metal content should be pretreated to avoid plant and soil con-
tamination. Table 6.5 shows the wastewater constituents that have potential adverse
effects on crops [25]. Forestland irrigation is more suited to cold weather operation,
since soil temperatures are generally higher, but nutrient removal capabilities are
less than for most field crops.

6.5.2 Applications

Slow rate systems produce the best results of all the land treatment systems.
Advantages of sprinkler application over gravity methods include [34]:

Table 6.5 Potential adverse effects of wastewater constituents on crops. Source: U. S. EPA [25]
Constituent level
Problem and related No Increasing Severe
constituent problem problems problems Crops affected
Salinity (ECW), <0.75 0.75–3.0 >3.0 Crops in arid climates only (see
mmho/cm Table 9.4)
Specific ion toxicity from root absorption
 Boron, mg/L <0.5 0.5–2 2.0–10.0 Fruit and citrus trees—0.5–1.0 mg/L;
field crops—1.0–2.0 mg/L;
grasses—2.0–10.0 mg/L
 Sodium, <3 3.0–9.0 >9.0 Tree crops
adj-SARa
 Chloride, mg/L <142 142–355 >355 Tree crops
Specific ion toxicity from foliar absorption
 Sodium, mg/L <69 >69 – Field and vegetable crops under
 Chloride, mg/L <106 >106 – sprinkler application
Miscellaneous
 NH4-N + NO3-N, <5 5–30 30 Sugarbeets, potatoes, cotton, grains
mg/L
 HCO3, mg/L <90 90–520 >520 Fruit
 pH, units 6.5–8.4 4.2–5.5 <4.2 and Most crops
> 8.5
a
Adjusted sodium adsorption ratio
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 297

(a) More uniform distribution of water and greater flexibility in range of applica-
tion rates
(b) Applicability to most crops
(c) Less susceptibility to topographic constraints
(d) Reduced operator skill and experience requirements
Underdrainage provides a means of recovering renovated water for reuse or for
discharge to a particular surface water body when dictated by senior water rights
and a means of controlling groundwater. The system also provides the following
benefits:
(a) An economic return from the use of water and nutrients to produce marketable
crops for forage
(b) Water and nutrient conservation when utilized for irrigating landscaped areas

6.5.3 Limitations

The slow rate process is limited by [2]:

(a) Soil type and depth
(b) Topography
(c) Underlying geology
(d) Climate
(e) Surface and groundwater hydrology and quality
(f) Crop selection
(g) Land availability
Crop water tolerances, nutrient requirements, and the nitrogen removal capacity of
the soil-vegetation complex limit hydraulic loading rate [35]. Climate affects grow-
ing season and will dictate the period of application and the storage requirements.
Application ceases during period of frozen soil conditions. Once in operation, infil-
tration rates can be reduced by sealing of the soil. Limitations to sprinkling include
adverse wind conditions and clogging of nozzles. Slopes should be less than 15% to
minimize runoff and erosion. Pretreatment for removal of solids and oil and grease
serves to maintain reliability of sprinklers and to reduce clogging. Many states have
regulations regarding preapplication disinfection, minimum buffer areas, and con-
trol of public access for sprinkler systems.
The process requires long-term commitment of large land area, i.e., largest land
requirement of all land treatment processes [36]. Concerns with aerosol carriage of
pathogens, potential vector problems, and crop contamination have been identified,
but are generally controllable by proper design and management.
298 L. K. Wang et al.

6.5.4 Design Criteria

The design criteria for the slow rate system can be summarized as follows [2]:
(a) Field area, 56–560 acres/MG/day
(b) Application rate, 2–20 ft/year, 0.5–4 in./week
(c) BOD5 loading rate, 0.2–5 lb/acre/day
(d) Soil depth, 2–5 ft or more
(e) Soil permeability, 0.06–2.0 in./h
(f) Minimum preapplication treatment, primary
(g) Lower temperature limit, 25 °F
(h) Particle size of solids, less than one-third of the sprinkler nozzle diameter
(i) Underdrains, 4–8 in. diameter, 4–10 ft deep, 50–500 ft apart; pipe material,
plastic, concrete (sulfate resistant, if necessary), or clay

6.5.5 Performance

Effluent quality is generally excellent and consistent regardless of the quality of

wastewater applied [37]. Percent removals for typical pollution parameters when
wastewater is applied through more than 5 ft of unsaturated soil are:
(a) BOD5, 90–99 + %
(b) TSS, 90–99 + %
(c) Total N, 50–95% depending on N uptake of vegetation
(d) Total P, 80–99%, until adsorptive capacity is exceeded [38]
(e) Fecal coliform, 99.99 + % when applied levels are more than 10 MPN/100 mL
This treatment is capable of achieving the highest degree of nitrogen removal.
Typically, nitrogen losses due to denitrification (15–25%), ammonia volatilization
(0–10%), and soil immobilization (0–25%) supplement the primary nitrogen
removal mechanism by the crop [17]. The balance of the nitrogen passes to the per-
colate. Typical design standards require preservation of controlling depths to
groundwater and establishing nitrogen limits in either the percolate or groundwater
as it leaves the property site. Nitrogen loading to the groundwater is often the con-
trolling consideration in the design. For further detailed information on slow rate
infiltration systems, the reader is referred to Refs. [39–44].

6.5.6 Costs

The construction and operation and maintenance costs are shown in Figs. 6.8 and
6.9, respectively [2]. The costs are based on 1973 (Utilities Index = 149.36, USEPA
Index 194.2, ENR Index = 1850) figures. To obtain the values in terms of the present
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 299

Fig. 6.8 Construction cost

of slow rate system.
Source: U. S. EPA [2]

2020 US Dollars, using the Cost Index for Utilities (Appendix 1), multiply the costs
by a factor of 5.81 [24].
Assumptions applied in preparing the costs given in Figs. 6.8 and 6.9:
(a) Yearly average application rate: 0.33 in./day.
(b) Energy requirements: Solid set spray distribution requires 2100 kwh/year/ft of
TDH/MG/day capacity. Center pivot spraying requires an additional 0.84 × 106
kwh/year/acre (based on 3.5 days/week operation) for 1 MG/day or larger facil-
ities (below 1 MG/day, additional power = 0.84–1.35 × 106 kwh/year/acre).
(c) Clearing costs are for brush with few trees using bulldozer-type equipment.
(d) Solid set spraying construction costs include lateral spacing, 100 ft; sprinkler
spacing, 80 ft along laterals; 5.4 sprinklers/acre; application rate, 0.20 in./h;
16.5 gpm flow to sprinklers at 70 psi; flow to laterals controlled by hydrauli-
cally operated automatic valves; laterals buried 18 in.; mainlines buried 36 in.;
all pipe 4 in. diameter and smaller is PVC; all larger pipe is asbestos cement
(total dynamic head = 150 ft).
(e) Center pivot spraying construction costs include heavy-duty center pivot rig
with electric drive; multiple units for field areas over 40 acres; maximum area
per unit, 132 acres; distribution pipe is buried 3 ft deep.
(f) Underdrains are spaced 250 ft between drain pipes. Drain pipes are buried
6–8 ft deep with interception ditch along length of field and weir for control of
discharge.
(g) Distribution pumping construction costs include structure built into dike of
storage reservoir; continuously cleaned water screens; pumping equipment
with normal standby facilities; piping and valves within structure; controls and
electrical work.
300 L. K. Wang et al.

Fig. 6.9 Operation and maintenance cost of slow rate system. Source: U. S. EPA [2]

(h) Labor costs include inspection and unclogging of drain pipes at outlets and dike
maintenance.
(i) Material costs include for solid set spraying: replacement of sprinklers and air
compressors for valve controls after 10 years; for center pivot spraying, minor
repair parts and major overhaul of center pivot rigs after 10 years; high pressure
jet cleaning of drain pipes every 5 years, annual cleaning of interceptor ditch,
and major repair of ditches after 10 years; distribution pumping repair work
performed by outside contractor and replacement parts; scraping and patching
of storage receiver liner every 10 years.
(j) Storage for 75 days is included; 15 ft dikes (12 ft wide at crest) are formed from
native materials (inside slope 3:1, outside 2:1); rectangular shape on level
ground; 12 ft water depth; multiple cells for more than 50 acre size; asphaltic
lining; 9 in. riprap on inside slope of dikes.
(k) Cost of pretreatment, monitoring wells, land, and transmission to and from land
treatment facility not included.
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 301

6.6 Land Treatment: Overland Flow System

6.6.1 Description

Wastewater treatment using the overland flow system is relatively new. It is now
extensively used in the food processing industry. Very few municipal plants are in
operation and most are in warm, dry areas. A flow diagram of the system is shown
in Fig. 6.10. Wastewater is applied over the upper reaches of sloped terraces and is
treated as it flows across the vegetated surface to runoff collection ditches. The
wastewater is renovated by physical, chemical, and biological means as it flows in a
thin film down the relatively impermeable slope.
A secondary objective of the system is for crop production. Perennial grasses
(reed canary, Bermuda, redtop, tall fescue, and Italian rye) with long growing sea-
sons, high moisture tolerance, and extensive root formation are best suited to over-
land flow. Harvested grass is suitable for cattle feed. Biological oxidation,
sedimentation, and grass filtration are the primary removal mechanisms for organics
and suspended solids. Nitrogen removal is attributed primarily to nitrification/deni-
trification and plant uptake. Loading rates and cycles are designed to maintain active
microorganism growth on the soil surface. The operating principles are similar to a
conventional trickling filter with intermittent dosing. The rate and length of applica-
tion are controlled to minimize severe anaerobic conditions that result from over-
stressing the system. The resting period should be long enough to prevent surface
ponding, yet short enough to keep the microorganisms in an active state. Surface
methods of distribution include the use of gated pipe or bubbling orifice. Gated
surface pipe, which is attached to aluminum hydrants, is aluminum pipe with mul-
tiple outlets. Control of flow is accomplished with slide gates or screw adjustable
orifices at each outlet. Bubbling orifices are small diameter outlets from laterals
used to introduce flow. Gravel may be necessary to dissipate energy and ensure
uniform distribution of water from these surface methods. Slopes must be steep
enough to prevent ponding of the runoff, yet mild enough to prevent erosion and
provide sufficient detention time for the wastewater on the slopes. Slopes must have
a uniform cross slope and be free from gullies to prevent channeling and allow uni-
form distribution over the surface. The network of slopes and terraces that make up
an overland system may be adapted to natural rolling terrain. The use of this type of

Fig. 6.10 Flow diagram of land treatment using overland flow system. Source: U. S. EPA [2]
302 L. K. Wang et al.

terrain will minimize land preparation costs. Storage must be provided for nonoper-
ating periods. Runoff is collected in open ditches. When unstable soil conditions are
encountered or flow velocities are erosive, gravity pipe collection systems may be
required. Common preapplication practices include the following: screening or
comminution for isolated sites with no public access and screening or comminution
plus aeration to control odors during storage or application for urban locations with
no public access [45, 46]. Wastewaters high in metal content should be pretreated to
avoid soil and plant contamination.
A common method of distribution is with sprinklers. Recirculation of collected
effluent is sometimes provided and/or required. Secondary treatment prior to over-
land flow permits reduced (as much as two-thirds reduction) land requirements.
Effluent disinfection is required where stringent fecal coliform criteria exist.

6.6.2 Application

Because overland flow is basically a surface phenomenon, soil clogging is not a

problem. High BOD5 and suspended solids removals have been achieved with the
application of raw comminuted municipal wastewater. Thus, preapplication treat-
ment is not a prerequisite where other limitations are not operative. Depth to ground-
water is less critical than with other land systems. It also provides the following
benefits: an economic return from the reuse of water and nutrients to produce mar-
ketable crops or forage and a means of recovering renovated water for reuse or
discharge. This type of applications is preferred for gently sloping terrain with
impermeable soils.

6.6.3 Limitations

The process is limited by soil type, crop water tolerances, climate, and slope of the
land. Steep slopes reduce travel time over the treatment area and thus treatment
efficiency. Flatland may require extensive earthwork to create slopes. Ideally, slope
should be 2–8%. High flotation tires are required for equipment. Cost and impact of
the earthwork required to obtain terraced slopes can be major constraints. Application
is restricted during rainy periods and stopped during very cold weather [47]. Many
states have regulations regarding preapplication disinfection, minimum buffer
zones, and control of public access.

6.6.4 Design Criteria

The design criteria for the overland flow system can be summarized as follows [2]:
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 303

(a) Field area required, 35–100 acres/MG/day

(b) Terraced slopes, 2–8%
(c) Application rate, 11–32 ft/year, 2.5–16 in./week
(d) BOD5 loading rate, 5–50 lb/acre/day
(e) Soil depth, sufficient to form slopes that are uniform and to maintain a vegeta-
tive cover
(f) Soil permeability, 0.2 in./h or less
(g) Hydraulic loading cycle, 6–8 h application period, 16–181 weeks’ resting period
(h) Operating period, 5–6 days/week
(i) Soil texture clay and clay loams
Below are representative application rates for 2–8% sloped terraces:
in./week Pretreatment Terrace length, ft
2.5–8 Untreated or primary 150
6–16 Lagoon or secondary 120

Generally, 40–80% of applied wastewater reaches collection structures, lower

percent in summer and higher in winter (southwest data). Table 6.6 shows the
required pretreatment and allowed application and hydraulic rates [48].

6.6.5 Performance

Percent removals for comminuted or screened municipal wastewater over about

150 ft of 2–6% slope:
(a) BOD5, 80–95%
(b) Suspended solids, 80–95%
(c) Total N, 75–90%
(d) Total P, 30–60%
(e) Fecal coliform, 90–99.9%
The addition of alum (Al2(SO4)3), ferric chloride (FeC13), or calcium carbonate
(CaCO3) prior to application will increase phosphorus removals.

Table 6.6 Design loadings for overland flow systems. Source: U. S. EPA [48]
Preapplication treatment Application rate m3/h m Hydraulic loading rate cm/day
Screening/primary 0.07–0.12a 2.0–7.0b
Aerated cell (1 day detention) 0.08–0.14 2.0–8.5
Wastewater treatment pondc 0.09–0.15 2.5–9.0
Secondaryd 0.11–0.17 3.0–10.0
a
m3/h m × 80.5 = gal/h ft
b
cm/day × 0.394 = in./day
c
Does not include removal of algae
d
Recommended only for upgrading existing secondary treatment
304 L. K. Wang et al.

Little attempt has been made to design optimized overland flow systems with a
specific objective of nitrogen control. Their performance depends on the same fun-
damental issues: nitrification-denitrification, ammonia volatilization, and harvest-
ing of crops. When measured, overland flow systems designed for secondary
treatment often reveal less than 10 mg/L total nitrogen [49]. For further detailed
information on overland flow systems, the reader is referred to Refs. [12, 50–52].

6.6.6 Costs

The construction and operation and maintenance costs are shown in Figs. 6.11 and
6.12, respectively [2]. The costs are based on 1973 (Utilities Index = 149.36, EPA
Index 194.2, ENR Index = 1850) figures. To obtain the values in terms of the present
2020 US Dollars, using the Cost Index for Utilities (Appendix 1), multiply the costs
by a factor of 5.81 [24].
Assumptions applied in preparing the costs given in Figs. 6.11 and 6.12:
(a) Storage for 75 days included.
(b) Site cleared of brush and trees using bulldozer-type equipment; terrace con-
struction: 175–250 ft wide with 2.5% slope (1400 yd/acre of cut). Costs include
surveying, earthmoving, finish grading, ripping two ways, disking, land plan-
ning, and equipment mobilization.

Fig. 6.11 Construction cost of overland flow treatment system. Source: U. S. EPA [2]
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 305

Fig. 6.12 Operation and maintenance cost of overland flow treatment system. Source: U. S. EPA [2]

(c) Distribution system: application rate, 0.064 in./h; yearly average rate of 3 in./
week (8 h/day; 6 days/week); flow to sprinklers, 13 gpm at 50 psi; laterals 70 ft
from top of terrace, buried 18 in.; flow to laterals controlled by hydraulically
operated automatic valves; mainlines buried 36 in.; all pipe 4 in. diameter and
smaller is PVC; all larger pipe is asbestos cement.
(d) Open ditch collection: network of unlined interception ditches sized for a 2 in./h
storm; culverts under service roads; concrete drop structures at 1000 ft intervals.
(e) Gravity pipe collection: network of gravity pipe interceptors with inlet/man-
holes every 250 ft along submains; storm runoff is allowed to pond at inlets;
each inlet/manhole serves 1000 ft of collection ditch; manholes every 500 ft
along interceptor mains.
(f) O & M cost includes replacement of sprinklers and air compressors for valve
controls after 10 years and either biannual cleaning of open ditches with major
repair after 10 years or the periodic cleaning of inlets and normal maintenance
of gravity pipe and also includes dike maintenance and scraping and patching
of storage basin liner every 10 years.
(g) Costs for pretreatment, land, transmission to site, disinfection, and service
roads and fencing not included.
306 L. K. Wang et al.

6.7 Subsurface Infiltration

Subsurface infiltration systems are capable of producing a high degree of treatment;

with proper design, they can provide a nitrified effluent, and denitrification can be
achieved under certain circumstances. Keys to their success are the adequacy of the
initial gravel infiltration zone for solids capture and the following unsaturated zone
of native or foreign soils. Failure to provide an oxygenated environment by either
resting or conservative loadings can lead to failure. Denitrification under gravity
loading is likely to be small, but may be improved through pressure/gravity dosing
concepts of liquid application to the trenches [53].
Subsurface infiltration wastewater management practices are embodied in the
horizontal leach fields that routinely serve almost one-third of the US population
that use more than 20 million septic tanks in their individual non-sewered establish-
ments and homes [2]. In recent years, they have also been advanced for collective
service in small isolated communities.

6.7.1 Description

A septic tank followed by a soil absorption field is the traditional on-site system for
the treatment and disposal of domestic wastewater from individual households or
establishments. The system consists of a buried tank where wastewater is collected
and scum, grease, and settleable solids are removed by gravity separation and a
subsurface drainage system where clarified effluent percolates into the soil. Precast
concrete tanks with a capacity of 1000 gallons are commonly used for house sys-
tems. Solids are collected and stored in the tank, forming sludge and scum layers.
Anaerobic digestion occurs in these layers, reducing the overall volume. Effluent is
discharged from the tank to one of three basic types of subsurface systems, absorp-
tion field [53], seepage bed [53, 54], or seepage pits [55]. Sizes are usually deter-
mined by percolation rates, soil characteristics, and site size and location.
Distribution pipes are laid in a field of absorption trenches to leach tank effluent
over a large area (Fig. 6.13). Required absorption areas are dictated by state and

Fig. 6.13 Septic tank absorption field. Source: U. S. EPA [2]

6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 307

local codes. Trench depth is commonly about 24 in. to provide minimum gravel
depth and earth cover. Clean, graded gravel or similar aggregate, varying in size
from ½ to 2½ in., should surround the distribution pipe and extend at least 2 in.
above and 6 in. below the pipe. The maintenance of at least a 2 ft separation between
the bottom of the trench and the high water table is required to minimize groundwa-
ter contamination. Piping typically consists of agricultural drain tile, vitrified clay
sewer pipe, or perforated, nonmetallic pipe. Absorption systems having trenches
wider than 3 ft are referred to as seepage beds. Given the appropriate soil conditions
(sandy soils), a wide bed makes more efficient use of available land than a series of
long, narrow trenches.
Many different designs may be used in laying out a subsurface disposal field. In
sloping areas, serial distribution can be employed with absorption trenches by
arranging the system so that each trench is utilized to its capacity before liquid flows
into the succeeding trench. A dosing tank can be used to obtain proper wastewater
distribution throughout the disposal area and give the absorption field a chance to
rest or dry out between dosings. Providing two separate alternating beds is another
method used to restore the infiltrative capacity of a system. Aerobic units may be
substituted for septic tanks with no changes in soil absorption system requirements.
In areas where problem soil conditions preclude the use of subsurface trenches
or seepage beds, mounds can be installed (Fig. 6.14) to raise the absorption field
above ground, provide treatment, and distribute the wastewater to the underlying
soil over a wide area in a uniform manner [2, 56, 57]. A pressure distribution

Fig. 6.14 Septic tank mound absorption field. Source: U. S. EPA [2]
308 L. K. Wang et al.

network should be used for uniform application of clarified tank effluent to the
mound. A subsurface chamber can be installed with a pump and high water alarm to
dose the mound through a series of perforated pipes. Where sufficient head is avail-
able, a dosing siphon may be used. The mound must provide an adequate amount of
unsaturated soil and spread septic tank effluent over a wide enough area so that
distribution and purification can be effected before the water table is reached.
The mound system requires more space and periodic maintenance than the con-
ventional subsurface disposal system, along with higher construction costs. The
system cannot be installed on steep slopes, nor over highly (120 mm/in.) imperme-
able subsurface. Seasonal high groundwater must be deeper than 2 ft to prevent
surfacing at the edge of the mound [2]. An alternative to the mound system is a new
combined distribution and pretreatment unit to precede the wastewater application
to the subsurface infiltration systems [58]. The new system is based on pumping of
septic tank effluent to one or more units filled with lightweight clay aggregates. The
wastewater is distributed evenly over the 2.3 m2 surface of the pretreatment filter.
The filter effluent is then applied to the subsurface infiltration system.

6.7.2 Applications

Subsurface infiltration systems for the disposal of septic tank effluents are used
primarily in rural and suburban areas where economics are favorable. Properly
designed and installed systems require a minimum of maintenance and can operate
in all climates.

6.7.3 Limitations

The use of subsurface effluent disposal fields is dependent on the following factors
and conditions [2]:
(a) Soil and site conditions
(b) The ability of the soil to absorb liquid
(c) Depth to groundwater
(d) Nature of and depth to bedrock
(e) Seasonal flooding
(f) Distance to well or surface water
A percolation rate of 60 mm/in. is often used as the lower limit of permeability. The
limiting value for seasonal high groundwater should be 2 ft below the bottom of the
absorption field. When a soil system loses its capacity to absorb septic tank effluent,
there is a potential for effluent surfacing, which often results in odors and, possibly,
health hazards.
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 309

Table 6.7 Required areas of subsurface infiltration absorption fields. Source: U. S. EPA [2]
Percolation rate, mm/in. Required area per bedroom, ft2
1 or less 70
3 100
5 125
10 165
15 190
30 250
45 300
60 330

6.7.4 Design Criteria

Absorption area requirements for individual residences are given in Table 6.7. The
area required per bedroom is a function of the percolation rate; the higher the rate,
the smaller is the required area [2].
The design criteria for the mound system are as follows [2, 56, 57]: design flow
75 gal/person/day, 150 gal/bedroom/day; basal area based on percolation rates up to
120 mm/in.; mound height at center approximately 3.5–5 ft; pump (centrifugal) that
must accommodate approximately 30 gpm at required total dynamic head (TDH).
Properly designed, constructed, and operated septic tank systems have demon-
strated an efficient and economical alternative to public sewer systems, particularly
in rural and sparsely developed areas. System life for properly sited, designed,
installed, and maintained systems may equal or exceed 20 years.

6.7.5 Performance

Performance is a function of the following factors [2]:

(a) Design of the system components
(b) Construction techniques employed
(c) Rate of hydraulic loading
(d) Area geology and topography
(e) Physical and chemical composition of the soil mantle
(f) Care given to periodic maintenance
Pollutants are removed from the effluent by natural adsorption and biological pro-
cesses in the soil zone adjacent to the field. BOD, TSS, bacteria, and viruses, along
with heavy metals and complex organic compounds, are adsorbed by soil under
proper conditions. However, chlorides and nitrates may readily penetrate coarser,
aerated soils to groundwater.
310 L. K. Wang et al.

Leachate can contaminate groundwater when pollutants are not effectively

removed by the soil system. In many well-aerated soils, significant densities of
homes with septic tank-soil absorption systems have resulted in increasing nitrate
content of the groundwater. Soil clogging may result in surface ponding with poten-
tial aesthetic and public health problems. The sludge and scum layers accumulated
in a septic tank must be removed every 3–5 years.
For further detailed information on subsurface infiltration systems and all other
natural systems for treating agricultural wastes, the readers are referred to additional
references [59–72].

Glossary of Emerging Natural Waste Systems [69, 70]

Evapotranspiration system Evapotranspiration (ET) system is a means of on-site

wastewater disposal that may be utilized in some localities where site conditions
preclude soil absorption. Evaporation of moisture from the soil surface and/or
transpiration by plants is the mechanism of ultimate disposal. Thus, in areas
where the annual evaporation rate equals or exceeds the rate of annual added
moisture from rainfall and wastewater application, ET systems can provide a
means of liquid disposal without danger of surface or groundwater contamination.
Overland flow land treatment system Wastewater treatment using the overland
flow system is relatively new. It is now extensively used in the food processing
industry. Very few municipal plants are in operation and most are in warm, dry
areas. Wastewater is applied over the upper reaches of sloped terraces and is
treated as it flows across the vegetated surface to runoff collection ditches. The
wastewater is renovated by physical, chemical, and biological means as it flows
in a thin film down the relatively impermeable slope. A secondary objective of
the system is for crop production. Perennial grasses (reed canary, Bermuda, red-
top, tall fescue, and Italian rye) with long growing seasons, high moisture toler-
ance, and extensive root formation are best suited to overland flow. Harvested
grass is suitable for cattle feed. Biological oxidation, sedimentation, and grass
filtration are the primary removal mechanisms for organics and suspended solids.
Nitrogen removal is attributed primarily to nitrification/denitrification and plant
uptake. Loading rates and cycles are designed to maintain active microorganism
growth on the soil surface. The operating principles are similar to a conventional
trickling filter with intermittent dosing. The rate and length of application are
controlled to minimize severe anaerobic conditions that result from overstressing
the system. The resting period should be long enough to prevent surface ponding,
yet short enough to keep the microorganisms in an active state. Surface methods
of distribution include the use of gated pipe or bubbling orifice.
Rapid rate land treatment system Rapid rate infiltration was developed approxi-
mately 100 years ago and has remained unaltered since then. It has been widely
used for municipal and certain industrial wastewaters throughout the world.
Wastewater is applied to deep and permeable deposits such as sand or sandy
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 311

loam usually by distributing in basins or infrequently by sprinkling and is treated

as it travels through the soil matrix by filtration, adsorption, ion exchange precip-
itation, and microbial action. Most heavy metals are retained on the soil; many
toxic organics are degraded or adsorbed. An underdrainage system consisting
of a network of drainage pipe buried below the surface serves to recover the
effluent, to control groundwater mounding, or to minimize trespass of wastewa-
ter onto adjoining property by horizontal subsurface flow. To recover renovated
water for reuse or discharge, underdrains are usually intercepted at one end of
the field by a ditch. If groundwater is shallow, underdrains are placed at or in the
groundwater to remove the appropriate volume of water. Thus, the designed soil
depth, soil detention time, and underground travel distance to achieve the desired
water quality can be controlled. Effluent can also be recovered by pumped wells.
Slow rate land treatment system Slow rate land treatment system represents
the predominant municipal land treatment practice in the United States. In this
process, wastewater is applied by sprinkling to vegetated soils that are slow to
moderate in permeability (clay barns to sandy barns) and is treated as it trav-
els through the soil matrix by filtration, adsorption, ion exchange, precipitation,
microbial action, and plant uptake. An underdrainage system consisting of a
network of drainage pipe buried below the surface serves to recover the efflu-
ent, to control groundwater, or to minimize trespass of leachate onto adjoining
property by horizontal subsurface flow. To recover renovated water for reuse or
discharge, underdrains are usually intercepted at one end of the field by a ditch.
Underdrainage for groundwater control is installed as needed to prevent water-
logging of the application site or to recover the renovated water for reuse. Proper
crop management also depends on the drainage conditions. Sprinklers can be
categorized as hand moved, mechanically moved, and permanent set.
Water hyacinth aquaculture treatment system Aquaculture or the production of
aquatic organisms (both flora and fauna) under controlled conditions has been
practiced for centuries, primarily for the generation of food, fiber, and fertilizer.
The water hyacinth (Eichhornia crassipes) appears to be the most promising
organism for wastewater treatment and has received the most attention. Other
organisms, such as duckweed, seaweed, midge larvae, alligator weeds, and a host
of other organisms, are also used. Water hyacinths are large fast-growing float-
ing aquatic plants with broad, glossy green leaves and light lavender flowers. A
native of South America, water hyacinths are found naturally in waterways, bay-
ous, and other backwaters throughout the South. Insects and disease have little
effect on the hyacinth, and they thrive in raw, as well as partially treated, waste-
water. Wastewater treatment by water hyacinths is accomplished by passing the
wastewater through a hyacinth-covered basin, where the plants remove nutrients,
BOD5, TSS, heavy metals, etc. Batch treatment and flow-through systems, using
single and multiple cell units, are possible. Hyacinths harvested from these sys-
tems have been investigated as a fertilizer/soil conditioner after composting, ani-
mal feed, and a source of methane when anaerobically digested.
Wetland aquaculture treatment system Aquaculture-wetland systems for waste-
water treatment include natural and artificial wetlands as well as other aquatic
312 L. K. Wang et al.

systems involving the production of algae and higher plants (both submerged
and emergent), invertebrates, and fish. Natural wetlands, both marine and fresh-
water, have inadvertently served as natural waste treatment systems for centu-
ries; however, in recent years, marshes, swamps, bogs, and other wetland areas
have been successfully utilized as managed natural “nutrient sinks” for polish-
ing partially treated effluents under relatively controlled conditions. Constructed
wetlands can be designed to meet specific project conditions while providing
new wetland areas that also improve available wildlife wetland habitats and the
other numerous benefits of wetland areas. Managed plantings of reeds (e.g.,
Phragmites spp.) and rushes (e.g., Scirpus spp. and Schoenoplectus spp.) as well
as managed natural and constructed marshes, swamps, and bogs have been dem-
onstrated to reliably provide pH neutralization and reduction of nutrients, heavy
metals, organics, BOD5, COD (chemical oxygen demand), TSS, fecal coliforms,
and pathogenic bacteria.

Appendix 1: US Yearly Average Cost Index for Utilities [24]

Year Index Year Index

1967 100 1995 439.72
1968 104.83 1996 445.58
1969 112.17 1997 454.99
1970 119.75 1998 459.40
1971 131.73 1999 460.16
1972 141.94 2000 468.05
1973 149.36 2001 472.18
1974 170.45 2002 486.16
1975 190.49 2003 497.40
1976 202.61 2004 563.78
1977 215.84 2005 605.47
1978 235.78 2006 645.52
1979 257.20 2007 681.88
1980 277.60 2008 741.36
1981 302.25 2009 699.70
1982 320.13 2010 720.80
1983 330.82 2011 758.79
1984 341.06 2012 769.30
1985 346.12 2013 776.44
1986 347.33 2014 791.59
1987 353.35 2015 786.32
1988 369.45 2016 782.46
1989 383.14 2017 803.93
1990 386.75 2018 841.84
1991 392.35 2019 866.18
6 Agricultural Waste Treatment by Water Hyacinth Aquaculture, Wetland Aquaculture… 313

Year Index Year Index

1992 399.07 2020 867.71
1993 410.63 2021 893.02a
1994 424.91 2022 918.91a
a
Projected future cost index values

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Chapter 7
Production and Applications of Crude
Polyhydroxyalkanoate-Containing
Bioplastic from the Agricultural
and Food-­Processing Wastes

Volodymyr Ivanov, Yung-Tse Hung, Viktor Stabnikov,

Robert Lee-­Kong Tiong, and Anatoliy Salyuk

7.1 Biodegradable Plastics

The nonbiodegradable petrochemical plastics are permanently accumulated in the

environment. Only in the USA, the quantity of plastics in municipal solid waste
(MSW) in 2010 was 27 million tons [1]. A significant portion of these materials is
incinerated or landfilled, which both are unsustainable and environmentally
unfriendly solutions. Therefore, there is considerable interest in the development of
biodegradable plastics. Their additional advantage is that they are producing from
renewable sources so their production will increase environmental and economic
sustainability.

V. Ivanov · R. L.-K. Tiong

School of Civil and Environmental Engineering, Nanyang Technological University,
Singapore, Singapore
e-mail: [email protected]; [email protected]
Y.-T. Hung (*)
Department of Civil and Environmental Engineering, Cleveland State University,
Cleveland, OH, USA
e-mail: [email protected], [email protected], [email protected]
V. Stabnikov
Department of Microbiology and Biotechnology, National University of Food Technologies,
Kiev, Ukraine
e-mail: [email protected]
A. Salyuk
Department of Biochemistry and Ecological Control, National University of Food
Technologies, Kiev, Ukraine
e-mail: [email protected]

© Springer Nature Switzerland AG 2022 317

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_7
318 V. Ivanov et al.

However, the cost of bioplastics produced by conventional biotechnologies is

several times higher than the cost of petrochemical-based plastics. With the rise in
oil prices, the cost of petroleum-based plastics has come more into line with that of
the biodegradable alternatives. There are known predictions that the market for such
biodegradable plastics as starch and cellulose derivates, polylactic acid (PLA), and
polyhydroxyalkanoates (PHAs) will grow by about 20% a year but the reduction of
the bioplastic production costs using cheap raw materials and technological innova-
tions is still essential for the bioplastic industry and applications.
Polyhydroxyalkanoates (PHAs) are polyesters accumulated in bacterial biomass
as a storage compound that can be used by a cell as intracellular carbon, energy, and
reducing power reserve material. It is well known that PHAs are accumulated under
excess of carbon and energy sources or shortage of oxygen and limitation of growth
by low concentrations of such nutrients as sources of nitrogen, phosphorus, and oth-
ers that are used for biomass synthesis. The most important polymers are poly-­3-­
hydroxybutyrate (PHB) with monomer formula (-OCH(CH3)-CH2-C(O)-) and
polyhydroxyvalerate (PHV) with monomer formula (-OCH(CH2CH3)-CH2-C(O)-).
PHAs are accumulated in the form of granules inside the cells of many bacterial
species, for example, representatives of the bacterial genera Acinetobacter,
Alcaligenes, Alcanivorax, Azotobacter, Bacillus, Burkholderia, Delftia, Klebsiella,
Marinobacter, Pseudomonas, Ralstonia, and Rhizobium.
The content of PHAs can be up to 80% of dry biomass. Accumulated PHAs can
be extracted from bacterial biomass and used in practice as bioplastic with a melting
temperature of 160–180°C, a tensile strength of 24–40 MPa, and an elongation at
break of 3–142%. The last property, elasticity of the bioplastic, depends on the con-
tent of PHV in PHAs. These properties are comparable with the properties of
petroleum-­based thermoplastics. The most common commercial PHAs consist of a
copolymer PHB and PHV together with a plasticizer/softener and inorganic addi-
tives such as titanium dioxide and calcium carbonate. The chemical and physical
properties of PHAs can be found in numerous reviews [2–11].
The production of biodegradable plastic PHAs can provide many benefits to the
industry and to the environment. However, known technologies of PHA production
have three essential disadvantages:
–– Use of aseptic culture of selected or genetically modified strains that requires
high expenses for the sterilization of equipment and medium, as well as for the
maintenance of aseptic conditions during biosynthesis of bioplastic
–– Use of relatively expensive nutrients such as pure mineral salts and pure (defined)
sources of carbon and energy
–– Use of expensive, often flammable and toxic organic solvents or energy- and
reagent-consuming methods for the extraction of PHAs from bacterial cells
The following options for raw materials, biotechnology of production, and appli-
cations of bioplastic can help to solve the problem of high cost of the bioplas-
tic PHAs:
1. Use of organic fraction of food-processing or agricultural wastes for bioplastic
production
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 319

2. Batch or continuous non-aseptic cultivation for the biosynthesis of bioplastic by

mixed bacterial culture
3. Concentration and extraction of bioplastic using chemical treatment, filtration,
centrifugation, or flotation for the production of crude bioplastic
4. Applications of crude bioplastic in the construction industry or in agriculture
The aim of this study is to analyze and examine the feasibility of these options
for low-cost bioplastic production and its applications.

7.2 Nutrients for Non-aseptic Bioplastic Production

Cheap sources of carbon and energy were considered for the production of PHAs in
several patents, reviews, and papers. These sources are mainly carbohydrates,
organic acids, and proteins. Their origin is as follows:
–– Food-processing wastes such as corn-steeped liquor, molasses, activated sludge,
starch, and starch-containing wastes; even palm oil mill effluent can be used to
produce PHAs [12, 13].
–– Agricultural wastes such as unbaled straw; corn cobs, stalks, and leaves; silage
effluent; horticulture residuals; farm yard manure; coconut fronds, husks, and
shells; coffee hulls and husks; cotton (stalks), nut shells; rice hull, husk, straw,
and stalks; sugarcane bagasse. Globally, 140 billion metric tons of biomass is
generated every year from agriculture, which is equivalent to approximately 50
billion tons of oil. So, as raw materials, biomass wastes have attractive potentials
for large-scale industries and community-level enterprises [14].
–– Municipal wastes such as activated sludge of municipal wastewater treatment
plant, sewage sludge, and reject water, which is liquid after separation and dewa-
tering of sewage sludge from anaerobic digester of excessive activated sludge.
Wild strains of bacteria, including Alcaligenes spp., Pseudomonas spp., recom-
binant strains of Alcaligenes eutrophus [15, 16], and a number of filamentous gen-
era can accumulate PHAs under limitation of growth with such essential nutrients
for growth as O2 (electron acceptor), N, P, Mg, K, or S, but at the same time there
are a lot of mutant or recombinant strains that do not require nutrient limitations for
the accumulation of PHAs [17, 18]. It is known that Alcaligenes eutrophus can
accumulate PHAs up to 80% of dry biomass under excess of carbon source and
limitation by P or N, and this stock can be oxidized after addition of P or N [17, 18].
There are some other storage compounds accumulating at nutrient limitation.
These storage compounds are extracellular and intracellular polysaccharides in pro-
karyotes and intracellular polysaccharides and lipids in eukaryotes [19, 20]. Mono-
and disaccharides, as well as easily hydrolyzing polysaccharides like starch and
glycogen that are used as a source of carbon and energy, are transformed preferably
to these storage compounds [19, 20]. Carbohydrates can be used for the production
of PHAs mainly in aseptic culture because of the following reasons: (1) almost all
320 V. Ivanov et al.

microorganisms can assimilate them; (2) during assimilation of carbohydrates, the

major storage compounds will be extracellular and intracellular polysaccharides in
prokaryotes or intracellular polysaccharides and lipids in eukaryotes. Excess of car-
bon and energy source could lead to the growth of glycogen-accumulating or slime-­
producing bacteria, but this problem can be easily overcome by acidification of the
medium with a mixture of volatile fatty acids, which are readily converted to PHAs.
Meanwhile, organic acids, if they are sole carbon and energy source for the
growth of bacteria, are accumulating preferably as PHA storage compounds.
Therefore, organic acids must be used for non-aseptic cultivation of mixed culture
to ensure selective conditions for the growth of PHA-accumulating bacteria. In case
when carbohydrates of organic wastes are the major source of carbon and energy,
these components should be converted to organic acids for the cultivation of PHA-­
accumulating bacteria under non-aseptic conditions.
Hydrolysis of polysaccharides and anaerobic acidogenic fermentation are the
most acceptable bioprocesses for this transformation (Fig. 7.1).
There are many bacterial species able to hydrolyze polysaccharides, including
cellulose and hemicellulose, and then ferment them to organic acids, hydrogen, and
carbon dioxide as end products. Microorganisms participating in these processes
depend on the sources of carbon and energy. For example, during acidogenic fer-
mentation of cellulose, such representatives of the genera Clostridium,
Ruminococcus, Butyrivibrio, Acetivibrio, Fibrobacter, Eubacterium, and
Bacteroides could be dominating in population of acidogens [21]. The most typical
material balance of this process can be shown by the following equation of acido-
genic fermentation in rumen (molar ratios of volatile fatty acids (VFAs) were taken
from [22]):

C6 H12 O6  0.82 H 2 O  1.13 CH 3 COOH  0.35 C2 H 5 COOH 

0.26 C3 H 7 COOH  1.67 CO 2  2.47 H 2 , (7.1)

where C6H12O6 is a monomer of cellulose and CH3COOH, C2H5COOH, and

C3H7COOH are acetic, propionic, and butyric acids, respectively.
Biotransformation of carbohydrates to VFA and hydrogen is most suitable in the
utilization of organic wastes for PHA production because a huge diversity of

Fig. 7.1 Microbial groups Biopolymers of wastes

and substances of (polysaccharides, proteins, nucleic acids)
acidogenic fermentation
Hydrolytic bacteria

Monomers
(monosaccharides, amino acids, nucleotides)
Fermentative acidogenic
bacteria
Organic acids, H2
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 321

substances in organic waste can be transformed to VFA, which are selective and
most favorable substances for the biosynthesis of PHAs under non-aseptic condi-
tions. In this case, organic wastes can be converted to PAHs using a two-stage sys-
tem including production of fermentative organic acids and biosynthesis of PHAs
from these organic acids [23]. Different organic acids and polyols added to the
medium are used by cells for copolymerization and enhancement of the PHA syn-
thesis. For example, even a small concentration of caproic acid in the medium sig-
nificantly increased the percentage of PHAs in bacterial biomass.
The system could contain two separate bioreactors, in order to satisfy the differ-
ent physiologies and metabolic activities of the two types of microbes. One bioreac-
tor is used for acidogenesis of organic wastes, and a second one for a mixed culture
of PHA-producing bacteria. The fermentative acids should preferably be transferred
from the first reactor to the second reactor without causing a solid mixing between
the two reactors. VFA can be transferred through a membrane into a reactor where
the acids can be utilized to produce PHAs.
However, in case when different organic wastes are used for fermentation, the
remaining organic dissolved substances and particles can reduce the quality of pro-
duced PHAs. To solve this problem, it could be better to extract all volatile organic
compounds and hydrogen from acidogenic bioreactor using recycling of biogas
with absorption of VFA and using these VFA in the second bioreactor.
It is a well-known use of hydrogen for the production of PHAs [8, 24]. For
example, it was proposed a process for converting organic materials, such as organic
wastes, into a bioplastic through thermal gasification of the organic material into
carbon monoxide and hydrogen, followed by photosynthetic bacterial assimilation
of the gases into cell material [25, 26]. However, this gas is used by photosynthetic
bacteria only and under anaerobic conditions. Some bacteria can transform gaseous
hydrogen and carbon monoxide into PHAs [8, 24]. It is possible to perform acido-
genic fermentation with sufficient yield of hydrogen [27] that will be used for the
production of bioplastic. However, hydrogen-oxidizing bacteria cannot utilize
simultaneously with hydrogen the wide range of organic compounds for the accu-
mulation of PHAs. So, hydrogen produced during acidogenic fermentation of
organic wastes could be a useful source for PHA production but has to be used
altogether with VFA produced from organic wastes.
This point was used in the US Patent Application (provisional patent) 61/967616
(24 March 2014) “Method for Production of Biodegradable Plastic from Organic
Wastes” [28]. The method for the production of biodegradable plastic including
polyhydroxyalkanoates (PHAs) from organic wastes in the non-aseptic bioreactors
differs from other methods because it includes four stages combined in a sequence
and a cycle: (a) the stage of anaerobic transformation of organic matter into volatile
fatty acids and hydrogen, which are supplied for stages (b) and (c); (b) the stage of
aerobic production of microbial biomass which is supplied to stage (c); (c) the stage
of microaerophilic accumulation of PHAs in produced microbial biomass and the
recycle of the part of this biomass to stage (d); and (d) the stage of selection of PHA-­
producing microbial biomass through starvation of biomass supplied from stage (c)
following with its recycling to stage (b).
322 V. Ivanov et al.

Inorganic nutrients, such as N, P, S, Fe, and microelements, can be also supplied

as the components of organic wastes. The ratio of the major inorganic nutrients and
the sources of N and P to organic carbon should be in the range from 75:5:1 and
125:5:1 [27]. The typical C:N:P ratio for anaerobic acidogenic fermentation is
100:5:1. N could be from amines, nitrates, and ammonium. P is usually from nucle-
otides and orthophosphates. Suitable sources of inorganic nutrients could be reject
water of municipal wastewater treatment plants (see below) and the mixtures of
food-processing and agricultural wastes containing N, P, S, Fe, and
microelements.

