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Course No. VMI- 211: Introductory Veterinary Microbiology (Credit hours: 2+1=3)

Theory
 Microbiology of unicellular organisms and their classification.
 Microbiology and structure of bacteria.
 Shape, size and arrangement of bacteria.
 Microbiological variations and classification of bacteria.
 Important bacterial, viral and fungal disease of animal.
 Source of infections. Methods of transmission of infections.
 Sterilization, disinfection, evaluation of disinfectants and antiseptics. Aseptic
handling of sterilization materials; disinfection of animals.
 Introduction, morphology, growth, nutrition, reproductive and classification of fungi.
 Classification, cultivation and replication of viruses.

Practical
 Microscopy and routines.
 Staining (simple & Grams), Acid fast.
 Lactophenol cotton blue
 Special staining : leishmenn, methylene blue staining.
 Glassware preparation.
 Sterilization.
 Evaluation of disinfectants, asepsis etc.
 Preparation of reagents media,
 Demonstration: Equipment and sterilization disinfection.
 Cultural characters
 Pathogenicity test and antibiogram
 Slide culture technique for fungus

Complied by: Dr. A .I. Dadawala & Dr. F .M. Parth

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Chapter 1:- Microbiology of unicellular organisms and their

classification
“Microbiology is a branch of biological science in which microorganisms are studied i.e bacteria,
virus, yeast and mold”.
 The morphology, chemical composition, physiology, nutrition and reproduction of these
organisms are studied.
 The disease produced by these organisms in man and animals are also studied.

What is a microorganism?
Microorganisms are the subject study of microbiology, a branch of science. A
microorganism can be one cell or a cluster of cells that can only seen by using microscope.
Microorganisms are organized into five fields of study: bacteriology, virology, mycology,
phycology, protozoology and parasitiology.

Pathogenic microorganism
Those microorganisms which can cause disease in animals or humans are referred to
as pathogenic microorganisms.

Non-Pathogenic microorganism
Most microorganisms found in nature are not harmful to humans, animals or plants.
Indeed, many bacteria and fungi make an important contribution to biological activities
which take place in soil, in water and in the alimentary tract of animals. For example, flora
are microorganisms found in our intestine that assist in the digestion of food and play critical
role in the formation of vitamins such as vitamin B and vitamin K. They help by breaking
down large molecules into smaller ones.

Bacteriology
Bacteriology is the study of bacteria.
 Bacteria are prokaryotic organisms. A prokaryotic organism is a one-celled organism
that does not have a true nucleus.
 Many bacteria absorb nutrients from their environment and some make their own
nutrients by photosynthesis or other synthetic processes.
 Some bacteria can move freely in their environment while others are stationary.
 Bacteria occupy space on land and can live in aquatic environment and in decaying
matter.
 They can even cause disease. Bacillus anthracis is a good example. It is the
bacterium that causes anthrax.

Virology
Virology is the study of viruses.
 A virus is a submicroscopic, parasitic entity composed of nucleic acid core
surrounded by a protein coat. Parasitic means that a virus receives food and shelter
from another organism and is not divided into cells. An example of a Virus is the
varicella-zoster virus, which is the virus that causes chickenpox in humans.

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Mycology
Mycology is the study of fungi.
 A fungus is eukaryotic organism, often microscopic, that absorbs nutrients from its
external environment. Fungi are not photosynthetic.
 A eukaryotic microorganism is a microorganism whose cells have a nucleus,
cytoplasm and organelles. These include yeast and some molds. Tinea pedis, better
known as athlete‟s foot, is caused by a fungus.

Importance of Microorganisms

Microbes, even though of very small size, they are very important. They take part in many natural
processes and help men and animal.
1. They utilize dead organic matter viz. dead animals and decaying plants and produce carbon,
nitrogen, sulphur etc. so recycling of natural elements are possible without any interruption.
2. Some bacteria fix the nitrogen from the atmosphere into root tubercle of green plants, which
use this utilizable nitrogen for their metabolism and synthesis of food.
3. Bacteria (and protozoa) are present in rumen of animals. Through their enzymes, they are
useful in the digestion of cellulose of grass. By this way ruminants can get energy from grass
due to cellulose digestion.
4. Bacteria and yeast take part in many chemical reactions of various industrial processes. They
ferment sugar and produce ethyl alcohol or organic acids. They are responsible for conversion
of milk into milk products like curd, cheese and yogurt.
5. The antibiotics, which are very useful in therapeutic purpose, viz. streptomycin, Tetracycline,
Erythromycin and Chloromycetin are produced from bacteria while penicillin produced from
fungi.

