NPTEL – Biotechnology – Microbiology

Module – 4: Microbial Nutrition

Lecture 1 – Microbial nutrient requirements and nutritional types of
microorganisms
In this lecture we shall be looking into the aspects of microbial nutrition related to
the nutrient requirements of microorganisms and the nutritional types.
To obtain energy and construct new cellular components, organisms, must have a
supply of raw materials or nutrients. Nutrients – are substances used in biosynthesis and
energy production.
Nutrient Requirements:
Microbial cell composition shows that 95% of cell dry weight is made up of a few
major elements: Carbon, oxygen, hydrogen, nitrogen, sulfur, phosphorous, potassium,
calcium, magnesium and iron.
Macronutrients or macro elements:
These are required by microorganisms in relatively large amounts. Carbon,
oxygen, hydrogen nitrogen, sulfurs and phosphorous are components of carbohydrates,
lipids, proteins and nucleic acids. The remaining four macro elements (K, Ca, Mg and Fe)
exist in the cell as cations.
K+ - is required for the activity by a number of enzymes, including those involved in
protein synthesis.
Ca2+ - contributes to the heat resistance of bacterial endospores. 15% of spore contains
dipicolinic acid and calcium.
Mg2+ - serves as a cofactor for many enzymes, complexes with ATP and stabilizes
ribosomes and cell membranes.
Fe2+ and Fe3+ - part of cytochromes and a cofactor for enzymes and electron-carrying
proteins.
Micronutrients or Trace elements:
These are manganese, zinc, cobalt, molybdenum, nickel and copper. These are
normally part of enzymes and cofactors, and they aid in the catalysis of reactions and
maintenance of protein structure.
Zn2+ - is present at the active site of some enzymes but is also involved in the association
of regulatory and catalytic subunits in E.coli aspartate carbomoyl transferase.
Mn2+ - aids many enzymes catalyzing the transfer of phosphate groups.

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NPTEL – Biotechnology – Microbiology
Mo2+ - required for nitrogen fixation.
Co2+ - is a component of Vitamin B12.
Besides macro and micro nutrients, some microorganisms may have particular
requirements that reflect the special nature of their morphology or environment. Diatoms
need silicic acid to construct their beautiful cell walls of silica. Bacteria growing in saline
lakes and oceans depend on the presence of high concentrations of sodium ion.
Microorganisms require a balanced mixture of all the above nutrients for proper growth.
Requirements for carbon, hydrogen and oxygen:
Carbon is needed for the skeleton or backbone of all organic molecules and
molecules serving as carbon sources normally also contribute both oxygen and hydrogen
atoms. One important carbon source that does not supply hydrogen or energy is CO2.
Autotrophs – can use CO2 as their sole or principal source of carbon. Many
microorganisms are autotrophic, and most of these carry out photosynthesis and use light
as their energy source. Some autotrophs oxidize inorganic molecules and derive energy
from electron transfer. Heterotrophs – are organisms that use reduced pre-formed
organic molecules as carbon sources. Ex. Glycolytic pathway produces carbon skeleton
for use in biosynthesis and also releases energy as ATP and NADH. Actinomycetes will
degrade amyl alcohol, paraffin and even rubber. Burkholderia cepacia can use over 100
different carbon compounds. Some microorganisms can metabolize even relatively
indigestible human-made substances such as pesticides. Indigestible molecules can be
oxidized and degraded in the presence of a growth promoting nutrient that is metabolized
at the same time, a process called Co-metabolism. The products of this breakdown can
then be used as nutrients by other microorganisms.
Nutritional types of microorganisms:
In addition to Carbon, hydrogen and oxygen all organisms require sources of
energy and electrons for growth.
Carbon sources:
Autotrophs - CO2sole or principal biosynthetic carbon source
Heterotrophs – reduced, preformed organic molecules from other organisms.
Energy sources:
Phototrophs – use light as their energy source.