7.3 Food-Processing and Agricultural Wastes

for Bioplastic Production

Almost all wastes of food-processing plants or from agricultural activities can be

anaerobically transformed to VFA and hydrogen, which can be used further for the
production of PHAs [29]. Fermented organic compounds containing in food pro-
cessing wastes include carbonates, liquid and solid lipids, microbial biomass, glu-
cose, saccharose, lactose, and starch. Food processing wastes include wastes from
potato processing and starch manufacturing, molasses from sugar industry, dairy
producing factories, fruits and vegetables processing plants, cheese whey, vinegar
or acetate-­containing waste, valeric acid or valerate-containing wastes, wastes of
slaughterhouses or meat-processing plants.
Lipids are one of the major pollutants in food-processing wastewater. Wastewaters
produced from edible oil refinery, slaughterhouse, wool scouring, and dairy prod-
ucts industry contain high concentration of lipids. Physicochemical treatment can
remove 90% of lipids, but final biological treatment is necessary because of remain-
ing emulsified and/or colloidal lipids. However, vegetable oils and fats will be trans-
formed not too deeply in acidogenic anaerobic bioreactor. Fats are hydrolyzed to
long-chain fatty acids (LCFA) and glycerol in the anaerobic digestion, but LCFA
are inhibitors of both acidogenic fermentation and methanogenesis mainly because
of their surface activity causing damage of cell membranes. Addition of calcium or
dissolved ferrous/ferric salts reduced the inhibitory effect of LCFA because of pre-
cipitation of LCFA as calcium or iron salt. Iron(II) was used to reduce the inhibition
caused by long-chain fatty acids to prokaryotes involved in anaerobic digestion.
Degradation of stearic acid, one of model compounds of LCFA, was improved in
the presence of divalent iron. The methane production rate was higher in the pres-
ence of iron (0.21 mL/L/h) as compared to control (0.17 mL/L/h) where iron was
absent. The methane yield was 0.1 L/g COD in experiment and 0.08 L/g COD in
control. Iron-containing clay was applied for degradation of vegetable oil. The
methane production was increased 1.5 times as compared to control receiving no
clay. The methane yield was 0.09 and 0.06 L/g COD in experiment and control,
respectively. The COD removal efficiency was 98%, 80%, and 77%, when the iron
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 323

dosage was in the ratio of 20, 40, and 80 mg COD/mg Fe, respectively. Acetic and
propionic acids were accumulated in reactors and inhibited the methanogenic pro-
cess when iron was not present or when the COD/Fe ratio was higher than 20.
However, no accumulations of acetic and propionic acids were observed, when the
ratio of COD/Fe was 20. The presence of iron(II) significantly improved the anaero-
bic digestion of fat [30, 31]. Iron(II) can be produced in the treatment system from
iron(III) hydroxide and iron-containing minerals. These results were confirmed by
other researchers [32, 33].
Water-saving process could be shredding of organic wastes and performance of
acidogenic fermentation in seawater. The complication of anaerobic acidogenic fer-
mentation of organic fraction of MSW in seawater is sulfate reduction that is using
organic acids formed by acidogenic bacteria for the production of toxic and bad-­
smelling dihydrogen sulfide:

CH 3COOH  SO 4 2  2CO2  H 2S  2OH 

. (7.2)

It could be possible to diminish sulfate reduction using Fe(III) compounds like

iron ore to stimulate iron bioreduction that is competing with sulfate bioreduction
[34, 35]:

CH 3 COOH  8Fe 3  2H 2 O  2CO2  8Fe 2   8H  . (7.3)

Fatty acids and hydrogen for PHA synthesis can be produced from food-­
processing wastes or agricultural wastes using fermenting bacteria from the genera
Acetobacter, Bacteroides, Clostridium, Citrobacter, Enterobacter, Moorella,
Propionibacterium, Ruminococcus, Thermoanaerobium, and many others. Initial
inoculation of the bioreactor for acidogenic fermentation can be made using sewage
sludge of municipal wastewater treatment plants (MWWTP), anaerobic sediments,
wet soil, or manure. There must be anaerobic conditions in the acidogenic reactor,
and oxidation/reduction potential should be from −50 mV to −400 mV.
The mass ratio of supplied carbohydrates, producing acids, and proteins, produc-
ing alkali, has to be in optimal region close to 3 to maintain near-neutral pH. However,
if the content of wastes does not ensure self-maintenance of pH, for example, due to
the high content of carbohydrates, an application of rotating drum bioreactor and
addition of limestone or dolomite powder can ensure near-neutral pH stability in the
acidogenic fermentation reactor. VFA produced in this reactor can be used as
remaining culture liquid or can be separated from the culture liquid using mem-
brane filtration or enhanced evaporation with recycling biogas as shown in Sect. 7.3.
Acidogenic fermentation of organic food-processing or agricultural wastes is a
preferable process than methanogenesis because acidogenesis is significantly a
faster process than methanogenesis. Methanogenesis requires long time of the waste
treatment: hydraulic retention time (HRT) in anaerobic digester is about 10–20 days
food-processing wastes and about 20–40 days for cellulose-containing wastes and
manure [36]. Biotransformation of the same wastes to VFA and hydrogen, which
324 V. Ivanov et al.

can then be used for the production of bioplastic, can be performed with HRT in
anaerobic acidogenic reactor about 3–5 days.
Sugarcane or sugar beet molasses can be also used for PHA production by mixed
microbial culture through acidogenic fermentation at first stage. At higher pH, ace-
tic and propionic acids were the main products, while pH from 5 to 6 favored the
production of butyric and valeric acids. The yield of PHA from carbon of VFA
ranged from 37 to 62% [37].
Acidogenic fermentation of organic wastes for the synthesis of bioplastic should
be done on centralized anaerobic digestion (CAD) plants with multiple digesters of
about 2000–4000 m3 [36]. Similar to anaerobic methanogenic digestion of organic
wastes [38], CAD plants can accept animal manures, together with other waste aris-
ing from local food-processing plants, abattoirs, breweries, and municipal sewage
sludge. Regarding the agricultural use of manures, sewage sludge or organic frac-
tion of MSW, it is recommended pre-hygienization treatment at 50°C for several
hours or at 70°C for 1 h [36]. The agricultural and food-processing wastes com-
monly used in AD plants include (1) cattle and swine manures and slurries, (2)
poultry manure (with or without litter), (3) abattoir wastes, (4) potato and other
vegetable processing residues, (5) maize and cereals, (6) silage effluent, (7) dairy
processing residues, (8) brewery residues, (9) fish processing wastes, and (10) can-
ning wastes and wastewaters [36].
The efficiencies of continuous acidogenic fermentation of food waste by rumen
microorganisms were 71 and 82% at dilution rates 3 d−1 and 1 d−1, respectively. The
final product contains mainly acetic acid (2.5 g/L) and propionic, butyric, and vale-
ric acids with concentrations approximately 1.5 g/L each [39]. Liquid after acido-
genic fermentation can be directed to aerobic bioreactor for biomass synthesis and
then to microaerophilic reactor for PHA accumulation in biomass. Anaerobic acido-
genic fermentation of organic wastes and biosynthesis of biomass and PHAs can be
performed in one reactor with separated anaerobic and aerobic ones [23].
The remaining organic and inorganic particles of agricultural and food-­processing
wastes that are used for acidogenic fermentation and production of bioplastic from
VFA can reduce the quality of produced PHAs. To solve this problem, a chain of
membrane anaerobic and aerobic bioreactors can be used, similar to the described
technology [40]. Another technological solution to avoid pollution of bioplastic or
inhibition of microbial mixed culture with the toxic components of wastes is the
recycling of biogas from anaerobic bioreactor with absorption of VFA and their sup-
ply to the stage of biomass and PHA production (Fig. 7.2).
This recycling of biogas can maintain pH in anaerobic acidogenic reactor due to
permanent removal of VFA and CO2 from culture liquid of the bioreactor. The recy-
cling of the biogas through absorber of VFA and CO2 will increase the concentration
of H2 in biogas. Therefore, at some concentration of hydrogen, determined by auto-
matic device, the removal of the part of biogas will be used to supply hydrogen to
the microaerophilic bioreactor for the accumulation of PHAs (Fig. 7.2). It is a well-­
known use of hydrogen for the production of PHAs [8, 24].
During acidogenic fermentation, the pH is dropped and the concentration of
organic acids is increased. The microbial communities of acidogenic fermentation
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 325

Biogas

(CO2 +H2)

Cellulose-
containing Stirring
waste acidogenic
reactor

Alkaline
solution
Biogas
pump Production of
bioplastic

Fig. 7.2 Maintenance of pH during acidogenic fermentation with recycle of biogas with adsorp-
tion of VFA

are active at high concentrations of organic acids, for example, at least of 20, 5, and
15 g/L of acetate, butyrate, and propionate, respectively [41]. Therefore, the pH of
fermented organic waste can be dropped below 5.5 during acidogenic fermentation
[42]; meanwhile, the optimal pH for acidogens is above 6.0 [43]. There are several
ways to maintain optimal pH during acidogenic fermentation of organic waste:
1. For fast and effective acidogenic fermentation, pH must be controlled automati-
cally by measuring and titration with alkali (NaOH). However, it will require
reagent and the system of pH control, so the cost of products could be higher
than the acceptable value.
2. The digestibility of carbohydrate-rich wastes can be improved by co-digestion
with the wastes containing high amounts of protein [44, 45]. Thus, the additional
source of protein containing waste, mixing of carbohydrate- and protein-­
containing waste could balance the concentration of protons results during
anaerobic digestion of carbohydrates and hydroxide ions released during ammo-
nification of protein.
3. VFA can be removed from the reactor of acidogenic fermentation using recycle
of biogas, containing CO2 + H2, and absorption of VFA from biogas using alka-
line solution (Fig. 7.2).
4. This recycle of biogas ensures pH maintenance and high mass transfer of VFA
from acidogenic bioreactor to the tank for VFA collection and storage. This is
similar to the use of biogas recycle for the removal of ammonia from the metha-
nogenic fermentation reactor [46, 47].
326 V. Ivanov et al.

The empirical formula of VFAs after acidogenic fermentation is CH2O0.57 (m.w.

is 23.1) [48]. Considering that the growth yield (Yb) from organic acids is 0.5 g of
dry biomass/g of organic acid, the material balance of biomass synthesis from VFA
of reject water could be written as:

CH 2 O0.57  x1O2  x 2 CO2  x 3 CH1.8 O0.48  x 4 H 2 O, (7.4)

where CH1.8O0.48 (“m.w.” is 23.1) is the empirical CHO formula of microbial

biomass without PHAs [49].

=
Yb 0=
.5 x 3 21.5 / 23.1, so x 3 = 0.54 (7.5)

1  x 2  x 3  balance of C  , so x 2  0.46 (7.6)

2  x 3 1.8  2x 4  balance of H  , so x 4  0.54 (7.7)

0.57  2x1  2  0.46  0.48  0.54  2  0.54, so x1  0.84. (7.8)

So,

CH 2 O0.57  0.84 O2  0.46CO2  0.54 CH1.8 O0.48  0.54 H 2 O. (7.9)

The material balance of PHB (empirical formula is CH1.5O0.5, “m.w.” is 21.5)

synthesis from VFA after acidogenic fermentation of biomass (empirical formula is
CH2O0.57) could be written as:

CH 2 O0.57  x1 H 2  CH1.5 O0.5  x 2 H 2 O, (7.10)

so from the balances of O and H x2 = 0.07 and x1 = 0.18

CH 2 O0.57  0.18 H 2  CH1.5 O0.5  0.07 H 2 O. (7.11)

The equations of the production of bacterial biomass with PHB are as follows:

CH 2 O0.57  0.84 O2  0.46CO2  0.54 CH1.8 O0.48  0.54 H 2 O, (7.12)

c CH 2 O0.57  c 0.18 H 2  c CH1.5 O0.5  c 0.07 H 2 O, (7.13)

1  c  CH 2O0.57  0.84 O2  c0.18 H 2  0.54 CH1.8O0.48 

cCH1.5O0.5  0.46CO 2   0.54  00.7c  H 2 O. (7.14)

The content of PHB in biomass, P, %, can be determined by the following

equation:
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 327

P  100·c CH1.5O0.5 /  0.54 CH1.8O0.48  c CH1.5O0.5  

2150c / 12.5  21.5c  , so c  0.58 P / 100  P  . (7.15)

The yield of biomass with PHB   0.54 CH1.8O0.48  cCH1.5O0.5  / 1  c  CH 2 O0.57 

12.5  21.5c  / 1  c  23.1   0.54  0.93c  / 1  c  g of PHB / g of consumed VFA.
(7.16)

The yield of PHB  cCH1.5 O0.5 / 1  c  CH 2 O0.57 

0.93 c / 1  c  g of PHB / g of consumed VFA. (7.17)

The yield of biomass with PHB and yield of PHB as the functions of PHB con-
tent in biomass are shown in Table 7.1.
The amount of PHAs, which can be accumulated by activated sludge of aerobic
wastewater biotreatment systems, is from 15 to 62% of dry biomass weight. The
addition of fatty acids to the medium enhances the synthesis of PHAs. These PHAs
have useful functions and properties. The maximum accumulation of PHAs in acti-
vated sludge can be up to 37.4% in case when the C:N ratio will be increased to 144,
and the maximum specific polymer production yield was 9.3% of consumed glu-
cose at an optimum C:N ratio of 96 [50]. Accumulation of PHAs by activated sludge
can be stimulated by the changes of the aerobic conditions to anoxic [51–55].
Sludge acclimatized with wastewater supplemented with acetate could accumulate
PHAs up to 30% of sludge dry weight, and the production of PHAs was stimulated
when the pH was kept at 8 or 9 [56, 57]. However, activated sludge, acclimatized in
the microaerophilic aerobic process, accumulated PHAs up to 62% of dry activated
sludge [58]. Essential conditions for the accumulation of PHAs, using mixed cul-
ture or activated sludge, are transient feeding of raw wastewater and the presence of
electron donor and acceptor in the raw wastewater [59]. Activated sludge fed by
VFA at alkaline pH accumulated up to 56% of PHAs in dry biomass probably due
to permanent removal of phosphate and ammonium as struvite, NH4MgPO4·6H2O
[60]. Activated sludge with addition of valeric acid produced mainly poly(3-
hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with the mole fraction
3-hydroxyvalerate (3 HV) up to 54% and a melting temperature of 99°C instead of
178°C for PHB [61].

Table 7.1 Yields of biomass and PHB at different contents of PHB in biomass
P (content of PHB in Growth yield of biomass with PHB, Yield of PHB, g/g of
biomass, % w/w) c g/g of consumed VFA consumed VFA
0 0.00 0.54 0.00
20 0.14 0.59 0.29
40 0.37 0.64 0.25
60 0.87 0.72 0.43
80 2.32 0.81 0.65
328 V. Ivanov et al.

Accumulation of PHB is possible also by the cultivation of fast-growing rhizobia

Sinorhizobium meliloti, Rhizobium leguminosarum bv. viciae, R. leguminosarum
bv. phaseoli, and R. leguminosarum bv. trifolii in sludge and in wastewater. Maxima
HB yields were 7 or 11% w/w after 60 h of cultivation on sludge or 35 h of cultiva-
tion on slaughterhouse wastewater, respectively [17]. Growing rhizobia on sludge
could be a way for bioplastic production from the wastes. However, feeding of
activated sludge with VFA altogether with mineral nutrients could be the best way
for the production of bioplastic from the organic wastes (Fig. 7.3).

Solid wastes
(agricultural, food residuals)
Fresh water
or seawater
Shredding and extracon of
organics

Separaon of liquid fracon Agriculture/ Food Processing

Liquid organic wastes Liquid organic wastes

Biogas
Acidogenic fermentaon
(hydrogen + CO2
+ VFA)

Biogas Absorpon of
Separaon of solids
(Hydrogen) VFA and CO2

Liquid aer anaerobic digester: Hydrogen Soluon of

soluon of VFA and nutrients for PHAs VFA for PHAs
for PHAs producon producon producon

Fig. 7.3 Preparation of nutrients for bioplastic production from wastes

7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 329

7.4 Batch and Continuous Biosynthesis of PHA Bioplastic by

Mixed Culture

Aseptic cultivation of selected or genetically recombinant strain of bacteria requires

thermal sterilization of materials and equipment as well as specialized equipment.
The cost of aseptic cultivation is several times higher than the cost of non-aseptic
cultivation. Therefore, non-aseptic cultivation of mixed microbial culture, which is
able to accumulate PHAs, could be a low-cost technology for the industrial produc-
tion of PHAs.
The major points of using mixed bacterial culture for PHA production were con-
sidered in several patents, reviews, and papers and are shown below:
(a) Activated sludge, a well-known mixed culture, is able to store PHAs as carbon
and energy storage material under unsteady conditions arising from an intermit-
tent feeding regime and variation in the presence of an electron acceptor.
(b) Activated sludge accumulates PHAs to around 20% of dry weight under anaer-
obic conditions, but the content can be increased to 62% in a microaerophilic-­
aerobic sludge process. Oxygen management is crucial to conserving reducing
power, as excessive aeration rates decrease the PHA yield and allow higher
biomass growth.
(c) When compared with a pure culture accumulating up to 80% of cell dry weight,
the merits of PHA production in mixed culture would be an enhanced economy,
a simpler process control, no requirement of aseptic processing, and use of
wastes. Nitrogen deficiency in wastewater is essential for the synthesis of PHAs
using mixed culture [62].
It is expected that PHA-producing mixed culture will comprise representatives
of the genera Acinetobacter, Alcaligenes, Alcanivorax, Azotobacter, Bacillus,
Burkholderia, Delftia, Klebsiella, Marinobacter, Pseudomonas, Ralstonia,
Rhizobium, and others. Initial inoculation can be made using aerobic forest or gar-
den soil suspension or activated sludge of MWWTP. Accumulation of PHAs by
mixed microbial cultures occurs usually under transient conditions of carbon and
energy sources, known respectively as aerobic dynamic feeding and anaerobic/aero-
bic process. In these processes, PHA-accumulating organisms, which are quite
diverse in terms of phenotype, are selected by the dynamic operating conditions
imposed to the reactor. The stability of these processes during longtime operation
and the similarity of the polymer physical/chemical properties to the one produced
by pure cultures were demonstrated. The accumulation of PHAs in mixed culture
can be implemented at an industrial scale [63, 64].
A multistep cultivation process can be used for the production of bioplastic
PHAs from organic raw materials and wastes as it was claimed in the US Patent
Application 61/967616 (24 March 2014) “Method for Production of Biodegradable
Plastic from Organic Wastes” [28]. The method for the production of biodegradable
plastic includes production of polyhydroxyalkanoates (PHAs) from organic wastes
330 V. Ivanov et al.

in the non-aseptic bioreactors. This can effectively reduce raw material and cultiva-
tion cost. This system includes following stages:
1. A process for producing VFA from anaerobically digested wastes is combination
of wastes or addition of limestone/dolomite powder to maintain the neutral pH
in the anaerobic reactor.
2. A separation of the liquid fraction after the anaerobic digester.
3. Continuous separation of VFA and hydrogen from the anaerobic digester, extrac-
tion, and use of VFA.
4. PHAs comprising the combination of the selection and culturing steps, which
includes culturing PHA-producing microbes in a culturing zone and feeding said
PHA-producing microbes into a selection zone. It includes a process for produc-
ing PHAs comprising the step of culturing PHA-producing microbes in a culture
media containing hydrogen gas. It includes a process for extracting PHAs com-
prising the step of degradation of microbial cells into PHA granules and cell
debris and the step of separation of the PHA granules from the cell debris.
The batch biosynthesis of bioplastic is simpler than the continuous one but could
be less productive than the continuous process in large-scale applications. At the
first stage of the batch process, mixed culture is growing in as rich as possible
medium under intensive aeration and optimal temperature and pH but to ensure the
fast growth of microbial biomass with the highest growth yield. After some period
of cultivation, some essential nutrients, such as the sources of N or P, are consumed,
and accumulation of PHAs is started [17]. Continuous systems of cultivation, for
example, a two-stage chemostat with two reactors for the growth of bacteria and for
the production of PHAs, are giving highest PHA accumulation [17]. A batch pro-
cess was used by Imperial Chemical Industries for the large-scale production of
PHB. A continuous process was used by the Austrian Company Chemie Linz GmbH
for the pilot production of PHB. The productivity in the last case was about 1 kg of
PHB/day/m3 of the bioreactor [17].
The growth of biomass must be performed at concentrations of oxygen higher
than 1 mg/L. Using mixed culture, the growth parameters of conventional activated
sludge can be used: 0.5 g BOD/g of dry biomass/day and 0.8 g of biomass/g of BOD
consumed [65], where BOD is “biological oxygen demand,” i.e., oxygen used for
biooxidation of organic matter. So, to produce 1 kg of biomass in a 10 L bioreactor
with a maximum oxygen transfer rate of 1 g/L/h (which is corresponding to a bio-
mass growth rate of 0.8 g of dry bacterial biomass/L/h), it is needed about 5 days.
Considering that the content of PHAs in dry bacterial biomass will be accumulated
to the level of 30%, the maximum rate of PHA production in mixed culture could be
at the level about 0.24 g of dry PHAs/L/h. So, 1 kg of PHAs in a 10 L bioreactor
could be produced for about 17 days. The process of PHA synthesis in the second
reactor of the continuous system or in the second stage of the batch process can be
initiated by decreasing dissolved oxygen concentration to 0.5 mg/L and by increas-
ing the concentration of VFA as well as supply of hydrogen gas, which is produced
during acidogenic fermentation.
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 331

Another technological approach for the production of PHAs is semicontinuous

cultivation of a mixed culture using a feast-famine cycle comprising a feast phase
and a famine phase in one bioreactor. This cycling process promotes not only accu-
mulation of PHAs in biomass but also selection of PHA-producing microorganisms
[66–68]. The continuous process can be used not only because of higher productiv-
ity but also because it can be used for permanent selection and maintenance culture
with highest productivity of PHA synthesis. In every mixed culture, for example, in
biological wastewater treatment, desired selection is ensured by the retention, recy-
cling, or transfer of some microbial groups between different reactors [20, 65]. That
is why the process for producing PHAs in continuous culture should comprise the
combination of the selection, cultivation, and PHA synthesis stages with the recy-
cling loop of PHA-accumulating cells as shown in Fig. 7.4.
The selector could be a bioreactor with intensive aeration but without supply of
nutrients so that cells with intracellular storage of carbon and energy will have
selective advantages for growth in such a bioreactor. This advantage will depend on
the retention time in the bioreactor selector. There must be sufficient long time of
starvation, approximately from 2 to 20 h, so that cells with accumulated PHAs will
grow and dominate in the microbial population but cells without intracellular stor-
age of PHAs will be suppressed in their growth or even die because of starvation.
The hydraulic retention time (HRT) in the bioreactor for the growth of biomass
should be in the range from 1 to 20 h, while the HRT in the bioreactor for PHA
accumulation should be several times longer.
Additional selection factor, high salinity of the medium, could be used in case
when seawater is used for the cultivation of PHA-producing mixed bacterial culture
and osmotic shock is used to disrupt cells of halophilic bacteria and to release PHA
granules in low-salinity fresh water. There are known halophilic bacterial species
that accumulate PHAs, for example, representatives of the genera Alcanivorax [69,
70] and Delftia [71]. We isolated similar strains from the water of the Dead Sea in
Jordan. These bacteria grew and accumulated intracellular PHAs in the medium

Nutrients and air

Bioreactor Bioreactor Bioreactor

to grow to produce
-selector
biomass PHAs

Biomass
enriched
with PHAs

Fig. 7.4 Continuous production of PHAs with selector

332 V. Ivanov et al.

with 10% of NaCl. Organic acids for the cultivation of PHA-accumulating mixed
halophilic culture also can be produced by acidogenic fermentation of organic sub-
stances dissolved or suspended in seawater.

7.5 Downstream Processes

A typical rod-shaped bacterial cell has a diameter of about 1–2 μm and a length of
2–5 μm and is covered with one or two membranes and thin or thick cell wall in
Gram-negative or Gram-positive bacteria, respectively [20]. PHA granules in bacte-
rial cell have diameters from 0.2 to 0.7 μm and are surrounded by a membrane [3].
So, to release the PHA granules from bacterial cell, the cell wall must be mechani-
cally broken or (bio)chemically lysed.
According to the present state of the art, PHA-containing biomass is processed:
(1) by extraction of PHAs from dried biomass with organic solvents following the
separation of solution by centrifugation or decantation; (2) cell walls and mem-
branes are degraded chemically by oxidants and surfactants and then granules of
PHAs are concentrated by centrifugation or flotation; (3) cell walls and membranes
are degraded enzymatically and then granules of PHAs are concentrated by cen-
trifugation or flotation; and (4) cell walls and membranes of halophilic bacteria can
be broken by osmotic shock. Different methods for isolation and purification of
bacterial PHAs were described in detail in several reviews [3, 9, 72].
PHA solubilization by organic solvents occurs at low temperature [73] or cell
wall breaking, adjustment of pH to alkali value, adding of surfactant, separating of
coagulant from liquid, washing and drying of product [74]. Recovery and purifica-
tion of PHAs can be performed also by solubilization of the non-PHA cell mass in
an acidic solution, leaving a suspension of partially crystallized PHA granules,
adjusting the pH of the suspension to 7–11 and separating the PHA solids from the
dissolved non-PHA cellular mass, resuspending the PHA solids in a bleaching solu-
tion for decolorization, and drying the resulting PHA solids [75].
Different chemical pre-treatments can be used before and/or after enzymatic cell
disruption to enhance extraction of PHAs [76]. It is well known that oxidation of the
cell wall with hydrogen peroxide or other oxidants such as chlorine or chlorine-­
containing oxidation agents can be used for the recovery of PHAs. For example, one
of the simplest ways could be mixing of biomass with bleach (solution of 5% of
sodium hypochlorite with pH 13) so that the final concentration of hypochlorite will
be about 1.2% and to pH about 11, and after 1 h of stirring, pH should be adjusted
with HCl to 7. The suspension could be treated by continuous or batch centrifuga-
tion or flotation for separation of cell walls and PHA granules.
All known methods of PHA extraction suffer from high cost or environmental
pollution and are difficult to be industrialized. To avoid disadvantages of chemical
treatment, the proteolytic enzymes can be used [77]. The generated solid and liquid
phases are separated by filtration or centrifugation. Instead of pure enzymes, it
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 333

could be possible to use crude enzymes or even intact microorganisms hydrolyzing

bacterial cell walls. For example, there are known fungi, representatives of the gen-
era Absidia, Agaricus, Aspergillus, Chaetomium, Fusarium, Neurospora,
Penicillium, Phanerochaete, Phialophora, Pleurotus, Rhizoctonia, Trichoderma,
and many others that are able to lyse bacterial cell walls. These microorganisms
should be grown in a separate bioreactor using biomass remainder after separation
or extraction of PHAs from biomass. When extracellular cellulolytic activity will be
at the sufficient level, bacterial biomass containing PHAs can be treated with intact
fungal suspension or its filtrate containing extracellular cellulolytic enzymes.
Disruption of biomass of halophilic bacteria, containing PHAs, can be performed
just mixing with fresh or distilled water to break cells and to release PHA granules
due to osmotic shock. Then suspension can be treated with fine dispersed air or by
dissolved air flotation (DAF) at pH 3 to concentrate PHA granules in foam.
Separation of the PHA granules of PHAs from lysed or disrupted cells can be
done using flotation, centrifugation, or membrane filtration. Probably, a low-cost
technology could be flotative separation of PHA granules from destroyed cells. The
bubbles of air have the hydrophobic surface, so at the pH about 3.0–3.5, the differ-
ence in hydrophobicity of the surface of PHA granules and cell walls ensures the
preferable adhesion of PHA granules to the air bubbles [78]. This flotation process
is affected by air bubbles, cells, and PHA granule interactions as well as dynamics
of liquid and foam, sizes of the particles, hydrophobicity, and surface charge (zeta
potential), which depends on pH of aqueous solution. Finally, a PHA purity of 86%
(w/w) can be obtained using flotation separation and concentration [78].
However, flotative separation using just supply of air in the flotation tank with
bacterial suspension after cell disruption has to be performed with small-diameter
air bubbles and intensive aeration to have the sufficient specific surface for PHA
granule adhesion and a fast foam formation rate for its continuous removal.
Additionally, for the effective batch or continuous flotation, there must be automatic
control of the level of liquid and liquid supply rate, as well as the level of foam and
foam removal rate in the flotation tank to ensure proper separation of biomass and
PHA granules. The simpler and more reliable way for PHA granule separation and
concentration is dissolved air flotation (DAF) in batch mode. Usually, it involves
supply of compressed air into the tank with bacterial biomass for dissolved oxygen
saturation at excessive pressure, for example, at 5 atm, and then supply of this sus-
pension to the flotation tank with atmospheric pressure. The smallest air bubbles are
releasing in the flotation tank due to the difference of gas solubility at the excessive
or atmospheric pressure. These small-size air bubbles adsorb most hydrophobic
substances at their surfaces and float forming the foam. The foam, containing con-
centrated hydrophobic substances, is removed and collected. PHA granules are
dried and then used as bioplastic. There are known many designs of DAF facilities
for experimental and industrial applications.
334 V. Ivanov et al.

7.6 Crude Bioplastic for Construction

and Agricultural Applications

Potential market of bioplastics includes packaging materials, catering products,

consumer electronics, medical materials, agriculture and horticulture (biodegrad-
able mulch foil), toys, and textiles (en.european-­bioplastics.org/market). PHA bio-
plastic has been used in the medical applications in skin substitutes, and drug
delivery microspheres because of PHA biocompatibility and biodegradability [79].
However, PHAs producing by non-aseptic cultivation from waste materials using
mixture cultures of bacteria cannot be used for biomedical, food packaging, or
catering applications because chemical and physical properties of bioplastic are not
controlled in non-aseptic cultivation and there may be present pollutants from
wastes and microorganisms.
The type of application depends also on the mechanical properties of bioplastic.
PHA mechanical properties depend very significantly on the chain length of the
monomer. For example, PHB is stiffer and more brittle than for polypropylene, but
copolymerization with hydroxyvalerate (PHB-co-PHV) makes bioplastic much
more flexible [4]. This copolymer can be used for packaging material like films and
bottles [17]. However, PHB applications are limited by its thermal degradation dur-
ing molding and stiffness of bioplastic. For aseptic cultivation of genetically modi-
fied strains of PHA producers, it is possible to select specific medium and conditions
for the production of PHAs with the desired mechanical properties, but for the non-­
aseptic cultivation of mixed culture, the chemical content of accumulated PHAs will
be determined mainly by the spectrum of fatty acids in the medium.
The major advantage of PHAs for applications is biodegradability of bioplastic
by soil and aquatic microorganisms. These microorganisms produce PHA depoly-
merases and other enzymes finally transforming PHA-made items to carbon dioxide
and water for about 1.5 months in anaerobic sewage, 1.5 years in soil, and 6.5 years
in seawater [4, 80, 81]. Dead bacterial biomass with PHAs contains also polysac-
charides of the cell wall, proteins, polynucleotides, and phospholipids, where the
contents are about 15%, 50%, 25%, and 10% of dry biomass without PHAs, respec-
tively, and the biodegradation rates are higher than those of PHAs. Therefore, from
the point of view of biodegradability in soil, there is no sense to extract PHAs from
biomass but to use dry biomass with PHAs as crude nanocomposite from the gran-
ules of PHAs and interlayers of cellular biopolymers.
Speculatively, such nanocomposites should be more flexible and better biode-
gradable than extracted PHAs. There are known many natural biocomposites, where
brittle nanocomponents like hydroxyapatite crystals in the bone or aragonite crys-
tals in the pearls are composed of flexible biopolymer nanocomponents, usually
nanolayers or nanoaggregates of protein molecules. Production of similar nanocom-
posite material from PHA-containing dry bacterial biomass will permit to exclude
an expensive technological stage of PHA extraction and diminish additionally the
cost of crude PHA-containing biodegradable material.
7 Production and Applications of Crude Polyhydroxyalkanoate-Containing Bioplastic… 335

However, there are two potential problems in the applications of such crude PHA
nanocomposite. The first problem is the temperature of malting for PHAs, which is
in the range of 160–180°C [4–8]. The melting temperature of PHB is close to its
thermal decomposition temperature Td-10% [82]. Thermal decomposition tempera-
tures of proteins, polysaccharides, and polynucleotides are also close to this value,
i.e., all biopolymers have poor thermal stability at temperature of PHA melting.
Natural antioxidants, which present in biomass, can reduce the rate of thermal
destruction of biopolymers [83]. Protein itself can be considered as thermoplastic
material but with additions of plasticizers, which inhibit the formation of cross-
linking that can result in the formation of thermoset material from extruded protein
[84]. Therefore, the molding of composite crude bioplastic material should be for as
short process as possible to diminish the thermal decomposition of PHAs and other
biopolymers of bacterial biomass.
There is a clear trend in the construction industry for using biodegradable materi-
als and biopolymers [85–88]. One area of applications of nanocomposite bioplastic
from bacterial biomass containing PHAs is the production and use of biodegradable
construction materials. Actually, almost all construction activities started up from
using the most abundant biodegradable polymeric material, wood, and other
cellulose-­containing natural materials. Currently, biodegradable construction mate-
rial can diminish the area of land used for landfilling because they are degraded very
quickly in soil or in the landfill.
For example, this sustainable, biodegradable bioplastic can be used for insulation
walls and partitions, construction of nonstructural (internal) elements such as sepa-
rating walls, and for manufacturing of geotextiles, drainage pipes, silt and dust
fences, and different kinds of the temporarily constructions. Plastic foam and foam
insulators that are used in the construction industry produce hazardous nonbiode-
gradable wastes after demolition of the buildings or temporal constructions. The
non-biodegradability of these plastics limits their construction applications. The
bioplastic could be used for new, green, sustainable construction materials because
this easily biodegradable material can be being landfilled, composted, or even left
on the site of use without excavation.
Other examples of potential application of crude nanocomposite from bacterial
biomass containing PHAs are construction silt and dust fences that can be landfilled
for fast biodegradation or composted as biomass. Sustainability of biodegradable
construction materials is due to (1) production of bioplastic from renewable sources
or even from organic wastes and (2) fast biodegradability of this material under the
conditions of landfill or composting, so negative effect of construction waste on the
environment will be minimized. There could be a big market for biodegradable
bioplastic foam construction material, which does not require incineration after
demolition.
There are also a lot of potential agricultural applications of nanocomposite bio-
plastic from bacterial biomass containing PHAs. One potential application is dark
plastic mulch, which suppresses weeds, reduces water evaporation from soil, and
warms soil for earlier planting. Millions of hectares of arable land are cultivated
under plastic mulch. However, nonbiodegradable plastic mulch requires
336 V. Ivanov et al.

labor-consuming annual removal from the field and disposal or energy-consuming

and environmentally unfriendly recycling of the used film. The advantage of the
film from biodegradable nanocomposite bioplastic from bacterial biomass contain-
ing PHAs is that this used mulch material can be left for natural biodegradation on
the field.
Another agricultural application of nanocomposite bioplastic from bacterial bio-
mass with PHAs is manufacturing of slow-release fertilizers using bioplastic coat-
ing or embedding of fertilizers in bioplastic granules, bars, or films. For example,
fertilizer-embedding bioplastic bars can be used for oil palm plantations for slow
release of fertilizers during 0.5–2 years of bioplastic biodegradation in soil.

7.7 Conclusions

It is possible to produce low-cost crude bioplastic using mixed microbial cultures

under non-aseptic conditions of cultivation. Organic acids from liquid wastes or
produced by acidogenic fermentation of solid or liquid organic wastes must be the
dominating source of carbon for the biosynthesis of crude bioplastic.
There will be a lot of environmental benefits from production and use of crude
bioplastic from organic fraction of solid municipal wastes: (1) reduction of the
amount of municipal solid wastes to be incinerated, (2) reduction of the amount of
ash to be landfilled, and (3) use of seawater for municipal solid waste separation that
will save fresh water consumption.
Organic fraction of municipal solid and liquid wastes, as well as agricultural and
food-processing wastes, can be used for the production of low-cost crude bioplastic
with or without extraction of PHAs from microbial biomass. However, this crude
bioplastic can be used only for specific applications in the construction industry or
in agriculture. The applications in the construction industry could be the bioplastic
foam, the foam insulators, the silt and dust fences, and the temporary constructions.
The applications in agriculture could be the dark plastic mulch and different carriers
for slow-release fertilizers.

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Chapter 8
Optimization Processes of Biodiesel
Production from Pig and Neem
(Azadirachta indica A. Juss) Seeds Blend
Oil Using Alternative Catalysts from Waste
Biomass

T. F. Adepoju and Yung-Tse Hung

8.1 Introduction

The important factors that affect the economic growth of a country are human
resource, natural resources, capital formation, technological development, and
social and political factors. Among the listed factors, natural resources come from
nature, and this has become an important factor contending with human resource.
As human growth increases, the needs for energy increase. Therefore, no country
can achieve its economic growth and social stability without accessibility and opti-
mality of energy.
Meanwhile, over 80% of energy utilization of a country comes from fossil fuel
and its derivatives (natural gas, coal, and oil), and almost three-quarters of it origi-
nate from combustion of fossil fuel derived from carbon emission.
Recent reports by International Energy Statistics (IES) revealed that there has
been an alertness to improve the biofuel production in the European Union
Membership Countries and United States starting from 2019 to 2030. Countries
such as China (the most populous country in East Asia), India (the second most
populous country, and seventh largest country by area located in south Asia), Brazil
(largest country in both South American and Latin America, the world fifth largest

T. F. Adepoju
Chemical/Petrochemical Engineering Department, Akwa-Ibom State University,
Ikot Akpaden, Nigeria
e-mail: [email protected]; [email protected]
Y.-T. Hung (*)
Department of Civil and Environmental Engineering, Cleveland State University,
Cleveland, OH, USA
e-mail: [email protected]; [email protected]; [email protected]

© Springer Nature Switzerland AG 2022 341

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_8
342 T. F. Adepoju and Y.-T. Hung

country by area and fifth most populous), and ASEAN (comprises ten countries in
southern Asia, which promotes intergovernmental cooperation and facilitates eco-
nomic, political, security, military, educational, and sociocultural) countries have
increased the permitted feedstock base for biofuel and introduced subsidies to
expand production capacity. Europe however has injected millions of euros in order
to meet up with safety data sheet (SDS) level and is currently leading producer of
biofuel follow by Brazil. Therefore, globally, working towards a secure, sustainable
future for all through biofuel production is the technology roadmap.
For biofuel production, there exist more than 350 potential oil-bearing crops,
excluding animals’ fat and algae [1]. In China, popular biofuel feedstocks include
sugar beet, sugarcane, tuber crops, sweet sorghum, Jatropha curcas, etc., while in
India, the major feedstocks are palm oil, edible oil from oilseed crop, Jatropha oil,
sugarcane, maize, sugar beet, and cassava [2]. In Brazil, the feedstocks include beef
tallow, cottonseed oil, waste frying oil, pork lard, chicken fat, palm oil, etc. [3]
while in the ASEAN countries, the first-generation biofuel feedstocks include sug-
arcane, wheat, corn, cassava, oil palm, soybean, rapeseed, and Jatropha [4]. Europe
also used the feedstocks for the production of oil-based residue, lignocellulose,
crops, algae, waste gases, etc. To continue to lead the world production in biofuel
for 2020–2030, Europe identifies four primary sources that could provide additional
biomass and supportive growth of bio-based industries, namely, agricultural resi-
dues, forest biomass, waste, and non-food crops [5].
Pig fat (lard), obtained from pig meat called pork, is the most commonly avail-
able waste, owing to the fact that pork meat is the most commonly consumed red
meat worldwide (due to high in protein and rich in minerals and vitamin), especially
in Eastern Asia, but its consumption is forbidden in certain religions (Islam and
Judaism). High-quality protein is the main nutritional component of pork meat,
making it useful for muscle growth and maintenance. However, pig contains vary-
ing amount of fat (lard) ranging from 10 to 32% depending on the level of trimming,
but the fat is mainly composed of saturated and unsaturated fats, present in approxi-
mately equal amounts. Surprisingly, this fat causes harm to man, cattle, and other
domestic animals when consumed. The risks in consumption include trichinosis,
bovine spongiform, diarrhea, roundworm infestation, heart disease risk, bladder
cancer risk, mad cow diseases, gastroenteritis, and many more [6]. Hence, it is
needed to channel the fat as a feedstock for biofuel (biodiesel) generation.
The use of pig fat and other animal fats as a feedstock for biodiesel production
have been reported by various researchers. Chinyere et al. [7] reported 96% of bio-
diesel yield in optimization of the methanolysis of lard oil in the production of
biodiesel with response surface methodology using KOH. Chung et al. [8] produced
81.3% fatty acid methyl ester (FAME) content in a transesterification reaction of
duck tallow with methanol using NaOH catalyst, while Anildo Jr. et al. [9] reported
a total 83% conversion in a synthesis and characterization of ethylic biodiesel from
animal fat wastes using KOH. Animal fat wastes for biodiesel production was
reviewed by Vivian et al. [10], while Jishy and Sankar [11] gave a report on produc-
tion of biodiesel from chicken fat, pork fat, and combination of the two feedstocks
using KOH. The use of mixtures of waste frying oil and pork lard to produce
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 343

biodiesel using NaOH was also reported by Joana et al. [12]. Encinar et al. [13]
reported a total of 89.0 wt.% ester content in a study of biodiesel production from
animal fats with high free fatty acid content using H2SO4/NaOH.
Azadirachta indica belongs to the mahogany family Meliaceae, usually known
as Neem, Nimtree, or India Lilac [14]. Its origin is from India and the Indian sub-
continent which includes Nepal, Pakistan, Bangladesh, and Sri Lanka. It can also be
found growing in tropical and semi-tropical regions, Islands in the southern part of
Iran as well as Africa in Nigeria. Neem is considered a weed in many countries,
including some parts of the Middle East, and most of Sub-Saharan Africa including
West Africa and Indian Ocean states. Naturally, it lives well in similar environments
to its own, but its weed potential has not been fully assessed [14]. Neem leaves are
dried in India and placed in cupboards to prevent insects eating the clothes and also
while storing rice in tins [15]. The tender shoots and flowers of the neem tree are
eaten as a vegetable in India, a soup-like dish called Veppampoo charu. Products
made from neem trees have been used in India for over two millennia for their
medicinal properties. Neem seed contains 25–45% oil which is non-edible and has
greater disadvantages when used. In small children it is highly toxic and can lead to
death, in pregnant woman, it can cause miscarriages, in man, it can lead to infertil-
ity, and low blood sugar [16]. It is useful as a raw material in industries such as
cosmetics, resin, soap, honey, neem blossom, fertilizer, and gum making.
Meanwhile, the use of mixture of oils in different proportions (70:30, 50:50,
25:75, 40:60) have been exploited for biodiesel synthesis, and the reports showed
high yield of biodiesel when compared with the results obtained with single oil [12,
17–20], but all of these reports utilized homogeneous catalyst (KOH/NaOH) for
biodiesel production apart from the work reported by Falowo et al. [17], where
nanoparticles from elephant ear tree pod (Enterolobium cyclocarpum) husk was
used as heterogeneous bio-base catalyst in biodiesel production intensification via
microwave irradiation-assisted transesterification of oil blend.
However, the use of homogeneous catalysts for biodiesel synthesis produces
good results, but not with its own shortcomings. The disadvantages include high
cost price, toxic effects, inability to recycle, hygroscopic nature, time consuming in
washing process making it difficult to separate glycerol from biodiesel and utility
wastage.
Heterogeneous catalyst can be obtained from biomass waste, animal bones, acti-
vated carbon supported catalyst, and waste coral [21–24]. Direct burning of wastes
does not decompose easily and substantial furnace modifications are usually
required if they are to be used as firewood or fuel oil [25]. Heterogeneous catalyst
however comprises many advantages including non-toxic easily recycle and reuse,
non-hygroscopic, and availability with low or no cost.
Considering the large amounts of the solid waste that kola nut husk (KNH) and
palm kernel shell husk (PKSH) constituted to the fruits, there is need for the process
of recycling/recovery. It has been reported that using fermented kola nut husk
(FKNH) before furnace modifications increases the calcium, potassium, phospho-
rous, and magnesium contents [26] enough to produce energy.
344 T. F. Adepoju and Y.-T. Hung

Therefore, this study synthesizes biodiesel from pig fat oil—neem oil blend
using derived catalyst of the mixture of palm kernel shell husk (PKSH) and fer-
mented kola nut husk (FKNH) as heterogeneous catalyst. Detailed characterization
of the catalyst developed was carried out using SEM, FTIR, BET, and XRD. Process
parameter optimization was investigated with a view to determine the optimum
yield of biodiesel from miscible oil blend.

8.2 Materials and Methods

8.2.1 Materials

8.2.1.1 Pig Fat, Neem Oil, Palm Kernel Shell Husk, and Kola Nut Husk

Freshly harvested pig fat was obtained from slaughter house at Ete Market, Mkpat
Enin L.G.A., Akwa Ibom State, Nigeria. Neem oil (5 L) was purchased from
National Cereals Research Institute (NCRI), Abuja, while FKNH was freely
obtained from a local market in Ikot Abasi Local Government Area, Akwa Ibom
State, Nigeria, and PKSH was obtained from gasifier plant.

8.2.1.2 Chemicals

99.5% pure methanol, 36 wt.% HCl, HPLC grade n-hexane, chloroform, H2SO4
acid, ethanol acetic acid, etc., were of analytical grade and need no further purifica-
tion obtained from Finelib Nig. Ltd.

8.2.2 Methods

8.2.2.1 Oils Preparation

The method used by Anildo Jr. et al. [9] was adopted but with little modifications.
To a flask of 5000 mL capacity, 2 kg of pig fat was washed with 1000 mL of diso-
dium carbonate (1 mol/L) and was mechanically stirred for 25 min. The mixture
was centrifuged for 10 min at a temperature of 15 °C using propylene tubes. The
supernatant was separated by filtration, and 50 g of anhydrous disodium sulfate was
added, stirred for another 10 min, and then centrifuged again for 5 min at a tempera-
ture of 15 °C. The pure pig fat oil (PFO) obtained was kept in a clean jar. Neem oil
(NO) was heated at a temperature of 30 °C for 20 min, filtered to remove dirt, and
then kept in a clean jar for further processing.
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 345

8.2.2.2 Oil Blends and Their Characterization

Since neem oil is a non-edible oil with high free fatty acid [27] and pig fat oil is with
low acid value [7, 8]; to achieved a mixture of the oil with low acid value, low vis-
cosity, and low density, oils were blend in a ratio of neem oil:pig fat oil (v/v) as
10:90 (NO10), 20:80 (NO20), 30:70 (NO30), 40:60 (NO40), 50:50 (NO50), 60:40 (NO60),
70:30 (NO70), 80:20 (NO80), and 90:10 (NO90), respectively, to enhance biodiesel
synthesis using derived heterogeneous catalyst.
The blended oils were properly mixed at 35 °C in magnetic for easy miscibility
considering the instability in fat nature. Each resulting mixture was examined for its
acid value, density, and viscosity. The mixture with low acid value, density, and
viscosity was used for biodiesel synthesis. Other properties of the oil mixture were
further determined using association of official analytical chemists [28, 29]. The
constituent of the volatile matter, long and branched chain hydrocarbons, esters and
alcoholic acids, and others was determined using complete Agilent 5973 N gas
chromatography–mass spectrometry instrument control and data storage. 10:90
(NO10) is used as an abbreviation for 10 mL of neem oil:90 mL of pig fat oil and
NO0 is an abbreviation used for 100 mL of neem oil.

8.2.2.3 Catalyst Preparation

A 500 g KNH was washed with distilled water twice in a cleaned bucket to remove
unwanted materials. Solid state fermentation was carried out for 10 days on the
sample. The fermented sample (FKNH) was oven dried to constant weight and then
milled to powder of 0.5 mm particles size for easy calcination. 500 g of PKSH pow-
der obtained from gasifier plant was made through the mesh size 0.5 mm and was
mixed in the same ratio (1:1) with the FKNH powder; the mixed powder was cal-
cined at 800 °C for 3 h in a muffler furnace at standard atmospheric pressure. The
calcined powder obtained after calcination was characterized using scanning elec-
tron microscopy (SEM), energy dispersive spectroscope (EDS), X-ray diffraction
(XRD) analysis equipped with Kά and Cu radiation source, accelerated at 20 mA
and 30 kV, Fourier transform infrared spectroscopy (FTIR), and BET isothermal
adsorption. The calcined mixed based catalyst was used for transesterification of
mixed oil to biodiesel.