History of Microbiology

 During 17th to 18th centuries, most important facts related to microbiology were discovered.
 Antony Van Leeuwenhoek: In 1683, he developed simple microscope and examined the
material collected from the teeth under it. He observed small sized organisms and named
them „Animalcules‟. In 1920, a compound microscope was prepared with 900 times
magnification of the organism.
 Louis Pasteur: Father of Microbiology: He worked on microbial fermentation. During this
work, He observed that there is relationship between microorganisms and chemical changes
took place in organic infusion. In 1857, he proved the formation of lactic acid due to
fermentative actions of bacteria in sugar solution. He confirmed that production of acetic acid
and butyric acid in sugar solution due to fermentative actions of specific microorganisms.
 Robert Koch: Father of Bacteriology. In1876, He clearly demonstrated that bacteria
produce the disease. He confirmed that Bacillus anthracis was the cause of Anthrax in sheep.
A few years earlier i.e between 1863 to 1868, Davine demonstrated rod shaped object in
blood and organs of animals that died due to anthrax. Koch cultivated the anthrax organisms
taken from spleen of diseased animal in serum media. He observed three different forms of
organism. In Impression smear, it looks like rod shaped, in culture it seems chain form and in
outside the living conditions, it is spore form. Koch injected anthrax culture to experimental
animal and it died showing typical symptoms and anthrax organisms were found in the
blood and organs. This experiment provides the relationship between organisms and disease
and genesis of Koch‟s postulates.

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After the discovery of anthrax organisms by Koch in 1876, most of other bacteria causing
important human diseases like plague, typhoid, cholera etc. were discovered by various
scientist. Due to landmark research in Bacterial biology, Robert Koch is considered as the
“father of bacteriology”.
 Edward Jenner: Father of Immunology. First time introduced successful immunization
against small pox by using related but mild live virus of cow pox.
 Karl Landsteiner: Discovered Human Blood group A, B, AB and O groups.
 Kohler & Mailstein: Developed Hybridoma Technique.
 Ruska & Knoll: Discover Electron Microscope.
 James Phipps: First Human Guinea Pig.
 Paul Ehrlich: German Scientist known as Father of Chemotherapy. He proposed Side
Chain Theory. He also reported acid Fast Stain.

BRANCHES OF MICROBILOGY

Microbiology is a wide subject. It contain different branches due to various type of bacteria
found in diverse habitats like food, milk, water, soil, plant, sewage etc.
1. General bacteriology: Study of the general characteristic like morphology, metabolism,
nutrition, physiology and reproduction of bacteria.
2. Systemic bacteriology: Classification and arrangement of bacteria into various orders,
families, genera and species as per their characters.
3. Industrial Microbiology: Study of industrial use of bacterial metabolic activity like in
alcoholic fermentation in wine industry, manufacture of organic acid, tanning of leather,
curing of tobacco, silage production and manufacture of antibiotics.
4. Agricultural bacteriology: Study of bacteria related with agriculture. It is further divided
into soil bacteriology and plant bacteriology. Soil bacteriology is the study of bacteria present
in soil, which are important in maintaining fertility of soil. Plant bacteriology is the study of
bacteria on plants.
5. Food Bacteriology: Study of bacterial actions and its outcomes in food industry viz. food
preservation by canning.
6. Dairy Microbiology: Study of usefulness of the bacteria in the preparation of functional
fermentative dairy products. This subject also includes bacteria, which are harmful as well as
causing spoilage of dairy products.
7. Sanitary bacteriology: Study of bacteria, which is injurious to health of man and animals
and causing diseases.
8. Medical Microbiology: Study of bacteria , virus ,yeast and fungi injurious to health of man
and animals and which disease in them .( Human and veterinary Microbiology)

