Introduction to Microbiology

Microbiology

 Study of microscopic (living )

things
 E.g.
viruses, bacteria, algae, protists,
fungi
History of Microbiology

 1590 - First compound light

microscope Janssen
 History
 1 6 7 6 – first observation of
bacteria
“animalcules”
Anton Von Leeuwenhoek
 History
 1796 - First vaccine (smallpox)
Edward Jenner
 History
 1857 - Germ Theory of Disease
Louis Pasteur
 History
 1867 Antiseptic Surgery
Joseph Lister
 is tK rcyh
 188 o ’s
Postulates of
Disease Transmission
Robert Koch
History
 1885 - Vaccine against

Rabies
Louis Pasteur
 History

 1929 Discovery of Penicillin

(first antibiotic)
Alexander Fleming
 History
 1938 - First Electron
Microscope

 The electron microscope is

capable of magnifying
biological specimens up to
one million times. These
computer
enhanced images of 1.
smallpox, 2. herpes simplex,
and 3. mumps are magnified,
respectively, 150,000,
150,000 and 90,000 times.
 History

1953 Structure of DNA Revealed

Watson & Crick
 History

1954 Polio Vaccine

Jonas Salk
 Recent History
 Genetic engineering
 Cloning
 Human Genome Project
 Biotechnology
 Who knows what is next?
Sizes of Microbes

 Virus - 10 → 1000 nanometers *

 Bacteria - 0.1 → 5 micrometers **
-
(Human eye ) can see .1 mm (1 x 10
3
m)

* One billionth or 1 x 10 -9 m
** One millionth or 1 x 10 -6 m
 Tools of Microbiology

 Compound light Microscope

- live specimens
- 1,000 mag. or less
 Electron Microscope
- non-living specimens
- > 1,000 X mag.
 Incubator - keep microbes warm
for growth
Techniques of Microbiology
 Staining - to better see
structures
 Microbial Culture - growing
Container for microbe culture
- usually Petri dish
 Culture media
- Food for the microbes
- E.g. Agar - (from red algae)
- Others such as nutrient broths
 Pure Culture Techniques

1. Inoculation
2. Isolation
3. Identification
.
. Microbes are in the air we
breathe, the ground we walk on,
the food we eat—they're even
inside us. We couldn't digest
food without them—animals
couldn't, either. Without
microbes, plants couldn't grow,
garbage wouldn't decompose
and there would be a lot less
oxygen to breathe.
Classification of Microorganisms:
Microbes can be classified into four major groups:
1- Protozoa 2- Bacteria.
3- Fungi. 4- Viruses.
1- The Protozoa: These are unicellular organisms with protoplasm
differentiated into nucleus and cytoplasm.
Diameters in the range of 2- 100 μm.
The most important groups of medical protozoa are:
A-Amoeba: Entamoeba species. Mode of Motility: pseudopodia.
a

B- Mastigophora: Mode of Motility: the Flagella.

Gastrointestinal flagellates: Giardia intestinalis
Urogenital flagellates: Trichomonas vaginalis

Tissue and blood flagellates: Trypanosoma, Leishmania .

C- Ciliophora: motile by cilia.

Example: Balantidium coli.

D- Sporozoa: intracellular infection.

Example: Plasmodium that cause Malaria.

2- The bacteria:
Bacteria are unicellular prokaryotic microorganisms that multiply
by binary fission.

Bacteria can be classified according to morphology, arrangement,

and staining reaction into the following groups:
1- Filamentous bacteria: Streptomyces: antibiotic producers.
2- True bacteria:
Cocci: Gram positive: Staphylococcus, Streptococcus.
Gram negative: Neisseria.
Bacilli: Gram positive: Bacillus, Clostridum, Corynebacterium.
Gram negative: Enterobacteriaceae, Brucella.
3- Spirochetes: Slender flexuous spiral bacteria.
Borrelia, Treponema, Leptospira.
4- Mycoplasma: The Smallest bacteria that lack of a rigid cell wall.
5- Rickettsiae and Chlamydiae: intracellular parasites.
3- The Fungi:
These are saprophytic or parasitic organisms possessing relatively rigid
cell walls.

Medical fungi can be divided into:

1- Mould: Branching filaments; hyphae, mycelium. Usually 2 to 10 μm in width.
Example: Epidermophyton, Trichophyton,
Microsporum, Aspergillus.
2- True Yeasts: these are ovoid or spherical cells that reproduce asexually by
budding and sexually with formation of spores.
Example : Cryptococcus spp.

3- Dimorphic fungi:
Produce a vegetative mycelium in artificial
media, but are yeast like in infected lesions.
Example: Histoplasma.

4- Yeast- like fungi: Example: Candida ( Pseudomycelium).

4- The viruses:
Viruses consist of DNA or RNA enclosed in a simple protein shell known
as a capsid.

General properties of viruses

They are very small in size, from 20-300 m.

They contain one kind of nucleic acid (RNA or DNA) as their genome.

They are metabolically inert

They are obligate intracellular parasites.

They are only seen by electron microscope.

Depend on the parasitized cell for survival and multiplication

a
a
Structure of bacterial cells:
Size, Shape, and Arrangement of bacterial cells:
Morphology and arrangement of bacterial cells are criteria used for
classification of bacteria into following groups:
1. Cocci (Singular: coccus).
2. Rods (bacilli), (Singular: rod, bacillus).
3. Vibrios (Singular: vibrio).
4. Spirilla (singular :Spirillum)
5. Spirochetes. (Singular: Spirochaete).

1. Cocci:
These are round or oval bacteria measuring about 0.5- 1.0 micrometer in

diameter. When they multiplying, cocci may form pairs, chains,

or irregular groups.
a

Cocci in pairs are called diplococci, for example, meningococci

and gonococci.
Cocci in chains are called streptococci, for example Streptococcus pyogens.