8.2.2.4 Characterization of Calcined Powder

After calcination of the powder, the calcined sample was cooled at room tempera-
ture, and then characterized using scanning electron microscopy (SEM), to examine
the surface morphology of the catalysts, energy dispersive spectroscope (EDS) to
determine the elemental analysis of the samples and the quantitative composition of
the catalysts, X-ray diffraction analysis (XRD) equipped with Kά and Cu radiation
source, accelerated at 20 mA and 30 kV, to establish the angular scanning electron
346 T. F. Adepoju and Y.-T. Hung

performed in the range of 10o < 2ɸ < 70o at a speed of 2 °C min−1 [25], and Fourier
transforms infrared spectroscopy (FTIR), to check the presence of functional group
and verify the presence of characteristic absorption bands of major elements pres-
ent. The surface area and the basicity of the catalysts were examined using BET
isothermal adsorption and Hammett indicator method.

8.2.3 Biodiesel Production

8.2.3.1 Esterifying the Oil Mixture

Considering the oil mixture, it was observed that the low viscous (21.50 mm2/s)
blended oil was obtained at 60:40 (NO: PFO), with acid value of 4.10 mg KOH/g
oil (free fatty acid (FFA) = 2.05) and low density of 852 kg/m3. This oil blend used
for acid treatment stage is referred to as esterification. The method used can be
found elsewhere [27]. The free fatty acid (FFA < 1.5) required for successful trans-
esterification was obtained following the procedures already adopted by Adepoju
et al. [30].

8.2.3.2 Transesterification of Esterified Oil Mixed to Biodiesel

The method used by Kostic et al. [31] was adopted with little modifications. The
transesterification of the esterified mixed oils to biodiesel was carried out in a three
necked reactor at the catalyst amount 2–5 g, reaction time of 50–80 min, methanol/
oil ratio of 3–9, and reaction temperature of 50–65 °C. Measured derived catalyst
amount was added to a known methanol in 250 mL flask, thermostated at 65 °C for
20 min, and then the predetermined mass of preheated mixed oil (thermostated at
60 °C for 1 h) was poured into the reactor and the reaction was monitored for a
certain period. At the end of the reaction, the catalyst was separated by decantation
and the biodiesel phase was separated from alcohol phase by separating funnel. The
leach catalyst in the biodiesel was removed by washing with a mixture of 1.5 g
sodium carbonate and 50 mL methanol, thermally heated at 65 °C for 3 h in a mag-
netic shaker. The mixture was filtered and then washed with ionized water twice for
20 min as earlier reported by Alba-Rubio et al. [32]. The biodiesel was separated
through gravity settling and then dried over anhydrous sodium sulfate, which was
separated by filtration to obtain mixed oil biodiesel (MOB). The yield of biodiesel
was obtained using volume ratio of biodiesel produced to volume of oil used. The
catalyst was collected for reuse at the end of the reaction, but there was reduction in
the fourth and fifth cycle. Hence, the catalyst reusability was stopped after third
usage. These processes were repeated based on experimental runs generated by
deign expert.
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 347

8.2.3.3 Experimental Design and Statistical Analysis

A three levels four factors was adopted for experimental design of transesterifica-
tion of mixed oil to biodiesel, namely, catalyst amount (g), reaction time (min),
reaction temperature (°C), and MeOH/OMR (mL/mL) (Table 8.1). Central compos-
ite design was used to produce a total of thirty (30) experimental runs so as to give
room for a well-designed experiment, more interactive effects among the variables,
higher number of runs to increase the axial and center points, and greater room for
duplication.
For statistical analysis, the regression parameter and test of significance were
used to confirm the level of contribution of each variable on the response (MOB).
Also, the experimental results were compared with predicted value generated by
design expert via a straight line plot of predicted against actual value (supplemen-
tary file). Process optimization was carried out by determining the significance of
probability test value (p-value < 0.05), F-distribution (f-test), variance inflation fac-
tor (VIF), and the degree of freedom (df). The adequacy of the model was confirmed
by determining the adjusted R-square Radj. 2
 
and predicted R-square Rpred. 2

val- 
uee. A graph of three-dimensional plots with contour lines was used for interactive
effects among the variables.
The model fitting equation that correlates the relationship between the response
variable and the independent variables is as given in Eq. (8.1).
k k k
MOB   0   i X i   ii X i2   ij X i X j  T (8.1)
i 1 i 1 i j

where MOB is the response, γ0 is the intercept, γi is the linear coefficient, γii is the
interaction coefficient, γij is the quadratic coefficient terms, Xi, Xj are the four fac-
tors, and ϵ is the residual error.

8.2.3.4 Physicochemical Properties of MOB

Physicochemical properties of the pure MOB were determined by standard meth-

ods: viscosity (ASTM D6079), density (ISO 17828:2015), moisture content (EN
ISO 12937:2000), iodine value (EN 14111), acid value (ASTM D664), cetane

Table 8.1 Three-level four-factor central composite design for transesterification of mixed oils
Levels
Variable Units Symbol -1(low) 0 +1(high) -alpha +alpha
Catalyst amount (g) X1 2 3 4 1 5
Reaction time (min) X2 50 60 70 40 80
Reaction temp (°C) X3 50 55 60 45 65
MeOH/OMR (mL/mL) X4 3 5 7 1 9
Where MeOH/OMR = methanol/oil molar ratio
348 T. F. Adepoju and Y.-T. Hung

number (EN ISO 5165), higher heating value (ASTM D2015 [33]), oxidative stabil-
ity (EN 14112), cold filter plugging (ASTM D6371), carbon residue (ASTM
D4530), flash point (ASTM D93), cloud point (ASTM 2500), pour point (ASTM
97), and fatty acid methyl esters (FAME) (EN 14103:2003).

8.3 Results and Discussion

8.3.1 Physicochemical Properties and Fatty Acid Composition

of Oils and Blend

Table 8.2 shows physicochemical and fatty acid compositions of mixed oils. For
comparative purposes, the mixture of the oil blend results are also presented. At
40 °C, the blended oil has low density and viscosity; this can be attributed to proper
mixing, blend ratio, and the method adopted for extraction of oil from PFO. Low
density and viscosity play very important roles in oil production, transportation

Table 8.2 Properties and fatty acid compositions of oil and its blend
Parameters Pig fat oil Neem oil Blend (NO60:PFO40)
Properties
Density (kg/m3) at 40 °C 860 890 852
Viscosity at 40 °C/(mm2/s) 23.95 24.80 21.50
Moisture content (%) 0.024 0.031 0.011
%FFA (as oleic acid) 0.425 7.33 2.05
Acid value (mg KOH/g oil) 0.85 14.66 4.10
Saponification value (mg KOH/g oil) 201.6 199.16 198.30
Iodine value (g I2/100g oil) 59.36 107.38 88.90
Peroxide value (meq O2/kg oil) 77.00 4.40 34.30
HHV (MJ/kg) 40.27 42.88 56.94
Cetane number 59.81 46.33 45.83
Mean molecular mass 277.78 281.18 282.40
Flash point (°C) 205.00 207.00 208.00
Cloud point (°C) −5 −5 −5
Pour point (°C) −15 −15 −15
Fatty acid compositions (%)
Myristic acid (C14:0) 1.40 1.30 1.80
Palmitic acid (C16:0) 22.60 23.10 18.20
Palmitoleic acid (C16:1) 1.80 2.70 3.10
Stearic acid (C18:0) 13.60 11.30 21.00
Oleic acid (C18:1) 43.10 50.60 40.42
Linoleic acid (C18:2) 15.70 8.70 14.40
Linolenic (C18:3) 1.50 2.10 1.40
Others 0.30 0.20 0.10
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 349

through the pipeline, and oil recovery processes [34]. The acid value obtained for
neem oil (NO) is higher than that obtained for the blended oil (BO), but the value
obtained for the PFO is less than that obtained for BO. The high acid value could be
attributed to the percentage of NO present in the blend, storage duration of NO
before purchase, and different agro-climatic conditions [35]. The iodine values
obtained for NO (107.38 mg KOH/g oil) and BO (88.90 mg KOH/g oil) were high
than that obtained for PFO (59.36 mg KOH/g oil). This observation supports the
report earlier given by Anyaogu et al. [36], according to which the PFO is low in
conjugated linoleic acid (CLA) and slightly richer in unsaturated acids. The oil is
mainly composed of unsaturated and saturated fats, presently in approximately equal
amounts. The peroxide values of NO and BO are less than that obtained for PFO,
which showed that the NO and BO have high of rancidity and usually stable [37].
The high heating value and the high cetane number value proved that the BO
serves as good feedstock for energy production with great combustibility. The flash
point of BO takes into account the ignition ability of the BO. Oil is considered to be
flammable and there is a risk of danger if its flash point is less than 60 °C. The value
of 208 °C obtained for BO in this study showed that the BO is a safe feedstock and
easy to handle with no risks. Cloud point is the temperature at which the wax begins
to separate when oil is chilled to a low temperature; the value of −5 °C obtained in
this study with low pour point showed that the BO is stable and of low wax forma-
tion at low temperature. The fatty acid compositions of mixed oil as presented in the
table indicated that the oil is mainly made of saturated and monounsaturated acids.

8.3.2 Catalyst Characterization and Elemental Analysis

Figure 8.1a shows the angular SEM images through the developed catalyst from
PKSH and FKNH at different wavelengths of 50, 100, and 200 μm performed in the
range of 10o < 2ɸ < 70o at a rate of 2 min−1. Observation from the images showed
the powder was of various shapes, non-uniform in size, and less porous as the

Fig. 8.1 (a) SEM magnification of developed catalyst taken at resolution of 350×, 500×, and
1500×. (b) FTIR results of calcined kola nut husk powder (CKNHP) at 800 °C for 3 h
350 T. F. Adepoju and Y.-T. Hung

Fig. 8.1 (continued)

magnification increases. Furthermore, the structure based on resolution showed a

glossy nature, permeability, spongy nature of the particle present, and sintering of
small mineral aggregates and agglomerated particles responsible for the spongy
nature. This observation can be likened to the abundance of calcium and oxygen
after calcination which proved that no carbon remains after the heat treatment that
resulted in gaseous formation of CO2 via decomposition of calcium carbonate.
Infrared spectroscopy (FTIR) is an amazingly useful materials analysis tech-
nique, assisting in detecting both organic and inorganic materials. It measures a
sample’s absorbance of infrared light at different wavelengths to decide the mate-
rial’s structural and molecular composition. The FTIR works to convert the raw data
from the broad-band light source to obtain the absorbance level at each wavelength.
The FTIR results of developed catalyst calcined at 800 °C for 3 h is displayed in
Fig. 8.1(b) with the infrared spectrum, which plots the intensity of infrared spectra.
The peaks (absorbance bands) correspond with the various vibrations of the sam-
ple’s atoms when it is exposed to the infrared region of the electromagnetic spec-
trum. The spectrum stretching above 1500 cm−1 in the infrared spectrum indicated
the presence of two functional groups CH2 and C=O, while the spectrum stretching
below 1500 cm−1 in the infrared spectrum indicated the presence of two functional
groups aldehyde/ketone and ester. The wavelength band observed in the figure from
711.9–1397.8 cm−1 stretches indicated the presence of aldehyde/ketone and ester
while the spectrum stretching from 1796.6–2516 cm−1 indicates the presence of two
functional groups CH2 and C=O, and when the wave length above 2526 cm−1 sug-
gest the absence of amide, catalytic acid and the presence of O-H.
Table 8.3 shows the elemental analysis results of the developed catalyst obtained
using energy dispersive X-ray (EDX-ray). At the temperature of 550 °C, the main
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 351

Table 8.3 Energy dispersive X-ray (EDX-ray) result of elemental analysis of the developed
catalyst
Constituents
Elements Symbol % Compositions
Calcium CaO 71.2
Potassium K2O 14.1
Sodium Na2O 3.0
Phosphorus P2O5 2.2
Magnesium MgO 6.3
Iron FeO 1.3
Zinc ZnO 1.2
Other <1.0

Table 8.4 Properties of sample of catalysts activity calcined at 800 °C for 3 h

N2-AA TPV BS (μmole.g−1) BSD Bio
Catalysts (m2 g−1) (cm3 g−1) %CaO 400 < BS < 500 > 500 TBS (μmol/m2) (wt.%)
CPKSH 1.00 0.003 57.50 20 92 112 112.00 80.34
CFKNP 1.10 0.003 62.58 28 102 130 118.18 85.78
CMCP 1.20 0.003 71.20 42 116 158 131.67 94.52
N2-AA nitrogen adsorption analysis, TPV total pore volume, BS basic site, TBS total basic site, BSD
basic site density

composite is calcium carbonate; when there is an increase in thermal temperature to

650 °C, decomposition of calcium carbonate to form calcium (II) oxide started
occurring using a grain model [38]. It was observed that the thermal decomposition
of calcium carbonate to calcium II oxide occurred at a higher temperature than that
observed by [27, 30]. This observation may be related to the presence of some func-
tional groups present in the kola nut plant during growth period which occurred as
a result of carbon growth inhibition. Owing to the abundance of calcium and potas-
sium in the developed catalyst, other metals (Na, Mg, Zn, P, Fe) are less discoursed,
but could also help in the transesterification reaction process [39].
Table 8.4 represents sample catalysts indicating the BET surface, total pore vol-
ume, basicity, and percentage of CaO converted through N2 adsorption–desorption
isotherm Brunauer-Emmett-Teller (BET) analysis. Observation from the results
shows the calcined mixed catalyst powder (CMCP) has high total basic site (TBS)
and high basic site density (BSD) than the individual catalysts (calcined palm kernel
shell husk (CPKSH) and calcined fermented kola nut husk (CFKNH)). However,
the BSD and the TBS of CFKNH are higher than that of CPKSH. However, the table
also reflects the result of biodiesel yields; at the same process conditions (run 27)
tested for each catalyst, the yields of biodiesel based on the nature of catalyst
showed that the CPKSH, CFKNH, and CMCP catalyst have high basicity for con-
version of mixed oil to biodiesel, but the CMCP produced highest yield (95.52%
wt.), due to the percentage of CaO in the catalyst and high basic site density. The
352 T. F. Adepoju and Y.-T. Hung

value of biodiesel yield (85.78% wt.) obtained when CFKNP was used (80.34%
wt.) was higher than the value obtained when CPKSH was used. This could be due
to fermentation process adopted which increases the surface area after thermal treat-
ment, thereby increasing the rate of reaction during transesterification.

8.3.3 Conversion of Oils to Biodiesel

8.3.3.1 Oil Blend

The results of mixture of oils (PFO and NO) in proper ratio are displayed in
Table 8.5. The initial acid value of NO was 14.66 mg KOH/g oil, having a density
of 902 kg/m3 at 40 °C and viscosity of 25.50 mm2/s at 40 °C, while PFO has the
initial acid value of 0.85 mg KOH/g oil, having a density of 870 kg/m3 at 40 °C, and
viscosity of 23.95 mm2/s, respectively. The results of the blend of the oil are dis-
played in the table. It was observed that the low viscous oil (21.50 mm2/s) with
moderate low density (852 kg/m3) was obtained at NO60; the oil is free-flow oil, and
with light hydrocarbon and low wax content.

8.3.3.2 Esterification of BO

The acid value of the low viscous BO/MO (4.10 mg KOH/g oil) was reduced to
minimum value (0.586 mg KOH/g oil) within the reaction time of 1 h, MeOH/OMR
of 4:1, and H2SO4 of 1.00 (v/v). Other methanol-to-oil ratio could further reduce the
acid value, but the ratio used in this study is sufficient for successful conversion of
esterified BO to a resulting mixed oil biodiesel (MOB) [30].

Table 8.5 Acid values, densities, and viscosities of different oil blend
Acid value (mg KOH/g Density at 40 °C (kg/ Viscosity at 40 °C
Blends oil) m3) (mm2/s)
100: 0 (NO100) 14.66 902 25.50
10:90 (NO10) 13.24 890 23.00
20:80 (NO20) 12.82 882 23.10
30:70 (NO30) 11.54 878 23.40
40:60 (NO40) 9.65 875 22.50
50:50 (NO50) 6.60 870 22.10
60:40 (NO60) 4.10 852 21.50
70:30 (NO70) 3.26 854 23.40
80:20 (NO80) 3.01 850 23.40
90:10 (N090) 2.98 850 22.10
0:100 0.85 870 23.95
(PFO100)
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 353

8.3.3.3 Statistical Optimization of Base-Catalyzed Transesterification

For statistical optimization of base-catalyzed transesterification of BO to biodiesel,

build information was developed, design expert version 12.0.3.0 was used, the study
type was response surface, the design type was central composite, the design model
chosen was quadratic while the build time was 15 min, the subtype was randomized,
and runs generated were 30, and these runs were carried out based on four-factor
three-level variable and the results are presented in Tables 8.6 and 8.7. From the
table, the highest MOB yield of 98.05 (wt. %) was obtained at catalyst amount of
2.0 (g), reaction time of 70 min, reaction temperature of 60 °C, and MeOH/OMR of
7:1 (mL/mL). The results in the table were optimized, based on the constraints

Table 8.6 Transesterification of mixed oil (MO) to mixed oil biodiesel (MOB)
Std. runs X1 X2 X3 X4 MOB (wt. %) PMOB (wt. %) Residual
1 −1.000 −1.000 −1.000 −1.000 83.83 83.81 0.0158
2 1.000 −1.000 −1.000 −1.000 78.58 78.65 −0.0717
3 −1.000 1.000 −1.000 −1.000 89.50 89.50 −0.0017
4 1.000 1.000 −1.000 −1.000 82.00 89.50 −0.0017
5 −1.000 −1.000 1.000 −1.000 88.15 88.23 −0.0817
6 1.000 −1.000 1.000 −1.000 84.63 84.57 0.0558
7 −1.000 1.000 1.000 −1.000 90.16 90.14 0.0158
8 1.000 1.000 1.000 −1.000 84.08 84.15 −0.0717
9 −1.000 −1.000 −1.000 1.000 88.09 88.17 −0.0783
10 1.000 −1.000 −1.000 1.000 86.71 86.66 0.0492
11 −1.000 1.000 −1.000 1.000 93.93 93.92 0.0092
12 1.000 1.000 −1.000 1.000 90.01 90.08 −0.0683
13 −1.000 −1.000 1.000 1.000 96.22 96.15 0.0692
14 1.000 −1.000 1.000 1.000 96.00 96.15 −0.1483
15 −1.000 1.000 1.000 1.000 98.05 98.13 −0.0783
16 1.000 1.000 1.000 1.000 95.84 95.79 0.0492
17 −2.000 0.000 0.000 0.000 92.71 92.69 0.0225
18 2.000 0.000 0.000 0.000 85.25 85.19 0.0625
19 0.000 −2.000 0.000 0.000 84.08 84.03 0.0525
20 0.000 2.000 0.000 0.000 89.39 89.36 0.0325
21 0.000 0.000 −2.000 0.000 87.65 87.62 0.0325
22 0.000 0.000 2.000 0.000 97.80 97.75 0.0525
23 0.000 0.000 0.000 −2.000 80.25 80.22 0.0292
24 0.000 0.000 0.000 2.000 96.27 96.21 0.0558
25 0.000 0.000 0.000 0.000 94.32 94.49 −0.1667
26 0.000 0.000 0.000 0.000 94.52 94.49 0.0333
27 0.000 0.000 0.000 0.000 94.52 94.49 0.0333
28 0.000 0.000 0.000 0.000 94.52 94.49 0.0333
29 0.000 0.000 0.000 0.000 94.52 94.49 0.0333
30 0.000 0.000 0.000 0.000 94.52 94.49 0.0333
PMOB predicted mixed oil biodiesel
354 T. F. Adepoju and Y.-T. Hung

Table 8.7 Predicted and validated MOB and its variable conditions
Predicted MOB (wt.%) catalyst reaction reaction
yield amount time temperature MeOH/OMR
98.05 (wt.%) 2.179 g 57.45 min 59.91 °C 5.9:1 (mL/mL)

Fig. 8.2 Variable level of contribution to the response in solution of 81 outcomes

(variable factors), 81 solutions (Fig. 8.2) were randomly provided, and the optimum
predicted yield of MOB was 98.05 (wt. %) at catalyst amount of 2.179 (g), reaction
time of 57.45 min, reaction temperature of 59.91 °C, and MeOH/OMR of 5.9:1
(mL/mL). The predicted value was validated in triplicate and an average MOB yield
of 98.03 (wt. %) was obtained. This showed that the base-catalyzed transesterifica-
tion of mixed oil to MOB is successive and the CaO derived from PKSH and FKNH
proved to be a suitable heterogeneous catalyst for the reaction process.
Furthermore, the results of experimental MOB yield and the selected factors
were fitted into second-order polynomial through regression coefficients and sig-
nificance of response surface quadratic equation. The results of second-order poly-
nomial equations obtained are presented in coded factor in Eq. (8.2) and actual
factor in Eq. (8.3).
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 355

Final equation in terms of coded

MOB  wt.%   94.52  1.88 X 1  1.33 X 2  2.53 X 3  4.00 X 4

0.5837 X 1 X 2  0.3763 X 1 X 3  0.9138 X 1 X 4  0.9438 X 2 X 3
0.0163 X 2 X 4  0.8912 X 2 X 3  1.39 X 12  1.95 X 22  0.4510 X 32  1.57 X 42 (8.2)

Final equation in terms of actual

MOB  wt.%   121.51391  3.52813 X 1  3.68069 X 2  2.95221X 3  0.403854 X 4

0.058375 X 1 X 2  0.075250 X 1 X 3  0.456875 X 1 X 4  0.018875 X 2 X 3  0.000812 X 2 X 4
  0.089125 X 3 X 4  1.38729 X 12  0.019485 X 22  0.018042 X 32  0.391823 X 42
(8.3)

The coefficient is the direct measure of influence of each variable on the response
(MOB). Following the second-order polynomial equation, negative coefficient
decreases the response while a positive coefficient increases the response based on
Eq. (8.2). All the linear variables have positive influence on the response; however,
X4 (4.000) with F-value = 48539.79 and p-value < 0.0001 is the most positively
significant variable among the selected variables (Table 8.8). Furthermore, the
model F-­value of 8034.35 implies the model is significant. There is only a 0.01%
chance that an F-value of this large could occur due to noise. Further analysis
proved that all the selected variables considered for MOB production via methano-
lysis of developed catalyst were remarkably significant except the interaction of
X2X4 (with p-value = 0.4760) that was non-significant term. The lack of fit F-value
of 0.4151 with p-value = 1.28 implies the lack of fit is not significant relative to the
pure error. There is a 77.68% chance that a lack of fit F-value could occur due to
noise. Non-­significant lack of fit is good [40]. Based on fit statistics and model com-
parison statistics, the coefficient of determination (R2) of 0.9999 obtained showed
the proportion of the variances in the dependent variable that is predictable from
independent variable.
Predicted R2 of 0.9994 was in reasonable agreement with the adjusted R2 of
0.9997. Meanwhile the difference is less than 0.200 with high adequate precision
greater than 4, which is required for a fit model.
Further process optimization was evaluated through the plot of predicted vs.
actual MOB yield; a straight line passing through the points indicated that the pre-
dicted values agreed with the experimental results, and the differences is negligible
(supplementary file: SF2). Also, the interaction among the variables was tested
through the three-dimensional plots. Fig. 8.3(a–f) shows the interactions among the
selected variables on the MOB yield. A plot of interaction between the reaction time
and catalyst amount on MOB yield is shown in Fig. 8.3(a); Fig. 8.3(b) depicts a plot
of interaction between reaction temperature and catalyst amount on MOB yield,
while a plot of interaction between reaction time and MeOH/OMR on MOB yield is
showed in Fig. 8.3(c). Fig. 8.3(d) shows a plot of interaction between MeOH/OMR
and reaction temperature on MOB yield, and Fig. 8.3(e) is the plot of interaction
356 T. F. Adepoju and Y.-T. Hung

Table 8.8 Test of Significance for Every Regression Coefficient for FAME production
Source Sum of squares df Mean square F-value p-value
Model 889.10 14 63.51 8034.35 <0.0001
X1 84.38 1 84.38 10674.37 <0.0001
X 42.61 1 42.61 5391.06 <0.0001
X3 153.93 1 153.93 19473.27 <0.0001
X4 383.68 1 383.68 48539.79 <0.0001
X1X2 5.45 1 5.45 689.77 <0.0001
X1X3 2.27 1 2.27 286.55 <0.0001
X1X4 13.36 1 13.36 1690.07 <0.0001
X2X3 14.25 1 14.25 1802.86 <0.0001
X2X4 0.0042 1 0.0042 0.5345 0.4760
12.71 1 12.71 1607.86 <0.0001
X12
52.79 1 52.79 6678.32 <0.0001
X 22
104.14 1 104.14 13175.02 <0.0001
X32
5.58 1 5.58 705.94 <0.0001
X 42
Residual 67.38 1 67.38 8523.77 <0.0001
Lack of Fit 0.1186 15 0.0079
Pure Error 0.0852 10 0.0085 1.28 0.4151
Cor Total 0.0333 5 0.0067
Fit statistics
R2 0.9999
Adjusted R2 0.9997
Predicted R2 0.9994
Adeq. precision 309.8089

between MeOH/OMR and catalyst on MOB yield, while a plot of interaction between
the reaction temperature and reaction time on MOB yield are depicted in Fig. 8.3(f).
From the plots, it was noticed that there were mutual interactive effects of vari-
ables on the MOB yield; however, Fig. 8.3b, c and e plots showed perfect interactive
effects than Fig. 8.3a, d, f. The interactive effects observed in Fig. 8.3(f) is less than
that observed in Fig. 8.3(a and d). These showed that with respect to quadratic equa-
tion of second order, single variables played significant roles, making positive con-
tribution to MOB yield than the quadratic and the interactive variables.

8.3.3.4 Properties of MOB

The characteristics of mixed oil biodiesel (MOB), obtained from the transesterifica-
tions, of mixture of the oils via derived CaO base catalyzed derived from PKSH and
FKNH, along with the data reported for the biodiesel standard ISO 17828, EN
14112; 14,214, and ASTM D97; D664; D2500; D4530; D6079; D6751
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 357

Fig. 8.3 (a–f) Three-dimensional plots of interaction among the selected variables
358 T. F. Adepoju and Y.-T. Hung

Table 8.9 Physicochemical and fatty acid composition of MOB

Parameters This study Biodiesel standard
Physicochemical properties
Density (kg/m3) at 40 °C 825.20 880 ISO 17828
Viscosity at 40 °C/(mm2/s) 2.20 1.9–6.0 ASTM D6079
Moisture content (%) <0.001 <0.03 ASTM D6751
%FFA (as oleic acid) 0.21 0.25 max ASTM D664
Acid value (mg KOH/g oil) 0.42 0.5 max ASTM D664
Iodine value (g I2/100g oil) 47.20 120 max EN 14214
Peroxide value (meq O2/kg oil) 73.50 – Jishy and Sankar [11]
HHV (MJ/kg) 41.00 – Chinyere et al. [7]
Cetane number 64.62 51 min EN 14214
Mean molecular mass 296.92 – Chinyere et al. [7]
Diesel index 75.86 40.40 min ASTM D6751
API 39.97 36.95 min ASTM D6751
Aniline point 136.65 331.0 max ASTM D6751
Flash point (°C) 120.00 120 min EN 14214
Cloud point (°C) +5 −3 to −12 ASTM 2500
Pour point (°C) +3 −15 to 16 ASTM 97
Cold filter plugging point (°C) +2 +5 max ASTM D6371
Oxidative stability (h) 4 3 min EN 14112
Carbon residue (% m/m) 0.028 0.05 max ASTM D4530
Fatty acid properties (%)
Palmitic acid (C16:0) 22.80 – –
Palmitoleic acid (C16:1) 1.90 – –
Stearic acid (C18:0) 14.00 – –
Oleic acid (C18:1) 45.20 – –
Linoleic acid (C18:2) 13.80 – –
Linolenic (C18:3) 2.10 – –
Others 0.20 – –

specifications are presented in Table 8.9. Observation shows that the properties of
produced MOB satisfied the biodiesel specifications standard. Main biodiesel phys-
icochemical properties including density, viscosity, acid value, cetane number,
higher heating value, diesel index, and API are within the limits recommended by
biodiesel standard. The peroxide value, iodine value, saponification value, flash
point, pour point, and cold point in the MOB agreed with the values reported by
other researchers [7, 11]. Oxidative stability of biodiesel is one of the most impor-
tant properties of fatty acid alkyl esters and primarily affects the stability of bio-
diesel during extended storage, compromises fuel properties, and impairs fuel
quality and engine performance. The minimum recommended value is 3 h, but the
value of 4 h minimum obtained in this showed the MOB is highly stable within a
short period. Cold filter plugging point (CFPP) is the lowest temperature at which a
8 Optimization Processes of Biodiesel Production from Pig and Neem (Azadirachta… 359

given volume of diesel type fuel still passes through a standardized filtration device
in a specific time when cooled under certain conditions. A high CFPP will clog up
vehicle engines in cold temperature regions. The maximum recommended value is
5; the value of 2 obtained for CFPP of biodiesel in this study showed the diesel is of
good qualities with low temperature clogging effects. The carbon residue value
obtained was within the ASTM D4530 specification. Fatty acid compositions of the
MOB showed the presence of saturated and monosaturated fatty acid in the MOB.

8.4 Conclusion

The mixture of two oils produced low viscous oil at blend level of 60:40 (v/v)
NO:PFO. CaO-based catalyst derived from the mixture of PKSH and FKNH is
helpful in successful conversion of the blend oil to biodiesel. The predicted bio-
diesel yield of 98.05 (wt. %) at different variable conditions was in agreement with
the validated average optimum biodiesel of 98.03 (wt. %).
Based on XRD analysis result, CaO is the predominant constituent element pres-
ent in the mixture of two catalysts, but all other elements constitute to the comple-
tion of reaction processes. The qualities of MOB were within the specification limits
of biodiesel standard which indicates that the developed catalyst successfully con-
vert the mixed oil with 60:40 ratio of NO:PFO to biodiesel.

Acknowledgments Authors acknowledge the effort of Bassey Gauis, Udoma William, and Mrs.
Florence of Department of Chemical Engineering, Akwa Ibom State University.
Funding: This work receives no fund from University, Private organization, or Government body.
Conflict of interest: Authors declare no conflict of interest whatsoever.

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Chapter 9
Castor Oil: A Promising Source
for the Production of Flavor and Fragrance
Through Lipase-Mediated
Biotransformation

Suman Singh, Naziya Syed, Shivani Chaturvedi,

Ashween Deepak Nannaware, Prashant Kumar Rout, and Yung-Tse Hung

9.1 Introduction: Why Castor Oil?

Castor plant (Ricinus communis) belongs to the family Euphorbiaceae and is com-
monly known as Arandi in Hindi. Castor plants are cultivated in South America,
India, and Africa and are widely grown in tropical regions [1]. Brazil, China, and
India are the major castor-producing countries in the world [2, 3]. Over 90% of cas-
tor oil is exported by India. On the other hand, about 84% of castor oil is imported
by the United States, European Union, and China [4]. The oil obtained from castor
beans is nonvolatile fatty oil. It is a pale yellow semi-transparent liquid having a
high boiling point and density. Castor oil and its derivatives are generally used in
making soaps, paints, surface coating, inks, waxes, cold-resistant plastic, biodiesel,
and lubricants. The importance of castor oil in the pharmaceutical industry is grow-
ing in terms of its use as an anti-inflammatory and antimicrobial agent. It also has
good moisturizing properties, due to which it is regarded as safe for hair and skin [5].

S. Singh · N. Syed · A. D. Nannaware · P. K. Rout (*)

Phytochemistry Division, CSIR-Central Institute of Medicinal and Aromatic Plants,
Lucknow, Uttar Pradesh, India
e-mail: [email protected]; [email protected];
[email protected]; [email protected]
S. Chaturvedi
Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute
of Technology Delhi, New Delhi, India
e-mail: [email protected]
Y.-T. Hung
Department of Civil and Environmental Engineering, Cleveland State University,
Cleveland, OH, USA
e-mail: [email protected], [email protected], [email protected]

© Springer Nature Switzerland AG 2022 363

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_9
364 S. Singh et al.

Furthermore, castor oil is also considered as a major raw material for the indus-
trial sector in the development of various products like perfumes and flavors, receiv-
ing increased attention nowadays. Castor oil is recognized as a low-priced,
biodegradable, and low-toxic oil compared to other vegetable oils. Due to this rea-
son, it has attained an impetus in the food industry in terms of food additives, mold
inhibitors, and flavoring agents [5]. Thus, all these characteristics of castor oil drive
it toward high market growth and valued products.
Castor oil, rich in ricinoleic acid (RA), is generally used as a substrate for the
production of γ-decalactone through biotransformation using efficient microbial
catalysts. RA having a long-chain fatty acid composes around 85% of the fatty acids
in castor oil [6, 7]. Consequently, it is promising to use castor oil as an economical
way and in copious amounts for industrial production of γ-decalactone [8]. Recently,
the use of the biotransformation method to produce aroma compounds has achieved
significant global attention. This biotechnological conversion method is of low cost
to produce flavor compounds. Before the bioconversion process, castor oil is sub-
jected to the hydrolysis process to produce 85% of pure RA-enriched castor oil [9].
Several microorganisms such as Candida, Yarrowia lypolytica, Pseudomonas, and
Rhodotorula have been reported to potentially produce aroma compounds [10].
RA enters the mitochondria within the cells of microorganism and degrade to
4-hydroxy-decanoyl-CoA after four β-oxidation cycles, which subsequently
cyclizes to γ-decalactone in the biotransformation process [11].
The current review mainly focuses on the properties, market value of castor oil,
and biotechnology methods to make value-added products from castor oil.

9.2 Geographical Distribution of Castor Plants

Castor is a perennial plant adapted to grow in tropical climates with a height of

10–12 m, whereas in temperate regions it is an annual plant with a height of 1–3 m
[12]. Castor plants grow well in low humidity seasons and fertile, slightly acidic,
and sandy loam soil. The optimum temperature required to for their growth is about
20–25 °C, but a temperature of above 35 °C can badly affect the seed setting. Cold
temperature can trigger chilling injury reducing plant growth [13]. In ancient times,
nearly about 6000 years ago, castor plants were cultivated for their oil content in
Egypt [14]. Castor plants are native to the Ethiopian region of east African tropical
regions and considered to be a weed in the south western United States. In India, it
grows abundantly on wastelands [15]. India produced 54% while China 23.4% of
castor seeds, followed by Brazil, i.e., 11.9%, in 2000–2009. Other countries that
produce castor seeds are Mozambique, Thailand, Paraguay, Cambodia, Ethiopia,
Indonesia, Madagascar, Peru, Russia, Philippines, Haiti, South Africa, Tanzania,
Vietnam, Uganda, Syria, Pakistan, Ecuador, and Colombia [16]. The highest castor-­
producing states in India are Gujarat (7.02 lakh ha), Rajasthan (1.54 lakh ha),
Andhra Pradesh (0.33 lakh ha), Telangana (0.22 lakh ha), and Odisha (0.04 lakh ha).
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 365

Narayanpet (6973 ha), Wanaparthy (5567 ha), Mahabubnagar (5104 ha), and
Gadwal (2163 ha) are the major growing districts in Telangana.

9.3 Castor Oil Scenario

In 2020, the production of global castor market has reached to 740.5 kilotons
(https://www.imarcgroup.com/castor-­oil-­manufacturing-­plant). It is anticipated to
show stable growth during the next 5 years. Asia Pacific leads the global market in
the growth of castor oil due to the constant expansion in the field of pharmaceutical,
surface coatings, and cosmetics industries. The largest market for castor oil has
been acquired by China itself because of having large geographical area [17]. In the
current pandemic (COVID-19) situation, many castor oil-producing entities have
been closed down due to the lockdown imposed by the government of different
countries, which has badly affected the market growth.
Due to the rising consumption of castor oil, its demand has increased globally at
a rate of 7320 tons per year. Based on the increased demand, it is not sufficient to
meet the expected rate of castor oil production for supply in the current pandemic
situation. Therefore, as the previous trends have shown that castor oil costs and
demand will increase gradually and are predicted to reach 1.81 billion by 2020 in
the global market [18]. The increasing use of castor oil can be found due to its appli-
cation in biopolymer and biofuels industries, cosmetics, and pharma industries.
Some companies working in the production of castor oil and its derivatives in the
global market are Adani Wilmer Ltd., Gokul Agri international Ltd. (GAIL), Jayant
Agro Organics Ltd., Kandla Agro and Chemicals Pvt. Ltd. (KACPL), Hokoku
Corporation, Thai Castor Oil Industries Co., Liaoyang Huaxing Chemical Co. Ltd.,
Bom Brazil, ITOH Oil Chemicals Co. Ltd., and Kanak Castor Products Pvt.
Ltd. [19].
The international castor oil market was recorded as $1180 million in 2018 and is
projected to reach $1470 million by the end of 2025, rising at a CAGR (compound
annual growth rate) of 2.8% between 2019 and 2025, according to global reports.
India exports castor oil to Europe, Japan, China, and the USA [20]. Due to the trade
war between USA and China, the demand from China has been slowed down.
Moreover, due to the shortage in the supply of castor oil in 2019, the export rate is
less as compared to 2018, when the country had exported 5.5–6 lakh tons of castor
oil. In the local market, its price has grown by 27% to Rs 1150 per 10 kg from Rs
900 a year ago, although the local market is very small compared to overseas
demand [21].
The castor seed production increased during 2018–2019 because its prices
enhanced up to 30% to Rs 54,750 a quintal. However, due to the COVID-19 pan-
demic across the globe, the prices have dropped to below Rs. 40,000 a quintal. Since
then, the prices have improved after the increase of exportation up to 71,900 tons in
May. From the commercial point of view, having hydroxylated fatty acid, castor oil
is a vital component for the chemical industry worldwide [22].
366 S. Singh et al.

9.4 Composition, Properties, and Applications of Castor Oil

Different methods have been applied for the extraction of castor oil from castor
beans, such as mechanical pressing, solvent extraction, or a combination of both.
Depending on the geographical distribution, extraction methods, and varieties, cas-
tor seeds contain around 44–55% of oil by weight. The alterations in the castor oil
properties can be a result of the extraction methods, different varieties, seasonal
variations, and type of soil. For example, low iodine value, low acid value, and high
saponification value have been reported in cold-pressed castor oil compared to sol-
vent extracted oil. In addition, the physicochemical property and composition have
also been observed to be changed due to seasonal variations [23]. Resembling other
vegetable oils, castor oil also contains unsaturated and saturated fatty acids attached
to a glycerol [24, 25]. The castor oil mixture has a higher content of fatty acids,
ricinoleic acid of around 90%, and other constituents in small amounts, as given in
Table 9.1. The commercial value of castor oil has increased due to the presence of
the high content of ricinoleic acid (RA), which can be utilized in various applica-
tions in the chemical industry.
Castor oil is colorless to pale yellow thick liquid with a unique taste, having a
mild odor, and have a boiling point at 586 K [26]. The distinctive property of castor
oil is due to the occurrence of the hydroxyl group in ricinoleic acid, which makes it
complementary with plasticizers of a broad range of synthetic and natural resins,
polymers, waxes, and elastomers [27]. For example, the oil has high specific gravity
and viscosity; it has little solubility in aliphatic petroleum solvents while highly
soluble in alcohols [28]. The physicochemical properties of castor oil are given in
Table 9.2. In comparison to other oils, castor oil has the highest viscosity and is
reported to be different in other parameters like cloud point, flash point, etc. As
compared to standard engine oil (SAE 40), the values of thermal conductivity, pour
point, viscosity, and density of castor oil were found to be higher [32].
Castor oil and its derivatives play a vital role in the production of soaps, lubricat-
ing agents, coating, pharma products, paints, plasticizers, food, pesticide, perfum-
ery, purgative, disinfectant, and germicidal agents [33]. Its high ricinoleic acid
content allows its ready derivatization through the presence of the hydroxyl group.
Biodiesel is also produced through the transesterification process. Due to these mul-
tiple applications, it is considered an industrial crop. Industrial uses of castor oil are
given in Table 9.3.

9.5 Castor Oil Processing Techniques

for Aromatic Compounds

As compared to other vegetable oils, castor oil is considered the most suitable raw
material for industrial purposes to convert various high value-added products. Due
to the presence of the high amount of RA in castor oil along with the occurrence of
 9

Table 9.1 Chemical composition of castor oil [23]

Composition Percentage Chemical formula Structure
Ricinoleic acid 89.5 C18H34O3

Linoleic acid 4.2 C18H32O2

Oleic acid 3 C18H34O2

Stearic acid 1 C18H36O2

Palmitic acid 1 C16H32O2

Dihydroxystearic acid 0.7 C18H36O4

Linolenic acid 0.3 C18H32O2

Eicosanoic acid 0.3 C20H40O2

Castor Oil: A Promising Source for the Production of Flavor and Fragrance…
367
368 S. Singh et al.

Table 9.2 Physicochemical properties of castor, jatropha, and pongamia oil

Physical properties Castor oil [29] Jatropha oil [30] Pongamia oil [31]
Density (Kg/m3) 899–955 889.27–910 924
Viscosity kinematic 40 °C (mm2/s) 15.25 33.86 40.2
Thermal conductivity (W/m°C) 4.727 0.09 –
Oxidation stability (110 °C, h) 1.1 – –
Specific heat (kJ/kg/K) 0.089 4.73 –
Cloud point (°C) −13.4 2 3.5
Flash point (°C) 260 144.67–292 225
Pour point (°C) 2.7 2.67 −3
Refractive index 1.480 1.48 –
Lipid content (%) 43.30 – –
Moisture content (%) 0.20–0.30 – –
Acid value (mg/g) 2.07–14.80 3.38 5.40
Iodine value (mg/g) 58.39–84.50 187.3 87
% Free fatty acid 3.4–7.4 1.70 –
Saponification value (mg/g) 175–182 – –
Peroxide value (mg/kg) 10–158.6 – –
Fire point (°C) 256 – –
Smoke point (°C) 215 – –
pH 5.8–6.2 – –
Congealing temperature (°C) 18 – –

hydroxyl and carboxyl groups and double bonds, it has more potential and versatile
applications in various industries [14]. It is also used in the formation of aroma
compounds where the oil needs to undergo a biotransformation process to obtain
value-added aroma compounds by using microbial catalysts. Before the biotransfor-
mation method, the castor oil undergoes chemical hydrolysis to purify the RA for
maximum conversion of aroma compounds. Cosmetics and pharmaceuticals
accounted for over 25% of total market volume in 2013 and was the largest applica-
tion segment. Growing demand for bioingredient-based cosmetics would continue
to remain a key driver for this segment. Castor oil and its derivatives are used in a
number of industries for the production of a wide variety of products. It is utilized
as a raw material in the production of a number of chemicals, which are further used
in the fabrication of surfactants, soaps, cosmetics, surface coatings, pharmaceuti-
cals, perfumes, plasticizers, greases, lubricants, and rubber (Table 9.3). Its basic
derivatives, undecylenic acid and heptaldehyde, are used to produce various per-
fumery compounds [41]. In pharmaceuticals and cosmetics, it is used as an ingredi-
ent in various formulations. The demand of around 4% of flavors and fragrance
from castor oil per annum is estimated globally. In Asia/Pacific region, the demand
for flavors and fragrances is estimated to be rising at a rate of about 7% per annum
through 2008. The current research studies mainly focus on the development of
flavor and fragrance from natural sources and have a preference to utilize raw mate-
rials that are safe and biodegradable. In light of this fact, there is an excellent pos-
sibility for castor oil derivatives in the international market.
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 369

Table 9.3 Applications of castor oil and its derivatives

Products Application References
Fuel and biodiesel Fatty acid methyl esters of castor oil when blended Bello and Makanju
with diesel fuel work as biodiesel [34]
Polymer materials Castor oil was cross-linked and polymerized with Yenwo et al. [35]
sulfur or di-isocyanates to produce the vulcanized
and urethane derivatives, respectively; plasticizers,
coupling agents, plastic films, polyols
Lubricants Castor oil has been utilized to produce soaps, wax, Burt and Mealy [36];
hydraulic fluids, corrosion inhibitors, aircraft Lehrer et al. [37];
lubricants, jet engine and racing car lubricants, and Dwivedi and Sapre
greases in some studies [38]
Fertilizers Mixture of castor cake and castor husks used as Lima et al. [39]
fertilizer supported significant plant growth up to
the dose of 4.5% of meal
Coatings and paints Castor oil can be successfully dehydrated by Grummitt and Marsh
nonconjugated oil–maleic anhydride adducts to [40]
provide constructive paint or furniture oil
applications; plasticizer for coatings, lacquers, and
varnishes
Textile chemicals Dyeing support, nylon, synthetic fibers, resins, Ibeagha [5]
synthetic detergents, pigment wetting agents
Pharmacological Act as antidiarrheal agent, antihelmintic, Ibeagha [5]
and medicinal use antidandruff, cathartic, emollient, also used a drug
delivery vehicle
Cosmetics and Used in hair tonic, shampoos, emulsifiers, Ibeagha [5]
perfumery polishes, deodorants, lipstick
Food Used as flavorings, surfactants, viscosity reducing Ibeagha [5]
additives

9.5.1 Hydrolysis and Purification of Castor Oil to Form

Ricinoleic Acid

Some chemical and biochemical methods have been performed for the isolation of
RA from castor oil. RA and glycerol have been yielded upon hydroxylation of the
ester linkages in the triglyceride molecules. Under chemical methods, castor oil
hydrolysis reaction is usually performed by the gradual addition of 80% sodium
hydroxide in castor oil for 1 h, and fatty acids are formed. The fatty acids are
released by adding distilled water and acidified with concentrated hydrochloric
acid. Further fatty acids are extracted in ethyl acetate and dried over magnesium
sulfate. Clearing up of fatty acids is performed by addition of n-hexane (1:5 w/v)
and kept at −4 °C for 3 days in darkness. This reaction produces ricinoleic acid and
glycerol. The purity of RA is 88%, and the rest are the palmitic, stearic, oleic, vac-
cenic, linoleic, and linolenic acids.
370 S. Singh et al.