NAMING AND CLASSIFYING MICROORGANISMS

Carl Linnaeus developed the system for naming organisms in 1735. This system is referred
to as binominal nomenclature. Each organism is assigned two Latinized name because Latin or
Greek was the traditional language used by scholars. The first name is called the genus. The second
name is called specific epithet, which is the name of the species, and it is not capitalized. The genus
and the epithet appear italicized.
Sometimes an organismis named after a researcher, as in the case with Escherichia coli,
better known as (E. coli.). The genus is Escherichia, which is named after Theodor Escherich, a
leading microbiologist. The epithet or species is coli, which implies that the bacterium lives in the
colon.

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Organisms were classified into either the animal kingdom or the plant kingdom before the
scientific community discovered microorganisms in the seventeenth century. It was at that time
when scientist realized that this classification system was no longer valid.

 Carl Woese developed a new classification system that arranged organisms according to their
molecular characteristics and then cellular characteristics. However, it wasn‟t until 1978 when
scientists could agree on the new system for classifying organism, and it took 12 years after
this arrangement before the new system was published.
 Woese devised three classification groups called domain. A domain is larger than a kingdom.
These are:

Domains
1. Eubacteria: Bacteria that have peptidoglycan cell walls. (Peptidoglycan is the molecular
structure of the cell walls of eubacteria which consists of N-acetyglucosamine, N-
acetylmuramic acid, tetrapeptide, side chain and murein.)
2. Archaea: Prokaryotes that do not have peptidoglycan cell walls.
3. Eucarya: Organisms from the following kingdoms:
4. Kingdoms
 Protista: Examples- Algae, protozoa, slime, molds.
 Fungi: Examples- one-celled yeasts, multicellular molds and mushrooms.
 Plantae: Examples- moss, conifers, ferns, flowering plant, algae.
 Animalia: Examples- insects, worms, sponges and vertebrates.
There are five kingdoms of living things.

KINGDOM CHARACTERISTICS EXAMPLE

Monera Prokaryocyte Bacteria

Protista Eukaryocyte Protozoa

Fungi Eukaryocyte Fungi
Plantae Eukaryocyte Plants
Moss
Animalia Eukaryocyte Arthropods, Mammals & Man

Size of microorganism
Microorganisms are measured using the metric system. In order to give you some idea of the
size of a microorganism, let‟s compare a microorganism to things that are familiar to you.
A human red blood cell:- 100 micrometers (μm)
A typical bacterium cell:- 10 micrometers (μm)
A virus: - 10 nanometers (nm)
An atom: - 0.1 nanometers (nm)
Exercise:-

1. Explain branch/ scope of Microbiology.

2. Explain contribution of Louise Pasture/ Robert Koch.
3. Define:- 1. Microbiology 2. Veterinary Microbiology 3. Bacteriology 4. Virology 5. Mycology.
4. Explain Importance of Microorganisms.
5. Write in details about naming and classifying microorganisms.

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Chapter 2:- Microbiology and structure of bacteria

Morphology of Bacterial Cell
Microorganisms are divided in two classes, prokaryotic and eukaryotic. Prokaryotic organisms
are simple while eukaryotic organisms are more advanced. The principal differences between two are
as follows.
Prokaryotic Eukaryotic
Bacteria and virus Fungi and protozoa
No organized nucleus True organized nucleus
Nuclear membrane absent Nuclear membrane Present
Nucleolus absent Nucleolus absent or Present
ATP generating enzyme presents in Mitochondria function as ATP generating
Cytoplasmic membrane molecule
Reproduction only asexual By sexual and asexual

Cell: Cell is structural and functional unit of living organism. Cell consists of protoplasm, which is
enclosed by cytoplasmic membrane and then cell wall.

Bacteria: They are the microorganisms with prokaryotic cellular organization. They are unicellular
and seen as single organisms but may remain attached with each other to from clusters or chains.