Cocci in irregular groups are called Staphytococci, for example, Staphylococcus aureus.
2. Rods (bacilli):

These are stick-like bacteria with rounded, square, or swollen ends. They
measure 1- 10 micrometer in length by 0.3- 1.0 micrometer in width.
It may arranged in:
A- Chains, for example, Streptobacillus species.
B- Branching chains, for example, lactobacilli .
C- Mass together, for example, Mycobacterium leprae.
D- Remain attached at various angles resembling Chinese letters,
for example, Corynebacterium diphtheria.
3-Vibrios:
These are small slightly curved rods measuring 3-4 micrometer
in length by 0.5 micrometers in width.
Most vibriosare motile with a single flagellum at one end.
They show a rapid darting motility.
For example:
vibrio cholerae.
4-Spirochetes:
These are flexible, coiled, motile organism, 6-20 micrometer in length.
They progress by rapid body movements.
Spirochetes are divided into three main groups:
A- Treponemes. B- Borreliae. C- Leptospires.
BACTERIAL
Structures
Bacterial Structures
Bacterial Structures
Outside the Cell Wall
Capsule or Slime Layer.
Flagella.
Pili.
Special Bacterial
Structures
(Endospores)
Capsule (or slime layer)

 A gelatinous layer covering the entire

bacterium. It is composed of
polysaccaride, except in the anthrax
polymerized
bacillus, D-glutamic
which has a acid.
capsule of
 All bacteria secrete some sort of
glycocalyx, an outer viscous covering of
fibers extending from the bacterium. If it
appears as an extensive, tightly
bound accumulation of gelatinous
material adhering to the cell wall, it
is called a capsule. If the glycocalyx
appears unorganized and more
loosely attached, it is referred to as
a slime layer.

 The glycocalyx is usually a viscous

polysaccharide or polypeptide
slime.
Actual production of a glycocalyx often
depends on environmental conditions.
Importance of the Capsule (or slime
layer)
 A determinant of virulence of many
bacteria, since it limits the ability
of phagocytes to engulf the
bacteria.
Variants of encapsulated bacteria have
lost their ability of the phagocytes to
engulf the bacteria. Variants of
encapsulated bacteria have lost their
ability to produce the capsule are
usually nonpathogenic.

 The glycocalyx also enables some bacteria

to adhere to environmental surfaces
(rocks, root hairs, teeth, etc.), colonize,
and resist flushing.
Clinical Importance
of the Capsule

For example, many normal flora bacteria

produce a capsular polysaccharide matrix or
glycocalyx to form a biofilm on host tissue. As
an example, Streptococcus mutans,
a bacterium responsible for initiating
dental caries, breaks down sucrose into
glucose and fructose. It uses an
enzyme called glucosyltransferase to convert
the glucose to a sticky polysaccharide called
dextran that forms its glycocalyx and allows
the S. mutans to
adhere to the enamel of the tooth
and form plaque.
The S-Layer
 Many gram-negative and gram-positive
bacteria, as well a many Archaea possess
a regularly structured layer called
an S-layer attached to the
outermost portion of their cell wall. It
consists of a single molecular layer
composed of identical proteins or
glycoproteins
and in electron micrographs, has a
pattern resembling floor tiles. Although
they vary with the species, S-layers
generally have a thickness between 5
and
25 nm and possess identical pores with 2-
8 nm in diameter .
Significance of The S-Layer
 Protect bacteria from harmful enzymes,
from changes in pH, from the
predatory bacterium Bdellovibrio, a
parasitic bacterium that actually uses its
motility to penetrate other bacteria and
replicate within their cytoplasm, and from
bacteriophages.
 Function as an adhesin, enabling the
bacterium to adhere to host cells
and environmental surfaces, colonize,
and resist flushing.
 May contribute to virulence by protecting the
bacterium against complement attack
and phagocytosis.
 Act as a as a coarse molecular sieve.
Flagella
 Flagella are long , whiplike
appendages that move the bacteria
toward food and other attractants, a
process called chemotaxis. The long
filament , which acts as a propeller, is
composed of many subunits of a single
protein, flagellin, arranged in several
intertwined chains. The energy for
movement, the proton motive force, is
provided by adenosine triphosphate
(ATP), derived from the passage of ions
across the membrane. Bacteria possing
Flagella Arrangement
A Monotrichous (single flagellum)
e.g., Vibrio cholers
B Lophotrichous
(multiple flagella located at the same spot on the
bacteria's surfaces which act in concert to drive
the bacteria in a single direction)
e.g. Helicobacter pylori
C Amphitrichous
(single flagellum on each of two opposite ends
,only one flagellum operates at a time, allowing
the bacteria to reverse course rapidly by
switching which flagellum is active)
D Kophotrichous
(more than one flagellum on each of two
opposite ends)

E Peritrichous
(have flagella projecting in all directions)
e.g. Escherichia coli
Monotrichous Lophotrichous Peritrichous
Axial Filaments
(Spirochetes Motility)

 Spirochetes move by using a

flagellum-like structure called
axial filament, which wraps around
the spiral-shaped cell to produce
an undulating motion (snake
like movement).
Spirochetes Motility
Pili (Fimbriae)
 Hairlike filaments that extend from the
cell surface. They are shorter and
straighter than flagella and
are composed of subunits of protein,
pilin, arranged in helical strands.
They are
found mainly on Gram negative bacteria.
Importance of Pili
 Mediate the attachment of bacteria to
specific receptors on the human cell
surface, which is a necessary step in the
initiation of infection for some bacteria.
Mutants of Neisseria gonorrhoeae that do
not form pili are nonpathogens.

 A specialized kind of pilus, the sex pilus,

forms the attachment between the
male
(donor) and the female (receptor)
bacteria during conjugation (play a
role in DNA transfer)
Conjugation Adherence
 Special Bacterial Structures
(Endospores or spores)
 An endospore is a dormant, tough, and non-
reproductive structure produced by bacteria.

 The primary function of most endospores is to ensure

the survival of a bacterium through periods
of environmental stress. They are therefore
resistant to ultraviolet and gamma radiation,
desiccation, lysozyme, temperature, starvation,
and chemical disinfectants. Endospores are
commonly found in soil and water, where they
may survive for long periods.

 In contrast to eukaryotic spores, which are produced

by many eukaryotes for reproductive purposes,
bacteria will produce a single endospore internally.
 Bacterial endospores are highly
resistant structures formed
in response to adverse conditions by
two genera of medically important
Gram positive rods: the genus
Bacillus, which includes the agent
of anthrax (‫ﺍﻟﺠﻤﺮﺓ‬ ‫)ﺍﻟﺨﺒﻴﺜﺔ‬, and
the genus Clostridium, which
includes the agents of tetanus and
botulism.
Spore Formation
(Sporulation)
 Spore formation occurs when nutrients,
such as sources of carbon and nitrogen
are depleted. The spore forms inside
the parent vegetative cell (hence
the name endospore)
and contains: bacterial
DNA, a small amount of
cytoplasm, cell membrane,
peptidoglycan, very little water, and most
importantly, a thick keratin-like coat
that is responsible for remarkable
resistance of the spore to heat,
dehydration, radiation, and chemicals.
This resistance may be mediated by
Endospore
Germination
 Once formed, the spore has no
metabolic activity and can
remain dormant for many years. Upon
exposure to water and appropriate
nutrients, specific enzymes degrade the
coat; water and nutrients enter; and
germination into a metabolizing,
reproducing bacterial cell
(vegetative forms) occurs.