Another method to isolate RA from castor oil is the use of biocatalysts such as
lipase enzymes. Some lipase-producing microbes such as Geotrichum candidum,
Candida rugosa, and Pseudomonas cepacia have been reported to be used for
hydrolysis of castor oil [42]. In this study, the hydrolysis of oil was performed in a
reaction including oil (100 mg), 0.5 M phosphate buffer (pH 7) (600 μL), and about
2–5 mg of lipase enzyme at 30 °C at 500 rpm for 1–4 h in an incubator shaker. The
level of hydrolysis was assessed by titration method against 0.1 N NaOH solutions
(pH 12), taking the hydrolysis mixture in diethyl ether:ethanol:water solution
(3:3:2). In this reaction, P. cepacia lipase showed successful conversion of castor oil
to RA up to 27% in comparison to other microbial enzymes like C. rugosa and
G. candidum that are recorded to be 13%. Another study also exhibited that when
immobilized C. rugosa was applied for the lipolysis of castor oil, the yield of RA
was about 20–40% under controlled conditions such as pH, temperature, and the
quantity of enzymes and substrate [43]. One more remarkable study was done by
Piazza and Farrell [44] on hydrolysis of castor oil using ground oats (Avena sativa
L.) lipase, which has shown a conversion of 90% of RA.
Another green method using microwave-assisted extraction of RA from castor
oil has been described [45] wherein hydrolysis of castor oil was done by ethanolic
KOH solution kept in a microwave oven with a few pieces of ice. A water condenser
and a magnetic stirrer were connected in the microwave oven and heating for 19 min
was given continuously for the reaction, which has yielded 89% of RA. The above-­
mentioned methods for the separation of RA from castor oil were good for RA yield
while using different catalysts and heat sources.

9.5.2 Chemical Transformation of Castor Oil

Due to the presence of long-chain hydrocarbon, OH, and COOH groups and double
bonds in RA, there are lots of possibilities of modifications and transformations of
the castor oil into a number of value-added products. Some chemical reactions have
been performed, for instance, esterification [46] and amidation [24], leading to
transformation due to the presence of carboxylic group, while double bond allows
the transformation of the oil through a number of reactions like carbonylation [47],
hydrogenation [24], and epoxidation [48]. In addition, the presence of OH group
also drives the chemical reactions of dehydration [49] to remove the hydroxyl group
and acetylation [48] and alkoxylation [50] to enhance the unsaturation of the oil. A
new double bond formation occurs in the chain of ricinoleic acid upon catalytic
dehydration reaction, which results in good color maintenance, fast drying, and
water resistance for protective coatings [49]. Further heating at 249.85 °C in the
presence of NaOH, capryl alcohol (2-octanol) and sebacic acid (a 10-carbon dicar-
boxylic acid) are formed. Both capryl alcohol and sebacic acid have several applica-
tions [51]. A study has reported that when a co-solvent system involving ethanol
and isopropyl ether (35:65; v/v) applied to castor oil produced a purified RA (about
98.6% purity). The recovery of RA was recorded at about 70%, and the total yield
of 55.5 ± 2.5% was obtained [51].
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 371

9.5.3 Biotransformation of Castor Oil to Form Enriched

Flavored Products

The synthesis of aroma compounds can be done by the biotechnological methods

following two important techniques: one is to use biotransformation method, and
another is via de novo synthesis. Complex molecules are synthesized from simple
molecules (sugars, amino acids, nitrogen salts, minerals, etc.) using intricate meta-
bolic pathways of organisms, and this is defined as the de novo synthesis method.
Biotransformation refers to the formation of value-added products from structurally
similar substrates in a single reaction catalyzed enzymatically (microbial cells or
pure enzymes) [52]. Biotransformation or bioconversion is considered a greener,
cheaper, and commercial approach in comparison to de novo synthesis methods for
the production of bioflavors [53]. In 1950, microbial production of blue cheese-note
compounds was discovered the first time, and since then, numerous bio-based
aroma compounds have been developed during the time [54].
γ-Decalactone is a peach-like aroma compound extensively used in food and
beverages, which is the cause of the big interest in its biotechnological production
[55]. To enhance the accessibility of ricinoleic acid to cells, castor oil can be hydro-
lyzed by lipases [56], generating esters such as methyl ricinoleate [55]. The process
comprises the breakage of ricinoleic acid into 4-hydroxy-decanoic acid, a precursor
of γ-decalactone, which is achieved by the process of peroxisomal 𝛽-oxidation
enzymes [57].

9.5.3.1 Lipase-Mediated Biotransformation

A variety of microbial strains are able to convert substrates into valuable aroma
compounds, and the examples of some microbial strains involved in the bioconver-
sion process are given in Table 9.4, which demonstrate the potential of microbes for
the synthesis of aroma compounds. Some microorganisms such as Pseudomonas,
Candida, Rhodotorula, and Yarrowia lipolytica have been reported for higher pro-
ductivity of aroma compounds through biotransformation [10]. Lactones are well
recognized for their wonderful flavor and fragrance (like pineapple, peach, rasp-
berry, apricot, mango, coconut, papaya, grapes), which are produced industrially
hundreds of tons yearly [64]. γ- and δ-lactones are the highly valuable five- and
six-membered rings, respectively, with 12 carbons equal or less. It comprises com-
pounds like γ-decalactone/4-decanolide (peach-like) [65], 4-dodecanolide (fruity-­
coconut like) [66], 4-octanolide (sweet herbaceous coconut-like) [67], 5-decanolide/ᵹ
-decalactone (creamy-coconut peach-like) [68], 5-dodecanolide (fruit-oily peach-­
like) [69], and 6-pentyl-α-pyrone (6PP, strong coconut-like) [70]. Although lactone
can be synthesized chemically from keto acids, production of δ-decalactone (DDL)
and γ-decalactone (GDL) through the biotransformation method is growing cur-
rently due to their generally recognized as safe (GRAS) values reaching between
US$ 1400 kg−1 and US$ 6000 kg−1 [71]. However, the prices reduced up to US$ 300
372 S. Singh et al.

per kilogram due to the growing production of both lactone compounds over the
years [65].
Castor oil is generally utilized as a substrate to produce γ-decalactone by yeast
cells through β-oxidation cycles of fatty acids recurring in the cells [11, 72].
However, prior to entering the biotransformation process, the castor oil is hydro-
lyzed to get refined castor oil having 86% of ricinoleic acid as the major component
[9]. In the microbial cells, RA inserts inside the microbial cells, and after recurring
four β-oxidation cycles, RA breaks into 4-hydroxy-decanoyl-CoA, which subse-
quently cyclizes to γ-decalactone [11]. In a study, it has been observed under bio-
conversion reaction that γ-decalactone concentration was 52.9 and 62.4 mg/L from
2.5% castor oil and 1.5% RA, respectively [73].
Currently, nonconventional oleaginous yeast, Yarrowia lipolytica, is getting
more importance in the bioconversion of castor oil or RA into γ-decalactone. Six-­
member family of acyl-CoA oxidases (Aox) encoded by POX1 to 6 gene, which
plays a vital role in β-oxidation of fatty acids, is present in Y. lipolytica [74]. This
four-step procedure involves mainly two oxidation steps, one hydration, and another
cleavage reaction, which are catalyzed by three enzymes. At each cycle, the com-
pound confers two-carbon shorter metabolite and an acetyl group [75]. One more
study has reported about Aox family and revealed that Aox2 is long-chain specific
(C18 to C10), whereas Aox3 is short-chain specific (C10 to C4) [74, 76, 77], both
showing strong activity [60]. Furthermore, some other studies utilized modified
strains of Y. lipolytica W29 (POX2 overexpressed and POX3–5 disrupted) and were
competent to make more lactone without any degradation [60, 76].
It has been illustrated that extracellular lipases, specifically the endogenous
lipase of Y. lipolytica (W29) and extracellular lipases (Lipozyme TL IM), were able
to release ricinoleic acid from castor oil, and as a result, γ-decalactone produces
rapidly [56, 78]. However, this method is not much appropriate as per industrial
approaches since it is time- and cost-consuming. Therefore, to fill the gap of this
problem, overexpression of the Lip2 enzyme would improve the production rate of
γ-decalactone with no additional expenses. This was further investigated by Braga
et al. [59] that Y. lipolytica (JMY3010) has the ability to synthesize extracellular
lipase due to the presence of an additional copy of LIP2 gene necessary for the pro-
duction of γ-decalactone. From a biotechnological point of view, gene expression
studies during bioconversion of RA into γ-decalactone are still very less; here, a few
studies involving upregulation and downregulation or disruption of some genes that
play an important role in γ-decalactone production are given in Table 9.4.

9.5.3.2 Bioconversion of Ricinoleic Acid (RA) to γ-Decalactone (GDL)

The most essential determining factor to produce γ-decalactone and 3-hydroxy-γ-­

decalactone is β-oxidation pathway in microbial cells, which was first discovered by
Okui et al. [79]. It is four-step reaction sequences, yielding an acyl CoA, which has
two carbons less and an acetyl-CoA. This reaction recurred many times until the
total breakdown of the compounds. Lactonization can take place at the C10 stage
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 373

Table 9.4 Various genes involved in the improved production of γ-decalactone

Disrupted/
Overexpressed underexpressed
Strain Substrate gene gene γ-decalactone Reference
Y. lipolytica Castor oil Aox2p, Lip2p POX2–5 Increased the Braga
MTLY40-2P γ-decalactone et al. [58,
production rate 59]
Y. lipolytica Methyl POX2 POX3 No Guo et al.
ricinoleate re-consumption of [60]
γ-decalactone
Yarrowia – CRF1 POX3 γ-decalactone Guo et al.
lipolytica TA1 production [61]
Escherichia Peach fruit PpAAT1 – Catalyze the Peng et al.
coli formation of [62]
γ-decalactone
Mono Methyl Pox3 – Formation of Waché
disrupted ricinoleate γ-decalactone et al. [63]
Yarrowia
lipolytica

resulting in the formation of strong fruity notes [80]. Under anoxic conditions, the
activity of 3-hydroxy-acyl-CoA dehydrogenase is minimized, because regeneration
of its cofactor (NAD+) is not enough. When the regeneration of NAD is not enough,
lactone accumulation takes place because a reduction in the β-oxidation flux
decreases the requirement for NAD and thus the cofactor is not necessary any lon-
ger, a limiting compound for the pathway. This phenomenon also happens when the
aeration of cells is changed, and this accumulation can be a signal of upscaling
complexities in industry. 3-Hydroxy lactone is released when aeration is reduced
with the declination in the β-oxidation pathway flux, and as a result, NAD is again
regenerated in plenty amount and 3-hydroxy lactone does not accumulate [81, 82].
A variety of microbial cells have been isolated from different sources by various
researchers for the bioconversion of lactones from different substrates (Table 9.5).
A study on bioconversion of methyl ricinoleate into γ-decalactone was carried
out using Pichiu guilliermondii through the peroxisomal beta-oxidation pathway
[102]. This study showed that enzymes of peroxisomes and peroxisomal β-oxidation,
i.e., catalase and acyl-CoA oxidase activities, respectively, were stimulated by
methyl ricinoleate. The activity of acyl-CoA oxidase increased approximately 40
times when this yeast was transferred in glucose medium in which fatty acid methyl
ester act as the only carbon source. The acetyl-CoA molecules are afterward moved
from the peroxisomes to the mitochondria by the acetyl-carnitine-transferase sys-
tem. It has been reported that peroxisomal β-oxidation has a detoxification role
because the peroxisomal reaction occurs in the absence of a mitochondrial respira-
tory chain involved in fatty acid consumption [103]. On the other hand, due to the
accumulation of hydroxy fatty acids in the culture medium, β-oxidation process
appears to be obstructed. Current research studies have focused on gene function
that encodes acyl-CoA oxidase isozymes, oxygen mass transfer rates, lipid
374 S. Singh et al.

Table 9.5 Biotransformation of various substrates using different strains in lactone

Conversion
Strain Substrate Product yield (%) References
Waltomyces lipofer 10-Hydorxystearic γ-Dodecalactone 76.1 An et al. [83]
acid
Waltomyces lipofer 10-Hydorxystearic γ-decalactone 46 An and Oh
acid [84]
Rhodotorula Castor oil γ-Decalactone 33 Alchihab
aurantiaca et al. [85]
Yarrowia lipolytica Castor oil γ-Decalactone 22.7 Rabenhorst
HR145 and Gatfield
[86]
Saccharomyces Grape juice γ-Butyrolactone Not reported Carrau et al.
cerevisiae [87]
Yarrowia lipolytica Ricinoleic acid γ-Decalactone Pagot et al.
(castor oil) [88]
G. fragrans and Ricinoleic acid γ-Decalactone 600 mg L−1 Neto et al.
Geotrichum sp. (castor oil) [89]
Ceratocystis Olive press cake δ-Decalactone Not reported Lanza et al.
moniliformis (coconut), [90]
γ-decalactone
S. salmonicolor Castor oil γ-Decalactone 54.6 mg/L Lee et al.
hydrolyzate [91]
Sporidiobolus Ricinoleic acid γ-Decalactone 0.39 mg/L/ Dufosse
ruinenii day et al. [92]
Yarrowia lipolytica 10-Hydroxystearic γ-Dodecalatone 24.3 Farbood
acid et al. [93]
Mortierella isabellina Dodecanoic acid γ-Dodecalatone 21.4 Han and Han
[94]
Baker’s yeast 10-Hydroxystearic γ-Dodecalatone 22.5 Gocho et al.
acid (0.5) and oleic [95]
acid
Sporobolomyces Oleic acid γ-Dodecalatone <14.0 Haffner and
odorus Tressl [96]
L. saturnus CCMA Castor oil γ-Decalactone 512.5 mg/L, Pereira de
0243 (pH 5), Y. 214.8 mg/L Andrade
lipolytica CCMA et al. [97]
0242 (pH -6.0)
Saccharomyces Ricinoleic acid γ-Decalactone 19.5 Rong et al.
cerevisiae MF013 [98]
Candida boidinii Safflower oil γ-Docecelactone 25 Jo et al. [99]
Yarrowia lipolytica Castor oil γ-Decalactone 0.061 Farbood and
Willis [100]
Yarrowia lipolytica Castor oil γ-Decalactone 0.0613 Gomes et al.
[9]
Geotrichum fragrans Castor oil γ-Decalactone 0.012 Neto et al.
[89]
Penicillium roqueforti Soybean oil γ-Docecelactone 0.00036 Chalier and
Crouzet
[101]
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 375

Fig. 9.1 The biosynthetic pathway of γ-decalactone (C10) from RA (C18) and the enzymes
involved in the yeast β-oxidation cycles in the biotransformation process

metabolism, interactions of cell–substrate to increase the production of γ- decalac-

tone, and selection of overproducing mutants. β-Oxidation is a four-reaction cycle
that shortens the carbon chain by two after catalyzed in peroxisomes by the follow-
ing enzymatic activities: acyl-CoA oxidase, 2-enoyl-CoA hydratase, 3-hydroxy-
acyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase (Fig. 9.1). A 10-carbon
compound (4-hydroxydecanoic acid) is obtained from the 18-carbon chain in the
last reaction, which is then cyclized to γ-decalactone. The products obtained from
biotransformation reaction also fluctuate according to the precursor fatty acid: ricin-
oleic acid is degraded to an 8-carbon acid, whereas homoricinoleic acid degradation
is guided to synthesize an intermediate of 11-carbon. The uncertainty of hydroxyl
fatty acid oxidation relies both on the carbon chain length and hydroxy group posi-
tion [104].
Even though some attempts have been made so far, the products yields are too
low; therefore, additional studies are required to overcome the constraints observed
at present to make the biotechnological process more useful. The necessary step
toward higher production of aroma compounds is mainly done by focusing on the
optimization attempts on fermentation technologies and downstream processes.
376 S. Singh et al.

9.5.3.3 Optimal Conditions for Biotransformation

The biotransformation process is dependent on various parameters such as tempera-

ture, pH, time, enzyme amount (microbial cells), substrate concentration, and types
of additives for γ-decalactone production. The first parameter is carbon source (sub-
strate such as castor oil or ricinoleic acid) that acts as an activator of enzymes in the
pathway of γ-decalactone production. The second parameter is nitrogen sources
comprising peptone and yeast extract that are imperative to enhance cell growth and
biotransformation process. The last factor is pH, which should be optimized to
maintain the highest cell growth and γ-decalactone production.

9.5.3.3.1 Effect of Carbon Source

Synthesis of lipase, which is responsible for the bioconversion of castor oil into
lactones, can be induced by the carbon sources in the culture medium. Some exam-
ples of carbon sources are carbohydrates, glycerol, organic acids, oils or fats, and
other alcohols [105]. Some oils such as soybean oil, olive oil, oleic acid, and tribu-
tyrin have been observed to be effective in inducing lipase activity [106, 107]. For
instance, the use of oleic acid exhibited increased production of lipase and biomass
content in comparison to other fatty acids of different carbonic chains [108]. Another
study showed that vegetable oil possessing linoleic and oleic acids was found super-
lative in lipase production from Candida species [109]. Olive oil used as a carbon
source was also found to induce lipase activity by Y. lipolytica [110].

9.5.3.3.2 Effect of Nitrogen Source

Like carbon source, nitrogen source is also a chief factor influencing the synthesis
of lipase enzymes. Both the inorganic and organic nitrogen have an essential role in
the synthesis of the enzyme [109, 111]. Yeast extract, malt extract, peptone, amino
acids, urea, nitrate and ammonium salts, agroindustrial waste, such as water soy
flour and corn, are nitrogenous compounds generally used for lipase production
[111, 112]. The selection of nitrogen compounds mostly depends on the microor-
ganism used and the addition of other ingredients in the culture medium [112].
Nitrogen sources are the necessary component for cellular physiology because they
provide vitamins and amino acids as enzyme cofactors [113]. For instance, cell
growth and lipase synthesis were found to be increased by the addition of yeast
extract and tryptone. In a study, various nitrogen sources have been examined on the
growth of Y. lipolytica for lipase synthesis, and it was observed that mineral nitro-
gen did not show any significant effect on the growth of yeast possessing lipase
activity [114]. Tryptone N1 (a casein hydrolysate) showed maximum production of
lipase compared with other nitrogen-containing organic or mineral substrates [105].
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 377

9.5.3.3.3 Effects of Temperature

The temperature influences the growth parameters of microbes, such as the specific
growth rate and the adaptation time (lag phase), and affects the biosynthesis of pri-
mary and secondary metabolites [115]. It has been observed in a study that lipase
production by Y. lipolytica (681) was most influenced by the temperature [116]. The
optimum temperature for significant lipase activity was 29.5 °C.

9.5.3.3.4 Effect of pH

Microbial growth can be affected by the hydrogen ion concentration in a culture

medium indirectly, affecting nutrient accessibility or directly acting on cellular sur-
faces [115]. It is reported that under low pH, the synthesis of lipase by liquid sub-
strate fermentation, with diverse microorganisms, is highest at the end of cultivation
[117, 118]. Mooradi et al. [73] have optimized that under acidic pH, nitrogen source,
and increased amount of yeast extract and castor oil, the production of γ-decalactone
will be enhanced. Other researchers have also observed that acidic pH is more suit-
able for bioconversion of RA into 4-hydroxydecanoic acid and then γ-decalactone
[57, 85, 91, 119].

9.5.3.4 Significance of Bioreactors

Bioreactors are devices for biotransformation or any fermentation process in which

biological or biochemical reactions are performed under closely monitored and con-
trolled conditions. A number of different types of bioreactors have been used for
fermentation methods. Among these, two are commonly categorized as solid-state
or stirred-tank bioreactors with two different fermentation methods, such as SSF
and SmF, respectively. For research and development, both SmF and SSF have been
employed for the production of bioflavor compounds. Approaches for the develop-
ment of new types of reactors are going on continuously for special purposes [120].
In SmF type of bioreactor, mostly the reactions of microorganisms and substrate are
performed in a liquid medium and the products produced by microbial enzymes
reaction are recovered from the liquid medium and purified. Gas/air mixture
required for fermentation reaction is delivered to a culture medium in sterilized
environments. During the bioconversion process in bioreactors, optimized condi-
tions for pH, temperature, aeration, and foam sensors are needed. Mechanical agita-
tion is required for mixing and bubble dispersion. In addition, nutritional factors
such as carbon and nitrogen sources are also important for the growth and synthesis
of flavor compounds in lab as well as industrial scale [81, 121]. Several researchers
have studied the microbial conversion of bioflavor compounds by SmF-type biore-
actors [122–126].
378 S. Singh et al.

De novo synthesis or transformation methods are generally performed in solid-­

state fermentation (SSF) system for the aroma compound production using various
yeasts and fungi such as Kluyveromyces marxianus, Ceratocystis fimbriata,
Moniliella suaveolens, Trichoderma harzianum, Pityrosporum ovale, Ceratocytis
oniliformis, Aspergillus niger, and Rhizopus oryzae [127–131]. Furthermore, com-
patible combinations of yeasts and fungi, which cannot be cultured in submerged
fermentation (SmF), can yield many aroma-active compounds in SSF [132]. In SSF,
the fermentation process takes place in the absence of water on solid support gener-
ating important metabolites in the presence of microbes. SmF is a technique in
which microorganisms can grow in liquid broth consuming the nutrients to release
the desired metabolites into solution during fermentation. Although SSF has been
used since ancient times and has many biotechnological advantages, there have
been some limitations involved during scaling up in this technology, usually due to
the heterogeneous nature of the mass, substrate, and troubles in heat transfer [127,
132, 133]. Besides the development of digital imaging technologies for assessing
the growth kinetics in filamentous fungi in SSF, further improvement in mathemati-
cal modeling tools has been achieved to describe the scale-up studies [134].
Braga et al. [58, 59] have used stirred tanks and airlift bioreactors for the produc-
tion of γ-decalactone using castor oil in batch cultures of Y. lipolytica (W29). Airlift
bioreactors are generally considered for the developing processes on the basis of
aerobic cultures because of the requirement for high oxygen transfer rates. Air was
introduced at the bottom of the bioreactor using a five-hole sparger. γ-Decalactone
concentration (around 3 g/L) was twofold increased in the airlift compared to the
STR (stirred tank reactor). In a study, it has been reported that when castor oil (sub-
strate) was reacted with microbial enzymes, the production of GDL (11 g/L) was
recorded in 55 h [65].
For immobilized enzymatic reactions for the production of bioflavor in industry,
packed-bed bioreactors, especially operated in continuous mode, and fluidized bed
reactors are the most often utilized [127, 135]. In addition, studies have led to
designing new reactors such as immersion bioreactors, rotating drum bioreactors,
and mixed solid-state bioreactors, which would succeed over the troubles for large-­
scale productions [10, 131].
Fed-batch strategy is an interesting strategy to avoid degradation of γ-decalactone
and cut down the inhibitory impacts of ricinoleic acid on the cells [136]. Using
intermittent feed in fed-batch, aroma concentration of 6.7 g L−1 was obtained as
compared to 1.9 g L−1 in batch fermentation and the formation of the side product
3-hydroxy-γ-decalactone augmented concurrently to 10 g L−1 in the bioreactor.
However, in this system, the maintenance of an emulsion causes several restraints to
ensure the supply of fresh medium and remove an emulsion with the same charac-
teristics. Thus, the substrate addition by step-wise fed-batch (pulses) is a method of
avoiding this problem. In a step-wise fed-batch method, the productivity of
γ-decalactone of 0.043 g L−1 h−1 has been reported when methyl ricinoleate
(30 g L−1) was fed twofold to the bioreactor using Y. lipolytica W29 [136]. Braga
et al. [59] also attempted a step-wise fed-batch method with MTLY40-2P strain, in
which castor oil (60 g L−1) was supplemented in two pulses, leading to a twofold
increase in γ-decalactone synthesis (about 7 g L−1) as compared to batch mode.
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 379

9.6 Castor Oil: Advantages and Disadvantages

9.6.1 Advantages

Castor is regarded as the most important industrial and medicinal plant since thou-
sands of compounds can be extracted from it. These compounds have been reported
to have antimicrobial activity against diverse pathogenic bacteria [137, 138]. Castor
oil is extensively utilized in the industry with diverse applications [23]. The usage
of castor oil is safe from broad historical usage to industrial application. Some
ground-breaking technologies have been developed for the production of value-­
added castor oil chemicals and their derivatives. Food-grade castor oil is utilized in
the formation of food additives, flavorings, and mold inhibitors and in packaging
[139]. The products obtained by castor oil are not dangerous to the environment. It
is biodegradable, nontoxic, and agriculture oriented. The export of castor oil can
boost the economy of the country and also encourage the development of the agri-
cultural sector.
Castor cake can be used as organic manure, which prevents soil from exhausting.
Castor cake has a high content of N (6.4%), potash (1%), and phosphoric acid
(2.55%) and has moisture retention capacity. It also contains crude protein (20%),
ash (15%), and sugar (50%) [140]. Castor cake is used to control nematodes in soils
[141]. Ricin can remain in the soil for about 2 years after castor harvesting [142].
However, the effect of the residual toxin on flora and fauna in the soil is yet to be
determined.

9.6.2 Disadvantages

Castor contains toxic ricin in seeds. However, these toxic compounds can be uti-
lized to formulate drugs to treat many diseases globally [143]. Soil micro flora is
one of the most imperative factors that augment soil fertility in various ways.
Antimicrobial activity of castor was also observed against soil microbial commu-
nity, which in turn disturbs soil health and fertility during its cultivation in soil.
It has been observed that microbial diversity declined in soils when castor was
cultivated as compared to other plants [144]. There may be possibilities of the
occurrence of some residual inhibitors in the soil sensitive to certain fungal and
bacterial species affecting the growth of plants and soil health [145]. However, cer-
tain microorganisms have the ability to degrade and survive at high concentrations
of inhibitors. It has been observed in a study that Pseudomonas and Erwinia sp. can
efficiently degrade the protein in an in vitro assay. Consequently, these bacteria can
be utilized as biofertilizers for castor cultivation without damaging microbial diver-
sity in soil enhancing plant growth and soil health [144].
380 S. Singh et al.

9.6.3 Safety Assessment or Toxicity Study of Castor

The seeds of castor contain two types of proteins, R. communis agglutinin (RCA120)
and ricin (RCA 60), which are toxic to eukaryotic cells [146]. Ricin is a strong
cytotoxin with weak haemagglutinin properties, while RCA120 is less toxic and has
powerful haemagglutinin, which functions as an allergen causing a health hazard
during harvesting and processing [147]. Ricin remains in the meal after its oil
extraction. There are various methods for the detoxification of castor toxins [148,
149]. Ricin can be detoxified with the help of proteolytic enzymes, sodium ricinole-
ate, H2O2, KMnO4, autolyzed yeast or Azotobacter, halogens, ethanol, high tem-
perature, and UV irradiation. In addition, other methods like steam treatment, NaCl,
Ca (OH)2, formaldehyde, NH4OH, (NH4)2SO4, KMnO4, or urea can also be employed
for detoxification of ricin [150]. US Department of Health and Human Services
(NIH) has established safety measures for the usage of castor oil. In 1990, no harm-
ful side effects of castor oil have been noticed in the feed studies on rats or mice. In
90 days, studies on dietary concentrations showed no effect on survival or body
weight gains.

9.7 Concluding Remarks

Various types of aroma compounds are isolated from natural and synthetic sources
to meet the market demand. The aroma industry has a great interest in natural prod-
ucts, especially fragrance and flavor compounds; therefore, an alternative method
has been generated in the metabolic engineering field to obtain natural aroma com-
pounds. Microbial biosynthesis and biotransformations are considered safe and
most suitable technologies for the production of bioflavors and fragrances. This
method has a number of advantages over traditional methods because microbes can
be genetically and metabolically altered to improve the production of desired impor-
tant compounds.
Castor oil is a significant and potential nonedible crop, producing numerous
industrially important chemicals and products. Due to its lots of functional value in
agriculture, industry, pharmaceutical, and cosmetics sectors, castor oil is proved to
be a potential bio-based preliminary material increasing the economy of a country.
Castor oil has unique properties of the presence of a double bond, carboxylate
group, and hydroxyl group in the ricinoleic acid, which is useful in its easy
derivatization into vital industrial raw materials. Due to the manufacture of large-­
scale end products from castor oil and being a green bioresource for chemical trans-
formations, castor seed has global demand for enhanced production rate.
Castor oil conversion in aroma compound γ-decalactone by the biotransforma-
tion method using efficient microorganisms is a striking method, but the production
time is high and quantity is very low. Various approaches have been adopted to
overcome these problems by applying suitable microbial enzymes for the synthesis
9 Castor Oil: A Promising Source for the Production of Flavor and Fragrance… 381

of aroma compounds. Fed-batch fermentation method can be applied, choosing

suitable microorganisms for higher yields of lactone. This method is also useful to
overcome the inhibitory effect of the substrate in lactone production. Solid-state
fermentation is the most appropriate method in producing microbial metabolites in
large quantities, which also provides high-quality yields with improved product
characteristics accompanied by low economic costs. A much higher research thrust
on productivity improvement is needed for mitigating the demand–supply gap of
castor seeds. Besides breeding, efforts should be focused on three other prime
objectives: (1) improvement of castor oil quantity and quality, (2) development of
ricin-free castor, and (3) production of value-added aroma products from cas-
tor crop.
Next to these strategies, the application of bioprocess engineering has countless
benefits to attain higher yields and product concentrations by a comprehensive
understanding of the regulation of different pathways involved in aroma production.
However, so far, the products yields are very poor to make the biotechnological
method feasible, and more research studies are needed to overcome these limita-
tions. Furthermore, selection of mutant or new potential strains, fermentation meth-
ods, downstream processes, and up-scaling from lab to industrial levels need severe
studies to maximize the yield as well as lessen the limitations. The continuous
development of genetic engineering and systems biology tools, in association with
advanced strategies, will allow more bioflavors to be produced in this manner in
the future.

Acknowledgments The authors are grateful to the Directors of CSIR-CIMAP, Lucknow, and IIT
Delhi. The authors are thankful to CSIR India for funding the project PME/FTT-FTC/FC2020-23/
MLP-09/2020. The author (SS) would like to acknowledge the contribution of FTT project
MLP-009 for the funding. The author (NS) is thankful for the DST-INSPIRE fellowship
(IF:180344), and the author (SC) is grateful for the UGC WM PDF fellowship (F.15-1/2017/
PDFWM-2017-18-DEL-3915 (SA-II) from UGC, India).

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Chapter 10
Treatment and Minimization of Waste
in Baker’s Yeast Industry

Esra Can Doğan, Ayla Arslan, Nevim Genç, Levent Dağaşan,

and Yung-Tse Hung

10.1 Introduction

Yeasts have been used by humans to produce foods for thousands of years.
Nowadays, modern industries require very large amounts of selected yeasts to
obtain high-quality reproducible products and to ensure fast, complete fermenta-
tions [1]. Today, the overall production of yeast (≈3.5 Mt/year) is placed in around
300 industrial sites across the world [2].
Baker’s yeast, which is one of the main products in the preparation of bread mak-
ing, is manufactured through the aerobic fermentation of the selected strains of
Saccharomyces cerevisiae according to their special qualities relating to the needs
of the baking industry [3]. The water usage in the baker’s yeast industry is around
75 m3 water/ton of dry yeast. For industries using large quantities of water such as
baker’s yeast plants, it is essential to treat and reuse their wastewater. In the baker’s
yeast industry, the amount of wastewater is around 7.8 m3 wastewater/m3 molasses
[4]. The industrial production of baker’s yeast results in the discharge of large quan-
tities of dark-colored liquid wastes generally called weak vinasses. These wastes

E. C. Doğan · A. Arslan · N. Genç

Environmental Engineering Department, Engineering Faculty, Kocaeli University,
Kocaeli, Turkey
e-mail: [email protected], [email protected];
[email protected], [email protected]; [email protected], [email protected]
L. Dağaşan
SVL Food Biotechnology Consultancy, İstanbul, Turkey
e-mail: [email protected]
Y.-T. Hung (*)
Department of Civil and Environmental Engineering, Cleveland State University,
Cleveland, OH, USA
e-mail: [email protected], [email protected], [email protected]

© Springer Nature Switzerland AG 2022 391

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3_10
392 E. C. Doğan et al.

have a high organic content, are strongly nitrogenous, sulfate-rich, phosphorous

variable, and recalcitrant for biodegradation, and are highly colored due to the pres-
ence of melanoidins [3, 5, 6]. The dark brown color from baker’s yeast effluents
interferes with the absorption of sunlight, which reduces the natural process of pho-
tochemical reactions for self-purification of the surface waters [7]. Since the efflu-
ent generated from the baker’s yeast industry is highly polluted, it needs to be
treated before discharging into receiving water bodies in order to prevent important
environmental problems. In addition to pollution, increasingly stringent environ-
mental regulations are forcing the manufacturers to improve existing treatment and
also explore alternative methods of effluent management [8]. Land disposal options
generate problems with ground water pollution and are prohibited in some coun-
tries [9].
Industries have traditionally treated the waste products before discharging them
to the environment. Because the treatment occurs after the production of waste, this
type of treatment is called “end-of-pipe” treatment and this is being seriously ques-
tioned. The alternative solution, “waste minimization,” aims at reducing the pollu-
tion problem by dealing with it during the manufacturing process itself. There are
many ways by which waste minimization can be achieved. Improved housekeeping,
changing process technology, changing product, changing input material, recycling
process chemicals and raw materials, recovering by-product/waste, and reducing
input to the process have been proven successful in achieving waste minimization.
Yeast bread fermentation is closely related to alcoholic/brewery fermentation.
Therefore, we evaluated all fermentation with S. cerevisiae together. In this chapter,
we reviewed the processes of yeast production, treatment, disposal, reuse, and mini-
mization of wastewater/waste from the baker’s yeast industry. Here, biological,
physicochemical, and advanced treatment methods of wastewater are presented.
Also, the disposal/reuse options, waste, sludge, and by-products such as vinasses
are evaluated.

10.2 Baker’s Yeast Industry

10.2.1 Yeast

Yeasts belong to the group of living things called fungi. Yeast size can vary greatly
depending on the species, typically measuring 3–4 μm in diameter, although some
yeasts can reach over 40 μm. Yeasts are facultative anaerobes [10]. Yeast growth is
affected by a number of factors. These include the composition of the medium com-
monly sugar source, aeration (oxygen), agitation of the medium, pH (4.5–5.0) [1],
temperature (30 °C) [1], and period of propagation [10]. For their nutrition, yeasts
require a source of carbon for growth and energy, a nitrogen source for the synthesis
of protein and other nitrogenous materials, inorganic nutrients for the buildup of the
normal functioning and structure of the cell, and vitamins. In the absence of oxygen,
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 393

they can metabolize sugar into alcohol (ethanol) and carbon dioxide and low bio-
mass. In well-aerated conditions, the cells could be able to get enough energy and
convert sugar into carbon dioxide, water, and biomass. Bread, wine, sake, and beer
are made with the essential contribution of yeasts, especially from the species
S. cerevisiae [1]. S. cerevisiae is a species of budding yeast. It is the microorganism
behind the most common type of fermentation. It reproduces by a division process
known as budding [10]. The fermentations focus on a maximum biomass yield with
limited ethanol production, favoring the oxidative or aerobic metabolic yeast path-
way (Fig. 10.1) [11].
Until now, in the production of baker’s yeast, whether continuous, batch, or fed-­
batch bioreactors, different operating parameters have been controlled using mod-
ern control techniques so that maximum yields of production and economic profit
can be achieved. One of the most popular control techniques is the self-tuning

Fig. 10.1 Simplified growth pathways of Saccharomyces cerevisia [11]

394 E. C. Doğan et al.

method. Generalized minimum variance, pole-placement self-tuning, and general-

ized predictive control are known as self-tuning control techniques [12]. In the last
few decades, the yeast biomass production industry has contributed with many
advanced approaches to traditional technological tools with a view to studying the
physiology, biochemistry, and gene expression of yeast cells during biomass growth
and processing. This has provided a picture of the determinant factors for the com-
mercial product’s high yield and fermentative fitness. Cell adaptation to adverse
industrial conditions is a key element for good progress to be made in biomass
propagation and desiccation and toward the characterization of specific stress
responses during industrial processes to clearly indicate the main injuries affecting
cell survival and growth. One major aspect of relevance in the complex pattern of
molecular responses displayed by yeast cells is an oxidative stress response, a net-
work of mechanisms ensuring cellular redox balance by minimizing structural dam-
ages under oxidant insults. Different components of this machinery have been
identified as being involved in cellular adaptation to industrial growth and dehydra-
tion, including redox protein thioredoxin, redox buffer glutathione, and several
detoxifying enzymes such as catalase and superoxide dismutase, plus protective
molecules like trehalose, which play a relevant role in dehydration [1].

10.2.2 Substrates

The raw materials used as a substrate for industrial yeast biomass production are
usually agricultural, forestry, and food waste by-products [10]. Beet or cane molas-
ses are the main substrates used in yeast production plants. These materials were
selected for two main reasons: first, yeasts grow very well using the sugars present
in the molasses and second, they are economically interesting since they are a waste
product coming from sugar refineries without any other application. Usually, molas-
ses contain approximately 50% sugars, mainly sucrose [13], but the composition is
highly variable depending on the sucrose-refining procedure and on the weather
conditions of that particular year. Sucrose is extracellularly hydrolyzed by yeasts in
two monosaccharides, glucose and fructose, which are transported to and incorpo-
rated into the yeast metabolism as carbon sources.
In the last years, the price of molasses has increased because of their use in other
industrial applications such as animal feeding or bioethanol production, thus ren-
dering the evaluation of new substrates for yeast biomass propagation a trending
topic for biomass producers’ research. New assayed substrates include molasses
mixtures with corn steep liquor, different agricultural waste products, and other pos-
sibilities such as date juice or agricultural waste sources, also called wood molasses,
which can be a substrate only for yeast species capable of using xylose as a carbon
source [1].
Molasses could not supply all the essential nutrients for yeast growth. Therefore,
the addition of supplements such as (NH4)2SO4, urea, yeast extract or peptone as
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 395

Table 10.1 Percentage composition of beet and cane molasses [10]

Molasses type
Constituent Beet (%) Cane (%)
Total sugar as invert 48–58 50–58
Nitrogen 0.2–2.8 0.08–0.5
Total solid 78–85 78–85
P2O5 0.02–0.07 0.009–0.07
MgO 0.01–0.1 0.25–0.5
K2O 2.2–4.5 0.8–2.2
Carbon 28–34 28–33
SiO2 0.1–0.5 0.05–0.3
Al2O3 0.0005–0.06 0.01–0.04
Fe2O3 0.001–0.02 0.001–0.01
Total ash 4.0–8.0 3.5–7.5

nitrogen source, KH2PO4, H3PO4 as phosphorus source, other macro elements such
as calcium in the form of calcium salts, magnesium in the form of magnesium salts,
and microelements such as iron, zinc, copper, and manganese is necessary for maxi-
mizing biomass yield of S. cerevisiae or any other types of yeasts. Vitamins are also
required for yeast growth [10]. The aromatic nature of molasses is the result of
phenolic compounds [14, 15]. Table 10.1 shows the percentage composition of both
beet and cane molasses [10].
Aransiola et al. [16] investigated the bioconversion of raw cassava starch into
value-added products. This involved hydrolysis of extracted starch from freshly har-
vested cassava tubers using three different methods. The results show that the
hydrolysates obtained by hydrolysis of indigenous raw cassava starch via acid,
acid–enzyme, and enzyme–enzyme methods were all able to support the growth of
S. cerevisiae. The highest Dextrose Equivalent of 34 with a starch conversion effi-
ciency of 87.3% was achieved by the enzyme–enzyme hydrolysis method [16].
Sugar beets contain no coloring materials, but they do contain color-forming
substances. Color compounds in molasses are predominantly melanoidins (sugar–
nitrogen complexes), caramel substances, polyphenol–iron compounds, and, to a
lesser extent, plant pigments. The humic non-sugars (or polymerized forms of mela-
noidins) are claimed to be particularly relevant to the color of yeast produced. The
color of sugar solutions varies with pH, with the darker colors occurring at higher
pH levels [14, 15]. It is of great significance that baker’s yeast is not contaminated
with toxic substances like heavy metals, during its production or through contact
with unsuitable packaging materials. Contamination of baker’s yeast with lead (II)
may originate from molasses produced from sugar beet or sugarcane crops irrigated
396 E. C. Doğan et al.

with contaminated water or grown in areas adjacent to high lead (II) emission indus-
tries [17].

10.2.3 Production of Baker’s Yeast

The baker’s yeast production process flow can be divided into four basic steps,
namely molasses and other raw material preparation, culture or seed yeast prepara-
tion, fermentation and harvesting, and filtration and packaging [18]. Nowadays,
biomass propagation of wine, distiller’s, and brewer’s yeasts is usually achieved
using baker’s yeast plants. The procedure is designed as a multistage-based fermen-
tation, previously defined for the production of baker’s yeast using supplemented
molasses as growth media [1]. The flowchart of the baker’s yeast process is shown
in Fig. 10.2 [19]. Also, the source, distribution, and characteristics of generated
wastewater are shown in the process flow diagram.
The first stage is initiated with a flask culture containing molasses inoculated
with the selected yeast strain [1]. Yeast can grow in the presence or absence of air.
In bread dough, yeast grows very little under anaerobic conditions; instead, the
sugar that can sustain either fermentation or growth is used, mainly producing alco-
hol and carbon dioxide. This means that the baker who is interested in the leaving
action of the carbon dioxide works under conditions that minimize the presence of
dissolved oxygen. In contrast, under aerobic conditions, in the presence of a suffi-
cient quantity of dissolved oxygen, yeast grows by using most of the available sugar
for growth and producing only negligible quantities of alcohol. So, a yeast manufac-
turer that wants to produce more yeast cell mass works under aerobic conditions by
bubbling air through the solution in which the yeast is grown. The problem to the
yeast manufacturer, however, is not just as simple as adding air during the fermenta-
tion process. If the concentration of sugar in the fermentation growth media is
greater than a very small amount, the yeast will produce some alcohol even if the
supply of oxygen is adequate or even in abundance. Adding the sugar solution
slowly to the yeast throughout the fermentation process can solve this problem. The
rate of addition of the sugar solution must be such that the yeast uses the sugar fast
enough so that the sugar concentration at any one time is practically zero. This type
of fermentation is referred to as fed-batch fermentation [18].
At the end of the fermentation stage, the yeast is present as a suspension of cells
in a dark brown liquid containing the residues of the molasses. The yeast is removed
from the fermentation liquid by a process of washing and separating in centrifugal
separators, signaling the end of the fermentation and beginning of the downstream
processing stage. Downstream processing can be defined as the stages of processing
that take place after the fermentation or bioconversion stage. The yeast broth pro-
duced by fermentation, containing approximately 5% solids, can be manipulated
into two main types of baker’s yeast product and an additional intermediate saleable
product. These are cake yeast, granular yeast, and cream yeast, each of which
requires the downstream process to arrive at the desired product. At the end of the
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 397

Fig. 10.2 Schematic representation of baker’s yeast production [19]

fermentation, the fermenter/yeast broth is concentrated using a series of combined

centrifugation and washing steps into a yeast cream with a solids concentration of
approximately 20%. The yeast is then cooled to approximately 4 °C, an ideal tem-
perature to restrict the growth of any contaminating mesophilic microorganism.
Cream yeast is basically the liquid product and can therefore be transferred into
sterile tanks/containers and distributed to bakeries, where it is used to produce
yeast-based products. The other pathway further manipulates the yeast cream into
compressed or dried yeast. Granular yeast, also known as “Instant Dried Yeast,” is a
398 E. C. Doğan et al.

Table 10.2 Chemical composition of baker’s yeast [20, 21]

Chemical composition (% as dry matter)
Composition [20] [21]
Dry materials 30–33 –
Nitrogen 6.5–9.3 –
Proteins 40.6–58 42–46
Carbohydrates 35.0–45.0 30–37
Lipids 4.0–6.0 4–7
Minerals and various 5.0–7.5 7–8
amounts of vitamins
Nucleic acid – 6–8
Moisture – 2–5
Ash – N/A

form of compressed yeast. Stored cream/liquid yeast is passed through a filter, usu-
ally a filter press or rotary vacuum filter, which removes water, increasing its solids
content to approximately 30%. Salt may also be added to the cream yeast prior to
filtration to aid the removal of water. The filtered yeast is then dried using fluid-bed
dryers. The yeast is dry and generally does not require refrigeration as the low water
content reduces the risk of microbial contamination. Emulsifiers and oils can be
added at this point to texturize the yeast and aid the cutting process. The filtered and
the dried yeast can alternatively be used to make cake yeast. Cake yeast is another
form of compressed yeast and can be categorized as active dried yeast. It differs
from granular yeast in that rather than granulation, the dried yeast is extruded or cut
into blocks/cakes. Similar to granular yeast, cake yeast also contains about 30%
solids. Both types of compressed yeast are then packaged, typically vacuum packed
to reduce the risk of contamination by aerobic bacteria, and distributed to wholesal-
ers or traders [11]. The quality of baker’s yeast is often discussed in terms of micro-
biological purity and gas-producing activity [18]. The chemical composition of
baker’s yeast is shown in Table 10.2 [20, 21].

10.3 Wastewater Management

10.3.1 Wastewater Sources

Wastewater is generated in large quantities in the yeast industry. The wastewater

originated from the baker’s yeast industry can be classified into two groups: high
strength process wastewater and low-medium strength process wastewater. The for-
mer one is generated from the yeast separators and processes such as centrifuges
and rotary vacuum filters, whereas the latter one mainly constitutes the floor wash-
ing and equipment cleaning water and water from cooling or packaging [22–24]
The generated wastewater from both the yeast washing stages and separators
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 399

accounts almost 85% of the total wastewater [19]. A major part of the non-sugar
substances in the molasses is not assimilable by the yeast and is released unchanged
to the processing wastewater. These compounds represent the principal waste from
the yeast production process. Besides, the chemicals added during fermentation
(e.g., various salt antifoams, propionic acids, brine, etc.), yeast metabolites, and
residual yeast cells are in the wastewater [15, 25, 26].

10.3.2 Wastewater Characterization

Yeast production wastewater is a complex mixture. Most of the contaminants in the

wastewater are due to the use of molasses as a main raw material [27]. Molasses
spentwash contains nearly 2% of a dark brown recalcitrant pigment called melanoi-
din formed due to Maillard amino-carbonyl reaction [28].
Melanoidins are acidic, high molecular weight polymers with a complex struc-
ture and behave as anionic hydrophilic polymers [29]. The formation of melanoi-
dins comprises a set of consecutive and parallel chemical reactions between amino
compounds and carbohydrates during a nonenzymatic browning reaction [29–31].
The empirical formula of melanoidin is C17-18H26-27O10N. It is a product of nonenzy-
matic reaction between sugars and amino compounds. The molecular weight distri-
bution is between 5000 and 40,000 [28, 32].
Effluents generated from baker’s yeast industries are characterized by the low
levels of readily degradable sugars, acids, and high concentrations of biochemical
oxygen demand (BOD5), chemical oxygen demand (COD), total Kjeldahl nitrogen
(TKN), total dissolved solids (TDS), trimethylglycine, phenol, and sulfate. The
other major characteristics of this industry are the generation of wastewater with a
dark brown color in large quantities [33–36]. The average concentration of organic
pollutants by total COD is 25,000 mg/L, of which up to 33% is accounted for beta-
ine [36]. The wastewater characterization from baker’s yeast industries that has
been reported in the literature is shown in Table 10.3 [14, 25, 27, 35, 37–42].

