Bacteriology

Bacteriology is defines as study of bacteria,

Classification,
Bacterial growth
Normal Body Flora
Sterilization and Disinfection
 Introduction to bacteriology
• Bacteria are single-celled microorganisms that lack a nuclear membrane,
• Metabolically active and divide by binary fission.
• Medically they are a major cause of disease.
• Cause food spoilage
• Therefore the study of the bacteria is called bacteriology
Beneficial interactions with other organisms
• Maintain balance of environment (microbial ecology)
• Basis of food chain
• Nitrogen fixation
• Photosynthesis
• Digestion, synthesis of vitamins
• Manufacture of food and drink
• Genetic engineering
• Synthesis of chemical products
• Recycling sewage
• Bioremediation: use microbes to remove toxins (oil spills)
• Use of microbes to control crop pests
 Unique &distinguishing characteristics features of bacteria
• Unique & Distinguishing Characteristics of bacteria
• No nucleus: Unlike another eukaryotic cell, the bacteria cell is the
only cell which lacks a prominent nucleus within. This feature has
compelled the scientists to consider bacteria as primitive organisms,
i.e., earliest forms of life on earth.
• Presence of Cell wall: Bacteria has a cell wall which is distinct from
other cells having a cell wall. The wall is made of different substances
like glycoproteins, lipopolysaccharides and lipoproteins.
• Cell membrane: This is present immediately below the cell wall. This
has a small variation with that of plants and animals cell membranes.
In plants and animals, it is mostly a single or double layer (bilayer) of
lipids. But bacteria especially the gram-negative ones have an extra
membrane, i.e., an outer membrane called periplasm which lies
bellow cell wall but above the cell membrane
• Pilli: These are small and tube-like projections from the bacterial
surface. They are the organelles of sexual reproduction and are
involved in the exchange of genetic material as part of reproduction
between two bacteria.
• Food dependence: All the organisms either live on their own or other
resources. But bacteria have different modes of food procurement.
Some of them synthesize their food like plants by use of sunlight.
• Other derive food energy from chemical sources around them
(chemotrophs) — sulfur bacteria, hydrogen, bacteria, iron bacteria,
etc.
• Few of them are parasites which live on alive plants and animals. They
cause diseases to the host in doing so.
• Many bacteria are saprophytes which means, they live on dead and
decaying matter.
• While others are symbiotic. They live in a give and take relationships
with other animals. Ex: rhizobium in roots, E-coli in the intestine.
• Respiration: Bacteria do not have mitochondria like a typical animal and
plant cells. However, they derive energy by aerobic respiration. In the
higher cells, aerobic respiration occurs due to the glycolysis and
oxidative phosphorylation pathway in the mitochondria.

• Reproduction: Asexual reproduction: The bacteria cell undergoes to

produce two bacteria by Binary fission and also by endospore by
asexual
• Sexual reproduction: It is also by two distinct methods like
transformation, conjugation, and transduction. In transformation, one
bacteria release naked DNA strand into the surroundings. The other
bacteria take it up and incorporate into their genome.
Structure of bacteria cell
 Bacteria Classification
Criteria used in bacteria
classification:
Nomenclature
Readily observable properties
Metabolic reactions
Genetic relatedness
 1. Nomenclature
Binomial nomenclature : genus and species
e.g.:
 Escherichia coli
 Staphylococcus aureus etc.

Note!
 Genus name always begin with a capital letter
 Species name should start with small letter
 Both names are always italicized or underlined

Abbreviation: E. coli
S. aureus
2. Readily observable properties

1. Gram staining
 A simple laboratory procedure

 Employs the dye, crystal violet and alcohol

 Was developed in 1884 by the Danish bacteriologist Hans Christian

Gram.
On the basis of Gram staining
properties, bacteria are grouped as:
Gram-positive
Gram-negative
The determinant of such properties’ difference is in the
cell wall of the respective bacteria
Gram stain procedure
But gram staining results are not universally applicable because some
bacterial cells stain poorly or not at all.

Acid-fast stain

This is another important differential stain that

bind strongly only to bacteria that have a wax
material in their cell walls.