Compositions of Bacterial cell:

Mainly composed of two: water (75%) and 25% dry matter. Apart from this, dry matter
consist organic (90%) and inorganic dry matter (10%).
 T.B. organisms contain very high lipid i.e up to 35% of total organic dry matter.

75% water Inorganic acid (10%)

Bacterial Cell
Composition

25% Dry Matter Organic acid (90%)

including CHO (20%),
Protein (65%), Fat
(4%), Minerals (6%)

Morphological, Structures in bacteria in Cell

The bacteria cell has two types of structure viz , Essential structures and accessory
structures.
Prokaryotic Eukaryotic
Bacteria and virus Fungi and protozoa
No organized nucleus True organized nucleus
Nuclear membrane absent Nuclear membrane Present
Nucleolus absent Nucleolus absent or Present
ATP generating enzyme presents in cytoplasmic
Mitochondria function as ATP generating
membrane molecule
Reproduction only asexual By sexual and asexual
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Essential Structure Accessory structures

a. They are present in all type a. They are not present in all type
of bacteria . of bacteria
b. Bacteria cannot live without b. Bacteria can live without it.
it. Without it life of organisms
not possible.

Essential Structure – their properties and Function

Each structure has got important function in the metabolism of bacteria.

A. Protoplasm: It is vicious; jelly like material, Protoplasm contains nucleus, Ribosomes,
Mesosomes and cytoplasmic inclusions. Protoplasm is responsible for the metabolic activities
of cell like energy and nucleic acids.
B. Nucleus: The nucleus of Bacteria is also Known as nuclear body or chromatin body. The
nuclear material is dispersed in cell as double stranded DNA. The function of nucleus is to
transmit the hereditary characters to the progeny.
C. Ribosomes: Ribosomes are spherical granules and protein synthesis is takes place in
ribosomes.
D. Mesosomes: They are converted in shape and attached to cytoplasmic membrane. The
function of Mesosomes is to provide the site of attachment for bacterial chromosomes on
cytoplasmic membrane at the time of cell division
E. Cytoplasmic membrane: It is also known as membrane. Cytoplasmic membrane covers the
protoplasm and cell wall is the outermost cover. Cytoplasmic membrane is very thin, elastic
and semi permeable membrane which cannot seen with ordinary microscope but visible under
electron microscope. This membrane is made up of lipoproteins.
Function
1) The Cytoplasmic membrane is semi permeable and it controls the entry of the
nutrients and exit of waste products.
2) The enzymes of TCA cycles are present in the membrane.
3) If the membrane damaged then there is death of cell but the membrane is protected by
cell wall.
F. Cell Wall: The cell Wall protects the living matter i.e protoplasm and also protects the
cytoplasmic membrane. It is made up of polysaccharide and protein. The cell wall is strong
and rigid. It is also porous and so allows inside the bacteria macro molecules of nutrients.

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Function:
1. It is maintain the shape of bacteria.
2. It protects the bacteria from outside pressure , because if cell wall break water enter
in side ,protoplasm swell, cytoplasmic membrane bursts leading to lysis and death of
bacteria .
3. Cell wall also protects from cytoplasmic membrane against internal osmotic pressure.
4. Cell wall takes part in division of bacteria and two daughter cells are formed by
splitting of cell wall.

Accessory structures their properties and functions

In some bacteria special structure are present. This structures have some function in the
bacteria cell .More over due to presence this structure in the bacteria cell we can identify sic
bacteria.