10.3.3 Wastewater Treatment Processes

The processes that are applied in the treatment of baker’s yeast industry effluent are
the following:
1. Biological treatment (combined anaerobic–aerobic system with nitrification and
denitrification)
2. Physicochemical treatment (coagulation/flocculation, adsorption, evaporation)
3. Advanced treatment (membrane processes, advanced oxidation processes
(AOPs), electrochemical processes, wet-air oxidation, ultrasound process)
Table 10.3 Characterization of baker’s yeast wastewater
400

Reference [37] [38] [35] [27] [39] [25] [40] [14] [41] [35] [42]
Baker’s High-strength Baker’s Baker’s Baker’s Baker’s Baker’s
Molasses yeast wastewater of Yeast Yeast Yeast yeast yeast yeast yeast yeast
Parameter Unit wastewater effluent yeast plant effluent effluent effluent effluent effluent effluent effluent effluent
Total COD mg/L 105,851 6090 20,000– 28,194 14,400– 25,020 15,848 20,760 10,000– 25,000 70,000–
24,000 25,700 30,000 100,000
Soluble COD mg/L – 4980 – – – 23,420 15,193 – – – –
BOD5 mg/L 22,985 3780 8000 – – – – 14,974 7000– 12,000 18,340–
21,000 27,200
TKN mg/L 2326 274 2000 – – – 1196 – – – 1610–
2060
NH4+-N mg/L 120.5 132 – – – – 206 – – – –
Total-N mg/L – – – 2106 250–350 1470 – – 250– 1500 –
1200
Total-P mg/L – 3 30 55 17.3– 100 20.1 – – 100 108
48.2
PO4−-P mg/L – 2.28 – – – – 6.6 – 27 – –
SO42− mg/L – 484 4000 2449 3500– 2940 – – 100– 2900– 1700–
5300 2700 5700 3200
pH 4.6–5.5 6.45 4–5 – – – 6.2 6.0 6–8 – 4–5
VFA mg/L – – – – – 3917 – – –
Conductivity μS/cm – – – – – – – – – – –
Total solids mg/L – 35,510 – – – – – – – – –
Suspended solids mg/L – 585 500 2654 – – 835 – 50–1200 – 3800–
(SS) 7300
Volatile suspended mg/L – 475 – – – – 810 1545 – – –
solids (VSS)
E. C. Doğan et al.
 10

Reference [37] [38] [35] [27] [39] [25] [40] [14] [41] [35] [42]
Baker’s High-strength Baker’s Baker’s Baker’s Baker’s Baker’s
Molasses yeast wastewater of Yeast Yeast Yeast yeast yeast yeast yeast yeast
Parameter Unit wastewater effluent yeast plant effluent effluent effluent effluent effluent effluent effluent effluent
Alkalinity mg – 1475 – – – – 2349 – – – –
CaCO3/L
Trimethylglycine mg/L – – – – 3700– – – – – – –
4000
Color Pt-Co 38 – – – – – 14,000 – 11,000 – –
475 nm
Magnesium mg/L – – – – – – 30.7 – – – –
Ferrous mg/L – – – – – – 4.9 – – – –
Oil and grease mg/L – – – – – – – 10 – – –
Temperature °C 78–84 – – – – – – – 20–28 – –
Treatment and Minimization of Waste in Baker’s Yeast Industry
401
402 E. C. Doğan et al.

Studies on the water quality of a river contaminated with distillery effluent dis-
played high BOD5 values of 1600–21,000 mg/L within an 8-km radius. Adequate
treatment is therefore imperative before the effluent is discharged. Land disposal of
distillery effluent can lead to groundwater contamination. Deep well disposal is
another option but limited underground storage and specific geological location
limit this alternative. Constructed wetland was also applied as the tertiary treatment
step to treat the effluent from the conventional anaerobic–aerobic treatment system.
Sohsalam and Sirianuntapiboon [43] worked on the treatment by surface flow con-
structed wetland system of anaerobically treated molasses wastewater. The results
show that both removal efficiency and plant growth rate were increased with the
decrease of the organic loading rate.
Decolorization of effluent is the main problem in baker’s yeast industries.
Physicochemical treatment, adsorption, coagulation/flocculation, oxidation pro-
cesses, and membrane treatment have been examined with particular emphasis on
effluent decolorization. Though these techniques are effective for both color removal
and reduction in organic loading, sludge generation and disposal is a constraint in
coagulation/flocculation and adsorption [8]. The other treatment methods like the
evaporation of spentwash to produce animal feed have also been practiced.

10.3.3.1 Biological Treatment Processes

A medium-sized fermentation factory, which uses about 50 metric tons of molasses

per day, is estimated to generate about 1.5 tons of COD daily [13, 24]. Yeast pro-
cessing wastewater contains melanoidins, which are known to be lethal to many
microorganisms because of their antioxidant properties. Generally, biotreatment of
melanoidins containing wastewater has not given impressive results, and post-­
treatment is necessary [44].
Biological treatment, typically the combination of anaerobic and aerobic pro-
cesses, is normally effective in removing BOD from molasses wastewater. Removals
of COD range from 55% to 75% by anaerobic pretreatment. However, the brown
color remains due to the presence of melanoidin pigment [22, 24]. Conventional
anaerobic–aerobic treatment processes can accomplish the degradation of melanoi-
dins only up to 6 or 7% [8, 30]. Phenolic compounds responsible for beet molasses
wastewater color are partly removed (63% removal) during the aerobic–anoxic
treatment process, but the color removal accounts for only 8–23% [26].
The conventional treatment method is the anaerobic pretreatment stage with
upflow sludge blanket (UASB) reactors followed by an aerobic treatment with acti-
vated sludge (Fig. 10.3) [35].
Zub et al. [35] investigated that the application of a combined anaerobic/anoxic
system to a full-scale treatment plant supported biogas production up to 1300 m3/
day (Fig. 10.4). Baker’s yeast wastewater was treated in a mesophilic anaerobic/
anoxic continuous stirred tank reactor (CSTR) system. The average removal effi-
ciency in this system was 79% by total COD and 100% by SO42− in which the con-
centration of sulfides in the effluent did not exceed 50 mg/L [35].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 403

Fig. 10.3 The conventional treatment method is: anaerobic pretreatment stage with UASB reac-
tors followed by an aerobic treatment with activated sludge [35]

Fig. 10.4 The application of combined anerobic/anoxic system to a full-scale wastewater treat-
ment plant (WWTP) [35]

The characteristics of biologically pretreated baker’s yeast wastewaters are sum-

marized in Table 10.4 [14, 40, 41, 45].
There are lots of studies on removing the color from molasses’ effluents. In gen-
eral, microbial decolorization is an environment-friendly and low-cost alternative to
chemical decomposition process. Nevertheless, the problem still persists because
several organisms that have been shown to degrade melanoidin are not best suited
for treating molasses-containing wastewater [46]. The studies so far can be seen as
an early step toward solving the problem. Moreover, most of the microbial decolor-
ization studies required effluent dilution for optimal activity. While using microor-
ganisms, the use of media supplement creates extra load on the overall effluent
treatment process [8].
Biological treatments using specific microorganisms such as Coriolus [8, 47–
50], Aspergillus [50–52], Phanerochaete [53–55], Bacillus [37, 56–58], and
Lactobacillus [59] have been reported. A detailed list of microorganisms studied by
different researchers for decolorization of melanoidins is given in Table 10.5 [6, 8,
49–52, 55, 57–64].
Miyata et al. [65] investigated the microbial decolorization of melanoidin-­
containing wastewaters using activated sludge and the fungus Coriolus hirsutus.
404 E. C. Doğan et al.

Table 10.4 Characterization of biologically pretreated baker’s yeast effluents

Parameter Unit [45] [14]a [40]b [41]
COD mg/L 3700 ± 100 731 529–2006 2000–2500
Soluble COD mg/L – – 490–1052 –
TOC mg/L 1000 ± 70 – – –
BOD5 mg/L 300 ± 100 119 – –
TKN mg/L – – 171–432 –
NH4+-N mg/L – – 152–418 550–600
Ammonium mg/L 1000 ± 30 – – –
SS mg/L – – 20–50 –
VSS mg/L – 39 – –
Color Pt-Co – – 915–3740 3500–4000
400 nm Abs – – 0.515–1.995 –
455 nm Abs – – 0.253–1.995 –
475 nm Abs 2.4 ± 0.1 – – –
Total-P mg/L – – 2.2–6.8 –
pH – 8.7 ± 0.1 8 8.1–8.2 8.5
Alkalinity mg/L CaCO3 9000 ± 200 – 3450–8850 –
Sulfate mg/L 700 ± 80 – – –
Chloride mg/L 800 ± 50 – – –
Oil and grease mg/L – 8 – –
VFA mg/L – 10 – –
Conductivity μS/cm – – – 3200–3500
a
Average of 27 values
b
Three different samples

The results showed that the increase in manganese-independent peroxides and man-
ganese peroxides activities were considered to play an important role in the enhanced
ability of C. hirsutus [65].
In the case of pure culture experiments, Candida utilis and Trichoderma viridiae
each showed less than 65% reduction in COD, whereas C. utilis and A. niger
together resulted in 89% COD removal. This reduction was from sugarcane stillage-­
based media with an initial COD of 40–75 g/kg [8].
Treatment of fermentation wastewater by the use of Pseudomonas putida fol-
lowed by Aeromonas sp. in a two-stage bioreactor resulted in COD as well as color
reduction. P. putida produces hydrogen peroxide, which is a strong decolorizing
agent. Since the organism cannot use spentwash as a source of carbon, some glu-
cose supplement was provided [8].
Many effluents contain nitrogen in much higher concentrations than municipal
wastewaters. In some industrial wastewaters, nitrogen can be in the form of organic
nitrogen (urea, proteins, etc.) like in food industry effluents [66]. Wastewaters from
baker’s yeast industry effluent have a high organic contamination. The classical
biological treatments under aerobic and anaerobic conditions lead to a good effi-
ciency in removing the organic carbon, but nitrogen efficiency is variable. The
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 405

Table 10.5 Microorganisms employed for the decolorization of baker’s yeast effluent
Color
removal
Name Comments (%) Reference
Coriolus Two types of enzymes, sugar-dependent and 80 [60]
versicolor sugar-independent, were found to be responsible for
melanoidin decolorizing activity
Aspergillus The thermophilic strain adsorbed lower molecular 75 [50]
oryzae weight fractions of melanoidin and required sugars
for growth
Bacillus Addition of 1% glucose as a supplementary carbon 22 [57]
thuringiensis source was necessary
Acetobacter acetii The organism required sugar, especially glucose and 76.4 [58]
fructose for decolorization of MWWs
Lactobacillus Immobilized cells of the heterofermentative lactic 40 [59]
hilgardii acid bacterium decolorized 40% of the melanoidins
solution within 4 days aerobically
Phanerochaete Diluted anaerobically digested spentwash 80 [55]
chrysosporium
Coriolus hirsutus Decolorization of melanoidin pigment was reported 71–75 [8]
by extracellular peroxidase produced by C. hirsutus
Aspergillus niveus With a reduction of 75% COD 60 [51]
Aspergillus niger With a reduction of 95% COD 69 [51]
Geotrichum Fungus immobilized on polyurethane foam showed 80 [8]
candidum stable decolorization of diluted molasses solution
Citeromyces The yeast Citeromyces resulted in high and stable 75 [61]
removal efficiency in both color intensity and
organic matter (BOD 76%) for diluted spentwash
Aeromonas 57% COD reduction was observed after 72 h on the 55 [6]
formicans pre-digested distillery effluent
Xanthomonas Three different bacterial strains were used both in 75(with [49]
fragariae free form as well as after immobilization on calcium B. cereus)
Bacillus alginate beads for the treatment of 33% predigested
megaterium fermentation effluent
Bacillus cereus
TA2 and TA4 When used together to the treatment of 76 [62]
anaerobically treated distillery effluent, the
reduction in BOD was found to be higher, 80%
P. putida With a reduction of 44% COD in spentwash 60 [63]
Aspergillus niger With diluted synthetic melanoidin 72 [52]
50% biodigested effluent 80
Lactobacillus Decolorization of molasses wastewater was achieved 68.12 [64]
plantarum under both anaerobic and facultative conditions

biological processes involved in nitrogen removal from wastewaters of the baker’s

yeast industry are also dependent on physical and chemical conditions in which the
activated sludge microbiota work to mineralize to organic compounds or to
406 E. C. Doğan et al.

bio-­convert them into gases. Nitrogen removal through biological nitrification and
denitrification processes is generally classified as an advanced treatment pro-
cess [67].
Baker’s yeast industry produces a great amount of nitrogen compounds as a
result of the use of water in biotechnological processing. Removal of nitrogen from
baker’s yeast wastewater is a complex process, even for large wastewater treatment
plants. Conventional nitrification–denitrification systems can be applied to remove
nitrogen from the baker’s yeast industry effluents. When an anaerobic pretreatment
is operated, the anaerobic effluent has a low COD/N ratio, which can lead to incom-
plete denitrification. It is possible either to operate conventional nitrification–deni-
trification process with the addition of an external carbon source or to combine the
anaerobic digestion process with an aerobic nitrifying reactor and to recycle the
nitrified effluent in the digestor. If nitrification and denitrification must be performed
in the aerobic stage, it is necessary to run a bypass (fresh or acidified wastewater)
around the methane reactor to provide sufficient biodegradable COD for denitrifica-
tion. The biological processes involved in nitrogen removal from wastewaters of the
baker’s yeast industry are also dependent on physical and chemical conditions in
which the activated sludge microbiota work to mineralize the organic compounds or
to bio-convert them into gases [67].
The biotreated effluent of molasses wastewaters still contains high levels of
refractory organic matter, color, and inorganic salts. Ozonation can decolorize
molasses secondary effluent (≫95%) and does not lead to toxic by-products; how-
ever, it does not remove much COD (35% maximum). Refractory organics and
color in secondary effluent are removable by activated carbon adsorption, but this
needs a relatively high carbon usage. COD and color removal efficiency were
obtained to be 97% and 91% by adsorption, respectively [68].
Current biological treatment of baker’s yeast effluent involves combinations of
anaerobic digestion and aerobic systems that successfully reduced BOD to accept-
able limits but does not deal effectively with either the dark color or the associated
COD that remains and limits the reuse/recycling of the process water [7]. The color-­
imparting melanoidins are barely affected by conventional biological treatment
such as methane fermentation and the activated sludge process [8].
Pirsaheb et al. [69] used integrated anaerobic baffled reactor granular activated
carbon for pretreatment of baker’s yeast wastewater to discharging sewerage or
aerobic treating processes. Also, 94.6% COD, 93.7% BOD5, and 54% color removal
efficiencies were obtained with a loading COD of 15,000 mg/L. They reported that
the optimal operating condition of the reactor was 4 d for HRT and 50% of granular
activated carbon filling ratio. They also investigated methane production. The
methane-­yielding rate was calculated between 0.31 and 0.44 L/g COD removed,
and specific methanogenic activity was between 0.13 and 0.38 g COD/g VSS [69].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 407

10.3.3.2 Physicochemical Treatment Processes

Sugarcane molasses spentwash after biological treatment by both anaerobic and

aerobic methods can still have a BOD of 250–500 mg/L [70] and a dark color [3].
The color-imparting melanoidins are barely affected by conventional biological
treatment such as methane fermentation and the activated sludge process [71]. The
shortcomings of multistage biological treatment include operational difficulty and
occasional formation of hazardous by-products/secondary pollutants and intensifi-
cation of the color due to re-polymerization of colored compounds [24, 30, 72].
In this context, various physicochemical treatment options have been explored.
Physicochemical processes such as coagulation/flocculation, carbon adsorption,
and chemical oxidation have been applied for the decolorization of molasses
wastewater.

10.3.3.2.1 Coagulation/Flocculation

Coagulation has remained the most widely practiced method of removing particu-
late and organic matter in wastewater treatment. Conventional coagulants in waste-
water treatment are alum (Al2(SO4)3.14H2O), ferric chloride (FeCl3.6H2O), sodium
aluminate, aluminum chloride, and ferric sulfate. Conventional coagulants are basi-
cally salts of a strong acid (e.g., HCl or H2SO4) and a weak base (Al(OH)3 or
Fe(OH)3); thus, they are a mixture of a cation (from a base) and an anion (from an
acid) [28]. Chemical coagulants can destabilize particulates by four distinct mecha-
nisms: double layer compression, charge neutralization, enmeshment in a metal
hydroxide precipitate, and inter-particle bridging [30].
Experimental results indicate that ferric chloride was the most effective among
the conventional coagulant, achieving 89% COD and 98% color eliminations, while
aluminum sulfate was the least effective, giving COD and color reductions of 66%
and 86%, respectively. In addition to metal cations, counter-ions significantly influ-
ence the coagulation performance since Cl−-based metal salts attained better
removal efficiency than SO42−-based ones at the optimal coagulant dosage.
Coagulation of molasses effluent is a highly pH-dependent process, with better
removal efficiency achieved at lower pH levels. Rapid mixing intensity, rather than
rapid mixing time, has a relatively strong influence on the settling characteristics of
flocs formed. Lowering mixing intensity resulted in increasing settling rate but the
accumulation of floating flocs. When used as coagulant aids, synthetic polyelectro-
lytes showed little effect on the improvement in organic removal. On the other hand,
cationic polyacrylamide was observed to substantially enhance the settleability of
flocs as compared to anionic polyacrylamide [24].
Liang et al. [31] studied coagulation for removal of color and COD from molas-
ses effluent using ferric chloride. Under the optimum conditions, up to 86 and 96%
of COD and color removal efficiencies were achieved. Residual turbidity in the
supernatant was less than 5 NTU, and Fe3+ concentration was negligible because of
effective destabilization and subsequent sedimentation. The low-molecular-weight
408 E. C. Doğan et al.

fraction of melanoidins is more reactive than the high-molecular-weight fraction,

and an increase in the concentration of the lowest molecular weight organic group
is related to the capacity of charge neutralization. Charge neutralization and co-­
precipitation are proposed as predominant coagulation mechanisms under the opti-
mum conditions [31].
Zhou et al. [30] showed that when alum is applied in post-treatment of molasses
wastewater, the initial pH value should be above 7 to guarantee coagulation effi-
ciency. Charge neutralization is the predominant mechanism in the coagulation of
molasses effluent with alum. The residual turbidity can be drastically reduced, and
the percentage of COD and color removal further improved. The mixing condition
also has some effect on coagulation performance, and the optimal mixing rate dur-
ing the rapid mixing stage is 500 rpm when the residual turbidity is reduced to 14
NTU [30].
Inanc et al. [3] reported that coagulation with alum and iron salts was not effec-
tive for color removal. They explored lime and ozone treatment to anaerobically
digested effluent. The optimum dosage of lime was found to be 10 g/L resulting in
82% COD removal and 67% reduction in color in a 30-min period. These findings
are in disagreement with those of Migo et al. [71] who used a commercial inorganic
flocculant, a polymer of ferric hydroxysulfate with a chemical formula
[Fe2(OH)n(SO4)3], for the treatment of molasses wastewater. The treatment resulted
in about 87% decolorization for biodigested effluents; however, an excess of floc-
culant hindered the process due to an increase in turbidity and TOC content.
FeCl3 and AlCl3 were also tested for decolorization of biodigested effluent and
showed similar removal efficiencies. About 93% reduction in color and 76% reduc-
tion in TOC were achieved when either FeCl3 or AlCl3 was used alone. The process
was independent of chloride and sulfate ion concentration but was adversely affected
by high fluoride concentration. However, in the presence of high flocculant concen-
tration (40 g/L), the addition of 30 g/L CaO enhanced the decolorization process
resulting in 93% color removal. This was attributed to the ability of calcium ions to
destabilize the negatively charged melanoidins; furthermore, the formation of cal-
cium fluoride (CaF2) also precipitates the fluoride ions. Almost complete color
removal (98%) of biologically treated distillery effluent has been reported with con-
ventional coagulants such as ferrous sulfate, ferric sulfate, and alum under alkaline
conditions [73].
The best results were obtained using Percol 47, a commercial organic anionic
polyelectrolyte, in combination with ferrous sulfate and lime. The combination
resulted in 99% reduction in color and 87 and 92% reduction in COD and BOD,
respectively. Similar findings have also been reported by Mandal et al. [74].
Coagulation studies on spentwash after anaerobic–aerobic treatment have also
been conducted using bleaching powder followed by aluminum sulfate. The opti-
mum dosage was 5 g/L bleaching powder followed by 3 g/L of aluminum sulfate
that resulted in 96% removal in color, accompanied by up to 97% reduction in BOD
and COD. Nonconventional coagulants, namely wastewater from an iron pickling
industry rich in iron and chloride ions and titanium ore processing industry contain-
ing significant amounts of iron and sulfate ions, have also been examined [73]. The
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 409

iron pickling wastewater gave better results with 92% COD removal, combined
with over 98% color removal.
Prasad [28] studied the color removal from distillery spentwash through coagula-
tion using Moringa oleifera seeds. In the study, the effects of dosage, pH, and con-
centration of salts (NaCl and KCl) were investigated for an optimized condition of
color removal. The actual color removal at optimum conditions was found to be
53% and 64%, respectively, for NaCl and KCl salts [28].

10.3.3.2.2 Adsorption

Adsorption is the process of accumulating substances that are in solution on a suit-

able interface. Adsorption is a mass transfer operation in that a constituent in the
liquid phase is transferred to the solid phase. The adsorption process takes place in
four more or less definable steps: (1) bulk solution transport, (2) film diffusion
transport, (3) pore transport, and (4) adsorption (or sorption). Because the adsorp-
tion process occurs in a series of steps, the slowest step in the series is identified as
the rate-limiting step. The principal types of adsorbents include activated carbon,
synthetic polymeric, and silica-based adsorbents, although synthetic polymeric and
silica-based adsorbents are seldom used for wastewater adsorption because of their
high cost. Economical application of activated carbon depends on an efficient means
of regenerating reactivating the carbon after its adsorptive capacity has been
reached [75].
Among the physicochemical treatment methods, adsorption on activated carbon
is widely employed for the removal of color from organic compounds from waste-
water, molasses wastewater, melanoidin, and other specific organic pollutants.
Activated carbon is a well-known adsorbent due to its extended surface area, micro-
porous structure, high adsorption capacity, and high degree of surface reactivity [76,
77]. Adsorption of organic compounds is controlled by physical and chemical inter-
actions [78]. It was recently demonstrated that an activated carbon with a significant
distribution of both micropores and mesopores and a significant amount of macro-
pores that are assumed to act as conduits providing access to micro- and mesopores
have a good adsorption efficiency for melanoidins and colored compounds of
molasses spentwash after anaerobic digestion [79]. Decolorization of synthetic mel-
anoidin using commercially available activated carbon as well as activated carbon
produced from sugarcane bagasse was investigated by Bernardo et al. [80].
Significant decolorization was observed in packed bed studies on anaerobically
treated spentwash using commercial activated charcoal with a surface area of
1400 m2/g [81]. Nearly complete decolorization (>99%) was obtained with 70% of
the eluted sample, which also displayed over 90% BOD and COD removal. In con-
trast, other workers have reported adsorption by activated carbon to be ineffective in
the treatment of distillery effluent [74, 82]. Adsorption by commercially available
powdered activated carbons resulted in only 18% color removal; however, com-
bined treatment using coagulation–flocculation with polyelectrolyte followed by
adsorption resulted in almost complete decolorization [82].
410 E. C. Doğan et al.

Low-cost adsorbents such as pyorchar (activated carbon both in granular and

powdered form, manufactured from paper mill sludge) and bagasse flyash have also
been studied for application. Ramteke et al. [83] reported color removal up to 98%
with pyorchar. However, to achieve the same level of color removal, larger doses of
the indigenously prepared powdered and granular pyorchar were required in com-
parison to commercial activated carbon [83].
Graphene oxide nanosheets could be a cost-effective, efficient, and potential
adsorbent for melanoidin removal. The adsorption capacity of graphene oxide
nanosheets as an adsorbent for melanoidin removal from industrial molasses waste-
water is 18.31 g/g [84].
Mall and Kumar [70] compared the color removal using commercial activated
carbon and bagasse flyash, and 58% color removal was reported with 30 g/L of
bagasse flyash and 80.7% with 20 g/L of commercial activated carbon. Since the
bagasse flyash has high carbon content and the adsorbed organic material further
increases its heating value, the spent adsorbent can be used for making fire bri-
quettes [70].
Yet another adsorbent that has been examined is the natural carbohydrate poly-
mer chitosan derived from the exoskeleton of crustaceans. Lalov et al. [85] studied
the treatment of fermentation wastewater using chitosan as an anion exchanger. At
an optimum dosage of 10 g/L and 30 min contact time, 98% color and 99% COD
removal were observed.

10.3.3.2.3 Evaporation Systems

The high-strength wastewater coming from the yeast separators is treated by the
evaporation process to reduce COD and nitrogen loads from the industry [38]. An
evaporator in a chemical plant or a fermentation operation is a highly engineered
piece of processing equipment in which evaporation takes place. All evaporators are
fundamentally heat exchangers because thermal energy must be added to the pro-
cess, usually across a metallic barrier or heat transfer surface, in order for
evaporation to take place. Since the purpose of an evaporator is to concentrate a
dilute feed stream and to recover a relatively pure solvent, this separation step must
be defined. Figure 10.5 is a scheme for any evaporator.
The properties of the liquid feed and the concentrate are important factors to
consider in the engineering and design of an evaporation system. The heating sur-
face of an evaporator represents the largest portion of the evaporator cost; heat
transfer is the most important single factor in the design of an evaporation system.
Most evaporators consist of three main elements or parts: a heating unit
(calandria), a region for liquid–vapor separation (sometimes called a vapor head,
flash chamber, or settling zone), and a structural body to house these elements and
to separate the process and heating fluids. The simple largest variable cost factor in
making separation by evaporation is the cost energy. If crude oil is the ultimate
source of energy, the cost of over $126.67 per m3 ($20 per barrel) is equivalent to
more than $3.33 for one million kJ. Water has a latent heat of 480 kJ/kg at
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 411

Fig. 10.5 Scheme of evaporator

760 mmHg, absolute, so the energy required to evaporate 1 kg of water exceeds 0.16
cents. Therefore, the efficient utilization of energy is the most important consider-
ation in evaluating which type of evaporation system should be selected.
From the process viewpoint, the two parameters that should be regulated are the
concentration and flow rate of the bottoms product. If the composition of the feed
stream is constant, good control of the feed rate and the evaporation rate will give
the desired concentrated product at the proper production rate. Of course, the
method of control can depend upon the evaporator type and method of operation.
Flow rates are the largest single group of process measurements used for control,
and flow is the only process variable for which significant energy may be required
by the measuring device. Energy economy and evaporative capacity are the major
measures of evaporator performance. Other performance variables to be considered
include product quality, product losses, and decrease in performance as scaling,
salting, or fouling occurs [86].
At the end of the fermentation stage, the resulting broth contains 5–6% yeast
cells. The cells are separated by large-scale continuous-type separators and col-
lected as yeast cream liquid, while the cell-free fermentation broth (mainly weak
vinasse) is sent for evaporation. The cell-free fermentation broth (wash) is pre-­
heated to about 90 °C by heat exchange with the effluent (“spentwash”) and then
sent to the evaporators. Here, the liquor is heated by live steam and fractionated to
give about 60–55% dry matter. The condensed water from this stage, known as
“spentlees,” is usually pumped back to the wastewater system. The photographs of
weak vinasse and evaporations outlet products are shown in Fig. 10.6 [19].
Economic considerations frequently suggest the simplest answers to the question
of how to utilize stillage produced, specifically as either fodder or fertilizer. Such
uses, however, do not resolve the problem because of the large scale of yeast pro-
duction. Molasses-based vinasse is characterized by high potassium content and
therefore limits its potential as farm animal fodder. Another use, particularly in the
case of vinasse, consists in classifying stillage as wastewater and making it subject
to anaerobic biodegradation.
412 E. C. Doğan et al.

Fig. 10.6 The photographs of weak vinasse and evaporations outlet products [19]

10.3.3.3 Advanced Treatment Processes

Biological treatment of wastewaters eliminates important compounds of the organ-

ics in the waste. However, biochemical decomposition by conventional treatment
methods may not be enough to satisfy the discharge limits if these pollutants are in
the form of refractory organics. Thus, it can be necessary to use more effective pro-
cesses for the destruction of such contaminants [40].
The main problems in the treatment of yeast wastewater are the high concentra-
tion of COD in the effluent, color, odor, sulfate, and a high amount of excess sludge
generated in the wastewater treatment process. If a higher degree of purification is
required, biological treatment can be used in combination with other processes such
as advanced treatment processes, and these processes are detailed in the following
section.

10.3.3.3.1 Membrane Process

Membrane processes have found wide applications for the treatment of aqueous-­
based systems involving material recovery, reuse, and for pollution prevention [87].
The role of the membrane, as shown in Fig. 10.7, is to serve as a selective barrier
that will allow the passage of certain constituents and will retain other constituents
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 413

Fig. 10.7 Definition sketch for a membrane process [75]

found in the liquid. The influent to the membrane module is known as the feed
stream (also known as feedwater). The liquid that passes through the semi-­permeable
membrane is known as permeate (also known as the product stream or permeating
stream), and the liquid containing the retained constituents is known as the concen-
trate (also known as the retentate, reject, retained phase, or waste stream). The rate
at which the permeate flows through the membrane is known as the rate of flux,
typically expressed as kg/m2 day [76].
Based on the different driving forces applied, the range of separations can be
divided into various filtration processes (microfiltration, ultrafiltration, nanofiltra-
tion, and reverse osmosis), gas and vapor separation, pervaporation, and electro-
membrane processes (including electrodialysis, membrane electrolysis, and bipolar
membrane processes). Additionally, based on preferential wetting properties, porous
membranes have been used as a support for liquid membranes and for various con-
tactor applications (including membrane-based solvent extraction and gas absorp-
tion). These processes usually focus on the desired separation of a gas or liquid
mixture [87]. Membrane distillation (MD) is a thermally driven membrane process
in which a hydrophobic microporous membrane separates a hot and cold stream of
water [88].
The ability of the membrane depends on the size of pores, types of materials,
types of wastewater, solubility, and retention time. Permeability, flux, transmem-
brane pressure (TMP), and resistance are the parameters that also need to be consid-
ered. Membrane structure plays an important role in transporting mechanism
whether the structure is parallel or in series. The types of membranes used are dif-
ferent depending on the size of contaminants contacting during the treatment pro-
cess [89]. Microfiltration membranes having the largest pore size (0.1–0.2 μm)
among the categories can separate microbial cells for their subsequent recycling to
the bioreactor to ensure high cell concentration and thus high productivity.
Ultrafiltration membranes with an average pore size much less than that of the
microfiltration membranes can retain cells and proteins. Separation by microfiltra-
tion and ultrafiltration membranes is based on size exclusion, and for effective cell
harvesting, at least 100–300 kDa molecular weight cutoff (MWCO) value should be
ensured for this. Nanofiltration membranes between reverse osmosis and ultrafiltra-
tion membranes with an average pore size of 1 nm can separate cells, proteins,
414 E. C. Doğan et al.

nutrients, salts, and unconverted carbon sources from lactic acids. Reverse osmosis
normally known as nonporous membrane, where separation is based on solution
diffusion mechanism, can separate the same components as nanofiltration mem-
branes do but at much higher operating pressure than what is needed in nanofiltra-
tion from fermentation [90].
Considering the large diversity of membranes suited for technical applications, it
will be useful to introduce the membrane classifications as membrane materials,
membrane cross-section, preparation method, and the membrane shape [91].
Two types of materials that are commonly used to construct membranes are
polymeric and ceramic. The ceramic membrane is usually used by industrial waste-
water, which has a good performance in filtration compared to polymer due to its
high chemical resistance, inertness, and ease to clean. The polymer membrane
(porous membrane) has its own weakness that it can foul easily because of its
hydrophobic characteristic. However, the hydrophobic membrane weakness can be
improved by coating the membrane using hydrophilic polymer. There are two types
of operation: dead-end and cross-flow operation. Cross-flow is liquid flow parallel
toward a filter surface and transports suspended particles of membrane surface by
permeating flow due to pressure drop. Basically, this type of filtration is carried out
by using hollow fiber (HF), flat sheet (FS), or multi tubular (MT). Cross-flow filtra-
tion can reduce the formatting of the cake layer on the surface of the membrane
[89]. A membrane can be homogeneous or heterogeneous, symmetric or asymmet-
ric in structure, and solid or liquid; it can carry a positive or negative charge or be
neutral or bipolar. Transport through a membrane can be affected by convection or
by diffusion of individual molecules, induced by an electric field or concentration,
pressure, or temperature gradient. The membrane thickness may vary from as small
as 10 μm to a few hundred micrometers [92].
After membrane treatment, membrane concentrates containing high amounts of
recalcitrant organics that must be disposed of. To avoid this problem, the concen-
trates are recycled back to the biological treatment plant influent, but this would
cause a continuous increase of recalcitrant and toxic or inhibitory pollutants in the
biological treatment system.
In order to overcome such a drawback, it has been proposed that, before being
recycled back, membrane concentrates can be treated by ozone. After ozone treat-
ment, recalcitrant pollutants are transformed to more easily biodegradable pollut-
ants as well as degradation of toxic pollutants that inhibit biomass activity.
Membrane treatment systems with UF and/or RO for biologically pretreated high-­
strength fermentation industry effluents are very effective in the removal of COD,
color, NH3-N, and conductivity. The final disposals of the concentrates from the
membrane systems are major problems in these types of applications. Koyuncu
et al. [41] stated that the biodegradability of the concentrates from the RO systems
could be increased significantly by applying advanced oxidation with ozone and
ozone + H2O2. Additional BOD5 in the ozonated concentrates can be treated by the
existing aerobic biological treatment facilities [41].
There have been many applications of membrane processes for the treatment of
color and COD from the baker’s yeast wastewater. Mutlu et al. 2002 studied the
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 415

decolorization of baker’s yeast wastewater with membrane processes. Maximum

rejections obtained were 94%, 89%, and 72% for optical density, color, and COD,
respectively, when 0.8-μm microfiltration membranes and 400-D nanofiltration
membranes were used in series [93].
Pretreatment of spentwash with ceramic membranes prior to anaerobic digestion
is reported to have the COD from 36.000 to 18.000 mg/L [94]. The total membrane
area was 0.2 m2, and the system was operated at a fluid velocity of 6.08 m/s and
0.5 bar transmembrane pressure. In addition to COD reduction, the pretreatment
also improved the efficiency of the anaerobic process, possibly due to the removal
of inhibiting substances.
Kumaresan et al. [95] employed the emulsion liquid membrane technique in a
batch process for spentwash treatment. Water–oil–water type of emulsion was used
to separate and concentrate the solutes resulting in an 86 and 97% decrease in COD
and BOD, respectively.
Electrodialysis has been explored for desalting spentwash using cation and anion
exchange membranes resulting in 50–60% reduction in potassium content [96]. In
another study, Vlyssides et al. [97] reported the treatment of vinasse from beet
molasses by electrodialysis using a stainless steel cathode, titanium alloy anode,
and 4% w/v NaCl as an electrolytic agent. Up to 88% COD reduction at pH 9.5 was
obtained; however, the COD removal percentage decreased at higher wastewater
feeding rates. In addition, reverse osmosis (RO) has also been employed for fermen-
tation wastewater treatment. A processing effluent was obtained after anaerobic
digestion, followed by a hold-up in a tank maintained under aerobic conditions in a
RO system. Furthermore, 290 m3/day of RO treated effluent is mixed with 300 m3/
day of freshwater and used in the process for various operations like molasses dilu-
tion, make-up water for cooling tower, fermenter washing, etc. Another unit employs
disc and tube RO modules for direct treatment of the anaerobically digested spent-
wash. Permeate is discharged while the concentrate is used for biocomposting with
sugarcane pressmud. In the other study, Decloux et al. [98] have studied electrodi-
alysis to reduce the potassium concentration to prevent the crystallization and even
increase the final dissolved solids of the concentrated vinasses and have found the
process successful and the results encouraging.
Nataraj et al. [99] reported pilot trials on fermentation spentwash using a hybrid
nanofiltration (NF) and RO process. Both the NF and RO stages employed thin-film
composite membranes in spiral wound configuration. NF was primarily effective in
removing the color and colloidal particles accompanied by 80%, 95%, and 45%
reduction in total dissolved solids (TDS), conductivity, and chloride concentration,
respectively, at an optimum feed pressure of 30–50 bar. The subsequent RO opera-
tion at a feed pressure of 50 bar resulted in 99% reduction each in COD, potassium,
and residual TDS.
416 E. C. Doğan et al.

10.3.3.3.2 Oxidation Process

The goal of any AOP design is to generate and use hydroxyl free radical (HO•) as a
strong oxidant to destroy compounds that cannot be oxidized by conventional oxi-
dants. Although AOPs have many advantages, one common problem in all AOPs is
the high electrical energy demand for UV lamps, which causes high operational
costs [22]. Advantages of ozonation in treatment applications are removal of toxic-
ity, destruction of organic matter, and enhancement of the biodegradability of recal-
citrant wastewaters [25].
Chemical oxidation, especially ozonation, has already been demonstrated to be
an effective means of removing refractory and/or toxic chemicals from water and
wastewater. Ozonation reduces color as well as the organic matter content due to its
ability to react with unsaturated bonds (olefins-aromatics) that are responsible for
the coloration of wastewaters [45]. Ozone reacts with aqueous organic pollutants
found in water and wastewater via two different pathways, namely direct molecular
(pH ≤ 2) and indirect (pH ≥ 7) radical chain-type reactions. The ozonation reaction
pathway strongly depends on the characteristics of the wastewater to be treated, i.e.,
pH, concentration of ozone decomposition initiators, promoters, and scavengers in
the reacting medium.
However, ozonation alone still has low TOC and COD removal due to some
refractory small molecular organic compounds produced in an aqueous solution.
This fact has led to the research on how to enhance the efficiency of ozonation for
various applications, and many advanced oxidation processes (AOPs), such as O3/
H2O2, UV/O3, and catalytic ozonation, have been developed.
Homogeneous oxidation with the Fenton reagent occurs in the presence of fer-
rous or ferric ions with hydrogen peroxide via a free radical chain reaction that
produces hydroxyl radicals. It is considered to be a metal-catalyzed oxidation reac-
tion, in which iron acts as the catalyst. Process efficiency is closely related to the
solution pH whose optimal values are between 2 and 4 as well as the
COD:H2O2:catalyst ratio in the feed. Moreover, efficiency can be enhanced in the
presence of UV irradiation as more hydroxyl radicals are produced in the so-called
photo-Fenton reaction.
Heterogeneous semi-conductor photocatalysis using TiO2 as the photocatalyst is
an emerging technology with key advantages including operation at ambient condi-
tions as well as the fact that the catalyst itself is inexpensive, commercially available
at various crystalline forms and particle characteristics, nontoxic, and photochemi-
cally stable. From a mechanistic point of view, illumination of an aqueous TiO2
suspension with irradiation with energy greater than the band gap energy of the
semiconductor generates valence band holes and conduction band electrons. Holes
and electrons may either undesirably recombine liberating heat or make their sepa-
rate ways to the surface of TiO2, where they can react with species adsorbed on the
catalyst surface. Valence band holes can react with water and the hydroxide ion to
generate hydroxyl radicals, while electrons can react with adsorbed molecular oxy-
gen reducing it to superoxide radical anion, which, in turn, reacts with protons to
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 417

form peroxide radicals. Besides TiO2, ZnO and CdS have also been employed as
photocatalysts in water treatment [100].
Çatalkaya and Şengül [22] investigated decolorization and mineralization of
baker’s yeast industry effluents by photochemical advanced oxidation processes
(AOPs) utilized UV with H2O2 and photo-Fenton. The optimum H2O2 concentration
and irradiation time were found to be 5 mM and 50 min at pH 3 for UV/H2O2 pro-
cesses. In the photo-Fenton process application, maximum decolorization efficiency
(96.4%) was obtained at the optimum reaction conditions that were 100 mM H2O2
and 1 mM Fe2+ doses at pH 3 and 10 min of irradiation time [22].
Altınbaş et al. [40] studied the effectiveness of chemical oxidation by applying
ozonation, ozonation with hydrogen peroxide, and Fenton’s processes for decolor-
ization and residual COD removal of biological pretreated baker’s yeast industry
effluents. In Fenton’s oxidation studies, the removal efficiencies of COD and color
for a reaction time of 30 min for three different types of baker’s yeast industry efflu-
ents were found about 86 and 92%, respectively. Unit costs of Fenton oxidation for
the post-treatment of baker’s yeast industry effluents are in the range of $0.6–1.9 per
cubic meter depending on the initial COD levels of the waste [40].
Ozone destroys hazardous organic contaminants and has been applied for the
treatment of dyes, phenolics, pesticides, etc. [72]. Oxidation by ozone could achieve
80% decolorization for biologically treated spentwash with simultaneous 15–25%
COD reduction. It also resulted in improved biodegradability of the effluent.
However, ozone only transforms the chromophore groups but does not degrade the
dark-colored polymeric compounds in the effluent [72, 101]. Similarly, oxidation of
the effluent with chlorine resulted in 47% color removal, but the color reappeared
after a few days [74]. Ozone in combination with UV radiation enhanced spentwash
degradation in terms of COD; however, ozone with hydrogen peroxide showed only
marginal reduction even on a very dilute effluent [102].
Peňa et al. [45] showed that continuous ozonation was effective for the decolor-
ization of molasses wastewater. Operating with a hydraulic residence time of 45 min
and an applied ozone mass flow of 1.7 g/h, color and COD reductions were about
80% and 14%, respectively The results showed that color reduction was mainly due
to direct oxidation reactions between ozone and chromophore groups, whereas the
indirect reaction pathway contributed to the reduction of the organic matter con-
tent [45].
Altınbaş et al. [40] cited that the best COD and color removals at an ozonation
time of 80 min were observed as 43 and 96%, respectively, in ozonation processes.
Ozonation always requires a significantly high initial investment cost compared to
Fenton’s oxidation [40].
Blonskaja et al. [25] studied the treatment of yeast industry wastewaters using
ozone. They showed that the post-ozonation of biological treatment resulted in the
reduction of 30–49% COD and consumed ozone dosage ranged from 1.2 to 2.5 mg
ozone/mg removal COD [25].
Blonskaja and Zub [15] studied post-treatment of biologically treated wastewa-
ter from yeast factories. They reported that coagulants and ozone could be used in
the process of the post-treatment of effluents of yeast industry for the purpose of
418 E. C. Doğan et al.

decreasing the color and general concentration of pollutants, but these processes are
very expensive [15].
Inanç et al. [103] studied color removal from biological treatment effluent of
baker’s yeast industry with massive lime and ozone treatment. The optimum lime
dose for reducing the color to values around 1000 Pt-Co was found as 10.0 g/L,
while 0.9 g/L ozone was necessary to obtain the same residual color. Economical
evaluation has indicated that the cost of lime treatment was 1.3–1.4 US$/m3 while it
was 2.5 USD/m3 for ozone treatment. Color, COD, total phosphorus, and ammonia
removed were 84.3%, 89.5%, 99.9%, and 31.1% with 20 g/L lime, respectively. It
was possible to remove all the apparent colors and produce clear effluent at pro-
longed contact periods [103].
The industrial applications of ozonation are limited by its high costs and low
ozone utilization caused by the mass transfer rate of ozone [104].
Pala and Erden [14] studied Fenton oxidation using biologically pretreated bak-
er’s yeast effluent. The best Fe2+/H2O2 dosage was 1200 mg/L Fe2+/800 mg/L H2O2
at pH 4 and in a reaction time of 20 min for mineralization of dissolved organic
carbon (DOC) and COD. For these conditions, the maximum color removal effi-
ciency was obtained as 99%, and maximum DOC and COD removal efficiencies
were obtained as 90% and 88%, respectively [14].
Another option is photocatalytic oxidation, which has been studied using solar
radiation and TiO2 as the photocatalyst [105]. The use of TiO2 was found to be
highly effective as the destructive oxidation process leads to the complete mineral-
ization of effluent to CO2 and H2O. Up to 97% degradation of organic contaminants
was achieved in 90 min. Pikaev et al. [106] studied combined electron beam and
coagulation treatment of distillery slops from distilleries processing grain, potato,
beet, and some other plant materials. Humic compounds and lignin derivatives con-
stitute the major portion of this dark brown wastewater. The distillery wastewater
was diluted with municipal wastewater in the ratio of 3:4, irradiated with an electron
beam, and then coagulated with Fe2(SO4)3. The optical absorption in the UV region
was decreased by 65–70% after this treatment. The cost was found to be less than
the existing method wherein the effluent was transported about 20 km via pipeline
to a facility for biological treatment followed by sedimentation. The treatment cost
was 0.45–0.65 US$/m3, which dropped to 0.25 US$/m3 using the combined
electronic-­beam and coagulation method.
Can and Genç [107] studied the pretreatment of fermentation wastewater by pho-
tocatalytic oxidation. They showed the effects of photocatalytic oxidation process
on color removal and improvement in biodegradability by observing the BOD/COD
ratio. It was seen that the ratio increased from 0.11 to 0.18 at the end of the irradia-
tion time of 120 min [107].
The advanced treatment of biologically treated baker’s yeast wastewater for the
purpose of irrigation reuse was studied by Balcıoğlu et al. Baker’s yeast wastewater
was treated using combined ozonation with the membrane process. The wastewater
was treated with NF90 and BW 30 membranes after ozonation, and 96–98% color
and 56% COD removals were obtained with ozonation at pH 7.5 and 25 °C. The
treated wastewater was classified as class B considering pH, BOD5, SS, and fecal
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 419

coliform parameters. It was met the criteria of II class in terms of SAR parameter.
They reported that the concentrate obtained from the membrane process could be
circulated back to the existent unit of the primary sedimentation tank in the baker’s
yeast mill [108].