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NPTEL – Biotechnology – Microbiology
Chemotrophs – obtain energy from the oxidation of chemical compounds (either organic
or in organic)
Electron sources:
Lithotrophs – use reduced inorganic substances as their electron source.
Organotrophs – extract electrons from organic compounds.
Four major nutritional classes based on their primary sources of carbon, energy and
electrons is known.
Phtotolithotrophic autotrophs or photoautotrophs or photolithoautotrophs:
Source of energy – light energy
Source of electrons – Inorganic hydrogen/ electron
Carbon source - CO2
Example: Algae, purple and green sulfur bacteria and cyanobacteria.
Photoorganotrophic heterotrophy or photoorganoheterotrophy:
Source of energy – light energy
Source of electrons – organic hydrogen/ electron
Carbon source –organic carbon sources (CO2 may also be used)
Example: Purple and green nonsulfur bacteria (common inhabitants of lakes and streams)
Chemolithotrophic autotrophs or chemolithoautotrophy:
Source of energy – Chemical energy source (inorganic)
Source of electrons – Inorganic hydrogen/ electron donor
Carbon source - CO2
Example: Sulfur-oxidizing bacteria, hydrogen bacteria, nitrifying bacteria, iron-oxidizing
bacteria.
Chemoorganotrophic heterotrophs or chemoorganoheterotrophy:
Source of energy – Chemical energy source (organic)
Source of electrons – Inorganic hydrogen/ electron donor
Carbon source – organic carbon source
Example: Protozoan, fungi, most non-photosynthetic bacteria (including most pathogens)
The most common nutritional types are photolithoautotrophs and
chemoorganoheterotrophs. Bacteria Beggiatoa rely on inorganic energy sources and
organic (or sometimes CO2) carbon sources. These microbes are sometimes called

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Mixotrophic because they combine chemolithoautotrophic and heterotrophic metabolic
processes.
Requirements for nitrogen, phosphorous and sulfur:
Nitrogen is needed for the synthesis of amino acids, purines, pyramidines, some
carbohydrates and lipids, enzyme cofactors and other substances. Most phototrophs and
many nonphotosynthetic microorganisms reduce nitrate to ammonia and incorporate the
ammonia in assimilatory nitrate reduction. A variety of bacteria like many Cyanobacteria
and Rhizobiium can reduce and assimilate atmospheric nitrogen using the nitrogenase
systems. Phosphorous is present in nucleic acids, phospholipids, ATP, several cofactors,
some proteins and other cell components. All microorganisms use inorganic phosphate as
their phosphorous source and incorporate it directly. E.coli can use both organic and
inorganic phosphate. Organophosphates such as hexose 6- phosphate can be taken up
directly by transport proteins. Other organophosphates are often hydrolyzed in the
periplasm by the enzyme alkaline phosphatase to produce inorganic phosphate which is
then transported across the plasma membrane. When inorganic phosphate is outside the
bacterium, it crosses the outer membrane by the use of a porin protein channel. Sulfur is
needed for the synthesis of substances like the amino acids cysteine and methionine,
some carbohydrates biotin and thiamine. Most of them use sulfate as a source of sulfur
and reduce it by assimilatory sulfate reduction; a few require a reduced form of sulfur
such as cysteine.
Growth factors:
Many microorganisms have the enzymes and pathways necessary to synthesize all
cell components. Many lack one or more enzymes and hence require organic compounds
because they are essential cell components or precursors of such components and cannot
be synthesized by the organisms are called – growth factors. There are three major
classes of growth factors:
Amino acids – needed for protein synthesis.
Purines and Pyramidines – for nucleic acid synthesis
Vitamins – small organic molecules that usually make up all or part of enzyme cofactors,
and only very small amounts sustain growth.

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NPTEL – Biotechnology – Microbiology
Knowledge of the specific growth factor requirements of many microorganisms makes
possible quantitative growth response assays for a variety of substances. The observation
that many microorganisms can synthesize large quantities of vitamins has led to their use
in industry. Several water-soluble and fat-soluble vitamins are produced using industrial
fermentations.
Ribofalvin – Clostridium, Candida, Ashbya, Eremothecium
Coenzyme A – Brevibacterium
Vitamin B12 – Streptomyces, Propionibacterium, Pseudomonas
Vitamin C – Gluconobacter, Erwinia, Corynebacterium
β- Carotene – Dunaliella
Vitamin D - Saccharomyces
REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An

Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

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NPTEL – Biotechnology – Microbiology