This dye is used to identify all bacteria in the

genus Mycobacterium and the pathogenic
strains of the genus Norcardia
2. Morphologic features
 Another important factor in classification of bacteria include::
A) Shape
 There are three basic groups (cocci, bacilli, spirochaetes
 Some bacteria are variable in shape (pleomorphic)

1. Cocci (round shaped)

a) Cocci in clusters ( Staphylococcus)
b) Cocci in chains (Streptococcus)
c) Cocci in pairs with pointed ends (Strep. Pneumonia)
d) Cocci in pairs with kidney bean shape (Neisseria)
2.Bacilli (rod shaped)
a) with square ends (Bacillus spp)
b) With round ends (Salmonella)
c) Club shaped (Corynebacterium)
d) Fusiform (Fusobacterium)
e) Comma shaped (Vibrio)
Spirochaetes (spiral shaped)
a) Relaxed coil (Borrelia)
b) Tightly coiled (Trepanoma)
B) Arrangement of the cell
 Bacteria are found as a single
cells or irregular groups of two
or more cells
 3. Metabolic reactions
1. Oxygen requirement
Aerobic bacteria
Require oxygen for normal growth and development
Anaerobic bacteria
They can only grow in the complete absence of
oxygen
Facultative bacteria
They can grow in the presence or absence of oxygen
2. Carbohydrate utilization
Species can be identified by the variety of
carbohydrate substrates they use as energy
sources.
Some bacteria do not use glucose under any
condition
In oxidative metabolism
Oxygen is required to degrade glucose
In fermentative metabolism
Glucose is degraded anaerobically
3. Enzyme production
A) Detoxifying or oxidative enzyme may be produced
Catalase destroy hydrogen
peroxide
Oxidase transfers hydrogen
directly from a substrate to
oxygen
B) Proteolytic or toxic enzymes also may be
produced
1. Haemolysins lyse red blood cells
Alfa haemolysins lyse red cell
membranes
Beta haemolysins both lyse and
dissolve red cell membranes
2. Coagulase causes clotting of blood
plasma
4. Genetic relatedness

 Is based on similarities of nucleic acid

sequence
 and provides a more phylogenetic scheme
of bacterial classification than do the
other methods.
 Genetic classification is based on
the study of
1. DNA homology among bacteria
2. Nucleotide base composition
3. Exchange of genetic material
(ie transformation, conjugation)
btn. related organisms
 Bacterial structure
The basic components
 Structures that form the cell
envelope
 Cytoplasmic constituents
 External structures that project
through or cover the cell envelope
 Bacterial peptidoglycan
 Made up of peptides and
sugars (glycan)
 Provides support of the cell
 Allows cell to withstand media
of low osmotic pressure e.g.
water
 Cytoplasmic membrane
Has four important functions:
1. Active transport of molecules into cell
2. Energy generation by oxidative
phosphorylation
3. Synthesis of precursors of the cell wall
4. Secretion of enzymes and toxins
 Plasmids
 Extra-chromosomal , double stranded, circular
DNA
 Capable of replicating independently of bacterial
chromosome
 They can be integrated in bacterial chromosome
 Occur in both gram-negative and gram-positives
 Transmissible plasmids
 Can be transferred from cell to
cell by conjugation
 Contain a number of genes (some
responsible for synthesis of the sex
pilus and the rest for the protein to
be transferred
 Non transmissible plasmids
They are smaller
They do not contain the transfer genes
 Specialized structures outside the cell
wall

1. Capsule
 - Gelatinous layer covering entire bacteria
 - Composed of polysaccharide, EXCEPT
anthrax bacillus

 (polymerized D-glutamic acid)