 This differentiation process is not a

means of reproduction, since one cell
produces one spore that germinate into
one cell .
Germination
 Positions of Spores
• The position of spores in relation to the body
of the bacillus may be : central ,terminal , or
subterminal.

• They differ in shape they may be oval

or rounded. The position and shape of
spores are characteristic of the species and
may aid in the microscopic identification
of the
bacterium.
BACTERIAL Cell
Wall
Peptidoglycan
(Mucopeptide, Murein)
 The vast majority of bacteria have a cell wall
containing a special polymer called peptidoglycan.
 The cell wall lies outside the cell membrane, and the
rigid peptidoglycan is important in defining the
shape of the cell, and giving the cell mechanical
strength.
 The bacterial cell wall is a unique biopolymer in that
it contains both D- and L-amino acids. Its basic
structure is a carbohydrate backbone of
alternating units of N-acetyl glucosamine
(NAG) and N- acetyl muramic acid (NAM).
 The NAM residues are cross-linked with
oligopeptides.
 The terminal peptide is D-alanine . This is the onl y
biological molecule that contains D-amino acids and
it is the target of numerous antibacterial antibiotics
The Structure of
Peptidoglycan
Basic Structural Unit of
Peptidoglycan
Properties associated with
bacterial cell walls

Bacteria may be conveniently divided into

two further groups, depending upon their
ability to retain a crystal violet-iodine dye
complex when cells are treated with
acetone or alcohol. This reaction is
referred to as the Gram Reaction (
named after Christian Gram,
who developed the staining
protocol in 1884). It may seem
a very arbitrary basis on which to
build one's classification system. This
of
reaction, however, reveals fundamental
differences in the structure
 Cells with many layers of peptidoglycan
can retain a crystal violet- iodine
complex when treated with alcohol.
These are called Gram- positive
bacteria and appear blue-black or
purple when stained using Gram's
method. Gram-negative bacteria
have only one or two layers of
peptidoglycan and cannot retain the
crystal violet-iodine complex. These
need counterstaining with another
dye to be seen using Gram's
method. A red dye such as
Safranin red. They appear red when
stained using Gram's method.
Gram Positive Bacteria
 The cell wall of Gram-positive
bacteria lies beyond the cell
membrane and is largely made up
of pepidoglycan. There may be up to
40 layers of this polymer,
conferring enormous mechanical
strength on the cell wall. Other
polymers including teichoic and
lipotichoic acids also lie in
the cell walls of Gram-positive
bacteria. These act as surface
antigens.
Cont. Gram Positive Bacteria
Lipoteichoic acids (LTA)
 Lipoteichoic acids are found only in
Gram positive bacteria.
 These polysaccharides extend though
the entire peptidoglycan layer and
appear on the cell surface.
 As a consequence, these structures
can serve as antigenic
determinants.
Gram Negative Bacteria
 In contrast to Gram-positive cells, the cell envelope
of Gram- negative bacteria is complex. Above the cell
membrane is a periplasm. This area is full of
proteins including enzymes. One or two layers
of peptidoglycan lie beyond the periplasm. Gram-
negative bacteria are thus mechanically much
weaker than Gram-positive cells. Beyond the
peptidoglycan of the Gram-negative cell wall lies an
outer membrane. This has protein channels -
porins - through which some molecules may pass
easily. The outer side of the Gram-negative
outer membrane contains
lipopolysaccharide . This provides
the antigenic structure of the surface of Gram-
negative bacteria and also acts as endotoxin.
It is this that is responsible for eliciting the
Proteins (OMPs) act transporters through the
symptoms of Gram-negative shock if it gains access
to the bloodstream. Porins and Outer Membrane
Cont. Gram Negative Bacteria
Lipopolysaccharides (LPS)
(Endotoxin)
 Also known as lipoglycans, are large molecules
consisting of a lipid and a
polysaccharide joined by a covalent bond;
they are found in the outer membrane of
Gram negative bacteria, act as
endotoxins and elicit strong immune
responses.
 LPS is additionally an exogenous pyrogen
(external fever-inducing compound).
 With the Lipopolysaccharide being of crucial
importance to gram negative bacterial cells, it

substances.
is therefore a prime target for future
antimicrobial

substances.
 Structure of Lipopolysaccharide

It comprises three parts:

1. Polysaccharide (O) side chains.

2. Core polysaccharide (core oligosaccharide
in genus Neisseria).
3. Lipid A.
O - antigen

 The polysaccharide side chain is referred to as the O-

antigen of the bacteria. O side chain (O-antigen)
is also a polysaccharide chain that extends
from the core polysaccharide.
 The composition of the O side chain varies between
different Gram negative bacterial strains. The
presence or absence of O chains
determine whether the LPS is considered rough
or smooth.
Full length O-chains would render the LPS smooth
while the absence or reduction of O-chains would
make the LPS rough. Bacteria with rough LPS
usually have more penetrable cell membranes
to hydrophobic antibiotics since a rough LPS
is more hydrophobic.
 O side chains are easily recognized by the antibiotics
of the host, however, the nature of the chain can
easily be modified by Gram negative bacteria to
Core oligosaccharide

 Core oligosaccharide contains unusual

sugars (e.g. KDO, keto-
deoxyoctulosonate and heptose). The
core oligosaccharide is attached to
lipid A, which is also in part
responsible for the toxicity of Gram
negative bacteria.
Lipid A

 Lipid A contains unusual fatty acids('e.g.'