Module – 4: Microbial Nutrition

Lecture 2 – Uptake of nutrients by the cell
In this lecture we shall be dealing with the uptake of nutrients by microorganisms
and the different ways by which movement of materials takes place across the plasma
membrane.
Uptake of the required nutrients by the microbial cell is important. Since
microorganisms live in nutrient poor habitats, they must be able to transport nutrients
from dilute solutions into the cell against concentration gradient. Finally, they must pass
through a selectively permeable plasma membrane. Microorganisms use different
transport mechanisms like facilitated diffusion, active transport and group translocation.
Eukaryotic microorganisms do not employ group translocation but take up nutrients by
endocytosis.
Movement of materials across the plasmamembrane is mostly done by two processes:
Passive processes: Substances cross the area from an area of high concentration to an
area of low concentration without any expenditure of energy (ATP). Example, simple
diffusion, osmosis and facilitated diffusion.
Active process: The cell must use energy (ATP) to move substances from areas of low
concentration to areas of high concentration. Example, Group translocation.
Passive processes:
Passive or simple diffusion:Often called diffusion, is the process in which molecules
move from a region of higher concentration to one of lower concentration. The rate is
dependent on the size of the concentration gradient between a cell’s exterior and its
interior. Very small molecules such as water and oxygen and carbon dioxide move across
membranes by simple or passive diffusion. Larger molecules, ions, and polar substances
do not cross membranes by this method.
Osmosis: Is the net movement of solvent molecules across a selectively permeable
membrane from an area in which the solvent molecules are highly concentrated to an area
of low concentration until equilibrium is reached. In living systems the chief solvent is
water. The three types of solutions which are normally found are isotonic, hypotonic and
hypertonic.

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NPTEL – Biotechnology – Microbiology
Facilitated diffusion:The rate of diffusion across selectively permeable membrane is
greatly increased by using carrier proteins, sometimes called permeases which are
embedded in the plasma membrane. Because a carrier aids the diffusion process, it is
called as facilitated diffusion.Carrier proteins also resemble enzymes in their specificity
for the substances to be transported; each carrier is selective and will transport only
closely related solutes. Because there is no energy input, molecules will continue to enter
only as long as their concentration is greater on the outside. Two widespread major
intrinsic protein channels in bacteria are aquaporins that transport water and glycerol
facilitators which aid glycerol diffusion. The carrier protein complex spans the membrane
(Figure 1). After the solute molecule binds to the outside, the carrier may change
conformation and release the molecule on the cell interior. The carrier would
subsequently change back to its original shape and be ready to pick up another molecule.
The mechanism is driven by concentration gradients and therefore is reversible.
Examples. Glycerol is transported by facilitated diffusion in E.coli, Salmonella
typhimurium, Pseudomonas, Bacillus and many other bacteria. This is prominent in
eukaryotes where it is used to transport a variety of sugars and amino acids.

Fig. 1. Facilitated diffusion. The carrier proteins aid in the release of solute molecules from extracellular space to the
intracellular space.
Active transport:
Active transport is the transport of solute molecules to higher concentrations or
against a concentration gradient, with the use of metabolic energy input. It resembles
facilitated diffusion in the involvement of protein carrier activity, but differs in its use of
metabolic energy and in its ability to concentrate substances. One example of active
transport is binding protein transport systems or ATP-binding cassette transporters (ABC
transporters) are active in bacteria, archaea and eukaryotes.

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NPTEL – Biotechnology – Microbiology
ABC transporters: These transporters consist of two hydrophobic membrane spanning
domains associated on their cytoplasmic surfaces with two nucleotide-binding domains.
The membrane spanning domains from a pore in the membrane and the nucleotide
binding domains bind and hydrolyze ATP to drive uptake. ABC transporters employ
special substrate binding proteins, which are located in the periplasmic space of gram-
negative bacteria or are attached to membrane lipids on the external face of the gram
positive plasma membrane. These binding proteins, bind the molecule to be transported
and then interact with the membrane transport proteins to move the solute molecule
inside the cell (Fig 2). E. coli transports a variety of sugars (arabinose, maltose, galactose,
and ribose) and amino acids (glutamate, histidine, leucine) by this mechanism.

Fig. 2. ABC transporter system.