 Capsule and its importance
1. Specific identification of organism
2. Determines bacterial virulence (capsule limits
phagocytosis)
3. Capsular polysaccharides can be used in vaccines
as antigens
4. May play a role in bacterial adherence to human
tissue- an important initial step to infection
 2.Flagella
 Movement – towards nutrients and
other attractants (chemotaxis)
 Some have one flagella, some have
more than one
 Some have polar flagella, some
have flagella all over the entire
bacterial surface
 3. Pili (Fimbriae)
Hair like filaments extending to the
outer cell surface
1. Mediate attachment of bacteria to
specific receptors in the human cells
(initiation of infection).
2. Specialized pilus (Sex pilus), attaches
male (donor) and female (recipient)
during conjugation
 4. Glycocalyx
 Slime layer covers bacterial surface
like a film secreted by many
bacteria
 Allows bacteria to adhere firmly to
various structures
 e.g skin, heart valve, catheters,
surface of teeth
 (in Streptococcus mutans it plays an
important role in plaque formation)
 4.Spores
 Highly resistant structures formed
during conditions of nutritional
deprivation
 Formed by two genera of medical
importance (Bacillus & Clostridium)
 Spores can stay dormant for many
years
 Spores are extremely resistant to
heat and chemicals
 Highly resistant to many disinfectants
 Sterilization can not be achieved by
boiling
 Steam under pressure (autoclaving)
at 1210C, for 30 min
Bacteria growth
Growth phases
• When bacterium is inoculated into suitable media and incubated at a
suitable temp.and pH
• The culture passes through 4 different phases of growth
 Growth phases
Four major phases:
1. Lag phase
Vigorous metabolic activity
occurs, no cells divide.

Can last few minutes up to hours

1-4 hrs
Necessary enzymes and metabolic intermediates
are synthesized
2. Log (logarithmic) or
exponential phase

Rapid cell division

(Active growth)
3.Stationary phase
Death = Growth rate
Occurs when there is nutrient
depletion, production of toxic by
products
Bacterial growth slows down until
number of cells produced equal
number of cells that die, resulting
in a steady state
4. Death phase
marked decline in number of
viable cells
 Factors influencing bacterial growth
◦ Physical factors
 Temperature -growth best at body temp 37 C
 pH-7.2-7.6
 Oxygen
 Osmosis
◦ Nutritional factors
 Carbon Source
 Energy source
 Organic compounds
 Nitrogen Source
 Minerals
 Water- vehicle for entry of nutrients, metabolic reactions and
elimination of waste products
 Normal body flora

• Normal flora are the bacteria and microorganisms that are present
and normally live in certain areas of the body, such as the:
• skin,
• mouth,
• digestive tract,
• reproductive tract and respiratory tract.
• Some normal flora are important to certain body processes, such as
digestion.
• Other areas/organs of the body are sterile
• Bladder,
• kidneys,
• liver,
• spleen,
• adrenal glands,
• thyroid gland,
• parathyroid glands
• and brain
 Types of normal flora

• 1 The resident flora: this consist of relatively fixed types of

microorganism regularly found in a given area at a given age.
If disturbed it promptly reestablish itself
• 2 The transient flora: This consist of non pathogenic or
potentially pathogenic microorganisms that inhabit the skin
and mucous membrane for hours, days or ,weeks ,it is
derived from the environment ,does not produce diseases
and does not establish itself permanently on the surface. If
disturbed members of transient flora may
colonize ,proliferate and may produce diseases.
ROLE OF THE MICROBIAL NORMAL FLORA

• 1.Members of the resident flora in the intestinal tract

synthesize vitamin K and aid in the absorption of nutrients
2.On mucous membranes and skin, the resident flora may
prevent colonization by pathogens and possible disease
through “bacterial”
3.Vaginal lactobacilli maintain an acid environment which suppresses
growth of other organisms
4. Antigenic stimulation provided by intestinal flora help to ensure the
normal development of the immune system
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Normal flora at different areas in the
body
• Skin eg Staphylococcus epidemidis, Staphylococcus aureus(in small
no),Micrococcus spss, alpha hemolytic and non hemolytic Streptococci .etc
• Nasopharynx eg any amount of the following :Diphtheroids, non
pathogenic Neisseria spp, Alpha hemolytic Streptococol, S.epidemidis, non
hemolytic Streptococci etc
• GIT and rectum
eg various Enterobacteriacea except Salmonella, Shiggella,yersinia Vibrion
and Camplobactor .
Non dextrose fermenting gram negative rods,Enterococci,Alpha hemolytic
and non hemolytic Streptococci,Diphtheroids,…
• Genitalia
eg Corynebacteria species, Lactobacilus,alpha hemolytic and non
hemolytic Streptococci,non pathogenic Neisseria spp, plenty of
anaerobes, Enterococci,Enterobacteriacea,and other gram negative
rods, S.epidemidis,Candida albicans and other yeasts
factors that may be important in eliminating nonresident
microorganisms from the skin are
• the low pH,
• the fatty acids in sebaceous secretions, and
• the presence of lysozme.