A. Capsule: some bacteria are surrounded by covering known as capsule. Capsule is about 0.2
micron wide and consists mainly of water and has 2% wide solid (polysaccharides) and
(protein). Capsule can be seen by ordinary microscope by special staining method Hiss
Copper Stain and LAMB (Loffler’s Alkaline Methylene Blue) stain. This staining used for
the identification of various bacteria like Bacillus, Klebseilla and Pneumococci
Function:
 Capsule protect bacteria against phagocytosis because due to capsule, the
phagocytosis are repelled and cannot ingest capsulate bacteria.
 The capsule of Pneumococci has got different type of polysaccharide and
classification of pneumococci is done on type specific polysaccharide.
 In the diagnosis of the disease like anthrax, presence of capsule in blood smear on
the suspected animal is useful for diagnosis.
B. Flagella: Certain bacteria possess filamentous structure known as flagella are thin long, wavy
filamentous. They are so thin that they can be seen by microscope only after special
staining method, Leifsons stain which increase the thickness of flagella. The origin of
flagella is from protoplasm and then come out through cell wall. Flagella are the organ of
locomotion and motility of bacteria is due to flagella.
 Bacteria may have single flagellum or many flagellum and are either at poles or
surrounding the bacteria. According to the number and position of flagella they are
classified as follows.
Atrichous No flagella -----
Monotrichous One flagellum at one poll Eg. Vibrio cholerae
Amphitrichouos Flagella at both poles Eg. Alcaligenes faecalis
Lophotrichous A group of flagella at one pole Eg. Spirilla
Peritrichous Flagella surrounding bacteria Eg. Salmonella typhi

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Functions:
a. Motile bacteria can move and go to area where more nutrition
b. Motility may also help pathogenic bacteria in penetration of mucus secretions
and help in spreading through the body.

C. Fimbria: Some organisms possess filament like appendages known as Fimbria. Fimbrias are
shorter and not curved but straight. Their number is much more than flagella .They may act
as organ of adhesion.
D. Spore: Two bacterial organisms viz, Bacillus and clostridium produce highly resistang
structure, called spore or endospore. Due to spore, the organisms can survive in a dormant
state for long period of starvation or other unfavorable environments conditions.
 Demonstration of spore : The spore is highly refractile structure within the bacteria seen
in a slide stained by simple staining method like Gram‟s method. Special staining method
Schaeffer & Fulton is used for spore‟s identification.
 Position and shape of spore: The position and shape of spore is useful in identification of
bacteria.
A. No bulging: In Bacillus species the spores do not produce bulging of cell wall.
B. Bulging of cell wall : In clostridium species the spores are wider than the
bacteria and their is bulging of cell wall. The position and shape are as follows
 Oval, sub terminal ( snow shoe) Cl.septicum
 Oval, terminal ( Tennis racket) Cl.botulinum
 Spherical, terminal ( Drum stick) Cl.tetani
 Oval, central navicular or centron Cl.chauvoei
 Structure of spore: The structure of spore is complex, which consists of many layers. The
spore cytoplasm is surrounded by membrane and spore cortex. Then there is outer membrane,
spore coat and exosporium. Due to so many layers the living matter within is protected and
spore is resistant due to these multiple layers.
 Sporulation / Sporogenesis: It is a process in which the bacteria produces spore. So
sporulation is a formation of spore from vegetative cell. It is also called sporogenesis. The
bacteria produce spore under unfavorable conditions like
1. Accumulation of waste products.
2. Change in ph of culture media
3. Decrease or increase of oxygen
4. Exhaustion of nutrients.

Functions of spore:
1. As spores are resistant to heat, starvation, drying, chemicals and disinfectants, they can
survive during the crisis period and thus help bacteria to overcome such crisis again under
favorable conditions spore germinates to vegetative bacteria.

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2. Due to particular shape and position spores in the bacterium, you can identify spore bearing
bacilli in the smears from diseased animals. Thus spore helps in diagnosis of disease like
tetanus and black quarter.

Exercise:-

1. Write in detail Morphological structure of bacteria with suitable diagram.

2. Differentiate between Essential & accessory structure of bacteria.
3. Differentiate between Prokaryotic and Eukaryotic cell.

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Chapter 3:- Shape, size and arrangement of bacteria

Shape, size, arrangements and staining of bacteria:

The bacteria are having different shapes. They have fixed size. In some bacteria, after cell
division, they do not separate but remain attached and produce characteristics cell arrangements. All
these morphological characters (shape, size, arrangements) are useful in identification of bacteria in
stained smear.