myristic acid) and is embedded into the
outer membrane while the rest of the LPS
projects from the surface.
 Lipid A is a disaccharide with multiple fatty
acid tails reaching into the
membrane. This is the key in the toxicity.
 When bacterial cells are lysed by the immune
system, fragments of membrane containing
lipid A are released into the circulation,
causing fever, diarrhea, and possible fatal
endotoxic
shock (also called septic shock).
Cell Wall of Acid Fast
Bacteria
 Mycobacteria, e.g. Mycobacterium
tuberculosis, have an unusual cell wall,
resulting in their inability to be Gram
stained. These bacteria are said to be
“Acid-fast ” , since they resist
decolorization with acid-alchohol after
being stained with carbolfuchsin (red
dye), the initial stain, during the acid-
fast staining procedure (Zeihl-Neelsen
Stain). And thus they are stained red.
This property is related to the high lipid
content of the cell wall (mycolic acids).
Mycobacterium tuberculosis
(red rods in a blue
background)
Composition of the Acid-Fast
Cell Wall
 In addition to peptidoglycan, the acid-fast cell
wall of Mycobacterium contains a large amount
of glycolipids, especially mycolic acids. The
peptidoglycan layer is linked to
arabinogalactan (D-arabinose and
D - galactose) which is then linked to high-
molecular weight mycolic acids. The
arabinogalactan/mycolic acid layer is overlaid
with a layer of polypeptides and
mycolic acids consisting of free
lipids, glycolipids, and
peptidoglycolipids. Other glycolipids
include lipoarabinomannan and
phosphatidyinositol mannosides (PIM). Like
the outer membrane of the gram-negative cell
wall, porins are required to transport small
hydrophilic molecules through the outer
Cont. Composition of the Acid-Fast
Cell Wall
Mycoplasma
A genus of bacteria which naturally lacks
the cell wall. Without the cell wall, they
are unaffected by many
common antibiotics such as penicillin
or other
beta-lactam antibiotics that
target cell wall synthesis. They
can be parasitic or saprophytic.
Several species are pathogenic in
humans, including M. pneumoniae,
which is an important cause of
atypical pneumonia and other
respiratory disorders, and M.
genitalium , which is believed to be
involved in pelvic inflammatory diseases
L-form Bacteria
 Partial or complete loss of the bacterial cell wall
components results in cell wall deficient ( CWD) or
L-form bacteria.
 Cell wall deficient forms of bacteria were first
described in 1883 by Malassez and Vignal. They were
named “ L” for Lister
Institute.
 When a bacterium has no cell wall present
(complete removal of the cell wall), it is defined
as a protoplast.
 Bacteria that have some cell-wall remaining (partial
removal of the cell wall) are termed
spheroplasts.
 Spheroplasts and protoplasts are osmotically
fragile, and will lyse if transferred to a hypotonic
solution.
BACTERIAL
METABOLISM
 Metabolic processes are concerned with all those
biological or chemical reactions which can
be carried out by microorganisms.

 Since microorganisms are capable of carrying out

chemical reactions, the processes involved
must follow the basic lows of
thermodynamics and catalysis.
(A) Catabolism
 In catabolism, two processes take place :-

1. Oxidation or destruction of the large

substrates into smaller units ( building units )
such as amino acids, monosaccharide, fatty
acids, etc …. These units will then be used in
biosynthesis.

2. Energy will be elaborated and will be stored

in adenosine triphosphate ( ATP ) molecules.
(B) Anabolism
 In anabolism, the following takes place :

1. The building blocks produced by catabolic processes

are converted to activated monomers and coenzymes This
step requirs energy in the form of ATP.

2. The activated monomers are then polymerized

into macromolecules (e.g. polyeptides,
polynucleotides, polyasccharides, …. ). Energy in the form of
ATP is also required in this step.

3. The polymerized molecules are then transported to

the appropriate area of the cell and essembled to form the
cell
structures. The net result will be cell growth.
 Regulation of metabolism
 A large number of microbial enzymes participate in
the conversion of carbon and energy sources
into small building units and liberation of energy
i.e. ( catabolism ). These enzymes are
generally synthesized insid the microbial cell
irrespective of the presence of their substrates
and are so called “ constitutive enzymes “ .
 Some other enzymes, usually concerned in
the breakdown of a particular
substrate, are not produced by the
organism except in the presence of the
substrate itself or other compounds of
a close chemical structure ( inducers );
these enzymes are called “ induced
enzymes “ and this is of great economic
importance to the cell.
 The majority of enzymes are in fact produced only
when needed and in the mount needed. This is
generally referred to as enzyme induction i.e.
formation of an enzyme only upon stimulation which
is triggered by a need. Once the need
disappears, synthesis of the enzyme stops. The trigger
is often the product of the reaction catalyzed by the
enzyme .

 when this reaches a certain concentration ( above

what the cell needs ) this product represses further
synthesis of the enzyme and the phenomenon is
called end product repression.
 The enzymes catalyzing biosynthesis ( i.e. anabolism )
are subject to control by feedback inhibition
and repression just as the enzymes
concerned in catabolism of specific compounds are
controlled by induction.

 In feedback inhibition other end product in the

pathway inhibits the action of the first enzyme in that
pathway .

 In “ repression “, the end-product if it already present

in the growth medium or when it is formed
in sufficient quantities.
 The product influences the extent of activity of the
enzyme. High concentration of the
product immediately inhibits the activity of
enzyme molecules already synthesized and thus
the rate of product formation decreases. This
phenomenon is called end product inhibition .
 Both end product inhibition and
repression act only on the first
specific enzyme reaction in a
sequence of divergent reactions so
that the product does not inhibit
formation of other products in the
same pathway.
High energy
transfer
compounds
 It is essential for the life of bacteria that the energy
released from exergonic reactions be uesd to derive
endergonic of coupling exergonic reactions with
endergonic rections.

 The common reactants of greatest use to the cell are

those capable of transferring large ammounts of free
energy, called high energy-transfer compounds.

 A variety of such compounds exist in cells. For example

: ATP, GTP, UTP , PEP ( phospho - enol pyruvic acid ).

 ATP is by far the most important. All the compounds

listed before can transfer their energy directly or
indirectly to ATP synthesis.
Energy Released
From ATP By
Hydrolysis :

OVERALL REACTION
 ATP + H2O → ADP + H3PO4; ΔG = -7300 cal .
ADP is also a high – energy transfer compound, since
its hydrolysis yields a large quantity of energy.
 ADP + H2O → AMP + H3PO4; Δ G = 7300 cal .
AMP however is a low – energy compounds;
its hydrolysis liberates only a small quantity of energy.

 AMP + H2O → Adenosine + H3 po 4; Δ G = - 2000 cal.