Eukaryotic ABC transporters are sometimes of great medical importance. Some tumor
cells pump drugs out using these transporters. Cystic fibrosis results from a mutation that
inactivates an ABC transporter that acts as a chloride ion channel in lungs.

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NPTEL – Biotechnology – Microbiology
Bacteria also use Proton gradients generated during electron transport to drive
active transport. The lactose permease of E.coli transports a lactose molecule inward as a
proton simultaneously enters the cell. Such linked transport of two substances in the same
direction is called Symport (Fig. 3). E.coli also uses proton symport to take up amino
acids and organic acids like succinate and malate.

Fig. 3. Comparison of the three different types of transport mechanisms.

A proton gradient also can power active transport indirectly, often through the formation
of a sodium ion gradient. In E. coli, sodium transport system pumps sodium outward in
response to the inward movement of protons. Such linked transport in which the
transported substances move in opposite directions is called Anitport. The sodium
gradient generated by this proton anitport system then drives the uptake of sugars and
amino acids. E.coli has at least transport systems for the sugar galactose.

Fig. 4. Antiport mechanism - linked transport of two substances in opposite direction.

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NPTEL – Biotechnology – Microbiology
Group Translocation: In active transport, solute molecules move across a membrane
without modification. Many prokaryotes also take up molecules by group translocation, a
process in which a molecule is transported into the cell while being chemically altered.
For example, Phosphoenolpyruvate: Sugar phosphotransferease system (PTS). It
transports a variety of sugars while phosphorylating them using phosphoenolpyruvate
(PEP) as the phosphate donor.
PEP + Sugar (outside) → Pyruvate + Sugar-P (inside)
In E. coli and Salmonella typhimurium, it consists of two enzymes and a low molecular
weight heat stable protein (HPr). HPr and enzyme I (EI) are cytoplasmic. Enzyme II (EII)
is more variable in structure and often composed of three subunits or domains. EIIA is
cytoplasmic and soluble. EIIB also is hydrophilic but frequently attached to EIIC, a
hydrophobic protein that is embedded in the membrane. A high energy phosphate is
transferred from PEP to enzyme II (EII) with the aid of enzyme I (EI) and HPr. Then a
sugar molecule is phosphorylated as it is carried across the membrane by enzyme II (EII).
Enzyme II (EII) transports only specific sugars and varies with PTS, whereas enzyme I
(EI) and HPr are common to all PTS’s (Fig. 5).

Fig. 5. Group translocation: Bacterial PTS transport.

PTS’s are widely distributed in prokaryotes. Aerobic bacteria lack PTS’s. Genera
Escherichia, Salmonella, Staphylococcus and other facultative anaerobic bacteria have
phosphotransferase systems; some obligate anaerobic bacteria (Clostridium) also have
PTS’s. Many carbohydrates are transported by these systems. E. coli takes up glucose,
fructose, mannitol, sucrose, N-acetylglucosamine, cellobiose and other carbohydrates by
group translocation.

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NPTEL – Biotechnology – Microbiology

Iron Uptake:
All microorganisms require iron for use in cytochromes and many enzymes. Iron
uptake is made difficult by the extreme insolubility of ferric ion (Fe3+) and its derivatives,
which leave little free, iron available for transport. Many bacteria and fungi have
overcome this difficulty by secreting siderophores. Siderophores – are low molecular
weight molecules that are able to complex with ferric ion and supply it to the cell. These
are either hydroxamates or phenolates-catecholates. Ferrichrome is a hydroxamate
produced by many fungi; enterobactin is the catecholate formed by E.coli.
Microorganisms secrete sidereophores when little iron is available in the medium. Once
the iron-siderophore complex has reached the cell surface, it binds to a siderophore
receptor protein. The iron is either released to enter the cell directly or the whole iron-
siderophore complex is transported inside by an ABC transporter. In E.coli, the
siderophore receptor is in the outer membrane of the cell envelope; when the iron reaches
the periplasmic space, it moves through the plasma membrane with the aid of the
transporter. After the iron has entered the cell, it is reduced to the ferrous form (Fe2+).
Iron is so crucial to microorganisms that many use more than one route of iron uptake to
ensure an adequate supply.
REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An

Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

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NPTEL – Biotechnology – Microbiology