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NORMAL FLORAL OF THE INTESTINAL TRACT

• At birth the intestine is sterile, but organisms are soon

introduced with food
• In breast-fed children, the intestine contains large
numbers of lactic acid streptococci and lactobacilli.
There aerobic and anaerobic, gram-positive, nonmotile
organisms (e.g., Bifida bacterium produce acid from
carbohydrates and tolerate pH 5.0.
• In bottle-fed children, more mixed flora exists in the
bowel, and lactobacilli are less prominent

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• As food habits develop toward the adult pattern, the
bowel flora changes. Diet has a marked influence on
the relative composition of the intestinal and fecal
flora.
• In the normal adult, the esophagus contains
microorganisms arriving with saliva and food. The
stomach’s acidity keeps the number of microorganisms
at a minimum (103 – 105/g of contents) unless
obstruction at the pylorus favors the proliferation of
gram-positive cocci and bacilli
• The normal acid pH of the stomach markedly protects
against infection with some enteric pathogens, e.g.,
cholera. . As the pH of intestinal contents becomes
alkaline, the resident flora gradually increases.
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• In the adult duodenum, there are 103-106 bacterial per gram of
contents; in the jejunum and ileum 105-108 bacteria per gram; and
in the cecum and transverse colon, 108-1010 bacteria per gram.
• In the upper intestine, lactobacilli and enterococci predominate,
but in the lower ileum and rectum, there are about 1011 bacteria
per gram of contents, constituting 10-30% of the facel mass. In
diarrhea, the bacterial content may diminish greatly, whereas in
intestinal stasis the count rises.
• 1-4% are facultative aerobes (gram-negative coliform bacteria,
enterococci, and small numbers of proteus, Pseudomonas,
lactobacilli, Candida, and other organisms).
• Intestinal bacteria are important in synthesis of vitamin K,
conversion of bile pigments and bile acids, absorption of nutrients
and breakdown products, and antagonism to microbial pathogens

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NORMAL FLORA OF THE URETHRA
The anterior urethra of both sexes contains small
numbers of the same types of organisms found on the
skin and perineum. In normal voided urine these
organisms appear about 100_10,000per ml

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NORMAL FLORAL OF THE VAGINA

• Soon after birth, aerobic lactobacilli appear in the

vagina and persist as long as the pH remains acid
(several weeks). When the pH becomes neutral
(remaining so until puberty), a mixed flora of cocci and
bacilli is present
• At puberty, aerobic and anaerobic lactobacilli reappear
in large numbers and contribute to the main tenance of
acid pH through the production of acid from
carbohydrates, particularly glycogen. This appears to be
an important mechanism in preventing the
establishment of other, possibly harmful
microorganisms in the vagina
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• After menopause, lactobacilli again diminish in number
and a mixed flora returns. The normal vaginal flora
often includes also group B hemolytic streptococci,
anaerobic streptococci (peptostreptococci), Prevotella
species, clostridia, Gardenerella virginals, Urea plasma
urealyticum, and sometimes Listeria or Mobiluncus
species.

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 microbial and host factors leading
to pathogenicity of normal flora

• If forcefully removed from the restrictions of that

environment and introduced into the blood stream or
tissues, these organisms may become pathogenic.

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cont
• Bacteroides are the commonest resident bacteria of the
large intestine and are quite harmless in that location.
• If introduced into the free peritoneal cavity or into pelvic
tissues along with other bacteria as a result of trauma,
they cause suppuration and bacteremia .
• Members of normal flora are important causes of hospital acquired
infection when patients are exposed to invasive treatments.
• Patients suffering burns are at risk.

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References
• Jawetz,Melnick and Adelbergs Medical Microbiology23rd edition
• Medical microbiology by Cedric Mims et al Third edition