 Several types of chemical reactions are involved in

energy production, but OXIDATION - REDUCTION
is probably the commonest.
OXIDATION: It is the loss of electrons.
REDUCTION: It is the gain of electrons.
* Frequently, oxidation reactions are dehydrogenation
(reactions involving the loss of hydrogen atoms ), since
a H atom consists of a proton + an electron, a
compound which loses a H atom has essentially lost
an electron and therefore has been oxidized.
* In each reaction, a pair of substances is involved
: One is the reduced form } → Fe2+
One is the oxidized form } → Fe3+
* Each pair of substances is referred to as OXIDATION
REDUCTION
( O / R ) SYSTEM
 One O/R system may tend to absorb electrons from another
O/R system; i.e. the first system will oxidize the second.
This power ( the tendency to absorb electrons ) is expressed
by the STANDARD REDUCTION POTENTLAL or
ELECTROMOTIVE
POTENTLAL.

 ( E0) OF anO/R system ( expressed in volts ): the more positive

the E0 the greater the oxidizing ability of the
system consequently, any O/R system can oxidize any other
system has low E0 value, but not more E0 value. Such
relationships are very important in understanding the orderly
sequence in which biological oxidations occur.

 When one O/R system oxidized another, ENIERGY is released if

the voltage difference ( E0 ) between the two system is large
an
 amount of free energy sufficient to drive the synthesis of
ATP maybe liberated.
Carbon
Requirements
 Microorganisms are classified into 2 groups according
to carbon requirements :

 Lithotrophic ( or autotrophic ) : which are able to

utilize CO2 as the main source of carbon. They obtain
their energy either by oxidation of simple inorganic
compounds ( and hence are called chemosynthetic
autotrophs) or from sunlight ( and hence are called
photosynthetic autotrophis ).

 Organotrophic ( or heteretrophic): which require

complex organic compounds as a source of carbon.
This group of microorganisms also obtain their energy
from oxidation of suitable organic compounds.
Energy
Requirements
 Only chlorophyll - containing organisms can
utilize sun light as a source of energy. A
few bacteria do, and since they derive the
energy required for synthetic reactions from
photons. They are described as photosynthetic
while most bacteria and all fungi are
chemosynthetic since they derive energy
from chemical bonds of inorganic
compounds or more commonly, of organic
compounds.
 Chemosynthetic bacteria: derive energy from
chemical bonds by oxidizing (releasing electrons )
from nutrients in their environment.

 Different types of bacteria my oxidize specific

substances, e.g. hydrogen bacteria which convert
hydrogen to water, methane bacteria which oxidize
methane to CO2 nitrifying bacteria oxidize ammonia
to nitrite or nitrate, sulplur bacteria which oxidize
H2S to S or to sulphate. However, the most common
are the bacteria which oxidize various organic
common are the bacteria which oxidize various
organic compounds most notably sugars.
 In aerobic respiration glucose is oxidized through
krebs cycle and electron transport system to
give Co2 and H2 oand ATP ( each 1 mole of glucose
give 38 mole of ATP .
C6H12 o6 + 6 o2 → 6 Co2 + 6 H2o + 38 ATP

 This is called chemotrophic metabolism in which

chemotrophic microorganisms oxidize glucose in
a series of enzyme catalyzed reactions such
as glycolysis, krebs cycle and electron
transport system. The end result of these
oxidation is the production of energy.
 Fermentative metabolism utilizes organic
compound as both electom donors and
electron acceptors. Many bacteria use pyruvate (
glycolytic
and Embden - Meyehof pathways ) in
secondary fermentation processes.

 Mixed acid fermentation which is common among

most members of Enterobacteriacaea
is combination of lactate, acetate and formate,
in addition Co2, H2 and ethanol are produced.
 The glyolysis results in yield of 2 mol of ATP per 1
mol of glucose fermented.
 Phototrophoic microorganisms : Which obtain
energy from sun ( photosynthetic organisms
). The electron doner is water being split by
photon ‘s energy to oxygen and electrons, which
are used to reduce carbon dioxide ( the terminal
electron) acceptor.
 Several groups of bacteria ( green and purple
bacteria) can perform photosynthesis.
The
general equation for bacterial photosynthesis
2 H2A + Co2 → ( CH2o ) x + 2A + H2o
In presence of light and green pigment (
bacterial chlorophy II ).

 In bacterial reaction, H2A represent general

term for a reduced inorganic sulfur
compound or organic compound ; e.g. If H2A
stands for H2S, A is then elemental sulfur.
According to carbon
and energy
requirements
microorganisms are
classified into :
Energy production by aerobic respiration (
the cytochrome system ) :

 The respiratory chain ( or cytochrome system

or electron-transport system ) is a sequence
of oxidation reaction. The function of
this sequence is to accept electrons from
reduced compounds and transfer them to
oxygen, with the resulting formation of
water. At several steps in the chain, the
difference in E0 values is great enough to
permit sufficient energy to be liberated for
ATP synthesis.
 Sufficient energy for ATP synthesis is liberated
at three points along the chain . This is
called oxidative phosphorylation. The
respiratory chain of bacteria is associated
with the cell- membrane. The electromotive
potential ( E0 ) of the O/R system increased
gradually.
 Models of Energy Generation in
Biological System :

Energy generating reactions involve,

formation of high energy bonds, usually
phosphates, and
we can thus refer to it as phosphorylation.
This occures in two types :
(1) Substrate Phosphorylation

 Where a phosphotylated substrate

donates the phosphate directly to
ADP forming ATP without electron
donation.
(2) Oxidative Phosphorylation
Where ADP is phosphorylated by
inorganic phosphate. This process is catalyzed
by enzymes called dehydrogenase which transfer
the released electrons from the
substrate through intermediate series of
conzymes ( cytochrome enzymes ) in case of
aerobic bacteria until the electrons reached the
final electron acceptor by electromotive force.
 In microbial cells many net mediate coenzymes
acting as electron acceptorse.g. :-

 Nicotinamide – adenine – dinucleotide ( NAD )

 Nicotinamide – adenine – dinucleotide – phosphate (
NADP)
 Flavine – adenine – dinucleotide ( FAD )
 Riboflavine phosphate.
 Various cytochromes
 NAD→ NADH
 NADP→ NADPH
Microorganisms are
Classified into three major
classes according to the
nature of the terminal
electron acceptor in the
oxidative system;
these are :-
1. Aerobic respiration microorganism : where the final
electron acceptor is molecular oxygen.