Module – 4: Microbial Nutrition

Lecture 3 – Culture media, Isolation and cultivation of pure cultures
In this lecture we shall be looking into the aspects of culturing microorganisms,
the different types of culture media, isolation of pure cultures.
In the laboratory, bacteria are normally grown or cultured in either liquid medium,
in flasks, bottles or large culture vessels called fermenters or solid medium in Petri dishes
which are round, normally plastic or glass dishes. Introduction of microbes into or onto
these media is called inoculation. The nature of the medium depends on the
microorganism’s natural environment because its nutrient requirements reflect its natural
surroundings. The culture medium must contain all the nutrients that the microorganism
requires for growth like water, a source of energy, carbon, nitrogen, essential inorganic
ions and a number of trace elements. Bacteria that can synthesize all they require from
the basic ingredients are called Prototrophs – and most microorganisms that survive in
the outside environment can do this. Microbes that have become adapted to life in a
situation rich with nutrients such as human body may require other growth factors like
vitamins, amino acids or nitrogenous bases to be provided. These are called auxotrophs.
The media that are used in microbiology laboratories to culture bacteria are
referred to as artificial media or synthetic media, because they do not occur naturally;
rather they are prepared in the laboratory. There are number of ways of categorizing the
media, one way is to classify based on whether the exact contents of the media are known
or not.
Chemically defined media: Is one in which all the ingredients are known; and was
prepared in the laboratory by adding a certain number of grams of each of the
components (Carbohydrates, amino acids, salts etc). Particularly photolithoautotrophs
such as Cyanobacteria and eukaryotic algae can be grown on relatively simple media
containing CO2 as a carbon source (often added as sodium carbonate or bicarbonate),
nitrate or ammonia as a nitrogen source, sulfate, phosphate and a variety of minerals.
Chemoorganoheterotrophs can be grown in a defined media with glucose as a carbon
source and an ammonium salt as a nitrogen source.
Complex media: contains undefined ingredients and the exact contents are not known.
Complex medium may be sufficiently rich and complete to meet the nutritional
requirements of many different microorganisms. They contain undefined components like
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peptones, meat extract and yeast extract. Peptones – are proteins hydrolysates prepared
by partial proteolytic digestion of meat, casein, soyameal, gelatin and other protein
sources (Carbon, energy and nitrogen). Beef extract – aqueous extracts from lean beef
and contain amino acids, peptides, nucleotides, organic acids and minerals and
vitamins.Yeast extract – from brewer’s yeast and contain an excellent source of B
vitamins, nitrogen and carbon compounds. Three commonly used complex media are
1. Nutrient broth – Peptone (5.0g/L), Beef Extract (3.0g/L)
2. Tryptic soya broth – Tryptone (enzyme digest of casein), Peptone (enzyme digest of
soybean meal), glucose, NaCl and K2HPO4
3. MacConkey Agar – Pancreatic digest of gelatin, casein, peptic digest of animal tissue,
bile salts mixture, NaCl, neutral red, crystal red and agar.
These media are routinely used for cultivation of bacteria in the laboratory and
particularly useful for cultivation of bacteria whose growth requirements have not been
defined.
Culture media can also be categorized as liquid or solid. Liquid media (also called broths)
are contained in tubes, flasks and fermenters. Solid media are prepared by adding agar to
liquid medium and then pouring the media into tubes or Petridishes where the media will
solidify. Agar is a complex polysaccharide that is obtained from red marine algae. Other
solidifying agents are gelatin, silica. A 1% or 2% agar can be used to solidify, but 1.5% is
the most commonly used.
Enriched Medium: Is a broth or solid medium containing a rich supply of special
nutrients that promotes the growth of fastidious organisms (that have complex nutrition
and environmental requirements). It is usually prepared by adding extra nutrients to a
medium called nutrient agar. Blood agar (nutrient agar + 5% sheep red blood cells): It is
bright red in color and distinguishes between the hemolytic and non-hemolytic bacteria
(Streptococci and other pathogens)
Chocolate agar (nutrient agar + powdered hemoglobin): It is brown in color and is
considered more enriched than blood agar as hemoglobin is more readily accessible in
chocolate agar. Pathogens like Neisseria gonorrhoeae and Haemophillus influenza,
which will not grow on blood agar, can be cultured.