10.3.3.3.3 Electrochemical Processes

Electrochemical oxidation processes are environmentally friendly emerging meth-

ods for the decontamination of wastewaters contaminated with toxic and persistent
herbicides, pesticides, chlorophenols, nitrophenols, polychlorinated biphenyls,
pharmaceuticals, etc. Electrochemical reactions are heterogeneous ion transfer
reactions that ionic compounds left from the electrolyte to be oxidized or reduced
over anode or cathode.
Electrocoagulation (EC) involves the generation of coagulants in situ by dissolv-
ing electrically either aluminum or iron ions from respectively aluminum or iron
electrodes. The metal ion generation takes place at the anode, and hydrogen gas is
released from the cathode. The hydrogen gas would also help float the flocculated
particles out of the water. The electrodes can be arranged in a mono-polar or bi-­
polar mode. The materials can be aluminum or iron in plate form or packed form of
scraps such as steel turnings, millings, etc. The nascent Al3+ or Fe2+ ions are very
efficient coagulants for particulates flocculating. The advantages of electrocoagula-
tion include high particulate removal efficiency, compact treatment facility, rela-
tively low cost, and possibility of complete automation [109].
The supply of current to the electrocoagulation system determines the amount of
Al3+ or Fe2+ ions released from the respective electrodes. The quality of the treated
water depends on the amount of ions produced (mg) or charge loading, the product
of current, and time (Ah). For aluminum, the electrochemical equivalent mass is
335.6 mg/(Ah). For iron, the value is 1041 mg/Ah [109]. The value of the required
Al3+ power consumption for removing color is 0.04–0.1 mg Al3+ and 10–40 Wh/m3
and for preliminary purification is 0.1–0.2 mg Al3+ and 40–80 W h/m3, respec-
tively [110].
Electroflotation (EF) is a simple process that floats pollutants to the surface of a
water body by tiny bubbles of hydrogen and oxygen gases generated from water
electrolysis. Therefore, the electrochemical reactions at the cathode and anode are
hydrogen evolution and oxygen evolution reactions, respectively. The pollutant
removal efficiency is largely dependent on the size of the bubbles formed. In an EF
reactor, usually, an anode is installed at the bottom, while a stainless steel screen
cathode is fixed at 10–50 mm above the anode. Such an electrode arrangement can-
not ensure quick dispersion of the oxygen bubbles generated at the bottom anode
into wastewater flow, affecting flotation efficiency [109].
In the literature, there are a lot of studies interested in electrochemical technolo-
gies, including EC and EF; however, there are only a few studies on the electro-
chemical treatment of fermentation industry wastewaters.
420 E. C. Doğan et al.

In a study by Vlyssides et al. [97], a number of experiments have been conducted

in a laboratory-scale pilot plant using Pt/TiO2 anode in the presence of sodium chlo-
ride as a supporting electrolyte. Influent COD of 72,000 mg/L has been reduced to
8000 mg/L in effluent with an 89% COD removal. According to the authors, treat-
ment efficiency depends on the catalytic activity of the anodes used, the COD load-
ing rates, and the pH of the solution.
Manisankar et al. [111] have researched the effect of halides in the electrochemi-
cal treatment of distillery effluent using anodized graphite plate anodes and graphite
cathodes. The effect of pH and current density on the treatment has been studied.
Sodium fluoride, sodium chloride, and sodium bromide have been chosen as elec-
trolytes, and their influence has been studied by the authors. Complete decoloriza-
tion has been reached in all cases. A maximum of 93% of BOD reduction, 85% of
COD reduction, and 98% absorbance reduction have been obtained in the presence
of sodium chloride as a supporting electrolyte.
Electrochemical oxidation over anodes made of graphite, Pt, TiO2, IrO2, PbO2,
several Ti-based alloys, and, more recently, boron-doped diamond electrodes in the
presence of a suitable electrolyte (typically NaCl) has been employed for the decon-
tamination of various organic-containing effluents. Two mechanisms are responsi-
ble for organic matter electrochemical degradation, namely (a) direct anodic
oxidation where the pollutants are adsorbed on the anode surface and destroyed by
the anodic electron transfer reaction and (b) indirect oxidation in the liquid bulk,
which is mediated by the oxidants that are formed electrochemically; such oxidants
include chlorine, hypochlorite, hydroxyl radicals, ozone, and hydrogen perox-
ide [100].
Gengec et al. studied electrocoagulation for removals of color, COD, and TOC
from baker’s yeast effluent in a batch electrocoagulation reactor using aluminum
electrodes. The maximum color, COD, and TOC were 88%, 48%, and 49% at 80 A/
m2, pHi 4, and 30 min for anaerobic effluents and 86%, 49%, and 43% at 12.5 A/m2,
pHi 5, and 30 min for anaerobic–aerobic effluents, respectively. The operating costs
for anaerobic and anaerobic–aerobic effluents at the optimized conditions were
0.418 €/m3 and 0.076 €/m3, respectively [7].

10.3.3.3.4 Wet-Air Oxidation Process

Wet-air oxidation (WAO) is a technology used to treat the waste streams that are too
dilute to incinerate and too concentrated for biological treatment. It can be defined
as the oxidation of organic and inorganic substances in an aqueous solution or sus-
pension by means of oxygen or air at elevated temperatures and pressures either in
the presence or absence of catalysts. According to this method, the dissolved or
suspended organic matter is oxidized in the liquid phase by some gaseous source of
oxygen, which may be either pure oxygen or air. Typical conditions for WAO are
150–320 °C for temperature, 2–15 MPa for pressure, and 15–120 min for residence
time; the preferred COD load ranges from 10 to 80 kg/m3. WAO destroys toxics in
industrial wastewater by breaking down complex molecular structures into simpler
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 421

components such as water and carbon dioxide, without emissions of NOx SO2, HCl,
dioxins, furans, and fly ash. It is reported that the WAO process is capable of a high
degree of conversion of toxic organics with more than 99% destruction rate; how-
ever, some materials are not oxidized completely to carbon dioxide and water;
instead, some intermediate compounds are formed, which represent a quarter of the
original mass of organic matter [110].
By using homogeneous catalysts (Cu, Fe, Cu-Mn-Fe, Fe/H2O2, etc.) and hetero-
geneous catalysts (Co/Bi, Sn/Bi, Zn/Bi, Ni/Bi, Bi(OH)3 or Ru-Rh, Pt-Pd, Ru,
Mn-Zn-Cr, etc., and rare metals attached on alumina (Pt/Al2O3, Pd/Al2O3, Ce/
Al2O3)), it is possible to reach higher reaction rates at lower temperature and pres-
sure. Complete oxidation of organic materials can be achieved by applying severe
operating conditions; however, these conditions necessitate high operating costs.
Operating costs can be decreased appreciably by using the process as a step of pre-
treatment where partial oxidation is achieved at moderate conditions prior to con-
ventional biological treatment. When this process is considered a pretreatment, the
inhibition–toxic effect of degradation by-products to the biological system has to be
recognized. The aqueous phase formed as a consequence of the wet oxidation pro-
cess contains low-molecular-weight organic compounds, inorganic acids, and inor-
ganic salts. The residual of most of the pollutants in the aqueous phase is an
advantage of this process [112].
A combination of WAO and adsorption has been successfully used to demon-
strate the removal of sulfates from distillery wastewater. Studies were done in a
counter-current reactor containing 25-cm base of small crushed stones supporting a
20-cm column of bagasse ash as an adsorbent [113]. The wastewater was applied
from the top of the reactor, and air was supplied at the rate of 1.0 L/min. The treat-
ment removed 57% COD, 72% BOD, 83% TOC, and 94% sulphates. WAO has
been recommended as part of a combined process scheme for treating anaerobically
digested spentwash [114]. The post-anaerobic effluent was thermally pretreated at
150 °C under pressure in the absence of air. This was followed by soda-lime treat-
ment, after which the effluent underwent a 2-h wet oxidation at 225 °C. Also, 95%
color removal was obtained in this scheme.

10.3.3.3.5 Ultrasound Process

Various researchers have reported several biological, physicochemical, and phy-

toremediation approaches for the treatment of yeast industry wastewater [115].
However, all these studies have only concentrated on the removal of color/melanoi-
din from the yeast industry and not on the holistic issue of biodegradability enhance-
ment. A new technology such as cavitation, which has found several applications
[116–118], can be effectively utilized as a pretreatment option for refractory/recal-
citrant wastes. Cavitation is defined as the formation, growth, and subsequent col-
lapse of microbubbles or cavities in extremely little time intervals as micro to
milliseconds, releasing large magnitudes of energy over a small region but at mul-
tiple spots in the reactor [119]. Although ultrasonic cavitation is a costly option
422 E. C. Doğan et al.

owing to its ineffective spatial distribution of cavities on a large scale and less effec-
tive transducer outputs at higher operating frequencies [115], an alternative cavita-
tion phenomenon generated by manipulating the liquid flow pattern termed
hydrodynamic cavitation is reported to be more energy efficient over acoustic cavi-
tation for some application and even can find large-scale applications [115, 120].
In hydrodynamic cavitation, cavities are formed by passing the liquid through
the constriction/geometry provided in lines such as venturi and orifice plate. When
the pressure at the throat or vena contracta of the constriction falls below the vapor
pressure of the liquid, the liquid flashes, creating a number of vaporous cavities that
subsequently collapse when the pressure recovers downstream of the mechanical
constriction. The effects of cavity collapse are in terms of the creation of hot spots,
releasing highly reactive free radicals, surface cleaning and/or erosion, and enhance-
ment in local transport (heat, mass, and momentum) rates. The collapse of bubbles
generates localized “hot spots” with a transient temperature of the order of 10.000 K
and pressures of about 1000 atm. Under such extreme conditions, water molecules
are dissociated into OH• and H• radicals. These OH• radicals then diffuse into the
bulk liquid medium, where they react with organic pollutants and oxidize/mineral-
ize them. The two main mechanisms for the degradation of pollutants using hydro-
dynamic cavitation are the thermal decomposition/pyrolysis of the volatile pollutant
molecules entrapped inside the cavity during the collapse of the cavity and sec-
ondly, the reaction of OH• radicals with the pollutant occurring at the cavity–water
interface. In the case of nonvolatile pollutants, the main mechanism for the degrada-
tion of pollutants will be the attack of hydroxyl radicals on the pollutant molecules
at the cavity–water interface and in the bulk fluid medium. The mechanical effects
are also significant. In some cases, the intensity of shockwaves generated by the
collapsing cavity can break molecular bonds, especially the complex large molecu-
lar weight compounds. The broken-down intermediates are more amenable to OH•
attack as well as biological oxidation, which can further enhance the rate of oxida-
tion/mineralization of the pollutants in subsequent biological treatment [117].
The literature reports on the application of hydrodynamic cavitation are very
scanty and limited only to synthetic wastewater containing only specific pollutants
such as textile dyes, pharmaceutical drugs, and pesticides [115, 121]. The use of
hydrodynamic cavitation reactors in real industrial wastewater treatment applica-
tions has been rarely investigated. Chakinala et al. [115] have tried hydrodynamic
cavitation (HC) in conjunction with the advanced Fenton process for the treatment
of real industrial wastewater and reported that hydrodynamic cavitation is very
effective as a pretreatment to biological oxidation for the effluent samples consid-
ered in their work.
The best and most practical utilization of cavitation in an energy efficient manner
can be made by using it as a pretreatment in combination with any other advanced
oxidation system or conventional biological treatment systems. Hydrodynamic cav-
itation (HC) was evaluated as a pretreatment for complex wastewater such as bio-
methanated distillery wastewater. The biomethanated distillery wastewater was
subjected to an HC reactor, and the effect of various process parameters was assessed
and optimized for maximizing COD/TOC reduction and enhancing the
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 423

biodegradability index of the wastewater. The associated color reduction has also
been achieved.
In another study, Sangave and Pandit [122] employed sonication of distillery
wastewater as a pretreatment step to convert complex molecules into a more utiliz-
able form by cavitation. Samples exposed to 2 h ultrasound pretreatment displayed
44% COD removal after 72 h of aerobic oxidation compared to 25% COD reduction
shown by untreated samples. These results are contrary to those of Mandal et al.
[74], who concluded ultrasonic treatment to be ineffective for distillery spentwash
treatment.

10.3.4 Case Studies: Wastewater Treatment Plants

and Related Regulations

Combined anaerobic and aerobic treatment processes are commonly applied to the
baker’s yeast industry. Anaerobic first stage is also a two-phase system: acid pro-
duction phase and methane production phase. The anaerobic treatment system has
several units including a buffer tank, an effluent pumping station, an acid reactor,
two methane reactors, vacuum degasifiers, lamella separators, a gas storage tank, a
boiler system, and a flare of emergency. Anaerobic reactors are constructed as
UASB reactors. Lamella separators and vacuum degasifiers are added to prevent the
washout of the biomass from the system. The aerobic second stage is designed and
operated as an extended aeration-activated sludge system with a special selector
unit at the beginning. The main treatment units of the aerobic stage are the selector
compartment, the aeration basin with four equal aerated cells connected in series,
and the final sedimentation tank with a sludge recirculation facility. Surface aerators
are used for aeration. In these processes, high-strength wastewaters are fed to the
anaerobic reactors, while low-strength wastewaters are directly fed to the aerobic
process. The major factor affecting COD removals is high initial and microbial inert
COD [3]. The typical wastewater treatment plant scheme of the baker’s yeast indus-
try applied to Turkey is given in Fig. 10.8.
The “İzmit Industrial and Domestic Wastewater Treatment Plant” is a plant
where raw domestic wastewaters and pretreated industrial wastewaters of various
sectors such as tire, drug, chemical, and yeast industries that have been discharged
to collectors are treated biologically [123]. Wastewater discharge limits for this col-
lector are shown in Table 10.6 [124]. Wastewater discharge limits are provided by
such yeast industry with conventional biological treatment. In order to improve the
performance of the treatment plant, studies on the addition of evaporator and mem-
brane units are carried out.
In Turkey, industries are obliged to treat their effluent to the standards specified for
the category where they belong to the environmental legislation and as described in
the Water Pollution Control Regulations (12.31.2004/25687), before being discharged
into the receiving medium. Discharge standards for the yeast industry are given in
Table 10.7 [125]. It is difficult to ensure the standard of color parameter given in
424 E. C. Doğan et al.

Fig. 10.8 The another typical wastewater treatment plant scheme of baker’s yeast industry [11]

Table 10.6 Pretreatment limit values for Kocaeli Municipal Wastewater Treatment Plant [124]
Parameters Limit values Parameters Limit values
BOD (mg/L) 250 Copper (mg/L) 2
COD (mg/L) 800 Lead (mg/L) 3
SS (mg/L) 350 Nickel (mg/L) 5
Total nitrogen (mg/L) 40 Zinc (mg/L) 5
Total phosphate (mg/L) 10 Mercury (mg/L) 0.5
Detergent (mg/L) 5 Silver (mg/L) 5
Arsenic (mg/L) 10 Cyanide (mg/L) 10
Antimony (mg/L) 3 Phenol (mg/L) 10
Tin (mg/L) 5 Sulfur (mg/L) 2
Barium (mg/L) 3 Temperature 40 °C
Cadmium (mg/L) 2 pH 6–9
Chrome 5 – –

Table 10.7 Effluent standards of baker’s yeast industry in Turkey [125]

Table 5.2: Sector: Food industry (yeast
production)
Parameter Unit Composite sample (2 h) Composite sample (24 h)
Chemical oxygen demand (COD) (mg/L) 1200 1000
Suspended solids (SS) (mg/L) 200 100
Oil and grease (mg/L) 60 30
pH 6–9 6-9
Color (Pt-Co) 280 260
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 425

regulation with conventional treatment process at the baker’s yeast industry. It is

reached to this limit with applying advanced treatment process as post-treatment.
Another full-scale wastewater treatment plant, RomPak Baker’s Yeast Company,
Pascani, Romania, was investigated by Ifrim et al. [67]. The maximum flow of the
WWTP is ~4000 m3/day. The effluent of the baker’s yeast wastewater treatment
plant meets the requirements of NTPA 002/2002 normative being sent to the munic-
ipal plant. The researchers determined a strategy for nitrogen removal from baker’s
yeast wastewater in order to increase the yield and rate of nitrogen compound bio-
conversion. According to this strategy, the effluent of the baker’s yeast wastewater
is mixed with the recycled sludge from the clarifier in the selector (Fig. 10.9). Then,
nitrification and denitrification exist in the tank, which is an assembly of concentri-
cally tanks. In the middle, there is the denitrification zone, and in the outside, there
is the nitrification area. The volume of these areas is 960 m3 for the denitrification
zone and 2880 m3 for the nitrification zone. The best efficiency, almost 90%, is
obtained on nitrification–denitrification processes, and they have stated that the
advanced biological treatment is completely ecological, and it does not need chemi-
cals for pretreatment processes [67].
Because of the high amount of wastewater pollutants, which should be dis-
charged into the sewerage network, a pretreatment is necessary for clearing partially
industrial effluents so treated waters comply with either standard conditions for
direct wastewater discharge into the local sewerage networks and wastewater treat-
ment plants (NTPA 002/2005) or standard load limits of pollutants from industrial
wastewater discharge into the natural receptors (NTPA 001/2005). Nevertheless, by
the usage of wastewater treatment into a single anaerobic stage, the purified waste-
waters are not able to comply with standard discharges requirements for baker’s
yeast industries. For these reasons, the effluents of the anaerobic treatment process

Sludge recycling

Clean water

From factory Clarifier

Buffer
Selector

tank

Nitrification Nitrification
Denitrification Denitrification
Degazificator

Fig. 10.9 Biological treatment plant of baker’s yeast wastewater [67]

426 E. C. Doğan et al.

should be further treated by the other treatment technology in order to fulfill the
NTPA 002/2005 requirements [33].
Industrial fermentation plays an important role in Taiwan, where sugarcane is
one of the most important crops. Water quality items and limits for effluent stan-
dards for enterprises, sewage systems, and building sewage treatment facilities are
given in Table 10.8 for Taiwan [126].

10.4 Sludge Management

Wastewater treatment generates sludge, which in turn must be either disposed of or

used. Sludge management begins with sludge generation and continues through
sludge processing and ultimate disposal [127]. Sludge treatment and disposal opera-
tions on a local or regional basis need careful planning to ensure that the strategy
undertaken is environmentally acceptable, reliable, and cost-effective [128].
The product-specific waste from the food industry is characterized by its high
proportion of organic material. The disposing of this waste can be difficult for bio-
logical stability and the potential growth of pathogens, high water content, rapid
autoxidation, and changes due to enzymatic activity [129].
Generally, the baker’s yeast industry is equipped with a full wastewater treatment
plant comprising an anaerobic pretreatment phase and an aerobic post-treatment
phase. The anaerobic phase of the treatment generates very little excess sludge. The
aerobic treatment, however, results in the generation of relatively large amounts of
excess sludge. The physical and chemical characteristics of a typical baker’s yeast
industry sludge are provided in Table 10.9 [130].
Generally, the heavy metal content of baker’s yeast industry sludge is lower than
the maximum allowable heavy metal content for agricultural use. The agricultural
use of such sludges with organic carbon and high nitrogen content is preferred envi-
ronmentally and ecologically [131].
Anaerobic sludge is a potential source of organic matter, nutrients, and minerals
and may be useful as an agricultural soil supplement. Freshly digested sludge is

Table 10.8 Effluent characteristics and limits in Taiwan [126]

Effluent Effluent
Applicable scope characteristics limits
Fermentation industries (brewing industry; MSG production Biological oxygen 50
industry, wine or liquor, alcohol and vinegar production demand (BOD)
industries; soy sauce production industry; and antibiotic and (mg/L)
organic solvent manufacturing industries) Chemical oxygen 150
demand (COD)
(mg/L)
Suspended solids 50
(mg/L)
True color 550
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 427

Table 10.9 Physical and chemical characteristics of a typical baker’s yeast industry sludge [130]

Parameter Typical value (*)

pH 8–8.5
Total moisture content (%) 78–84
Total volatile solids (%) 8–11
Total nitrogen (%) 6.5–7
Total phosphorus, as P2O5 (%) 3–3.5
Lead (mg/kg) 47
Cadmium (mg/kg) 11
Chromium (mg/kg) 18
Copper (mg/kg) 61
Nickel (mg/kg) 45
Mercury (mg/kg) 0.4
Zinc (mg/kg) 56
(*) The metal concentrations are based on the dry weight

unstable under normal environmental conditions as it is biodegradable, has an

unpleasant odor, and contains various noxious or corrosive gases such as NH3 and
H2S. Therefore, it must be stabilized before it can be adequately disposed of in the
natural environment. All of these problems can be overcome by composting, which
is an obvious solution to this problem, where all unwanted by-products can be
reduced to an acceptable level.
There are a few management alternatives for the direct disposal of digested
sludge. With a lack of other options, mechanically dehydrated sludge can be dried
to 90% with the use of biogas and utilized as an alternative solid fuel in various
industrial kilns using various methods of energy production. The net calorific value
of dry sludge is approximately 10–12 MJ/kg, with an ash content of approx.
35–45% [132].
The excess sludge withdrawn from a biological treatment system usually con-
tains 93–97% water. In order to significantly reduce the amount of sludge to be
handled, the sludge is dewatered using equipment such as thickeners, centrifuges,
belt filter press, and filter press. Dewatered sludges usually contain 20–30% dry
matter. In some cases, sludge can be applied directly on agricultural land for the
purpose of irrigation and also to provide nutrients such as nitrogen and phosphorus.
When the sludge cannot be directly applied onto land, it must be treated to stabilize
it and then properly disposed of either on plant property or off-site at an appropriate
landfill, or composted for the purpose of being used as a soil conditioner.
428 E. C. Doğan et al.

In proper waste management, reuse/recovery is considered to be very high in the

hierarchy and composting is regarded as an appropriate management method that
results in a beneficial product where nutrients such as nitrogen and phosphorus are
returned to the soil for uptake rather than being unnecessarily burnt in an incinerator
or disposed at a landfill [130].

10.4.1 Description of Composting Operation

High-moisture (greater than 60%) organic wastes represent a rather unique manage-
ment problem. Direct application to land is possible, but such practice is usually
limited to rural areas where sufficient land is available. Composting can be particu-
larly effective in converting wet materials to a more usable or easily disposable
form. At the same time, composting can stabilize putrescible organics, destroy
pathogenic organisms, and provide significant drying of the wet substrate. All of
these advantages are obtained with minimal outside energy input: the major energy
resource being the substrate organics themselves. Furthermore, composting is a
flexible process: it can be viewed as a conversion process to produce a material suit-
able for reuse or simply as a stabilization and drying process to provide easier dis-
posal. Composting is also compatible with a wide variety of feedstocks [133]. Most
of the compost was produced from bio-waste (4.8 Mt), green waste (5.7 Mt), the rest
from sewage sludge (1.4 Mt), and mixed waste (1.4 Mt) [134].
Composting is the biological decomposition of organic material. The process can
be represented in general terms as follows:

Fresh organic waste  O2  stabilized organic residue  CO2  H 2 O  heat (10.1)

The extent of reaction is synonymous with the extent of decomposition (degree

of stabilization), and it is a determinant of product quality [135].
Aerobic composting is the decomposition of organic substrates in the presence of
oxygen (air). Anaerobic composting is the biological decomposition of organic sub-
strates in the absence of oxygen. The principal differences between these two pro-
cesses are summarized in Table 10.10 [136].

Table 10.10 Comparison of aerobic and anaerobic composting processes [136]

Characteristic Aerobic process Anaerobic process
Energy use Net energy consumer Net energy producer
End products Humus, CO2, H2O Sludge, CO2, CH4
Volume Up to 50% Up to 50%
reduction
Processing time 21–30 days 20–40 days
Primary goal Volume reduction Energy production
Secondary goal Compost production, waste Volume reduction, waste
stabilization stabilization
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 429

Dewatered Recycled compost

Distribution
Mixing Composting Screening Curing and and
Storage Marketing

Bulking material
Recycled bulking material

Fig. 10.10 The basic stages in composting of dewatered sludge [137]

Most sewage sludge composting operations are composed of two sources (sludge
and bulking agent), one process (composting), and one sink (utilization or disposal
of the end product). They represent the four main issues that must be considered in
any compost plant. The basic stages in any composting operation may include
dewatering, mixing, composting, curing, and screening (Fig. 10.10). Sludge must be
dewatered prior to composting, so it can be handled as a semisolid and not as a liq-
uid. The sludge is dewatered to approximately 25% solids and 75% moisture con-
tent. In the mixing stage, dewatered sludge is mixed with the bulking agent. The
objective is to avoid sludge compactness by creating spaces for air circulation and,
if needed, to provide enough extra carbon for a suitable C/N ratios [137].
Dewatered sludge cake is not a bulky material, and it lacks the porosity of materi-
als such as straw or refuse. Because of its plastic nature, sludge also tends to com-
pact under its own weight, which further reduces the void volume [133]. The
mixture of bulking agents and sludge is composted with three techniques. After
21 days of active composting, the material is further stabilized at a 30-day curing
period. The purpose of screening is to separate the finished compost from the bulk-
ing agent [137].
Wastewater treatment plant sludge hass high water content, low organic contant,
low porosity, and high viscosity. Therefore, for effective composting process, waste-
water treatment plant sludge must be mixed with leaves, sawdust, wood, straw,
compost amendment, or with the city life rubbish compost. Aerobic composting
processes of sewage sludge are mainly controlled by factors such as feed moisture
content, ventilation system, temperature, pH value, C/N, and conditioner. The
requirements for mixing composting sludge with raw material are moisture content
of 50–60%, pH 5–9, C/N 25–35/1, organic matter content of 20–80%, and 12–60 mm
size [138].
The integration of some organic wastes of the food industry in the composting of
sewage sludge might enhance the composting process and provide more aromatic
moieties for humic acid synthesis. In addition, the ratio of sludge:bulking
agent:organic food waste (i.e., initial C:N ratio) and the texture of the bulking agent
influence the composting reaction rate and the final compost quality [139].
430 E. C. Doğan et al.

The aeration provides oxygen, which is essential to avoid anaerobic conditions

and to carry out the aerobic decomposition of the organic matter. Because of that,
the aeration affects microbial activity, substrate degradation rate, and temperature
variation of the process. In general, the main aeration methods providing oxygen
during composting are physical turning of the mass, natural convection, and forced
aeration [140]. Using the forced-air (positive pressure) aeration that blows air from
the bottom of the composting container to diffuse air upward for aeration will main-
tain a favorable composting environment to reduce the composting time to about
28–35 days. A major disadvantage of this method is the emission of odorous gas to
the environment causing serious pollution. The negative-pressure aeration uses per-
forated pipes placed near the bottom of the composting reactor and connected to a
blower. When the blower is turned on to withdraw air from the reactor, negative
pressure is created in the reactor [141].
Oxygen feedback controllers have been used to enhance the composting process.
Several ranges of oxygen concentration in the exhaust gases have also been recom-
mended. It has been stated that oxygen levels between 5 and 20% (v/v) result in an
optimum range, a minimum of not less than 8% [140]. The oxygen uptake rate
controller is recommended for the airflow regulation in composting systems with
automatic control [137, 139].
Temperature controllers are based on the removal of excess heat accumulated in
the composting matrix. In most cases, by satisfying the needs of the temperature
criteria, the oxygen requirement is also met [140]. If the thermophilic phase period
of the composting process (T > 65 °C) has persisted for more than a few days, the
compost produced may be considered sanitized and free of pathogens [132].
Biodegradability of waste sludge from the yeast industry in in-vessel aerobic com-
posting was investigated [142]. The maximum temperature reached was 60 °C in
in-vessel composting (C/N ratio of the raw sludge at approximately 12.5).
Stabilization is an oxidation process of the sludge to a resistant form, not causing
any adverse environmental effect after the disposal of the organic fraction of sludge.
Composting is a stabilization process. The stabilization degree of treatment sludges
is not defined completely yet, but various stabilization indicators are given in the
literature. Some indicators that can be used in biological stabilization under aerobic
conditions are decreases in temperature, in organic content, and in oxygen uptake
rate, presence of nitrate, absence of ammonia and starch, odor reduction, and
increase of redox potential [123, 133, 143].
Degremont [144] describe the criteria that can be used for assessing stabilization
as follows: sludge respiration rate determines the stabilization degree (the limit is
0.10–0.15 kgO2/kg organic matter/day in endogenous phase); during aerobic stabi-
lization, dissolved oxygen in the sludge liquid should be 2 mg/L after 120 h continu-
ous aeration and loss dry matter should not exceed 10% by weight.
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 431

10.4.2 Composting Techniques

Composting is a bioremediation technique for the food industry waste such as fruit
and vegetable processing industry, meat and poultry, and fermentation industry.
There are several composting types with the same general stages but differ in capital
and operating costs and in the ways that they use to achieve the proper conditions
for bacterial growth and in the time required for completing their task. The methods
employed may be classified into three general categories [144, 145].
1. Windrow: In a windrow system, a mixture of dewatered sludge and the bulking
agent is constructed from 1 to 2 m high and 2 to 4.3 m at the base. The rows are
turned and mixed periodically during the composting period. Under typical
operating conditions, the windrows are turned a minimum of five times while the
temperature is maintained at or above 55 °C [146]. The conventional windrow
and aerated windrow processes are viable sludge-disposal options [147].
2. Aerated static pile: The aerated static pile system consists of a grid of aeration or
exhaust piping over which a mixture of dewatered sludge and the bulking agent
is placed. Material mixed of dewatered sludge and the bulking agent is compos-
ted for 21–28 days and is typically cured for another 30 days or longer. Typical
pile heights are about 2–2.5 m. A layer of screened compost is often placed on
top of the pile for insulation. Perforated pipe is commonly used for air supply
[146]. Feed mixture could be composted by either natural ventilation or intermit-
tent aeration [148]. Screening of the cured compost is usually done to recover
the bulking agent.
3. In-vessel: In-vessel composting is accomplished inside an enclosed container or
vessel. Mechanical systems are designed to minimize odors and process time by
controlling environmental conditions such as airflow, temperature, and oxygen
concentration [146]. Drying and destruction of volatile solids were highest with
high aeration rates and low process temperature. The highest composting rates
were achieved when temperatures were kept below 60 °C by high airflow
rates [149].
The characteristics of each method are given synoptically in Table 10.11 [145].
Different composting technologies are applied in industrial facilities. The method
selection is dependent on the investment and operation cost, the time required to
reach compost stability and maturity, the availability of land, and the origin of raw
materials. Among the available methods for composting, closed vessel technolo-
gies, such as the tunnel technology, can be defined as the most sophisticated and
environmentally controlled systems and are successfully implemented as high
capacity facilities in populated areas. In contrast, the open-air pile system is the
simplest and requires the lowest investment [140].
432 E. C. Doğan et al.

Table 10.11 Composting methods [145]

Composting
Method time Cost Usage Disadvantages
Windrow 2–6 months for Low Used mainly in Difficult control of
municipal solid combination with conditions,
waste in-vessel technology for temperature, water
curing the compost concentration odor
Aerated 6–12 weeks Medium Used for sewage sludges, Continued electrical
piles municipal solid waste, costs
yard wastes, and
industrial organic wastes
In-vessel Less than a High due to All types of waste High cost, intense and
week to installation skillful management
2 weeks costs

Table 10.12 Maturity indices [154]

Respirometry rates Very mature Mature Immature
mg CO2–C/gVS/day <2 2–8 >8

10.4.3 Compost Quality

The compost quality depends on the content of pollutants such as heavy metals,
persistent organic pollutants, pathogenic bacteria, and inert matter in the mature
compost. The properties of the standard compost leachate must also be considered.
Heavy metals and persistent organic pollutants accumulate during the composting
process and may cause problems upon utilization. The content of heavy metals and
persistent organic pollutants is determined by the quality of the input material,
which should be carefully controlled in sewage sludge or in the raw wastewater, and
by additional treatment and reduction at the source [132].
Nutrient loss, specifically N, can be a major problem in composting. The lower
initial C:N ratio had a major effect on N loss. Apart from N loss, P, K, and Na loss
can also be significant [150].
Fresh organic waste materials cannot be applied to the soil until they have been
sufficiently biostabilized because the application of immature organic materials to
soil may affect plant growth due to nitrogen starvation and production of toxic
metabolites [151]. During composting, germination and plant growth bioassays
showed the toxicity of immature compost. The reason for immature compost toxic-
ity suggested in the literature includes high concentrations of volatile organic acids,
high concentration of NH4+-N, oxygen depletion, or the presence of heavy metals
[152]. Methods such as chemical and physical analyses, microbiological assays,
plant bioassays, spectroscopy, and degree of humification were developed to assess
compost maturity [153]. The maturity of compost was evaluated with the maturity
indices (Table 10.12) [154].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 433

Table 10.13 Physical and chemical characteristics of a typical compost material [130]
Parameter Typical value
pH 7.2
Total moisture content (%) 60
Total nitrogen (%) 3
Total phosphorus, as P2O5 (%) 4

The degree of humification can be determined with spectrophotometric ratios.

The low values of spectrophotometric ratio reflect a higher degree of aromatic con-
densation and indicate a higher level of organic matter humification. The slight
increase observed in spectrophotometric ratios indicates a relative increase in com-
pounds with phenolic and benzene-carboxylic groups in the structure of the humic
substances [139].
Several important soil microbiological properties, such as substrate-induced res-
piration, potential ammonium oxidation, and nitrogen mineralization, were
improved after the application of both leftover material from biogas production and
composting. The genetic structure of the soil bacterial community appeared to resist
changes caused by the addition of organic waste [155].
Waste sludge obtained from the baker’s yeast industry is composted by using the
static aerated pile method. Composting was conducted under experimental condi-
tions using wood chips as a bulking agent, a sludge:bulking agent ratio of 1:1.5, and
an amount of air supply of 30 m3/h-ton dry sludge. The physical and chemical char-
acteristics of a typical compost material are presented in Table 10.13 [130].
Eriçyel [156] investigated the composting process to treat biosolid waste sludge
from the baker’s yeast industry. Two types of bulking agents (hazelnut husk and
shredded cornstalk) were used to lower the moisture content of the biosolids to
increase porosity and to add a source of carbon. At the end of the study, it was
proven that hazelnut husk and shredded cornstalk can be considered as a good bulk-
ing agent for the composting of biosolids. The optimal ratio for composting biosol-
ids from the yeast industry was 1:1:1 (biosolid:shredded cornstalk:hazelnut husk).
Furthermore, it was obviously shown that when compost was applied at high ratios
(10%), additional fertilizer was not needed [156].
Treatments frequently digest or compost their sewage sludge to reduce the level
of pathogens and odors. The degree to which a sludge is processed is very important
when placing sewage sludge in monofills or on surface disposal sites in order to
eliminate the spread of pathogenic diseases [127].
The degradation rate of aerobic digestion was higher than that of composting.
Genç et al. [131] stated that at the 13th day of the composting specific organic
434 E. C. Doğan et al.

nitrogen removal rate was 0.85 mg organic N/g VS/day while at the same time
3.54 mg organic N/g VS/day was measured in digestion. The specific organic car-
bon removal rate was determined to be 9.19 mg organic C/g VS/day on 13th day of
composting while 20 mg organic C/g VS/day in digestion [131].
In the composting facilities, exhaustive controls of the biological risks should be
carried out. These controls should include measurements in the compost (in its dif-
ferent phases from elaboration) and in the air (different zones of the facility) of the
concentrations of bioaerosols, paying special attention to the Gram-negative bacte-
ria and the fungus A. fumigatus. Moreover, the environmental concentrations of
VOCs should also be determined, and the personnel should be biologically moni-
tored with certain regularity [157]. Unpleasant odors may be released only during
the mechanical turning of the material. For the aerated piles, an air collection sys-
tem, fitted with a biofilter, is planned. Dust emissions from periodical mechanical
turnover are not considered critical when the compost is properly wetted; noise
emissions can be controlled by the selection of appropriate low noise equip-
ment [132].

10.4.4 Control of Nuisance Conditions

It is important to engineer appropriate systems for the control of nuisance condi-

tions during the design of a compost facility. Potential nuisances most often associ-
ated with composting are odors and dusts [133].

10.4.4.1 Control of Dust Formation

Dust control in a compost facility can be a major problem, particularly in semiarid

climates. Municipal sludge organics are characterized by a small-sized particle dis-
tribution. When sludge-based compost exceeds about 65–75% solids, the small-­
sized particles can be easily airborne if agitated. At higher moisture contents, dust
is generally not a significant problem. Therefore, dust conditions can potentially
occur any time dry compost is handled. This would include turning or agitation of
dry windrows, screening dry compost from bulking particles, loading of trucks, and
so on. Another potential source of dust is fugitive emissions that may occur from the
operation of mobile equipment on dry, unpaved surfaces [133].
The presence of bacteria and fungi is fundamental to the composting process.
Agitation of the composting material through shredding, turning, and screening
results in the liberation of these organisms into the air, commonly termed a bioaero-
sol. The concern regarding bioaerosols from composting activities arises because of
their potential to cause adverse health effects in employees and the public living in
close proximity to such facilities [158]. The airborne release appeared to be related
to the amount of microorganisms in the wastes, the design of the site, and the opera-
tional procedures used in the composting plants. Airborne microorganisms were
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 435

monitored at different composting facilities. A. fumigatus and mesophilic bacteria

were used as the principal monitoring parameters. The composting plants all showed
levels of both airborne microorganisms in the 103–105 cfu/m3 range in the operating
area, making it advisable for the staff to use protective masks [159].
Most of the control measures to avoid excessive dust formation are reasonably
straightforward and include the following [133]:
1. For open systems, turning or agitation should be discontinued when the material
exhibits noticeable dust formation, which should begin at about 65–70% solids.
2. Controlled aeration systems can be used to reduce the turning frequency.
3. Paved surfaces should be used whenever possible.
4. Mixing of wet cake and compost product in the open should be avoided when-
ever possible.
5. Operations with a high dust potential, such as mixing and screening, should be
conducted in an enclosed reactor or building.
6. Compost can be pelleted before drying or storage to increase particle size.

10.4.4.2 Odor Control

Control of odors is one of the most difficult problems in current sanitary engineer-
ing practice. Both inorganic and organic compounds can be malodorous. Hydrogen
sulfide (H2S) and ammonia (NH3) are the most common inorganic odorants. Organic
sources generally come from low-molecular-weight, more volatile organic com-
pounds (VOCs) [133]. The VOCs produced from composting typically include
nitrogen-based compounds, sulfur-based compounds, volatile fatty acids, hydrocar-
bons, trepans, esters, ethers, alcohols, and aldehydes/ketones [133, 159, 160].
Compost-generated VOCs can be evaluated in terms of two processes: the produc-
tion within the pile due to organic matter degradation and emission at the pile sur-
face after gas convection within the pile. VOC production within the pile was
different from emission at the pile surface. The total mass of VOC production was
1.09 g C/kg dry matter, which was 2.3 times as high as the total mass of emis-
sion [160].
The main sources for odorous emissions occur during material handling and dur-
ing processes where the material is moved. These include delivery of the biowaste,
pre-processing such as shredding and screening, and the composting process itself,
especially during turning and the final screening. A large portion of odorous emis-
sions can be avoided by ensuring optimal composting conditions. If the biowaste is
too wet and not well aerated, malodors often occur as a result of partly anaerobic
processes [162].
In a windrow system, frequent turning can help prevent anaerobic pockets in the
material. The traditional method to aerate a pile of compost depends on mechanical
equipment to turn over the material twice daily to once every other day. The odorous
emission is difficult to control with mechanical aeration [141]. If odor persists in an
aerated system despite adequate aeration, this may be due to compaction of the
436 E. C. Doğan et al.

materials and channeling of the air. A turn to remix the compost may solve the prob-
lem. In sludge composting, the choice of bulking agent might also be important, as
a range of particle sizes will help air movement through the heap [133].
Off-site emissions seem to be the main problem at open outdoor facilities. To
avoid these emissions, one option is to enclose at least parts of the process. The
aeration process provides operations to reduce emissions. Forced aeration would
increase fugitive emissions, whereas if vacuum-induced (negative-pressure) aera-
tion is practiced, the waste gas can be captured and treated. At enclosed composting
plants, off-gases are captured and treated [162].
A number of techniques are available to reduce the odor concentration in exhaust
gases collected during composting. Techniques such as absorption by scrubbing or
condensation, bioscrubbers, adsorption, oxidation by thermal, chemical, or biologi-
cal means, and use of masking agents are available to the designer [133, 161]. The
most relevant systems are bioscrubbers and biofilters [162].
Most exhaust gases collected during composting will be saturated with water
vapor (or nearly so) at temperatures above ambient. As these gases cool, condensa-
tion of water vapor will occur. Because of this, it is necessary to provide water traps
in ducting used to transport such gases. As condensation occurs, it is likely that
water-soluble species in the gas will be absorbed into the condensate.
Gas absorption is a unit operation in which one or more soluble components of a
gas mixture are dissolved in a liquid. Scrubbing with water or chemical solutions
(KMnO4, NaOCl, alkalies, or various acids) can affect a reasonably consistent
45–60% reduction in odor concentration [133]. Bioscrubbing is a process of bio-
logical waste gas treatment in which exhaust air is “washed” in an absorber with a
scrubbing liquid. The scrubbing liquid is subsequently drawn off and transferred to
an activation tank in which the constituents absorbed into the liquid are degraded by
microorganisms. The liquid is continuously cycled through the process [162]. A
biofilter is a fixed-bed reactor filled with biologically active packing material.
Microorganisms settled on the media feed on the organic compounds that are con-
tained in the waste gas. Biofilters usually are combined with wet scrubbers. The
scrubbers are used to humidify the air passing to the filter in order to avoid drying
of the filter material. Frequently used biofilter media are compost, peat, root wood,
bark, wood chips (normally used as a bulking agent), and various combinations
[163]. One important property of the media is the ability to store water [161, 163].
Adsorption involves contacting a fluid phase (gas or liquid) with a particulate phase
that has the property of selectively taking up and storing one or more solute species
from the fluid. Compost deodorization piles have accomplished measured odor
reductions averaging 50%, but with wide variations in removal efficiency [133]. The
produced unseparated compost is used as a biofilter to capture and biologically
oxidize the landfill gas (methane) and other organic compounds found in the landfill
gas [132]. In this system, a moistened-bulk solid medium, such as soil or composted
sludge, provides the contact surfaces for microbiological reactions to oxidize odor-
ants [146]. The air is passed through a heap of mature compost, which acts as a
biological filter. Odorous molecules dissolve in the film of water on the compost
particles, and residual micro-organisms from the composting process then break
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 437

down the compounds, effectively removing the odor problem [164]. A pilot-scale
active aeration reactor was studied for composting food wastes in an open-top con-
tainer aerated with negative pressure by vacuum. A biological filter bed filled with
mature compost was used to remove NH3 from the emission. Using the biological
filter to remove NH3, the emission contains less than 1 ppm of NH3 [141]. Oxidation
processes result in the oxidation of organic species to carbon dioxide and water, or
partial oxidation to other intermediate compounds. In masking agents, perfume
scents can be added to the gas stream to mask or combine with an objectionable
odor. Masking agents have limited application in compost facilities since more
effective techniques are available [133, 145].
The process of waste decomposition in composting facilities releases a variety of
odors, airborne particles, and bioaerosols. They cause infections or irritations to
humans, especially to sensitive or sick people. Park et al. [165] studied the simulta-
neous removal of odors, airborne particles, and bioaerosols in a composting facility
by dielectric barrier discharge. The removal efficiency of contaminants in the air
increased as the specific energy densities (SED) increased, with removal efficien-
cies of up to 80% and 76% being achieved for ammonia and amines, respectively.
The removal efficiency of the overall airborne particles was 75% when 113 J/L of
SED was employed, while the bioaerosols removal efficiency was 89% when 38 J/L
of SED was used [165].
A complete odor control system consists not only of the collection and treatment
of odorous gases but also their proper dispersal into the atmosphere. Design of sys-
tems for treatment and dispersal must be coordinated to assure that ground-level
ambient odor standards are not violated [133].

10.4.5 Land Application of Sludge

Food industry waste sludge is commonly applied to the land in a number of ways.
Applications of sludge to agricultural and nonagricultural lands are the following:
1. Land application to agricultural lands: The method of applying sludge to agricul-
tural land depends on the physical characteristics of the sludge and soil and the
crops grown. Liquid sludge may be applied with tractors, tank wagons, irrigation
systems, or special application vehicles. Liquid sludge may also be injected
under the surface layer of the soil. Dewatered sludge, on the other hand, is typi-
cally applied to cropland by equipment similar to that used for applying lime-
stone, animal manures, or commercial chemical fertilizer. Generally, the
dewatered sludge is applied to the land surface and then incorporated by plowing
or disking. When applied to pasture land, sludge is usually not incorporated into
the soil [127].
2. Land application to nonagricultural lands: Sludge is most often sprayed from
mobile equipment into established forest stands as a partially dewatered, but still
liquid, material. Other types of nonagricultural land application include sewage
438 E. C. Doğan et al.

sludge applied to public contact sites (e.g., parks, cemeteries, golf courses) and
reclamation sites. When sewage sludge is used to stabilize and re-vegetate land
at reclamation sites, typically large amounts of sludge are applied on a one-time
basis. This large amount is necessary to ensure that sufficient organic matter and
nutrients are introduced into the soil to support vegetation until a self-sustaining
ecosystem is established [127].
Physical site characteristics of concern include topography, soil permeability,
site drainage, depth to groundwater, subsurface geology, proximity to critical areas,
and accessibility [146].
Landfilling and land application of the sewage sludge are suggested to be the
most economical sludge disposal methods. Sludge amendment to the soil modifies
its physicochemical and biological properties. Land application of the sewage
sludge is becoming more popular due to the possibility of recycling valuable com-
ponents such as organic matter, N, P, and other plant nutrients. Organic matter added
to the soil as sewage sludge composts improved the soil properties, such as bulk
density, porosity, and water holding capacity. The chemical properties of sludge–
soil mixture not only depend on the properties of the soil and sludge and the applica-
tion rates of the mixtures, but also depend on their interaction and soil pH. Sewage
sludge amendment increased the soil microbial activity, soil respiration, and soil
enzymes activities. In general, it has been shown that the addition of sludge to agri-
cultural land increases the growth and production of crop plants. The increase of
crop yield as a result of sludge application often exceeded that of well-­managed
fertilized controls [166].
Prior to applying sludge to cropland, the following tests should be run: nitrogen
content and forms, phosphorus, potassium, heavy metals, percent solids, and tests
required by local health agencies. Prior to applying sludge to a particular site, the
soil should be tested for pH, cation exchange capacity, and phosphorus and potas-
sium availability. Crop selection would have to be consistent with local farming
practices and locally marketable. Site work includes drainage control, monitoring
wells, pH adjustment by lime addition, and field preparation. The application rate
calculation is frequently based mainly on the nitrogen requirements of the crop to
be grown. The computed rate is then adjusted, if necessary, to prevent excessive
heavy metal or phosphorus buildup [167].
Plant availability of heavy metals differed widely among the crop species. The
accumulation of Cd, Ni, and Zn in the plants showed the greatest increases com-
pared to their background levels. The Cu and Pb accumulation in the plants grown
on sludge-amended soils showed only small increases compared to those grown on
uncontaminated soil. Multiple regression analyses of various soil properties showed
that the surest way to control the accumulation of metals in food plants is by con-
trolling their concentrations in the soil [39].
The aim of Council Directive 86/278/EEC of the European Communities is to
regulate the use of sewage sludge in agriculture in such a way as to prevent harmful
effects on soil, vegetation, animals, and human beings. Values for concentrations of
heavy metals in soil to which sludge is applied, concentrations of heavy metals in
sludge, and the maximum annual quantities of such heavy metals that may be intro-
duced into soil intended for agriculture are given in Table 10.14 [168].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 439

Table 10.14 Limit values for concentrations of heavy metals in soil in sludge for use in agriculture
and the maximum annual quantities of heavy metals that may be introduced into the soil [168]
Annex 1A
Limit values for concentrations of heavy metals in soil (mg/kg of dry matter in a representative
sample of soil with a pH of 6–7)
Parameters Limit valuesa
Cadmium 1–3
Copperb 50–140
Nickelb 30–75
Lead 50–300
Zincb 150–300
Mercury 1–1.5
Chromiumc –
Annex 1B
Limit values for heavy-metal concentrations in sludge for use in agriculture (mg/kg of dry
matter)
Parameters Limit values
Cadmium 20–40
Copper 1000–1750
Nickel 300–400
Lead 750–1200
Zinc 2500–4000
Mercury 16–25
Chromiumc –
Annex 1C
Limit values for the amount of heavy metals that may be added annually to agricultural land,
based on a 10-year average (kg/ha/year)
Parameters Limit valuesa
Cadmium 0.15
Copper 12
Nickel 3
Lead 15
Zinc 30
Mercury 0.1
Chromiumc –
a
Member States may permit the limit values they fix to be exceeded in the case of the use of sludge
on land, which at the time of notification of this Directive is dedicated to the disposal of sludge but
on which commercial food crops are being grown exclusively for animal consumption. Member
States must inform the Commission of the number and type of sites concerned. They must also
seek to ensure that there is no resulting hazard to human health or the environment
b
Member States may permit the limit values they fix to be exceeded in respect of these parameters
on soil with a pH consistently higher than 7. The maximum authorized concentrations of these
heavy metals must in no case exceed those values by more than 50%. Member States must also
seek to ensure that there is no resulting hazard to human health or the environment and in particu-
lar to groundwater
c
It is not possible at this stage to fix limit values for chromium. The Council will fix these limit
values later on the basis of proposals to be submitted by the Commission, within one year follow-
ing notification of this Directive
440 E. C. Doğan et al.