2. Anaerobic microorganism : where the final electron

acceptor is an inorganic compound other than oxygen
( e.g . nitrate,sulfte, carbonate, … etc. )

3. Fermentative microorganism ( fermentation ) : where

the final electron acceptor is an organanic compound.
 Other Biosynthetic pathways :
 The enormous diversity in the
nutritional requirements of bacteria stems
from the biosynthetic abilities of different
groups of bacteria. Bacteria use various
catabolic pathways for biosynthesis, including
glycolsis pyruvate oxidation, and TCA cycle, The
glyoxylate bypass of the TCA cycle allows
bacteria to use acetate as a precursor of a
variety of 4-carbon dicarboxylic acids.
 Amino acid biosynthesis falls into
convenient grouping :

 Clutamate family : Clutamate and glutamine are

formed from reductive amination of a- ketoglutarate
and serve as the carbon skeletons for proline
and arginine.

 Aspartate family : Aspartate is synthesized by

transamination of oxaloacetate, gives rise to
asparagine, and can be reduced to lysine, methionine,
threonine, and isoleucine.

 Pyruvate family : Alanine, valine, isoleucine, and

leucine are formed from the initial transamination of
Pyruvate.
 Serine family : Serine, glycine, and cysteine are
derived from 3-p-glycerate; glycine resultes
from
the transfer of the 1-carbon fragment to
tertrahydrofolate.

 Histidine : is synthesized from the 5-carbon

backbone of phosphoribosylpyrophosphate ( PRPP
) and is unique in that its carboxy1 group is
not present on the starting compound but is
fromed later.
 Aromatic amino acids : Tyrosine, phenylalanine,
tryptophan are derived from shikimic acid.
Nucleotide biosynthesis :

a. Nucleotide biosynthesis in bacteria is identical

to that animal tissue.

Purine nucleotides are built on the ribose-p

chain from PRPP, glycine, and cabon.

Pyrimidine nucleotides are formed via a series of

carboxy1-containing intermediates, starting with
carbamy1 phosphate with ribose-p added late in
the sequence.
b. The principal macromolecules : DNA, RNA,
and polymers of the nucleotide and amino acid
building
blocks and are synthesized via a highly
regulated system in the cell cytoplasm.

The sequence of information transfer for

macromolecular synthesis involves replication ( DNA
to DNA ), transcription ( DNA to RNA ),
and translation ( RNA to protein).

DNA acts as the template necessary for synthesis of both

DNA and RNA .

RNA serves as the template for protein synthesis and

provides the core of ribosomes on which the proteins
are synthesized.
Oxygen Requirements
of
Microorganisms
Microorganisms are classified
according to
their requirements for oxygen into
four groups :
(1) Obligate aerobes : they fail to grow in absence of
oxygen since only oxygen can act a terminal electron
acceptor. They have the following reactions :

a. O2 + e → H2o + energy by cytochrome

oxidase system.

b. O2 + e → H2o2 + energy by flavoprotein system.

Since hydrogen peroxide is highly toxic to the cell
they also have.

c. H2o2 → H2o + o2 by catalase system.

(2) Micro-aerophilic : These grow in presence of little
amount of O2.

 They posses reaction (a) but the amount of

cytochrome oxidase is limited.

 They also posses reaction (b) but not (c) accordingly,

in high oxygen tension, both reaction (a) and
(b) operate and thus H2o2 accumulates and the cells
die.

 This happens in the top of the tube of liquid medium

in which they grow. At lower levels, reaction (a)
operates but (b) does not ( insuffecient oxygen
for both ) and the bacteria grow. At the bottom of
 the tube there is no oxygen and both reaction (a)
and (b)
fail hence they do not grow.
(3) Obligate anaerobes :
 They posses (b) but not (a) or (c) thus in
presence of any oxygen they accumulate H2o2
and thus die off instead they operate
reaction (d) where organic or inorganic
compounds are reduced by electrons released
from nutritive substrates, e.g. :
 Reduction of nitrate and sulfate.
(4) Facultative anaerobes : all of reaction (a), (b), (c) and (d)
operate and thus they grow in presence or absence of oxygen.

The toxicity of o2 results from its reduction by enzymes

found in the cell ( as flavoproteins ) to hydrogen peroxide
and the more toxic free radical superoxide ( o2 )
Aerobes and facultative anaerobes are protected from
these products by the presence of superoxide-dismutase
and catalase enzymes that catalyzes the reaction.

 20¯2 + 2 H suderoxide dismutase O2 + H2O2

------------ →

 2H2O2 catalase H2O + O2

------- →
 All obligate anaerobes lack both superoxid dismutaed
and catalase, enzymes

Many methods are available for exclusion of O2 in case

of anaerobic cultivation e.g. :
 Reducing agents such as sodium thioglycollate can
be added to liquid cultures.

 Tubes of agar can be sealed with a layer of paraffin.

 Culture vessel can be placed in a container from which

O2 is removed by evacuation or chemical means.
Nitrogen
Requirements
 Nitrogen is required by living beings for the synthesis
of amino acids ( and proteins ); purines and
pyrimidines ( and nucleic acid); vitamins and
coenzymes …. Etc.

 Some bacteria can utilize inorganic nitrogen

compounds ( ammonia, nitrates, nitrites ….. etc )
as the sole souree of nitrogen. These bacteria are
simple in their growth reqirements ( since it requires
only a few simple nitrogen compounds ) but
complex in its metabolism ( since it possesses a
large number of enzymes which continually
synthesize a large number of organic nitrogen
compounds ) and does not require
proteins; protein hydrolysates; amino acids; vitamins
and cofactors … etc in their food
 Some bacteria require e.g. only one vitamin or amino acid
and synthesize the rest of organic nitrogen
compounds from inorganic nitrogen. The growth of such
bacteria would
be proportional to the amount of the required vitamin
or amino acid in their food and hence are used for the
assay of such compound in pharmaceutical preparations.

 Other bacteria are more complex and usually require a

partially hydrolysed well-balanced protein diet.

 Others are strict in their requirement and need specific

materials in their food; e.g. Haemophilus influenze
which reiquires the “ X “ factor ( haematin ) , and / or
the “ V “ factor ( coenzyme I or II ) to grow.
 Such organic nitrogen compounds required for growth
are called growth factors; growth-promoting
compounds or accessory growth substances.