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NPTEL – Biotechnology – Microbiology
Types of Media:
Selective Media: Are designed to suppress the growth of unwanted bacteria and
encourage the growth of the desired microbes.
Examples: MacConkey agar inhibits growth of gram positive bacteria and thus is
selective for gram-negative bacteria. Phenylethyl alcohol (PEA) agar and Colistinalidixic
acid (CAN) agar are selective for gram positive bacteria.
Manitol salt agar contains a concentration of 7.5% NaCl that is quite inhibitory to most
human pathogens. Bile salts, a component of fecus, inhibit most gram positive bacteria
while permitting many gram negative rods to grow. This media is used for selecting
intestinal pathogens which contain bile salts. Dyes such as methylene blue and crystal
violet also inhibit certain gram positive bacteria (Staphylococcus can produce acid from
mannitol and turn the phenol red dye to bright yellow.
A medium containing acetate as a carbon source would be selective for organisms that
grow on acetate. Sabaraud’s dextrose agar is used to isolate fungi and cellulose for
cellulose digesting bacteria.
Differential media: Makes it easier to distinguish colonies of the desired organism from
other colonies growing on the same plate.
MacConkey agar is also a differential and selective medium to distinguish between
lactose fermenting organisms (e.g, E.coli) and other non-fermenters (e.g, Shigella sp.).
Lactose in the medium is fermented by E.coli producing acid which causes an indicator
dye to change color to red and colonies that do not ferment lactose are white. Dyes can be
used as differential agents because many of them are pH indicators that change color in
response to the production of an acid or base. MacConkey agar contains neutral red, a
dye that is yellow when neutral and pink or red when acidic.
Mannitol salt agar is used to screen for Staphylococcus aureus, and it turns the originally
pink medium to yellow due to its ability to ferment mannitol. In a sense, blood agar is
also differential because it is used to determine the hemolytic and non-hemolytic bacteria.
Blood agar is both differential and an enriched one. It distinguishes between hemolytic
and non-hemolytic bacteria. Hemolytic bacteria (eg., many Streptococci and
Staphylococci) produce clear zones around their colonies because of red blood cell
destruction.

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Isolation of pure cultures:
In natural habitats, microorganisms usually grow in complex, mixed
populations containing several species. One needs a pure culture, a population of cells
arising from a single cell, to characterize an individual species. Robert Koch introduced
pure culture techniques.
Spread plate and streak plate:
Spread plate - Mixture of cells is spread out on an agar surface so that every cell grows
into a completely separate colony, each colony representing a pure culture. A small
volume of around 30 to 300 cells (mixed) is transferred to the center of the plate and
spread evenly over the surface with a sterile bent-glass rod.
Streak plate – the microbial mixture is transferred to the edge of an agar plate with an
inoculating loop or swab and then streaked out over the surface in one of several patterns.
In both these techniques, successful isolation depends on spatial separation of single
cells.
The pour plate: Extensively used with bacteria and fungi. The original sample is diluted
several times to reduce the microbial population sufficiently to obtain separate colonies
when plating. The microbes are mixed with molten agar which has been cooled t 45ºCand
then poured into pertidishes. All these techniques require petridishes (special culture
dishes) after their inventor Julius Richard Petri (1887). They consist of two round halves,
the top half overlapping the bottom. Each bacterium replicates to form a colony that is
visible to the naked eye. A colony contains up to 109 copies of the original bacterium.
Colony morphology and growth:
Colony development on agar surfaces aids the microbiologist in identifying
bacteria because individual species often form colonies of characteristic size and
appearance. Structure of bacterial colonies also has been examined with the scanning
electron microscope. Generally the most rapid cell growth occurs at the colony edges.
Growth is much slower in the center and cell autolysis takes place in the older central
portion of some colonies. These differences in growth appear due to gradients of oxygen,
nutrients and toxic products within the colony. At the colony edge, oxygen and nutrients
are plentiful. Bacteria growing on solid surfaces such as agar can form complex and
intricate colony shapes. Nutrient diffusion and availability, bacterial chemotaxis and the

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presence of liquid on the surface all appear to play a role in pattern formation. These
patterns vary with nutrient availability and the hardness of the agar surface.
REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An

Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

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