Trace metals are trapped in the soil matrix, and nutrients are taken up by plants
and converted into useful biomass. The principal metal of concern is cadmium
because it can accumulate in plants to levels that are toxic to humans and animals
but below levels that are toxic to plants (phytotoxic) [146].
The soil conditioning properties of sludge are more significant than the nutrient
additions. Mismanagement of a land application system can result in public health
problems, odor nuisance, and/or soil destruction from excessive heavy metal
buildup. A land application system must be designed to provide maximum benefits
from the sludge without creating problems. The design should therefore include rate
determination schedules and methods to be used by operators [167].
The land application systems for sludges should be designed properly not to
exceed the maximum assimilation capacity of the land and the product growing on
it. Some factors like the scarcity or absence of suitable land area to apply sludge
directly and adverse climatic conditions for sludge application make the compost-
ing process suitable for obtaining a stabilized product that can be applied to land
easily in the sludge assessment [130].

10.5 Specific Subjects

10.5.1 Control of H2S in Biogas Generating from Anaerobic

Treatment of Fermentation Industry

Wastewaters containing organic matter and sulfate are generated by many industrial
processes that use sulfuric acid (food and fermentation industry) or sulfate-rich
feedstocks (sea food-processing industry). Also, the use of less oxidized sulfurous
compounds in industrial processes results in the generation of sulfate-rich wastewa-
ters. Sulfate concentration in the molasses fermentation industry is relatively high,
approximately 4000 mg SO42−/L. Although the COD/SO42− ratio is not very high,
inhibition problems in the anaerobic treatment due to H2S toxicity are mani-
fested [66].
Many processes have been proposed and employed to remove sulfide from gas
streams and sulfide-rich wastewaters. Can Doğan (2008) studied the effectiveness
of the control of H2S gas formed in biogas together with methane as a result of
anaerobic treatment of fermentation industry wastewater with the autotrophic deni-
trification process. High removal efficiencies obtained with low hydraulic retention
times revealed sulfide oxidation with the autotrophic denitrification process in
which nitrate or nitrite is used as an electron acceptor. In this study, loading rate and
molar loading ratios (NO3−/S2− and NO2−/S2−) did not affect the sulfide removal
efficiency. However, elemental sulfur formation as a by-product or sulfate forma-
tion as an end product as a result of denitrification were closely related to the load-
ing ratio [4].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 441

Removals of sulfide from wastewater of the fermentation industry were applied

by using continuous flow stirred tank reactor under anoxic conditions. The stoichi-
ometry of sulfide oxidation with nitrate is calculated assuming different end-­
products based on the thermodynamic approach and compared with experimental
yield values. The calculated maximum volumetric and specific sulfide oxidation
rates reached 0.076 kg S2−/m3 h and 0.11 kg S2−/kg VSS h, respectively [169].
Nitrite can be used instead of nitrate as the electron acceptor to remove sulfide in the
autotrophic denitrification process. When the nitrite to sulfide ratio was above 1.48,
the end product was mainly sulfate. Otherwise, as the value of the ratio was under
1.48, the distribution of end products was shifted to the mixture of sulfate and ele-
mental sulfur [170]. The results are obtained at industrially relevant conditions and
can be easily adapted to either the biogas cleaning process or to sulfide-containing
effluent streams.
Krapivina et al. [171] studied anaerobic mesophilic fermentation of sulfate-­
containing yeast industry wastewaters at the laboratory scale with anaerobic
sequencing batch reactors (ASBR). COD removal of 75–82% was achieved at vol-
ume loading rates up to 7.7–8.0 kg COD/m3day and at a COD/SO42− ratio of 8.0.
Also, the best results for sulfate removal (99%) were achieved in the CSRB, with
the concentration of sulfide in the reactor effluent being about 10 mg/L. In the large-­
scale experiments, the instability of the processes could create significant difficul-
ties in applying the ASBR technology for the treatment of yeast wastewaters. In
addition to the inhibition of the process, sulfide formation also costs major malodor
problems and corrosion of equipment during the experiment. Accumulation of sul-
fides was an indication that competition between methanogenous and SRB was won
by the latter [171].

10.5.2 Vinasse

The main discharge of yeast fermentation is called vinasses, and it consists of dif-
ferent organic compounds such as betaine, glycerol, and various reducing sugars.
Vinasse is a substance by fermentation of molasses. The characteristics of vinasses
depend mainly on the raw material used. Vinasse with high concentrations of solu-
ble solids can be obtained when sugarcane, sugar beet, grape, avages, or sweet sor-
ghum are used [172]. Dominant colored compounds are in molasses from the
degradation of sugars. Under conditions of high temperature and acidic or alkaline
pH, the nonenzymatic reactions of the hexoses may produce melanoidins, invert
alkaline degradation products, and caramels. The Maillard reaction occurs in the
presence of amine compounds and monosaccharides; brown products are created.
During the heating of the sugar with ammonium compounds, a series of heterocy-
clic compounds are formed. In recent years, the toxicological profile of these com-
pounds has been examined. The type, quantity, and characteristics of toxic colored
442 E. C. Doğan et al.

compounds of vinasse result in the potential utilization of this by-product being

highly limited [173].
Major organic components of sugarcane vinasse are glycerol, lactic acid, etha-
nol, and acetic acid, whereas sugar beet vinasses also contain glycerol and their
main compound is nitrogen-rich betaine, and cellulose and hemicellulose. Both
dilute and concentrated vinasse can be spread on agricultural fields or used as
organic fertilizer [174]. Vinasses can be used as fertilizer due to their nutrient con-
tent, mainly calcium and potassium, and their high organic material content.
Vinasses contain phytotoxic, antibacterial, and recalcitrant compounds. Highly col-
ored compounds lead to reduced sunlight penetration in rivers and lakes. Reductions
in soil alkalinity, manganese availability, and seed germination inhibition have been
reported by the use of these wastes in agriculture. Previous treatment of these wastes
is therefore needed before their final disposal. There are many proposals for physi-
cochemical and biological treatments for vinasses. The combination of different
processes where the first step was an anaerobic treatment followed by an aerobic or
physicochemical process gives better results in the removal of organic load and
color. Treated vinasses can be used in agriculture without the risk of polluting soil,
underground water, or crops [172].
The composition of compost derived from urban waste is often far from ideal for
plant nutrition, and vinasses applied to fields can be easily washed away. However,
the addition of vinasses to solid urban wastes before the fermentation steps can both
improve the compost formation process and allow problems associated with the
separate materials to be overcome [175].
Vinasses from sugarcane molasses were evaluated as a new source of sugarcane
wax. Nuissier et al. [176] showed that 1 L of alcohol produced in rum processing,
thus 20 L of vinasses, could lead to the recovery of approximately 3.4 g of crude wax.
Vinasse is a suitable feedstuff in the feeding of ruminants since most of its crude
proteins comprise amide-substance and are palatable to ruminants. The nutritional
value of vinasse as ruminant feed was reported. The use of vinasse as a feed additive
in poultry and pigs and the dosage used in ruminant diets were reported to show its
influence on animal performance [177]. Despite being a highly polluting effluent,
vinasse could be used in the production of single-cell proteins for feed supplemen-
tation due to its high carbon content. The generated microbial biomass by biological
treatment of vinasse with yeast presented a low anti-nutritional value and an average
protein content of 46.85% [178].
Molasses vinasses always have a high level of organic content such as crude
proteins, lactic acid, glycerol, cholesterol, amino acid, and reducing sugars (COD in
the range of 80,000–100,000 mg/L and BOD in the range of 40,000–50,000 mg/L)
and have strong odor and dark brown color, and they also contain nutrients in the
form of nitrogen (1660–4200 mg/L), phosphorus (225–3038 mg/L), and potassium
(9600–17,475 mg/L). Recently, many research teams focused on the study of pre-
paring nutriment fodder using molasses vinasses. However, the dosage of molasses
vinasses was also limited because unexpectedly high potassium content in molasses
vinasse could lead to diarrhea in bulls and pigs. Various techniques have been pro-
posed to reduce the potassium level in molasses vinasses [179].
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 443

Another alternative for vinasse consumption is combustion. Molasses spentwash

containing 4% solids can be concentrated to a maximum of 40% solids in a
quintuple-­effect evaporation system with thermal vapor recompression [178, 179].
The condensate with a COD of 280 mg/L can be used in fermenters. The concen-
trated other liquor is spray dried using hot air at 180 °C to obtain a desiccated pow-
der with a calorific value of around 3200 kcal/kg. The powder is typically mixed
with 20% agricultural waste and burnt in a boiler. The use of recirculating fluidized
bed (RCFB) incinerator is recommended to overcome the constraints due to the
stickiness of spentwash and its high sulfate content [180]. Combustion is also an
effective method of on-site vinasse disposal as it is accompanied by the production
of potassium-rich ash [181] that can be used for land application. Experience with
the treatment of vinasse indicates the following trends:
1. Anaerobic digestion, complemented by oxidative chemical treatments (e.g., ozo-
nation), is usually placed as a pretreatment.
2. Aerobic treatment alone and combined with ozonation, which have been directed
to remove phenolic compounds and color, have been successfully applied.
3. Physicochemical treatments such as Fenton, electro-oxidation, oxidants, and so
on., which are now mostly at the lab-scale stage, have demonstrated a significant
removal of recalcitrant organic compounds.
4. Fungal pretreatment with chemical treatment followed by oxidative (O3) or
anaerobic digestion seems to give attractive results.
5. Vinasses may be co-composted with solid organic wastes, particularly with those
from agricultural activities and agro-industry.
Unicellular protein, biohydrogen, and enzyme production from vinasses could
be promising ways to improve the economic feasibility of vinasses treatment. Raw
vinasses can be filtered to give two main streams: vinasses and solids rich in yeast
and microbial biomass. The latter can be processed to generate an alternative pro-
tein source for animal feed or other uses. Filtered vinasses could be subjected to a
variety of biotechnological processes in order to give diverse added-value bioprod-
ucts: special bioproducts, enzymes, protein (yeast, algal, bacterial, fungal), and soil
amenders by co-composting vinasses with agro-industrial and other wastes. In par-
allel, filtered vinasses could be used for methane and biohydrogen production either
as separate stages or as a series of biohydrogen-methane processes. Treated vinasses
from bioenergy stages can be directed to co-composting of agro-industrial and other
wastes or post-treated with AOP or biological processes before discharge or reuse in
irrigation in such a way to close the environmental circle [182].

10.5.3 VOC Emissions

The VOC emissions are generated as by-product of the fermentation process. The
two major by-products are ethanol, which is formed from acetaldehyde, and carbon
dioxide. These by-products form as a result of excess sugar present in the fermenter
444 E. C. Doğan et al.

or an insufficient oxygen supply to the fermenter. Under these conditions, anaerobic

fermentation occurs and results in the excess sugar being broken down to form alco-
hols and carbon dioxide. When anaerobic fermentation occurs, 2 moles of ethanol
and 2 moles of carbon dioxide are formed from 1 mole of glucose [183].
The two types of control measures that are currently employed at yeast manufac-
turing facilities are (1) process control and (2) add-on controls [181, 182].
The ethanol production rate is a function of the yeast growth rate, and both of
these parameters are related to the residual sugar concentration. By continuously
adding only the exact amount of molasses required by the fermentation, conditions
of excess sugar are eliminated, thus minimizing ethanol formation. The most com-
mon add-on control devices for controlling VOC emissions are wet scrubbers, car-
bon adsorbers, incinerators, condensers, and biological filtration. A combination
system could result in lower control costs and relatively equivalent emission reduc-
tions [183]. At facilities manufacturing dry yeast, VOCs may also be emitted from
the yeast dryers used to dry the yeast [184].

10.5.4 Biogas Production

Industrialization exerts considerable pressure upon natural resources, along with an

increased demand for energy. In addition, the waste generated by the industries is a
major environmental concern. Thus, it is imperative for highly polluting industries
to adopt a suitable waste treatment process for the clean disposal of high-strength
wastewater. Anaerobic digestion is one such technology, which is gaining wider
acceptance in the present scenario over aerobic treatment due to the production of
biogas, which can be further used for meeting a part of energy demand [185].
Anaerobic digestion is the most common method for the treatment of high-­
strength wastewater and stabilization of waste sludge from baker’s yeast manufac-
turing facilities.
The following four stages comprise the digestion process: hydrolysis, acidogen-
esis, acetogenesis, and methanogenesis. Various microorganisms are involved in
each step. During the first stage, a group of microorganisms secretes enzymes that
hydrolyze polymers to monomers to convert particulate materials into dissolved
materials. Subsequently, the acidogenic phase includes the action of a large and
diverse group of fermentative bacteria. These bacteria hydrolyze and ferment
organic materials and produce organic acids, CO2, and H2. In the third phase, aceto-
genic bacteria convert these monomers to acetic acids. The final step in biogas pro-
duction is performed by acetoclastic methanogens [186].
One factor influencing the efficiency of waste-activated sludge anaerobic diges-
tion is access to cell-bound organic matter. The hydrolysis of the organic matter is
the rate-limiting stage of the anaerobic process. In order to improve hydrolysis and
anaerobic digestion performance, several pretreatments have been examined, which
cause the lysis or disintegration of sludge cells. In pilot-scale experiments, process
performance and biogas production of a cascade of two methanogenic continuously
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 445

stirred tank (CSTR) reactors connected in series were compared to a conventional

one-step CSTR reactor treating sewage sludge. Results showed that the serial con-
figuration could improve biogas production by 9.5–40.1%. Biogas selectivity was
estimated to be 0.49 ± 0.06 m3biogas/kg degraded total volatile solids for the cascade
and 0.44 ± 0.02 m3biogas/kg degraded total volatile solids for the one-step process.
The process performance and biogas production from sewage sludge can be opti-
mized through serial digestion [187].
Among the microorganisms involved in digestion, methanogens are the major
microbiological group responsible for methane production. To study the microbio-
logical equilibrium in an anaerobic reactor, Traversi et al. [186] detected the metha-
nogen concentration during anaerobic digestion as an indicator of biogas production
capacity. A positive and significant correlation between the biogas production rate
and methanogen abundance was observed. The applied real-time quantitative PCR
method is suitable to describe microbiome into the anaerobic reactor; moreover,
methanogen concentration may have the potential for use as a digestion optimiza-
tion tool [186].
The treatment process of wastewater requires large amounts of energy. The major
energy consumption process in wastewater treatment plants is sludge stabilization.
Besides having high energy consumption, sludge can be used as a renewable energy
source. As a result of the anaerobic digestion process, it is degraded to produce
biogas, which consists of 65–70% CH4, 30–35% CO2, 1–5% H2, and 0.3–3%N2
with various minor impurities, notably NH3, H2S, and halides [188]. Substances
such as H2S and NH3 may inhibit the anaerobic digestion process or cause corrosion
problems in pipelines of plants or in the distribution network [189].
The collected gas from digesters is cleaned of H2S and H2O before it is com-
busted to generate steam or electricity. Typical digester gas is about 70% CH4, with
most of the rest of the gas being CO2. Since the energy content of pure methane is
35,800 kJ/m3 at standard temperature and pressure, the digester gas has a net heat
value of about 25,000 kJ/m3. The heat value of methane is roughly the same as natu-
ral gas, which has a heat content of approximately 37,000 kJ/m3 [190].
If the intended use is for power generation, the biogas must be scrubbed to
remove a number of impurities. After conditioning, the biogas can be used for
onsite power generation to heat homes or can be added to the national natural gas
grid. Biomethane produced from biogas generated by anaerobic digestion of
organic matter is an alternative gas source to that of natural gas. Of particular
interest is the possibility to inject biomethane, refined biogas with quality com-
parable to that of natural gas (CH4 concentration greater than 95%), which can
be used in place of fossil fuels in all its network applications and in transporta-
tion. To produce pipeline-­quality biomethane starting from the biogas generated
by the anaerobic digestion process, it is necessary to remove water, sulfur com-
pounds, halogenated organic molecules, carbon dioxide, oxygen, and met-
als [189].
The total energy generation potential from the anaerobic digestion of industrial
wastewater is estimated to be 2963 GWhc/a equivalent electric energy. This is equiv-
alent to a 565 MW power plant installation. Distilleries and sugar industries are
446 E. C. Doğan et al.

potential areas with a combined capacity of 832 GWhc/a equivalent electric energy
per annum [185].
The developed thermoeconomic analysis procedure and formulations based on
the specific exergy costing method are applied to an existing municipal wastewa-
ter treatment plant using actual operational plant data by Abusoglu et al. [188].
Thermoeconomic analysis helps allocate the costs to the plants’ main streams and
subcomponents. Activated sludge is digested in the anaerobic digestion reactors
to produce biogas with a 60% methane content. For each 1 m3 biogas produced in
the wastewater treatment plant, 68.26 kg of sludge with a dry matter content of
5.0% is digested. The total exergy rate of the biogas produced with a mass flow
rate of 0.212 kg/s is obtained as 6653 kW. The actual exergetic efficiency of the
wastewater treatment plant is determined to be 34%, which indicates that 66% of
the total exergy input to the plant, mainly by sewage and power consumptions, is
destroyed.
A lab-scale anaerobic hybrid (combining sludge blanket and filter) reactor was
operated to study anaerobic biodegradation of distillery spentwash [191]. The model
of Karhadkar et al. [192], as given by Eq. (1), was employed for the prediction of
biogas yield from the reactor. Accordingly, the biogas yield can be expressed as
follows:

Qgas  GQ  S0  S  (10.2)

where Q is the flow rate of influent, G is the conversion factor, So is the influent
COD concentration, S is the effluent COD concentration, and Qgas is the biogas
yield. The value of G was determined as 0.52 m3/kg COD experimentally. The
model predicts gas yield within ±5% of the experimental value.
Anaerobic digestion can convert a significant portion (>50%) of the COD to
biogas, which may be used as an in-plant fuel, and also saves the energy that would
be required for aeration using aerobic treatment. Sirbu and Begea [33] treated dis-
tillery wastewater by full-scale thermophilic (50–55 °C) anaerobic digestion. In this
study, more than 60% removal of COD was achieved with 76% of biogas compris-
ing methane, thus making it a valuable fuel [33].
Bioremediation of distillery spentwash by anaerobic digestion is an attractive
primary treatment due to its reputation as a low cost, environment friendly, and
socioeconomically acceptable technology. Anaerobic digestion of wastewater from
a distillery industry was studied in an upflow anaerobic fixed-film bioreactor using
different support materials. Among the various support materials studied, the reac-
tor having coconut coir could treat distillery spentwash at 8 d hydraulic retention
time with an organic loading rate of 23.25 kg COD/m3 day leading to 64% COD
reduction with a biogas production of 7.2 m3/m3 day having high methane yield
without any pretreatment or neutralization of the distillery spentwash [193].
In the UASB reactor, due to the excellent settling characteristics of this granular
biomass and the presence of a specially designed three-phase (biogas, water, and
biomass) separator device in the upper part of the UASB reactor, an excellent sludge
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 447

retention is assured in this reactor system. Performance of full-scale UASB reactors

treating distillery spentwash was evaluated. The plant was designed to handle
650 m3/day of distillery spentwash, having an average COD concentration of
112,400 mg/L with an HRT of 6 day. The biogas production was stabilized to the
range of 48,290–135,115 m3/week with 60% methane content [194].
The startup performance of upflow anaerobic hybrid reactors (UAHRs) and
upflow anaerobic sludge blanket (UASB) reactors in the anaerobic treatment of dis-
tillery spentwash has been studied. The maximum volumetric gas production of
149 L/m3 produced more in UAHR than in the UASB reactor during steady-state
conditions [195].
The carbon and hydrogen fixed in sludge can be converted into a clean, high
calorific value energy source as hydrogen through a cost-efficient bioprocess.
Biohydrogen production by dark fermentation of the waste biological sludge
obtained from the sedimentation unit of the aerobic wastewater treatment plants of
yeast industry showed that the maximum H2 yields 41 mmol/g total sugar con-
sumed [196].

10.6 Approaches on Management of Sources in Baker’s

Yeast Industry

The yeast industry is facing environmental challenges, such as increasing costs of

production and non-value adding expenses of waste effluent and process water
charges. A satisfactory solution to these multilayered challenges calls for an inte-
grated approach in order to accomplish effective waste management and water
conservation.

10.6.1 Water Conservation

Freshwater conservation is another concern in the yeast industry, and it is no longer

recognized as a free commodity. Innovative technologies, along with modifications
of existing technologies, are commercially available to reduce water consumption.
In general, reduction in industrial water and wastewater can be achieved through
one or a combination of the following measures:
1. Process modification or change in raw materials to reduce water consumption
2. Direct reuse of wastewater
3. In-plant reuse of recovered wastewater
4. Use of treated wastewater for nonindustrial purposes
In this perspective, improved water use in yeast factories can contribute to better
operational efficiency, improved economic competitiveness through reduced water
448 E. C. Doğan et al.

demand combined with savings in water and wastewater treatment costs, and low
environmental impact due to a decrease in surface and subsurface withdrawals as
well as less groundwater contaminant intrusion [197].
A step-by-step study of the water cycle in a company allows significantly reduc-
ing water consumption. Making the water balance is useful to detect unknown water
consumption and is the basis for further optimization. Water pinch or water scan
methods allow deciding which streams can be reused, directly or after regeneration.
Finally, the remaining wastewater can be included in an overall process scheme
aiming at zero discharge of wastewater. For yeast factories, all three steps have been
proven to be useful, although a zero discharge is not entirely obtained.
A systematic approach to water management helps keep the water issue under
control. This can be done by controlling water input, consumption, and output and
by gearing all water-related activities to one another. At the input side, the emphasis
is on searching alternative sources for process water. Surface water, if available, can
be an option for some companies; brackish water is a possible source on the condi-
tion that an adequate pretreatment method is used. Wet countries might investigate
the possibility of using rainwater, although this is usually only feasible when small
quantities are needed for low-quality purposes, such as sanitary water. A further
option is to regenerate wastewater and recycle it as process water. Wastewater often
requires an extensive treatment before it can be discharged. Further purification in
order to obtain a water quality fit for reuse is often a realistic option in terms of
additional treatment cost versus the benefits of using recycled water [198].
Three advantages are to be taken into account. The first one is recycled water,
which is a supplementary and reliable source of freshwater and can add to existing
sources or replace them. The second advantage is that the net volume of water con-
sumed decreases drastically, and the last one is that the volume of wastewater to be
discharged decreases.
Here we use three steps to optimize the water balance in a yeast plant. The first
step is to investigate the current water balance in detail. This seems to be very
straightforward but usually leads to complex measurements and calculations.
Information of how much water is consumed in which processes allows us to take
simple measures to save water. The second step is to combine water-consuming
processes and reuse water where possible for other purposes requiring a lower water
quality. Typical examples of possible candidates are cooling processes and tank
cleaning. The water pinch method offers a theoretical approach to do this; however,
practical impediments often need to be challenged. The final step is to regenerate
partial waste streams and re-introduce them into the process cycle. Because of the
complexity of most wastewaters to be treated, this requires one or more membrane
separations, often using pressure-driven processes such as ultrafiltration and
nanofiltration.
Water balance is a numerical account of how much water enters and leaves a
plant and where it is used within the plant. It should contain detailed information
about the amount of water used by each process. The water balance is a crucial
instrument to understand and manage water flows throughout the plant, to identify
equipment with water-saving opportunities, and to detect leaks. The setup of a water
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 449

balance requires a preliminary survey of existing data, an assessment of major gaps

in the available information, and a decision on how detailed the water balance
should be. This leads to choices concerning the amount of work and the technical
resources that have to be invested in the project.
In a typical yeast plant, the main source of process water is well water. This water
is brought from a depth of 60–360 m at a constant temperature of 17 °C. At peak
moments, more water is needed than the well water pumps can provide. Therefore,
the company will supplement its water needs with tap water. Before being used in
the production process, the water is treated in order to obtain the quality that is
needed for each specific application in the plant. On arrival in the yeast factory, all
water is first chlorinated and then stored in a reservoir. Also, 80% of the stored water
is directly used from this reservoir, without further treatment. This is the “cleaning
water.” It is mainly used for CIP (cleaning-in-place) cleaning procedures. About
20% of the available water is demineralized. To this end, the water first passes an
activated carbon column for dichlorination and removal of organic matter. Afterward,
the water is deionized by passage through anion and cation columns. Thus, process
water or deminwater is produced mainly for use in the fermentation process. About
8% of the deminwater is further treated by UV irradiation to obtain the sterile and
“ultraclean water,” which is mainly used in critical yeast processes.
In order to obtain more detailed information on the individual water-consuming
processes, supply pipes need to be traced and schemes need to be drawn to under-
stand groupings and connections of equipment in the water network. In many cases,
water is taken from existing pipes left from an earlier installation or simply tapped
from the water supply of a completely another facility. Thus, for constructing a
rough internal water balance, existing water meters provided only partial informa-
tion. Additional flow measurements need to be carried out for all subunits, including
CIP stations, and noncontinuous operations such as cleaning of tanks.
Reusing wastewater streams as an input for less demanding installations is a
second step toward a zero-discharge system. The reuse of wastewater is well docu-
mented in the literature. Two theoretical approaches can be considered: water pinch
techniques and water scan techniques, which provide a more spontaneous approach.
In order to apply these concepts, a water balance is essential. Both qualitative and
quantitative requirements should be specified for each individual water input at an
installation. As for the qualitative requirements, it is important that the minimum
quality specifications for each water flow should be stated. The water pinch method
implies the determination of the minimal water use by an installation, which is
needed to evacuate contaminants from a process, given an initial contamination
load. The combination of all these minimal water quantities, effluent characteristics,
and maximum allowed contamination loads for all processes in the plant then deter-
mines the overall minimal water use. This calculation is executed three times under
different assumptions:
1. Existing allowed contamination loads for water inputs are not changed.
2. More effluents become available for reuse if allowed contamination loads for
selected processes are relaxed.
450 E. C. Doğan et al.

3. All or selected effluents are regenerated before being used as an input. In this
way, several effluents are theoretically eligible for reuse, but significant invest-
ments are required. The analysis of the results also reveals which process is a
bottleneck for further reducing water consumption in the plant. Improving the
water use characteristics of this process will result in the most cost-effective
impact on the water use for the company as a whole [199].
The water pinch method requires that effluents of installations are redirected,
serving as an input for another process. New water treatment equipment may have
to be added. Frequently, individual effluent streams have to be mixed together in
order to obtain the required input characteristics. This typically requires extra pip-
ing, pumps, valves, etc. More practical, common sense methods were developed to
help reduce the water consumption in a production plant. These methods can be
called water scan techniques; they are not based on a theoretical framework but all
start from a water balance of the plant, upon which ideas are gathered on how to
manage the water system more efficiently, concentrating on reducing costs.
Good housekeeping and opportunities for simple direct reuse are the main atten-
tion points at this stage. Application of new techniques for water treatment or invest-
ments in less water-consuming technologies is considered. Opportunities for
water-saving practices are then classified based on their financial and technical con-
sequences. Taking available resources into account, the most promising projects are
chosen and a planning for their implementation is proposed.
It is obvious that regeneration of partial wastewater streams only decreases the
water volumes needed for the yeast production activities, but a zero discharge is not
yet feasible. The reason for this is that wastewater treatment generates side streams
such as sludge volumes, which would need to be reused in the strict sense of zero
discharge. Furthermore, yeast production is particular in the sense that materials are
brought into contact with food (yeast), which requires special safety regulations,
especially in terms of microbiological safety. One must also be aware of the psycho-
logical barrier of using wastewater in food products. This is an impediment to zero
discharge systems. Current improvements are more focused on partial water recov-
ery. In order to achieve this, the remaining wastewater fraction needs to be regener-
ated up to a level similar to the freshwater currently used in the company.

10.6.2 Waste Minimization

Waste minimization in the yeast industry faces certain unique challenges. First, the
yeast-making process itself was developed from very old classical practical activi-
ties. This limits the application of modern technologies to minimize waste in many
yeast factories. Second, as a general rule in the yeast industry, many poor practices
are often not classified as “poor,” but as common practices. This mindset renders the
implementation of waste management in the yeast industry a challenging task.
Third, industrial operations are a mixture of both batch and semi-batch processes.
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 451

These uncontrollable variations adversely affect resources to the point where waste
minimization is being compromised during actual production operations. These
limitations offer an explanation as to why waste management is practiced as end-of-­
pipe technologies in numerous yeast plants [200]. However, owing to rapidly grow-
ing global demand on manufacturing processes and final products to exert minimal
or no environmental footprints [201], the yeast industry has begun to experience
legislative pressure to become more efficient [199, 200]. Thus, the increasing
demand for the greening of industrial production processes and products, both from
customers and legislative authorities, coupled with rising operational and waste
treatment costs in the yeast industry, has started to move toward the adoption of
integrated waste preventative approaches, as opposed to the traditional reparatory
environmental engineering practices [203]. To effectively analyze waste generated
in the yeast industry, a systematic methodology was followed comprising waste
source identification, causative evaluation of waste, and qualitative derivation of
feasible waste minimization alternatives [204].
It is highly suggested to concentrate on the entire production process and con-
sider comprehensive waste minimization strategies for each waste stream, process,
or unit operation.
Process flowsheets were evaluated from the inception of the raw materials up to
the packaging of the products. The visual representation of the flowsheets facilitated
the identification of critical points within the production processes, where major
releases and discharges were likely to occur. This also helped prioritize the waste
streams in need of further investigation. The standard production process consists of
preparation, fermentation, clarification, dewatering, and drying. However, various
companies use different process routes, which significantly impact waste manage-
ment for both intrinsic and extrinsic wastes. In that sense, it is impossible to derive
all feasible waste minimization strategies. Such a wide spectrum of process routes
highlights the need for a systematic analysis to determine alternatives for eliminat-
ing or improving the handling of waste streams from the production processes
[202, 203].
Changing process technology is an important technique for reducing waste vol-
ume and strength. Some examples are the following:
1. Alternation in cleaning procedures such as using counter-current washing, recy-
cling of used water, and reducing the cleaning frequency. In the production pro-
cess, it is possible to reuse the last yeast wash water as the first wash water. The
BOD5 values of this washing water range from 200 to 500 mg/L. This water can
be reused again in the washing process after heat sterilization. It is suggested that
filtration followed by sterilization is likely to produce treated wastewater suit-
able for recycling as the next batch fermentation first water.
2. Employing new methods in production line cleaning in baker’s yeast industry.
Several feasibility studies indicated that the installation of an automatic CIP con-
trol system not only reduced the extent of waste treatment but also permitted
wastewater recycling [205].
452 E. C. Doğan et al.

3. Changing waste transport methods such as self-draining piping systems will

help reduce the amount of wash water used within the plant.
Changing products, which can serve the purpose of those that they substitute, can
bring waste minimization as in the cases of yeast cream as a final product, instead
of block or dry yeast products.
The utilization of glucose syrup instead of molasses as a fermentation medium is
a good example of changing input material. About 85% of the pollution load (COD)
of the yeast industry is due to molasses, and it originates from the fermentation
stages. Therefore, it is necessary to reduce the pollution load of these stages.
Molasses can be partially substituted (up to 50%) for glucose syrup to improve
waste minimization as well as a reduction in COD and color load in the effluent.

10.6.2.1 Minimizing Extrinsic Waste

Yeast production and its complementary products are accompanied by the genera-
tion of large quantities of waste streams, namely organic waste and wastewater.
Variations adversely affect resources to the point where waste minimization is being
compromised during actual production operations. These constraints offer an expla-
nation as to why waste management is practiced as “end-of-pipe” technologies in
numerous yeast plants, notably wastewater treatment and landfilling of solid wastes.
However, owing to the rapidly growing global demand for manufacturing processes
and final products to exert minimal or no environmental footprints, the yeast indus-
try has begun to experience legislative pressure to become more efficient. Thus, the
increasing demand for the greening of industrial production processes and products,
both from customers and legislative authorities, coupled with rising operational and
waste treatment costs in the yeast industry, has started to move toward the adoption
of integrated waste preventative approaches, as opposed to the traditional reparatory
environmental engineering practices [206].
To effectively analyze waste generated in the yeast industry, a systematic meth-
odology needs to be followed comprising waste source identification, causative
evaluation of waste, and qualitative derivation of feasible waste minimization
alternatives.
Several generic methods for developing waste minimization options at an indus-
trial scale have been discussed, and a case study on industrial food processing has
been reported by Van Berkel [204].
Waste minimization is addressed through the acquisition of process knowledge
of fermentation operations. Knowledge acquisition can be achieved through regular
observation of a variety of yeast production processes and unit operations.
Two approaches for evaluating process flowsheets to identify feasible strategies
for achieving waste minimization in any industrial process have now been estab-
lished [207]. The techniques are broadly classified as quantitative and qualitative
approaches, and the latter approach was employed in this case, owing to the general
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 453

lack of reliable plant data in production processes. Second, a qualitative approach

provided a feasible way of evaluating flowsheets of existing processes, unlike quan-
titative methods where large sets of data of high integrity are required to precisely
determine feasible waste minimization alternatives. Third, most data and informa-
tion in the yeast industry have been acquired through experience, and qualitative
techniques were found most suited in synthesizing process flowsheets for the iden-
tification of waste minimization opportunities.
It is necessary to concentrate on the entire yeast production process and consider
comprehensive waste minimization strategies for each waste stream, process, or
unit operation [208].

10.6.2.2 Recovery of Waste

Betaine recovery from the wastewater is a good example of “Resource recovery” in

the yeast industry. High-strength waste generated at the end of the fermentation
process contains both high- and low-molecular-weight organic loads. It was shown
that betaine can be separated from the weak vinasse by using an ultrafiltration pro-
cess with a membrane of the molecular cutoff size of 4000. This does not help
reduce the load of the waste treatment but also by-product recovery of a commer-
cially valuable chemical named “betaine.”
In the yeast industry, strength of wastewater is influenced by the organic load
content of the discharge. As much organic load as possible should be recovered to
reduce the strength of wastewater by installing appropriate technology. Examples of
resource recovery and reuse may be listed as follows:
1. Vinasse has been found to be quite suitable for animal feed when properly pre-
pared and treated. The animal feed can substitute molasses.
2. The recovered betaine is used as a valuable animal feed additive.
3. The sludge would be suitable as a fertilizer due to its nutrient content and can be
used as a source of nitrogen, potassium, and magnesium.
4. Methane can be produced as a by-product through anaerobic digestion. This can
be utilized for domestic purposes or in the industry itself.
Despite being a highly polluting effluent, vinasse could be used in the production
of single-cell proteins for feed supplementation due to its high carbon content. The
generated microbial biomass by biological treatment of vinasse with yeast presented
a low anti-nutritional value and an average protein content of 46.85% [178].
Through the application of the conceptual framework, several alternatives for
reducing, eliminating, or reusing extrinsic waste should be identified. It is remarked
that acceptable effluent quality and quantity can be achieved through the implemen-
tation of integrated waste and production management strategies.
The yeast production industry has been generating a large amount of yeast waste
(the main sub-product of yeast production and the liquid fraction obtained by
decanting) rich in organic matter that could be used as soil improvers or organic
fertilizers. Yeast waste can be applied to an acidic sandy soil as organic fertilizer
454 E. C. Doğan et al.

since it provides not only nitrogen and potassium to plants but also contributes to
increasing soil organic matter content. But the potential nitrate leaching and increase
of CO2, N2O, and CH4 have to be carefully monitored to avoid environmental prob-
lems [209].

10.6.2.3 Methodological Approach

To help frame the problem of waste minimization in yeast production, two critical
issues need to be clarified. On the one hand, it was important to establish an under-
standing of the product route from raw materials to the final product. The yeast
production route was established through interviews, actual plant observations, and
reviewing the literature. The final waste matrix was found to be a combination of
interactive factors. Examples of such factors are the type of technology used, reuse
and recovery of useful by-products, and the operating practices within a given fac-
tory. On the other hand, different production scenarios were examined as they had a
critical influence on the consumption of raw materials and effluent quantity and
quality.

10.6.2.4 Product Route Determination

Process flowsheets were evaluated from the inception of the raw materials up to the
packaging of the products. The visual representation of the flowsheets facilitated the
identification of critical points within the production processes, where major
releases and discharges were likely to occur. This also helped prioritize the waste
streams in need of further investigation. The standard production process consists of
preparation, fermentation, clarification, dewatering, and drying. However, various
companies use different process routes, which significantly impact waste manage-
ment for both intrinsic and extrinsic wastes. In that sense, it is impossible to derive
all feasible waste minimization strategies. Such a wide spectrum of process routes
highlights the need for a systematic analysis to determine alternatives for eliminat-
ing or improving the handling of waste streams from the production processes
[202, 203].

10.6.2.5 Inventory Tools

The identification and quantification of waste sources in the yeast industry were
accomplished using inventory tools [204]. The tools are classified as either product-­
oriented or process-oriented. In the fermentation industry, process-oriented tools,
i.e., material mass balance and process flow chart methods, are preferred. At any
process or unit operation, the material balances are carried out to identify the com-
ponents of the waste streams generated during the fermentation process.
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 455

The process flow chart method facilitated the identification of all possible sources
of waste generated at any stage of the fermentation process. The process was divided
into unit operations. Note that a unit operation in this context refers to an area of the
process, or a piece of equipment where input materials are processed and output
material streams are generated, which could either be a product, a by-product,
or waste.
The qualitative mass balance method was used to establish the material flow at
each level of the production process or unit operation. As a result, the method
enhanced the understanding of the relative significance of different sources and
causes of waste, as well as clarifying the composition of wastewater streams and
thus the sources of pollutants. The sequence of defining the problem until all pos-
sible waste minimization alternatives has been identified.

10.6.2.6 Waste Reduction: Methodological Evaluation of Waste

Minimization Strategies

Data collected on different stages of the fermentation process were obtained and
analyzed to identify waste minimization opportunities. Some of the data obtained
comprised flow rates of materials, the composition of generated waste effluent, and
volumes of yeast produced, as well as operating conditions and practices in different
processes. To ensure systematic identification of waste minimization strategies, a
structured methodology was followed. The methodology is composed of three-step
sequential approaches, namely waste source identification, qualitative evaluation of
waste causes, and finally the derivation of feasible alternatives for waste
minimization.

10.6.2.7 Waste Source Identification

Optimal formulation of waste minimization strategies requires unambiguous identi-

fication of all possible sources of waste, such as the inputs and consequent outputs
from a given unit operation or process in order to trace material balances at each
stage of the yeast production process. Using waste classification proposed by
Berglund and Lawson [210] and Douglas [211], the fermentation wastes were clas-
sified as intrinsic (process) or extrinsic (utility). The intrinsic wastes are inherent in
the fundamental process configuration, while on the other hand, the utility wastes
are a function of auxiliary aspects of the operation [210]. Waste identification pro-
cess was achieved through waste stream analysis and process analysis [212]. Note
that, in the context of waste minimization and particularly in deriving the reduction
strategies based on this classification, the two classes of waste types were found to
be dependent on each other in the yeast industry. In that sense, care was taken in
understanding the interactions and interconnections between the two waste types to
ensure that root causes of various waste streams were adequately established. For
the systematic identification of intrinsic waste sources, the production process was
456 E. C. Doğan et al.

broadly divided into several categories. After completion of each production stage,
water and cleaning chemicals were used for a wide range of activities. These activi-
ties may include, but are not limited to, cleaning, cooling, and sanitizing of equip-
ment. Using the above classification scheme for the process, different wastes,
by-products, or product losses were identified from various unit operations and pro-
cesses based on process flow path decomposition.

10.6.2.8 Causative Analysis of Waste

In the first stage of the conceptual framework, waste inventories and characteriza-
tion profiles provided valuable baseline data regarding the nature of pollutants gen-
erated in the yeast plants. However, before comprehensive strategies for waste
reduction or minimization could be formulated, it was crucial to understanding
when, how, and why different kinds of wastes were generated. Therefore, a caus-
ative analysis provided the understanding of the core influencing factors to the efflu-
ent quantity and quality, as well as the reasons for product and by-product losses.
Thus, understanding of the causality formed a sound basis. In addition, causality
facilitated the grasping of cause-and-effect relationships that govern the fermenta-
tion unit operations and processes that, in reality, are complex and multidimensional
and are invariably influenced by diverse factors. It was not possible to provide defin-
itive answers on causality and explicitly identify the differences that exist among
various causes of waste.

10.6.2.9 Causes of Waste Related to Input Material Characteristics

Generally, in the process industries, the feedstock of any process or unit operation
has certain properties such as toxicity or nontoxicity. Based on these properties, the
input materials may require special handling to reduce waste generation. In the case
of the yeast industry, the key raw material is molasses, which are neither toxic nor
hazardous. However, molasses have high organic content, and second, they contain
dark colors, which are an unwanted but nevertheless unavoidable component of the
input material. Hence, to minimize or eliminate cross-media pollution, molasses as
raw materials require proper and effective handling. The same principle also holds
in handling the inevitable by-products and the high volumetric fluid product. In
facilities where improper handling of the raw materials, intermediate products, by-­
products, or finished products occurs, the resultant effluent composition is charac-
terized by high organic content, high conductivity, and dark color. Such an effluent
can be highly disruptive to the environment. On the other hand, certain cleaning and
sanitizing chemicals are not environmentally benign, owing to their toxic and haz-
ardous properties. For instance, while chlorine and ammonia solvents are effective
cleaning and sanitizing materials, their toxicity and hazardousness have led to their
substitution in certain fermentation plants by more benign agents, such as hydrogen
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 457

peroxide, ozone, or hot steam. This also reduced the need for higher quantities of
rinsing water to remove the chemicals.

10.6.2.10 Causes of Waste Related to Technology

This category accounts for the causes of waste related to “technological-based” fac-
tors, such as a type of material used for equipment design, equipment sizes, piping
layout, and equipment efficiency, among others, which influence the quantity or
other characteristics of waste streams as a result of some equipment or unit opera-
tions changes. For example, low equipment efficiency or poor design generally
leads to increased waste generation; in addition, the technology used has a consider-
able impact on the effectiveness of managing and harnessing useful, but inevitable,
by-products generated at various unit operations and processes. For example, during
the pressing process where yeast and fermentation medium are separated, the use of
modern separators reduces yeast losses significantly in comparison to old-style low-­
speed separators. On the other hand, it was observed that the efficiency of equip-
ment used for cleaning and sanitizing showed a strong correlation with the quantities
of potable water and chemical demand in plant operations. For instance, if open
hosepipes were used, water and chemical consumption was found to be higher than
in an operation where high-pressure low-volume cleaners or CIP systems were used.

10.6.2.11 Causes of Waste Related to Process Execution and Management

In industrial chemical production processes, waste generation mechanisms can be

regarded as functional dependent variables [213]. In the food and beverage industry,
and particularly in the yeast industry, procedural, administrative, and institutional
practices are the key causes of waste generation. These practices are simply good
housekeeping and have a significant effect on waste profile in terms of volume,
composition, and dispersion to other environmental media. One distinctive feature
of these practices is their requirement for relatively simple in-plant changes regard-
ing the operating procedures or methods of handling wastes. Such changes lead to
the reduced waste or concentration of the contaminants in a waste stream.

10.6.2.12 Causes of Waste Related to Recovery and Reuse

The degree of effectiveness in waste management depends on the nature of the

industry. This is due to the uniqueness of the feedstock materials as well as the spe-
cific nature of the products and the intermediate by-products formed during the
manufacturing process. The yeast industry is not an exception to this rule and was
found to experience unique waste management constraints that can only be
addressed adequately through effective reuse, recycling, or recovery. The best waste
minimization strategy was to recycle the waste and by-products, reuse them in other
458 E. C. Doğan et al.

processes, recover them in order to be sold, or use as input materials in other indus-
tries. The recycling should be done based on the understanding that by-products and
waste cannot be recycled in the process(es) generating them in an attempt to pro-
duce the same product or perform the same function.
High health standard requirements for food-based products, as stipulated by the
industrial food production act, render by-products and recyclable wastes not easily
reusable in the process(es) generating them owing to the risk and uncertainties asso-
ciated with microbial contamination. From this perspective, products, by-products,
and waste recovery and recycling in other associated industries were viewed as the
most feasible waste management alternative.
The rinse water produced from cleaning and sanitizing of equipment, which is
not heavily contaminated with yeast, can be reused on cleaning floor surfaces or
pre-rinse water for heavily contaminated equipment.
Credible evidence also indicates that where reuse and recycling are implemented
ineffectively, or not at all, high effluent volumes are generated and the solids from
various processes may result in odors, high organic content in the wastewater
stream, and other catastrophic environmental consequences, such as sodicity and
salinity when the effluent is used for irrigation without proper handling [203].