 A few species of bacteria can utilize atmospheric

nitrogen and these are called nitrogen-fixing
bacteria and grow in symbiosis with the roots of
legumes. On the other hand, almost all bacteria
can utilize ammonia as a source of nitrogen and
require in addition, only whatever specific growth
factors they may fail to synthesize. However, most
bacteria can not obtain such ammonia by
reduction of other inorganic nitrogen compounds
and a few can not
obtain ammonia even by degradation of organic
nitrogen compounds.
Requirement
Of
Other Elements
 Sulfur is required since it is a constituent of certain
amino acids, e.g. cystine. Except for
microorganisms which require these amino acids,
sulfates fill the need for this element. From is
required for the synthesis of cytochrome.
 Many enzymatic reactions require certain heavy
metal ions such as Mg ( which is also especially
required for the important reactions in
which ATPparticipates ), Zn . Co . Cu . Mn . Mb.
And many other metal ions required in
trace amounts. In addition, sodium,
potassium and calcium ions are generally
required for the proper functioning of enzymatic
reactins. Large quantities of
phosphorous are required for the synthesis of
ATP, nucleic acids and lipids.
 All these elements can be utilized by
microorganisms when they are supplied as
inorganic salte. In environments used for the
cultivation of microorganisms ( microbiological
media ) it is cucustomary to include in addition to
water, carbon and nitrogen sources, significant
quantities of phosphates and magnesium sulfate.
For other elements, requirements are so small that
their need can be satisfied from the impurities
present in other constituents of the medium.
PH
Requirements
 The best growth for most bacteria is obtained when the pH of
the environment is close to neutrality ( 6.5 to 7.8 ). A
few species can tolerate acid environments, e.g.
species of Lactobcillus and Acetobacter and a few can
tolerate high pHe.g.
vibrio colera.

 Molds tolerate a wider range of pH ( 2-8.5) and tend to grow at

pH slightly below neutrality ( about 5.5) while yeasts
usually grow at the low pH of 3.5 to 4.5 ( range about 3 to 6.5).

 When microorganisms reproduce, they release waste products

that may change the PH of the environment. If the pH change
is extensive, their environment become unsuitable and this
can be avoided by adding buffers ( chemicals that resist
change in pH ).
 Food can be preserved by lowering their PII and so we avoided
spoilage because their low pH inhibit microbial growth.
Moisture
Requirements
 Water in the environment is required to dissolve foods and
wastes, as a medium for biochemical reactions and in way as
a source of hydrogen and oxygen. Only free and not
bound moisture is considered, e.g. a 4% aqueous agar
gel contain about 94% moisture but very few bacteria can
grow in it while milk, with about 87 % moisture is suitable for
bacterial growth since it contains more free moisture.

 Molds can generally grow in the least amount of moisture thus

can be seen on almost dry surface e.g. on leather, while
yeasts more moisture.

 Bacteria require much more moisture than fungi and among

them some require more moisture than others, e.g.
Neisseria gonorrhoeae and Mycobacterium tuberculosis
respectively. A dry environment results in the loss of the
cell ‘s water to its surrounding and this disrupt cellular
activities.
 The available water surrounding the organisms is exposed as
water activity (aw), the aw is the relative humidity of the
air space in the immediate environment as broth RH.
aw = RH/100 = 98.5/100 = 0.985

 Microorganisms commonly require water activities above 0.90

in order to grow and multiply The optimal is 0.95. Some
fungi can grow in 0.60.

 The water activity of foods can be used to predict how rapidly

food spoilage occur. Food with high water activity spoil
more rapidly. In some cases, the water activities of foods
can be artificially lowered in order to prevent food spoilage
e.g freeze- dried foods are resistant to spoilage by
microorganisms because the foods have been desiccated in
vacuum and so have very low water activity.
The Osmotic
Pressure
 Osmotic pressure is defined as the minimum amount of
pressure that must be applied to solution in order
to
prevent the flow of water across a membrane within
the solution .

 Moisture affects the osmotic pressure and the ionic

strength of the environment. In a hypotonic
medium, water passes freely through the cytoplasmic
membrane and build up some pressure inside the cell in
attempting to balance the osmotic pressure inside and
outside the cell. Unless the cell wall is intact and
strong the cytoplasmic membrane would eventually
rupture and the cell dies. This happens in the
presence of penicillin which prevents the formation
of cell walls.
 In an isotonic environment penicillin does not kill bacteria,
which can grow and multiply without their protective
cell walls.
 In a hypertonic environment water passes from the cell
and since the cell wall is rigid while the cytoplasmic
membrane is not, plasmolysis occurs with the
cytoplamsic membrane shrinking inward. This hinders the
growth of the cell and may cause death.
 Some bacteria and many yeasts and molds are osmophilic i.e.
can grow easily in very hypertonic environments with very
low moisture content.
 In addition there are certain occean bacteria which are
halophilic i.e. can grow in high sodium chloride
concentration while most bacteria and fungi can tolerate
only low salt concentration. Some bacteria can tolerate up
to 6.5 % or 9 % sodium chloride, this being a characteristic of
the species.
 Foods fruit preserves and salted fish are
resistant to spoilage by
microorganisms
because of their high osmotic pressure (
hypertonic ). Microorganisms present
are unable to grow because the
hypertonic environment draws out the cells
water, thus inhibiting metabolism.
Temperature
Requirements
 The optimum temperature for growth is that at
which the microorganisms multiplies fastest and
produces the highest yield.

 For each species there are also maximum and

minimum temperatures for growth, beyond
which the organism does not multiply.
According to Temperature
Requirements, Bacteria
are divided into :
1- Mesophilic bacteria :
Growth temperature ranges
between 10oC, and 42oC, the optimum
being 35o C to 37oC for pathogenic
bacteria and about 22-25oC for non-
pathogenic bacteria . Most pathogenic
bacteria are mesop[hilic.
2-Thermophilic bacteria :
o
Have an optimum temperature of 45
C and capable of growing at 65o C or
higher.
3-Psychrophilic bacteria :
Few bacteria have an
optimum temperature of 15oC and can
grow near zero o C. However in the
frozen state no microorganisms can
grow. Most fungi tend to have an
optimum temperature of 20-25o C
Microbial Enzymes
 Bacteria are such active metabolic units and
so they are especially rich in enzymes.

 Enzymes are functional proteins that

catalyse metabolic reactions enhancing
their rates without directly affecting
their equilibrium.

 They may act alone or may require one or

more of the following :-
- Prosthetic groups ( not easily dissociable
from the protein apoenzyme );
- Conzymes ( usually vitamins that are
easily dissociable from the protein )
- Various metal ions.
There are three bas ic groups of
bacterial enzymes:

synthetic, respiratory and hydrolytic.