10.6.2.13 Formulation of Waste Minimization Strategies

Within the theoretical framework, the development of alternative strategies was

aimed at eliminating, reducing, and controlling the causes of waste generation or
segregating useful materials from the waste streams. This was guided by the identi-
fication and analysis of both waste streams and the processes giving rise to these
streams [212] and consequently establishing their true causes through systematic
causative analysis. Alternative strategies were derived from personnel expertise and
information from the technical literature. The information and knowledge from this
causative analysis are crucial in identifying viable waste minimization strategies.
The waste minimization techniques were classified into the following categories:
technological modifications, input substitution, operational practices, and waste/
product recovery and reuse. Note that the strategies derived for a waste stream,
operation, or process do not imply that they are applicable to every yeast plant, and
in certain cases, the information and knowledge presented here are inappropriate to
some of the current practices in the yeast industry.
By using the type of waste generated as a unit of classification, three distinct
waste minimization classes were identified:
1. Strategies for minimizing fundamental waste
2. Strategies for minimizing extrinsic waste
3. Strategies for improvement of effluent quality
10 Treatment and Minimization of Waste in Baker’s Yeast Industry 459

10.6.2.14 Strategies for Minimizing Intrinsic Waste

In any industry, including the yeast industry, intrinsic waste poses the greatest chal-
lenge to be eliminated or reduced [210]. This is because most waste minimization
alternatives with the capability of achieving a reasonable reduction or elimination of
intrinsic waste ought to be technology based. As a result, high capital investment is
required for the acquisition, installation, and operating costs of the equipment.
Nevertheless, the advantages of reducing waste generation are diverse such as
reduction of waste treatment costs as well as increasing the yield per unit through-
put. Note that most common alternatives are in the category of “operating prac-
tices.” Technology-related alternatives have the greatest potential for reducing
intrinsic waste or enhancing the effectiveness of handling unavoidable waste.

Glossary

Active dried yeast It is the form of yeast that consists of coarse oblong gran-
ules, with live yeast cells encapsulated in a thick jacket of dry cells with some
growth medium.
Biomass Biomass is biological material from living or recently living organisms,
most often referring to plants or plant-derived materials.
Budding yeast Yeast with an asymmetric cell division process for the production.
Compressed yeast Compressed yeast contains about 70% water and 30% yeast
solids. The water content of the yeast cream is reduced to form by passing the
yeast cream through a rotary vacuum filtration unit.
Emulsifier An emulsion is a dispersion of droplets of one immiscible liquid within
another. Emulsifiers allow for a stable and homogeneous mixture of two liquids,
which do not normally mix.
Melanoidins Melanoidins are brown, high-molecular-weight heterogeneous poly-
mers that are formed when sugars and amino acids combine at high temperatures
and low water activity. They are also present in the wastewater of sugar refiner-
ies, necessitating treatment in order to avoid contamination around the outflow
of these refineries.
Mesophilic microorganism An organism that grows best in moderate temperature,
neither too hot nor too cold, typically between 20 and 45 °C. The term is mainly
applied to microorganisms.
Molasses Molasses is a viscous by-product of the refining of sugarcane or sugar
beet, and the syrup left from the final crystallization stage is called molasses.
Beet molasses is about 50% sugar by dry weight, predominantly sucrose, but
also contains significant amounts of glucose and fructose.
Mycelium Mycelium is the vegetative part of a fungus, consisting of a mass of
branching, thread-like hyphae.
Seed yeast Small amount of pure laboratory-grown yeast culture, which will be
used for the startup of the commercial fermentation vessels.
460 E. C. Doğan et al.

Spentlees The concentrated part of the aqueous distillery effluent stream.

Spentwash The aqueous distillery effluent stream.
Stillage The residue grain from the manufacture of alcohol from grain; used as a
feed supplement.
Strains A strain is a subset of a yeast species differing from other yeast of the same
species by some minor but identifiable difference.
Sugar beets Sugar beet is a plant whose tuber contains a high concentration of
sucrose, and it is grown commercially for sugar production.
Sugarcane Sugarcane predominantly grows in the tropical and subtropical regions.
It belongs to the grass family and is an economically important plant. The main
product is sucrose.
Trehalose Trehalose is a disaccharide formed by a 1,1-glucoside bond between
two α-glucose units.
Vinasses The residue in a still after distillation.
Yeast cream At the end of the fermentation, the yeast broth is concentrated using
a series of combined centrifugation and washing steps into a yeast cream with a
solids concentration of approximately 20%.

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Index

A Airlift bioreactors, 378

Accumulated PHAs, 318 Air pollution, 4, 5
Activated bio-filters (ABF) Air pollution control, 108, 111
applications, 257, 258 Alcaligenes eutrophus, 319
bio-cells, 257 Alcaligenes spp., 319
biological treatment forms, 256 Alkaline hydrolysis, 116
bio-media, 256 Alkaline precipitation, 116, 117
carbonaceous material removal, 257 Alkoxylation, 370
description, 256 Amidation, 370
design criteria, 258 Anaerobic acidogenic fermentation, 320,
microorganisms, 256 322, 324
performance, 258 Anaerobic digesters, 19, 23, 69
process flow diagram, 256, 257 Anaerobic digestion, 68, 175
variable loads, 256 biogas production by animal, 19, 21
Activated sludge, 125–127, 327 blanket reactors, 22
Activated sludge treatment system, 125, 127, characteristics, anaerobic digesters, 19
135, 140 development of biogas, 19
Active pharmaceutical ingredients (API), 146 energy conversion, 18
Actual surface cleaning programs, 289 fixed film digester, 22
Acyl-CoA oxidases (Aox), 372 ISPAD, 23
Additives, 56 livestock manure, 19
Advanced oxidation processes (AOPs), plug flow digester, 22
399, 416 processes, 18
Aerated lagoons, 127, 128 short fatty acids, 18
Aerated static pile system, 431 TPAD, 23
Aeration, 17, 35, 56 types of anaerobic digesters, 19
Aeration basin, 257 Anaerobic lagoons, 30
Aerobic bacteria, 177 Anaerobic reactors, 423
Aerobic bacterial growth, 246 Anaerobic sequencing batch reactors
Aerobic lagoons, 31 (ASBR), 440
Aerobic wastewater biotreatment systems, 327 Anaerobic treatment, 130–133
Aerosols’ effects, 212 Anaerobic windrow preprocessing, 181
Agricultural biotechnology, 80, 152 Anerobic/anoxic system, 403
Agricultural wastes, 64, 190, 319 Animal dung, 190
AgStar program, 19 Animal fat wastes, 342

© Springer Nature Switzerland AG 2022 471

L. K. Wang et al. (eds.), Waste Treatment in the Biotechnology, Agricultural and
Food Industries, Handbook of Environmental Engineering 26,
https://doi.org/10.1007/978-3-031-03591-3
472 Index

Animal Waste Management Software Tool water minimization, 450, 451

(AWM), 64 yeast size, 392
Antibiotic production process system, Barley-based diets, 55
137, 138 Base-catalyzed transesterification, 353
Antibodies, 85 Beet/cane molasses, 394
Antisense oligonucleotides, 85 Bench-/pilot-scale studies, 244
Aquatic flora and fauna, 283 Best available technologies (BAT), 101, 103,
Aromatic compounds, 54 146, 154
Artificial aeration, 282 Best Practicable Control Technology Currently
Aseptic cultivation, 329 Available (BPT), 101, 103, 105, 157
Azadirachta indica, 343 BET livestock waste treatment technology, 67
β-Oxidation, 372, 375
Biocatalysts, 80, 370
B Biochar (B), 190
Bacterial species, 320 Biochemical oxygen demand (BOD5), 399
Baker’s yeast Biocovers, 57
aerobic/anaerobic composting, 428 Biodegradable construction materials, 335
biogas production, 444–446 Biodegradable plastics
bread fermentation, 392 development, 317
casuative analysis, 455 market predictions, 318
chemical composition, 398 Biodiesel production, process parameter
composting methods, 431 optimization
contamination, 395 animal fat wastes, 342
definition, 391 catalyst characterization and elemental
effluents, 404, 405, 424, 426 analysis, 349–352
foods, 391 conversion of oils to biodiesel
H2S, fermentation, 440 esterification of BO, 352
heavy metals, 439 oil blend, 352
industrial production, 391 properties of MOB, 356–359
industries, 392 statistical optimization, base-catalyzed
industry sludge, 427 transesterification, 353–356
input material characteristics, 456 esterifying the oil mixture, 346
inventory tools, 454 experimental design and statistical
membrane process, 413 analysis, 347
methodological approach, 453 KOH/NaOH, 343
minimizing extrinsic waste, 451, 452 materials, 344
minimizing intrinsic waste, 458 methods
percentage composition, 395 oil blends and their
pretreatment limit values, 424 characterization, 345
process execution/management, 457 catalyst preparation, 345
product route determination, 454 characterization of calcined
production, 396–398 powder, 345–346
recovery/reuse, 457 oils preparation, 344
recovery waste, 452, 453 physicochemical and fatty acid
substrates, 394, 395 compositions, mixed oils, 348–349
technology-based factors, 456 physicochemical properties, MOB, 347
typical compost material, 433 process optimization, 347
VOC emissions, 443 regression parameter and test of
waste minimization strategies, 458 significance, 347
waste reduction, 454 transesterification of esterified oil mixed to
waste source identification, 455 biodiesel, 346
wastewater, 398, 400–401, 424, 425 Biodiesel yield, 342
water conservation, 447–450 Biofilters, 58
Index 473

Biofuel production, 342 growth, 82

Biogas, 18, 19, 21, 22, 68 medical market, 82
Biogas production, 19, 21 molecular biology, 82
direct liquefaction, 34 nonmedical markets, 82
gasification, 34 research activities and biotechnology firms
pyrolysis, 33 in the USA, 82
thermochemical processes, 33 Biotechnology terminologies
Bioinformatics, 81, 86, 153, 154 agricultural biotechnology, 80
Bioingredient-based cosmetics, 368 biotechnology, 80
Biological and natural extraction chemical synthesis-based pharmaceutical
(pharmaceutical), 152 industry, 80
Biological oxygen demand (BOD), 330 computational biotechnology, 81
Biological product extraction, 93, 94 gray biotechnology, 81
Biological sludges, 135, 136 green biotechnology, 80
Biological treatment, 63 industrial biotechnology, 80
Biomass production, 283 medical biotechnology, 80
Bion Environmental Technologies, 65, 67 military biotechnology, 81
Bioplastic biodegradation, 336 modern green environmental
Bioprocess engineering, 381 biotechnology, 81
Bioreactor, 321 Biotransformation, 323
bioconversion process, 377 Biotransformation method
description, 377 complex molecules synthesis, 371
digital imaging technologies, 378 vs. de novo synthesis methods, 371
gas/air mixture, 377 lipase-mediated, 371
nutritional factors, 377 optimal conditions (see Optimal
SmF, 377 conditions, biotransformation)
STR, 378 RA–GDL bioconversion, 372–375
types, 377 Blanket reactor, 19, 22
Bioremediation technologies, 87 Blended oils (BO), 345, 346, 348, 349,
Biosolids, 113, 125, 127, 135–137, 140, 176, 352, 359
178, 180–182, 189, 190 Blue biotechnology, 81, 153
Biosynthetic pathway, γ-decalactone, 375 BOD5 removal, 250
Biotechnological methods, 381 British Water Research Centre (WRC), 247
Biotechnology, 80, 152 Bubbling orifices, 301
core technique, 84 Bulking agent, 175
industrial uses, 83
materials
antibodies, 85 C
antisense oligonucleotides, 85 Calcined fermented kola nut husk
cytokines, 84 (CFKNH), 351
enzymes, 85 Calcined mixed based catalyst, 345
restriction enzymes, 85 Calcined palm kernel shell husk
viral vectors, 85 (CPKSH), 351
to medicine California Integrated Waste Management
diagnostics, 86 Board (CIWMB), 180
gene therapy, 86 Candida species, 376
vaccines, 86 CAPTOR and CAST full-scale study
to nonmedical markets, 87 experimental system aeration basins, 248
regulatory environment, 87–88 F/M loading, 248
Biotechnology industry Freehold plant, 248
agricultural applications, 83 initial results, 249, 250
fermentation process-based pharmaceutical modifications, 252, 253, 256
industry, 80 modified trains, 248
474 Index

CAPTOR and CAST full-scale study (cont.) CAST system, 255

performance, 255 CAST variation, 251
pilot-scale studies, 250–252 Catalytic dehydration reaction, 370
Simon-Hartley design, 248 Centralized anaerobic digestion (CAD), 324
USEPA recommendations, 255, 256 Century-scale climate changes, 232
CAPTOR in activated sludge treatment Chemical methods, 369
(CAST), 250 Chemical oxygen demand (COD), 10–12, 15,
CAPTOR pad cleaner, 249 21, 22, 27, 28, 50, 68, 399
CAPTOR pads, 251, 252, 255 Chemical synthesis, 89, 90, 95–96, 99, 104,
CAPTOR zones, 254 105, 124, 132, 153
Carbon adsorption, 123, 134 Chemical synthesis-based pharmaceutical
Carbon-biomass mixture, 242 industry, 80
Carbon dioxide, 217 Chemical synthesis-based pharmaceutical
Carbonylation, 370 wastewater treatment
Cardboard compost, 190 technologies, 147
Carrier-activated sludge processes (CAPTOR Chlorination, 115, 135
and CAST systems) Clean Air Act, 4, 5
activated sludge plant, 247 Class A pathogen standard, 189
advantages, 246 Clean Water Act (CWA), 5, 48, 64, 69
biomass approach, 247 Climate change, 226
biomass wastewater treatment, 246 Climate/climate control, 279
full-scale evaluation (see CAPTOR and Coagulation, 407
CAST full-scale study) COD removal efficiency, 322
pilot-plant study, 248 Cold filter plugging point (CFPP), 358, 359
process development, 247 Cold-pressed castor oil, 366
Castor oil Combinatorial chemistry, 85
advantages, 379 Combined anaerobic and aerobic treatment
applications, 363, 366, 369 processes, 423
biotechnological conversion method, 364 Composting, 35, 36, 39, 56, 63
characteristics, 364 Comprehensive Environmental Response,
composition, 366, 367 Compensation and Liability Act
consumption, 365 (CERCLA), 87
demand, flavors and fragrance, 368 Computational biotechnology, 81, 153
derivatives, 368 Computer program, 86
disadvantages, 379 Computer software
extraction methods, 366 for livestock management, 64
global market, 365 Concentrated animal feeding operations
industrial sector raw material, 363, 366 (CAFOs), 6, 65, 67, 68
microorganisms, 364 Conjugated linoleic acid (CLA), 349
nonedible crop, 380 Constructed wetlands, 282
properties, 366, 368, 380 advantage, 24
RA, 364 design, 27
safety assessment/toxicity study, 380 floating aquatic plant systems, 26
seed production, 365 FWS, 24, 26
Castor oil processing techniques nutrient removal, 284, 285
biotransformation method (see processes, 24
Biotransformation method) purpose, 23
chemical transformation, 370 types, 24
hydrolysis and purification, 369, 370 vegetated submerged systems, 26
Castor plant (Ricinus communis) Continuous flow system, 181
family Euphorbiaceae, 363 Continuous stirred tank reactor (CSTR)
geographical distribution, 364 system, 403, 444
major producers, 363 Conventional activated sludge reactor, 251
oil (see Castor oil) Conventional composting, 175
Index 475

Conventional cost-effective biological waste Drought, 227, 229

treatment technologies, 149 Drug development, 85–86
Conventional nitrification–denitrification Drug-related research, 85
systems, 405 Duckweeds, 283
Conventional thermophilic composting, 180 Dust, 112
Copolymer, 334
Copolymerization, 321
Cowpea mosaic virus (CPMV), 179 E
Crash boxes, 144 Earthworm (Eisenia foetida)
Cream yeast, 397 bactericidal enzymes, 179
Creams, 97 cadmium and zinc, 188
Crop residues, 190 effects, 176
Cross-flow filtration, 414 growth, 177
Crude bioplastics metabolic parameters, 177
agricultural applications, 335 sewage sludge decomposition, 176
bacterial biomass, 335 vermistabilization, 177
biodegradability, 334 virucidal enzyme system, 179
biodegradable materials and WES bioassay, 178
biopolymers, 335 worm cast, 177
industrial production, 334 Earthworm bioassay procedure, 178
insulation, 335 Earthworm marketing, 174
market, 334 Earthworm media, 190, 195
mechanical properties, 334 EC treatment, 196
non-biodegradability, 335 Effluent, 290, 306
PHA nanocomposite, 335 Effluent biomass solids, 251
thermal decomposition, 335 Effluent disinfection, 302
Cyclic staging, 283 Eisenia andrei, 189
Cytokines, 84 Eisenia eugeniae, 176
Electrochemical oxidation processes, 419
Electrocoagulation (EC), 419
D Electroflotation (EF), 419
Dark biotechnology, 153, 155 Emergent aquatic vegetation, 281
Dead animal composting, 35 End-of-pipe treatment, 107, 108, 113,
Decision-making process, 3 122–124, 135, 137
Decision support system (DSSs), 64 case histories and green environmental
Dehydration, 187, 370 technologies (see Green
Delta boundary, 223 environmental technologies)
δ-Decalactone (DDL), 371 in the fermentation pharmaceutical
Dendrobaena veneta, 177 industry
Denitrification, 11, 24, 41, 42, 58, 60, 69, Ansa, at Izmit, Turkey, 137
282, 292 raw materials and production
De novo synthesis/transformation process, 137–138
methods, 378 waste generation and
Dewatered sludge, 429 characteristics, 138–140
Dipole moment, 215 pollutants, 123
Direct liquefaction, 34 POTW, 123
Discharge of wastes, 92, 93, 98, 105, 140 pretreatment, 124
Discharge runoff control systems, 48, 49 primary treatment methods
Dispersed odors, 53 equalization and
Dissolved air flotation (DAF), 125, neutralization, 124–125
245, 333 in the pharmaceutical industry, 124
Dissolved organic carbon (DOC), 418 primary and secondary flotation
Dissolved organic material, 128 clarification, 125
Droplet formation, 256 screening and clarification, 125
476 Index

End-of-pipe treatment (cont.) F

residue treatment and waste Facultative lagoons, 32
disposal, 135–137 Fatty acid methyl ester (FAME), 342, 348, 356
secondary biological treatment Fecal indicator organisms, 62
activated sludge, 125–127 Fed-batch bioreactors, 393
aerated lagoons, 127, 128 Fed-batch strategy, 378, 381
anaerobic treatment, 130–133 Federal regulations, 4–6
trickling filters, 128–130 Feedlot runoff control system
secondary treatment facility, 124 clean water diversion, 48
tertiary treatment, 134 discharge runoff control, 48, 49
carbon adsorption, 134 full containment systems, 48
chlorination, 135 processes, 48
coagulation/flocculation/clarification runoff control, 48
system, 135 VFS system, 50–52
filtration, 134 Fermentation, 89–91, 93, 99, 105, 112, 124,
Enterococcus sp., 190 135, 137, 149, 154
Environmental biotechnology, 81, 153 Fermentation process-based pharmaceutical
Environmental Earthworm Projects, Inc., 182 wastewater treatment
Environmental regulations technologies, 148–149
for direct discharge, 101 Fermentation processes, 91–93
BAT, 103 Fermented kola nut husk (FKNH), 343–345,
BPT, 103 349, 354, 356, 359
NSPS, 103 Fermenting bacteria, 323
regulation for cyanide, 101 Filtration media, 290
USEPA regulation, 101 Filtration processes, 93
historical view, 105 First-generation biofuel feedstocks, 342
for indirect discharge Five-day biochemical oxygen demand (BOD5),
POTW, 104 10–12, 16, 18, 26, 29, 69, 278
PSES, 104 Flexible biopolymer nanocomponents, 334
PSNS, 105 Floating aquatic plant (FAP) systems, 24–26
for managing pharmaceutical Flotative separation, 333
wastes, 106–107 Food-grade castor oil, 379
Enzyme–enzyme methods, 395 Food-processing and agricultural wastes,
Enzyme-linked immunosorbent assay PHAs production
(ELISA), 179 acidogenic fermentation, 323, 324
Enzymes, 58, 85 activated sludge, 327
Escherichia coli, 190 AD plants, 324
Esterification, 370 biogas recycling, 324
Eudrilus eugeniae, 177 CAD, 324
Eutrophication, 15, 16, 69 fast-growing rhizobia, 328
EV20 treatment, 196 fatty acids and hydrogen, 323
Evaporator, 410 fermented organic compounds, 322
Evapotranspiration (ET) system, 286, 287 LCFA, 322
applications, 287 lipids, 322
beds, 286 mass ratio, 323
conditions, 286 organic and inorganic particles, 324
costs, 288, 289 PHB, 326, 327
description, 286 rumen microorganisms, 324
design criteria, 288 sugarcane/sugar beet molasses, 324
limitations, 287, 288 sulfate reduction, 323
liquid disposal, 286 VFA, 322, 326
on-site wastewater disposal, 310 water-saving process, 323
performance, 288 Food-processing wastes, 319
vegetation impacts, 286 Food wastes, 174, 190
Extracellular cellulolytic activity, 333 Forest wastes, 190
Index 477

Forestland irrigation, 296 Heavy metal bioavailability, 178

Free water surface (FWS) constructed Heavy metal uptake, 278
wetland, 24–26, 282 Herbicides, 283
Freehold’s activated sludge system, 248 Heterogeneous catalyst, 343
Fruit plant debris, 190 Heterogeneous semi-conductor
Fruit pomace, 190 photocatalysis, 416
Full infiltration, 290 High-quality protein, 342
Full-scale CAPTOR system flow pattern, 254 Homogeneous catalyst
Fungi, 333 (KOH/NaOH), 343
FWS constructed wetland treatment, 282 Hormone-like proteins, 84
Human health, 231
Hyacinth aquaculture system
G applications, 279
Garbage-eating red wiggler worms, 189 chemical phosphorus removal, 278
Gas absorption, 436 design criteria, 279, 280
Gases limitations, 279
in manure, 38 performnce, 280, 281
Gasification, 34 wastewater treatment, 278, 311
Gated surface pipe, 301 Hyacinth growth energy, 279
Generally recognized as safe (GRAS), 371 Hyacinth harvesting, 280
Gene sequencing, 86 Hydraulic residence times (HRTs), 252
Genes, γ-decalactone production, 372, 373 Hydraulic retention time (HRT),
Genetically modified organisms (GMOs), 80 323, 331
Genetic engineering, 381 Hydrodynamic cavitation (HC), 422
Global climate change, 206 Hydrogenation, 370
Global per capita carbon emission, 216 Hydrogen-oxidizing bacteria, 321
Global warming, 206, 209 Hydrolysis, 320, 369, 370
climate change adaptation, 220 3-Hydroxy-acyl-CoA dehydrogenase, 373
climatic and non-climatic origin, 221 3-Hydroxy-γ-decalactone, 378
sea level, 219 3-Hydroxy lactone, 373
surface temperatures, 219
Global warming potential (GWP), 210, 213
global warming, 214 I
IPCC, 213 Immobilized enzymatic reactions, 378
Kyoto Protocol, 213 Impermeable covers, 56, 57
Glossary of emerging natural systems, 310 Incinerating vent gas, 112
Glycogen-accumulating/slime-producing Increased temperatures on land
bacteria, 320 agriculture, 218
Gold biotechnology, 154 global average temperature, 218
Good operating practices, 111 weather, 217
Gray biotechnology, 81, 154 Induced blanket reactor (IBR), 22
Green biotechnology, 80, 155 Industrial biotechnology, 80, 155
Green environmental technologies Infiltrated livestock waste
case histories, 140–141 consequences, 3
by LIWT, 142–143 Infrared spectroscopy (FTIR), 350
Green waste (GW), 190 In-plant control, 107, 108, 111
Greenhouse effect, 209–211, 213 good operating practices, 111
Greenhouse gases (GHGs), 209, 233–235 material substitution, 108, 109
process modification, 109–110
recovering and recycling, 110
H reduction of air and dust
Harvested grass, 301, 310 problems, 111–113
Heat absorption segregation and concentration of
carbon dioxide, 214 wastes, 111
dipole moment, 215 waste exchange, 113
molecule, 215 water conservation and reuse, 110
478 Index

In-plant treatment, 108, 111, 113, 119, 124 Land application

cyanide destruction technologies nutrient content, 37
alkaline hydrolysis, 116 treatment processes, 38, 39
chemical oxidation, 114 waste management technique, 37
chlorination, 114–115 Land application, livestock wastes
ozonation, 115 handling manure
metal removal, 116 liquid manure, 40–42
alkaline precipitation, 116 semisolid manure, 40
chemical reduction, 117–119 solid manure, 38–40
sulfide precipitation, 117 rate of application, 42
in pharmaceutical industry, 113 time of application, 42
solvent recovery and removal, 119 Landfill gas (LFG), 230
advanced physicochemical treatment Leachate, 310
processes, 123 Leaf litter, 190
air stripping, 122 Lenox Institute of Water Technology (LIWT),
steam stripping, 120–122, 131 142–143, 151
Instant Dried Yeast, 397 Lime, 116
In-storage psychrophilic anaerobic digester Lipase-mediated biotransformation
(ISPAD), 23 extracellular lipases, 372
Integrated Swine Manure Management γ-decalactone, 372
(ISMM), 64 lactones, 371
IPCC global warming potential consensus, 214 Lip2 enzyme, 372
Iron-containing clay, 322 microbes, 371
“İzmit Industrial and Domestic Wastewater microorganisms, 371
Treatment Plant”, 423 nonconventional oleaginous yeast, 372
oxidations, 372
Lipase-producing microbes, 370
J Lipids, 322
Jatropha curcas, 342 Liquefaction, 34
Liquid manure, 40–42, 69
application, 57
K holding ponds, 46
Kirchhoff’s laws, 215 larger-sized drops, 40
Kola nut husk (KNH), 343 long-term storage, 44
Kyoto Protocol, 213 short-term basis, 44
storage, 45–46
subsurface application, 40
L surface application, 40
Lactones, 371 Liquid manure systems, 46
Lagoons, 28, 69 Liquid sludge vermistabilization (LSVS), 178
aerobic lagoons, 31 Liquor DO concentrations, 252
anaerobic lagoons, 30 Livestock manure, 179
covered anaerobic lagoons, 21 Livestock operations management, 59
effluent storage, 28 Livestock waste management, 2
facultative lagoons, 32 AWM, 64
maintenance, 29 computer software, 64
operators, 29 use of software, 64
pretreatment of wastewater, 29 Livestock wastes, 190
sizing, 29 storage (see Storage, livestock wastes)
sludge storage, 28 treatment processes, 30
treatment volume, 28 Livestock Water Recycling (LWR)
volumes, 28 system, 65–67
Land and ocean temperature, 218 Loading cycles, 292, 293
Index 479

Long-chain fatty acids (LCFA), 322 Modern green environmental

Low-cost crude bioplastic, 336 biotechnology, 81, 155
LSVS reactors, 178 Modified CAPTOR system, 254
Molecular weight cutoff (MWCO) value, 413
Mono- and disaccharides, 319
M Multistage hybrid green environmental
Market-driven strategies, 65 technologies, 151
Mechanical evaporators, 286 Municipal PACT facilities, 244
Medical biotechnology, 80, 155 Municipal sludge, 174, 186
Melanoidins, 399 Municipal sludges management,
Membrane distillation (MD), 413 earthworms, 176
Membrane filtration (MF), 245
Mesophilic, 18, 23, 69
Metal-contaminated soils/sediments, 179 N
Methane, 232 Nanocomposite bioplastic, 336
Methane gas, 231 Nanocomposites, 334
Methanogenesis, 323 National Ambient Air Quality Standards
Microbial biosynthesis, 380 (NAAQs), 4
Microbial enzymes, 370 National Pollution Discharge Elimination
Microwave-assisted RA extraction, 370 System (NPDES), 5, 6, 48, 69
Milankovitch cycles, 232 Natural antioxidants, 335
Military biotechnology, 81, 153, 155 Natural and constructed wetland systems, 281
Milk house wastewater Natural product screening, 85
ammonia, 15 Natural resources, 341
characteristics, 14, 15 Natural variability
constituents, 14 climate, 232
four-stage cleaning process, 15 temperatures, 232
phosphorus, 14 Natural ventilation, 59
sources, 13 Natural wetlands, 281
treatment methods, 16–18 Natural wetland systems, 282
water conservation, 18 Neem, 343
Mineralization, 11, 38 leaves, 343
Mixed liquor volatile suspended solids Neem oil, 345
(MLVSS), 126 Neem oil blend, 344
Mixed oil biodiesel (MOB), 346, 347, 355 Neutralization, 124, 127, 134
base-catalyzed transesterification of mixed New Source Performance Standards (NSPS),
oil, 354 101, 103, 105, 158–160
MeOH/OMR and reaction 1994 International Symposium, 179
temperature, 355 Nitrification, 11, 24, 38, 42, 52, 58, 60, 69,
methanol-to-oil ratio, 352 245, 292
optimum predicted yield, 354 Nitrification experiments, 252
physicochemical and fatty acid Nitrogen control, 283
composition, 358 Nitrogen elimination strategies, 292
physicochemical properties, 347 Nitrogen removal, 301
production via methanolysis, 355 Nitrogen sources, 376
properties, 356–359 Non-aseptic bioplastic production nutrients
transesterification, 353 agricultural wastes, 319
and variable conditions, 354 carbohydrates, 319
Mixed oils, 346 food-processing wastes, 319
Mixed powder, 345 inorganic nutrients, 322
Mixed solid-state bioreactors, 378 municipal wastes, 319
Mixture of oils, 343 nucleotides and orthophosphates, 322
Modern environmental biotechnology, 81 organic acids, 320
480 Index

Non-aseptic bioplastic production design criteria, 302, 303

nutrients (cont.) limitations, 302
organic dissolved substances and loading rates and cycles, 301
particles, 321 objective, 301
organic materials, 321 performance, 303, 304
organic wastes, 321 preapplication practices, 302
storage compounds, 319 slopes, 301
wild strains of bacteria, 319 sprinklers, 302
Nonbiodegradable petrochemical plastics, 317 wastewater treatment, 301, 310
Nonbiodegradable plastic, 335 Oxytetracycline, 137, 140
Non-PHA cellular mass, 332 Ozonation, 115
Nutrient content, 37 Ozone, 207
Nutrient sinks, 281

P
O PAC activated sludge systems, 245
Odor prevention PAC concentration, 244
animal nutrition management, 55 PAC physicochemical process, 246
livestock operations management, 59 PAC process systems, 244
manure treatment and handling, 56–57 PACT nitrification systems, 245
odor emission strategies for livestock PAC/WAR systems, 243
housing, 55 Palm kernel shell husk (PKSH),
waste treatment methods (see Waste 343–345, 349
treatment methods) Paper mill sludge, 183, 190
Odors, 30 Particulate matter (PM), 207
ammonia emissions, 54 Pathogen treatment methods, 63
anaerobic digestion, 53 Pathogens
causes, in livestock operations, 53 category, 62
dispersed odors, 53 in livestock industry, 62–63
dust particles, 53 survival, 62
emission strategies, 56 Penicillin, 93, 112, 135
hydrogen sulfate odor emissions, 54 Perennial grasses, 301
in pig manure, 54 Perforated plastic piping system, 286
sources, 54 Perionyx excavatus, 177
VFAs, 54 Permeable covers, 56, 57
Oil blends, 345 Petrochemical-based plastics, 317
Oil mixture, 346 pH neutralization, 284
Ointments, 97 PHA-accumulating bacteria, 320
Operation and maintenance (O & M), 23 PHA-containing biomass, 332
Optimal conditions, biotransformation PHA extraction suffer, 332
bioreactors, 377–378 PHA granules, 332, 333
carbon source, 376 PHA-producing bacteria, 321
nitrogen source, 376 PHA-producing microbial biomass, 321
pH, 377 PHA-producing mixed culture
temperature, 377 activated sludge, 329
Optimization, 283 batch biosynthesis, 330
Oregon Soil Corporation, 181 biological wastewater treatment, 331
Organic acids, 336 biomass growth, 330
Organic contaminants, 135 bioreactor, 331
Organic load distribution, 282 BOD, 330
Organic solvents, 92–94, 96, 111, 112, 123 dry bacterial biomass, 330
Overland flow system essential nutrients, 330
applications, 302 genera, 329
costs, 304, 305 HRT, 331
Index 481

intermittent feeding regime and Pharmaceutical wastewaters, 123, 149

variation, 329 Pharmaceutically active ingredients, 155
multistep cultivation process, 329 PHAs chemical pre-treatments, 332
non-aseptic bioreactors, 330 Pheretima hawayana, 177
osmotic shock, 331 PhoStrip process
PHA-accumulating organisms, 329 activated sludge, 264
technological approach, 331 applications, 265
vs. pure culture, 329 combined biological-chemical precipitation
PHA solubilization, 332 process, 264
PHA synthesis, 321 compatibility, 265
Pharmaceutical biotechnology, 156 construction cost, 266, 267
Pharmaceutical formulation, 96, 97 design criteria, 266
Pharmaceutical industry, 80, 88, 156 electrical energy requirement, 267, 268
pharmaceutical manufacturing, 89–90 municipal wastewater treatment, 265
products, categories, 88 operation and maintenance cost, 267, 268
SIC, 88 performance, 266
toxic pollutants, 88 phosphorus precipitation, 264
Pharmaceutical manufacturing, 89 process flow diagram, 264
category, 89 settled wastewater, 264
impacts on the environment, 100 Photosynthesis, 207
pharmaceutical ingredients, 89 Photosynthetic bacterial assimilation, 321
processes and waste generation, 90 Phragmites spp., 281
biological product extraction, 93–94 Phytoplankton, 207
chemical synthesis, 95–96 Pig fat (lard), 342
fermentation, 91–93 Pilot-scale CAPTOR BOD5 removals, 251
formulation, mixing and Pilot-scale CAST, 253
compounding, 96–97 Pilot-scale operating conditions, 252
plants, 90 Point sources of water pollution, 64, 69
products, 90 Polyhydroxyalkanoates (PHAs), 318
R & D processes, 97, 98 accumulation, 318
products, processes and activities, 89 bacterial species, 318
Pharmaceutical plants, 98, 99, 104, 109, 122, definition, 318
132, 140, 149, 150 nutrients (see Non-aseptic bioplastic
Pharmaceutical waste management production nutrients)
end-of-pipe treatment (see End-of-pipe production technologies,
treatment) disadvantages, 318
in-plant control (see In-plant control) properties, 318
in-plant treatment (see In-plant treatment) raw materials, 318
management program, 106 sources, 319
strategy, 107–108 Polylactic acid (PLA), 318
Pharmaceutical waste minimization Polymers, 318
case study Pork meat, 342
of Falls Memorial Hospital Powdered activated carbon (PAC), 134
(FMH), 145–146 Powdered activated carbon treatment
of Hennepin County Medical Center (PACT), 242
(HCMC), 143–144 activated sludge process, 243
Pharmaceutical wastewater treatment advantages, 243
technologies biological reactor, 243
BAT, 146 biological solids, 242
chemical synthesis-based, 147 definition, 242
conventional biological treatment design procedure, 244
methods, 146 economic analysis, 243
environmental technologies, 147 nitrification, 243
fermentation process-based, 148–149 process equipment, 245
482 Index

Powdered activated carbon treatment Regulatory environment, 87–88

(PACT) (cont.) Research and development (R & D), 97–98
process limitations, 246 Residuals, 285
types, 242 Resource Conservation and Recovery Act
underflow solids, 242 (RCRA), 87, 88, 106, 107
Practical management practices, 3 Respiration studies, 251
Pretreatment Standards for Existing Sources Restriction enzymes, 85
(PSES), 104, 105, 160–161 Reverse osmosis (RO), 415
Pretreatment Standards for New Sources Rhizobia, 328
(PSNS), 104, 105, 161–162 Rice straw, 190
Process to Further Reduce Pathogens Ricinoleic acid (RA), 364
(PFRP), 180 Ricinus communis agglutinin (RCA120), 380
Pseudomonas spp., 319 Ricinus communis ricin (RCA60), 380
Psychrophilic, 18, 23, 69 Rod-shaped bacterial cell, 332
Publicly owned treatment works (POTW), 99, RotoClone, 113
104, 111, 122, 123
Pyrolysis, 33
applications, 33 S
fast pyrolysis, 33 Saccharomyces cerevisia, 393
slow/moderate pyrolysis, 33 Safety data sheet (SDS), 342
Salinity intrusion
aquifers, 222
R Delta, 222
Radiative forcing (RF), 212, 213 freshwater diversions, 223
RA-enriched castor oil, 364 historical record, 223
RA–GDL bioconversion IPCC, 222
acetyl-CoA molecules, 373 plant pollen, 222
β-oxidation pathway, 372 saltwater intrusion, 222
biotechnological process, 375 Salmonella sp., 190
hydroxy fatty acids, 373 Salt accumulation, 229
lactones, 373, 374 Sawdust, 190
lactonization, 372 Schoenoplectus spp., 281
NAD, 373 Scirpus spp., 281
peroxisomal beta-oxidation pathway, 373 Second-stage nitrification process, 250
precursor fatty acid, 375 Sedimentation, 282
reaction sequences, 372 Self-mixed anaerobic digester (SMAD), 23
Rapid rate land treatment system Self-tuning control techniques, 393
applications, 291 Semisolid manure, 40, 69
basins/beds, 290 equipment for handling manure, 40
costs, 293, 294 storage, 45
description, 290 Sequencing batch reactors (SBR), 245
design criteria, 292 Sewage sludge amendment, 438
land-based technologies, 289 Sewage sludges, 179
limitations, 291 Short fatty acids, 18, 68
nitrogen control, 289, 291 Site-specific vegetation, 282
nitrogen removals, 290 Skin coffee (SC), 190
performance, 292, 293 Slopes, 301
preapplication treatment practices, 290 Slow rate land treatment system
wastewater treatment, 289 applications, 296, 297
Rational drug design, 85 costs, 298–300
Recombinant DNA technologies, 82 design criteria, 298
Red biotechnology, 156 limitations, 297
Refractory volatile materials, 243 municipal land treatment practice, 294, 311
Index 483

performance, 298 Subsurface flow system (SFS), 282

preapplication treatment practices, 296 Subsurface infiltration systems
sprinklers, 295 absorption systems, 307
underdrainage system, 294 anaerobic digestion, 306
vegetation, 295 applications, 308
Sludge, 176 conventional subsurface disposal
Sludge characteristics, 178 system, 308
Sludge conversion, 176 design criteria, 309
Sludge management disposal field, 307
anaerobic, 426 distribution pipes, 306
compost quality, 432, 433 limitations, 308
composting operation, 428–431 nitrified effluent and denitrification, 306
definition, 426 performance, 309, 310
dewatered, 427 pressure distribution network, 307–308
dust formation, control, 434 soil absorption field, 306
land application, 437, 438 soil absorption system requirements, 307
nuisance conditions, 434 subsurface trenches, 307
odor control, 435, 436 wastewater management practices, 306
Sludge management systems, 177 Sulfate bioreduction, 323
Sludge storage, 28 Sulfate-reducing bacteria, 54
Soil carbon, 208 Surface constructed wetlands, 24
Soil clogging, 310 Surface flow wetlands, 24
Solar energy, 208 Surface methods of distribution, 301
Solar radiation, 208
Solid manure, 38, 40, 44, 69
application, 40 T
equipment for handling manure, 38–40 Temperature, 283
storage, 44–45 Temperature increases, 219
Solid municipal wastes, 336 Temperature-phased anaerobic digestion
Solid separation, 56, 57 (TPAD), 23
Solid-state fermentation (SSF), 378, 381 Temperature trend, environment
Solvent extraction, 92, 93, 153, 154 carbon dioxide, 216
Specific energy densities (SED), 436 greenhouse gases, 215
Sprinklers, 295 land temperatures, 215
Standard Industrial Classification (SIC), 88 Terrestrial radiation, 208
State environmental agencies, 6–9 Tetracycline, 137, 140, 141
State regulations, 6 Thermal decomposition, 335
Steam stripping, 120–122, 131 Thermochemical processes, 33
Step-wise fed-batch method, 378 Thermophilic, 18, 23, 39, 69
Stirred tank reactor (STR), 378 Thermophilic composting pretreatment, 181
Storage, livestock wastes Three-chamber domestic wooden
cost for manure storage facilities, 44 vermicomposter, 189
facilities Tobacco mosaic virus (TMV), 179
liquid manure, 45–46 Total dissolved solids (TDS), 228, 399, 415
semisolid manure, 45 Total dynamic head (TDH), 309
solid manure storage, 44–45 Total Kjeldahl nitrogen (TKN), 399
long-term storage, 44 Total suspended solids (TSS), 278
short-term storage, 43 Toxicants, 187
storage area design, 47 Toxic/hazardous pharmaceutical
Stratosphere, 208 pollutants, 149
Stratospheric ozone, 207 Transmembrane pressure (TMP), 413
Submerged fermentation (SmF), 378 Trickling filters, 128–130
Submerged-flow constructed wetlands, 282 Trimethylglycine, 399
484 Index

Troposphere, 208 fundamental factors, 177

2020 cost index table, 312–313 LSVS process, 178
Two-stage system, 321 operational parameters, 176
operation and troubleshooting, 187, 188
pioneers, 183–184
U posttreatment, 181
Underdrainage, 290 process applications, 189–197
Upflow anaerobic hybrid reactors progess, outside the USA, 182
(UAHRs), 446 requirement, 175
Upflow anaerobic sludge blanket (UASB) resources, 184
digester, 22 schematic representation, 175
reactors, 446 sludge management technologies, 178
Upflow sludge blanket (UASB) reactors, 402 technical and economic feasibility, 176
US Environmental Protection Agency worm’s castings, 174
(USEPA), 4–6, 19, 60, 65, 226, Vermiconversion System, 180
244, 255 Vertical loop reactor (VLR)
US EPA’s PFRP requirements, 180 advantages, 260
US policy changes, 64–65 aeration, 259
applications, 260
costs, 264
V description, 259
Vegetable plant debris, 190 design criteria, 260
Vegetated infiltration basin (VIB), 52 dissolved oxygen profile, 259
Vegetated submerged bed (VSB), 282 energy requirements, 262, 263
Vegetated submerged systems, 24, 26 performance, 260, 261
Vegetation, 281, 295 preliminary treatment, 259
Vegetative cover, 288 required capacity, 259
Vegetative filter strips (VFS) system, 50–52 schematic representation, 259
Vegetative treatment area (VTA), 52 USEPA evaluation, 261, 262
Ventilation, 59 Vinasses, 441–443
Veppampoo charu, 343 Viral vectors, 85
Vermicompost Virucidal activity, 179
biochar, 190 Volatile fatty acids (VFAs), 54, 320, 321
biological control tool, 197 Volatile organic compounds (VOCs), 435
characteristics, 190, 196 Volatile solvents, 93, 94, 96, 97
coffee plant growth, 190 Volatilization, 38, 41, 54, 69
composting time, 196 Volumetric loading rates, 251
effects, 196
fertilizer, 196
properties, 196 W
Vermicomposting, 36, 37 Waste characterization
adoption, 186 fermentation processes and chemical
advantages, 179 synthesis processes, 99
agricultural wastes, 190–195 inorganic priority pollutants, 99
anaerobic digestion, 175 options for waste disposal, 99–100
biological system, 174 pharmaceutical wastes, 98
challenges, 179–182 toxic pollutants, 99
vs. conventional composting, 175 wastewaters with COD concentration, 99
definition, 174 Waste exchange, 113
design criteria, 188, 189 Waste generation, 88, 90, 107, 108,
disadvantages, 188 111, 137–139
earthworm species, 177 See also Pharmaceutical manufacturing
environmental engineering, 197 Waste management, 83, 106, 107
Index 485

in pharmaceutical industry, 107 treatment process, 399, 402

pharmaceutical (see Pharmaceutical waste ultrasound process, 421, 422
management) WAO, 420, 421
Waste minimization, 90, 106, 107, 109, Wastewater treatment plant (WWTP), 81, 154,
111, 113 155, 403
See also Pharmaceutical waste Wastewater treatment system, 142
minimization Water conservation, 18
Waste reduction project, 143 Water hyacinth (Eichhornia crassipes),
Waste saltwater, 226 278, 311
Waste treatment Water quality
anaerobic digestion (see Anaerobic absorb and process, 225
digestion) coastal state, 223
composting, 35, 36 desalting units, 226
constructed wetlands (see Constructed dissolved solids, 228
wetlands) drinking water, 224
lagoons (see Lagoons) fisheries, 227
vermicomposting, 36, 37 irrigated areas, 227
Waste treatment methods irrigation drainage, 228
anaerobic digestion, 58 physicochemical, 225
biofilters, 58 salinity intrusion, 223
enzymes, 58 TDS, 225
management of lagoons, 59 water supply, 225
mechanical filtration devices, 58 Weather, 209
Wastewater Weed residuals, 182
BOD5, 11 Weeds, 190
characteristics of swine waste, 12 Wet-air oxidation (WAO), 244, 420
COD, 11 Wet air oxidation/regeneration
manure characteristics per animal, 13 (WAR), 243
physical and chemical properties, 6, 10 Wetland aquaculture system
poultry lagoons, 12 applications, 283
swine lagoon analysis, 11 constructed wetlands, 282–283
swine waste characteristic, 14 design criteria, 284
Wastewater constituents, adverse effects, 296 limitations, 283, 284
Wastewater management performance, 284, 285
adsorption, 409 wastewater treatment, 281, 311
advanced treatment processes, 411 White biotechnology, 156
biological treatment process, 402, 403, Worm castings, 181
405, 406 The Worm Concern, 181
characterization, 399 Worm drowning, 187
coagulation/flocculation, 407, 408 Worm loss, 187
definition, 398
electrochemical oxidation processes,
419, 420 Y
electrolysis, 415 Yarrowia lipolytica, 372
evaporation systems, 410, 411 Yellow biotechnology, 156
membrane processes, 412–415
oxidation process, 415–418
physicochemical treatment process, 406 Z
plants, 423, 425 Zero-discharge system, 449
sludge, 426