(1) Synthetic enzymes :
These participate in
anabolism, or synthesis of
various molecules and
structures required by the
bacteria.
They are intracellular.
 (2) Respiratory enzymes :

 These are intracellular enzymes concerned

with the release of energy from nutrients in
the process of catabolism.

 If in oxidation, oxygen is the final oxidizer (

terminal electron acceptor ) respiration is
aerobic and if it is not, respiration is
anaerobic, ( often called fermentation) and
usually resulting in the formation of acids
and possibly gases ( mainly Co2 and
hydrogen ).
Aerobic Respiration
 This occur rarely by direct incorporation
of oxygen in the substrate, ( which is then
called an autooxidizable substance ), and
may require enzymes called oxidases.

 Examples of such substrates are certain

dyes, falvoproteins and cytochromes.

 Most often, oxidation occurs by removal of

electrons ( hydrogen) from the substrate
by enzymes called dehydrogenases and
depositing them directly or indirectly
onto molecular oxygen.
 The substrate may be oxidized completely
or incompletely.

 Thus glucose may be oxidized

completely to Co2 and water or
incompletely to various acids, alcohols,
aldehydes, ketones … etc. and
ammonia may be oxidized to nitrate
(complete) or to nitrogen or nitrite
(incomplete).

 From these examples it can be seen that

the substrate may be organic or
inorganic. Thus species of Nitrosomonas
oxidied ammonia with oxygen to nitrites,
and species of Nitrobacter oxidized
nitrites with oxygen to nitrates.
Anaerobic Respiration
  Electrons removed from the substrate
may be accepted by organic or
inorganic compounds but not by
oxygen.

 If it is an inorganic substance, the

oxidation- reduction reaction is
intermolecular i.e. the oxidizable
substrate and the electron acceptor are
two different molecules, e.g. glucose may
by oxidized to Co2 and water while nitrate
is reduced to nitrite, or sulfur oxidized to
sulfate ( with water ) while nitrate is being
reduced to free nitrogen.

 Among the most common inorganic

2
electron acceptors are the nitrates, nitrites
, sulfates and CO ( reduced to methane ) .

2
 In the case of organic electron
acceptors, the reaction may by
intermolecular or intramolecular.

 The latter being a reaction in which the

substrate also acts as an electron
acceptor, part of which being oxidized
while another part is being reduced, e.g.
two carbons of glucose are oxidized to CO2
while the rest of the glucose molecule
is reduced to ethanol in the
alcoholic fermentation of sugars by
yeasts. This is the process classically
termed “
fermentation or airless respiration”
Examples of anaerobic
respiration

 Sulfate reducers :
Here the bacteria use sulfate as a
final electron acceptor and thus
reducing it to sulfide.
So-2 + 8e + 8H + Sulfate + 4H2o
------→
redictase
Nitrate reducers:
2 No 3 + 12e + 12 H + Sulfate N2 + 6H2o
------→
redictase

Methane bacteria :
Co2 + 8e + 8H+ → CH4 + 2H2O
This bacteria is normal flora of the GIT of
animal, it is found in sewage. CH4
produced is used as fuel.
(3) Hydrolytic Enzymes :

 Certain nutrients are so complex that

they can not diffuse from the
medium into the protoplasm (
across the cytoplasimc membrane )
unless they are hydrolyzed to
smaller diffusible molecules by
hydrolytic enzymes that are secreted
by the organism into the medium and
hence are extracellular.
Some of the extracellular enzymes may
not serve food- hydroly zing enzymes
but basically to digest a dense medium
so that the bacteria can diffuse more
rapidly and thoroughly throughout the
medium, or to inactivate or inhibit
hostole bodies and structures in the
medium. This is common with
pathogens which secrete enzymes that
destroy the tissues or that kill various
defense bodies or substances ( e.g.

bacteria.
Some of the extracellular enzymes may
leucocytes) which the body sends out to
the site of infection to fight the

bacteria.
 The ability of bacteria to produce various
extracellular enzymes is characteristic of the
species( or even the strain ) and is enzymes that
act upon :
- Lopids and fats, by lipases ( mainly estrases ).
- Carbohydrates, by enzymes that hydrolyses
starch ( amylases ):
- Pectins ( pectinases ) : cellulose ( cellulasese )
disaccharides ( e.g. lactase : maltase and sucrase )
… etc. Bacteria which produce such enzymes
are called saccharolytic bacteria .
 proteins, by proteinases and polypeptidases that
produce polypeptides and amino acids
respectively. These are followed by deaminases and
disamidases which release ammonia, CO2 and
organic acids : and urease which decomposes
urea to CO2 and ammonia. Natural protein
hydrolysis ( proteolysis ) is called
putrification and is carried out by proteolytic
bacteria which are commonly anaerobic.
 Example of extracelluler enzymes enhance the
pathogenicity of which are :
bacteria

 Hyaluronidase : which dissolves

hyaluronic acid ( aa carbohydrate ).
 Collagenases : which dissolve collagen.
 Gelatinase : which dissolves gelatin.
 Fibrinolysin : which dissolves fibrin.
 Haemolysins : which dissolves RBC’s .
 Coagulases : which dissolves plasma .
 Leucocidins : which kill leucocytes .
Other Bacterial Products :
Pigments :

Some bacteria characteristically

produce specific pigments when
they are cultivated under specific
conditions. These pigments may by
intracellular and can not diffuse into the
medium and thus only the cells are
colored : or they may be extracelluar
and diffuse into the medium coloring
it and these my be soluble in
water only or may also dissolve
in one or more organic solvent .
 Toxins :

 These are poisons which are produced as a

result of the growth of the bacteria or may
be part of the bodies of the bacteria.

 They are usually responsible for the

characteristic lesions which pathogenic
bacteria produce in their hosts. They may
be extracelluar ( exotoxins ) or intracellalar
( endotoxins ). Many of the

 non- digestive extracellular enzymes

mentioned above may be also considered
bacterial toxins
 Pyrogenic materials :
 These are special endotoxins which produce a
febrile reaction upon injection in minute doses in
a subject.

 These pyrogens are apparently very potent

lipopolysaccharide complexes produced by
certain gram negative bacteria.

 They are dangerous when present in parenteral

preparations and may be removed by
adsorption on ion- exchange resins.

 They may be inactivated only by heating at

temperatures over 300 ºC but not by normal
sterilization processes.

 The best method of eliminating them from

Thank You