1. Cocci: sThe organisms are roughly spherical or round in shape. The size is measured in
micron (µ). The diameter of cocci is about 1µ. The cocci organisms, after multiplication, do
not separate and show different types of cellular arrangement.
 Diplococci: cocci in pairs of two. Eg. Neisseria,
 Tetrad: cocci in pocket of fours.
 Sarcina: cocci in cubical pocket of eight.
 Streptococci: cocci in long chains (5 to 6 bacteria).
 Staphylococci: cocci in cluster just like bunch of grapes.

2. Bacilli: Here, the shape is rectangular and organisms have both, length and width. There is
variation in size of various organisms and length varies from 3 to 8 µ.
 Single cells: Bacteria after division separates and seen in the smear as single organisms.
Eg. E.coli,
 Chains: Bacteria form long chains and look like bamboo stick Eg. Bacillus anthracis,
 Bundles: Bacteria are seen in irregular as bundles and are forming acute angles with each
other. Also Known as Chinese letter like arrangement. Eg. Corynebacterium
3. Curved or comma: The bacteria are even though bacillary in shape but slightly curved just
like comma. The size 3µ. Eg. Campylobacter organisms.

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4. Spiral: The spiral organisms are having coils just like screw and length is from 5 to 15 µ. Eg.
Borellia, Leptospira.
5. Filament: The organisms are very slender and long filaments i.e. thread like Eg.
Actinomyces.

 Pleomorphism: Bacteria show considerable variation in size, shape–swollen, spherical,

elongated, pear shaped (most readily seen in strepto bacillus, monilfomis, Yersinia pestis) in
presence of penicillum, glycine. Abnormal cells are recognized as degenerate or involution
forms. Some might be grows and revert to normal form in suitable condition due to defective
cell wall synthesis.
 Spheroblast: If cell wall would be removed/weakened from bacterial cell (while keeping) in
0.2 to 1 M sucrose, magnesium to prevent osmosis & lysis. Organism converted in viable
spherical bodies called spheroblast.
 Free protoplast: If cell wall is removed completely. If cell walls remain enclosed by intact
but residing cell wall it is called Free protoplast.
 L Forms (of bacteria) are abnormal growth that may arise spontaneously or by inhibiting cell
wall synthesis in bacteria of normal morphology. They are stable form and continue to
reproduce as L forms. They differ from parent bacteria in lacking rigid cell wall. Further
onwards, they are lacking in cell wall.
 But unlike protoplast, L form bacteria are capable of growing and multiplying to
produce colonies in fried egg. L form are non pathogenic.
 Temporary Morphological variation
 Size: Old culture E.coli.– Very short bacilli, while in Fresh sub culture: long bacilli seen
again,
 Stain: Old culture of gram positive organism become gram negative,
 Loss of accessory structure – B.anthracis contain capsule when grow in body, while in
culture, they are non-capsulated.

Exercise:-

1. Write in detail size, shape and arrangement of bacteria with suitable diagram.

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Chapter 4:- Microbiological variations and classification of

bacteria
FACTORS AFFECTING THE GROWTH OF MICROORGANISMS

The different types of microorganisms requires many different conditions for growth and
metabolism so as to obtain energy and for building of cellular materials.
The required factors are as follows:
 Moisture and dryness
 Temperature
 Nutrition
 Requirement of gases (Oxygen and carbon dioxide)
 Hydrogen ion concentration –pH
 Light
 Osmotic pressure
 Moisture and dryness: The protoplasm of bacteria contains 70 to 75% water so water is very
important for growth .All the culture media require for cultivation of bacteria are in water .
Some organisms like mycobacterium can grow on dry media also. Spores survive for many
years in dry condition
 Temperature : Different type of organisms have a definite growth range and that range
growth takes place .According to temperature requirement the organisms are divided in to
i. Psychrophiles organisms can grow at lower temperature is 0 to 30 Celsius. Some can
grow at -7 Celsius. Generally such organisms are present in snow, cold water and soil
psychrophillic organisms like achromobacter can cause the cause the spoil of
refrigerated food or food kept in cold storage
ii. Mesophillic organisms prefer somewhat higher temperature and range is 25 Celsius.
Pathogenic organisms which grow best at body temperature i.e 35.5 Celsius are
mesophilic organisms. Sapropheytes like pseudomonas aeruginosa prefer room
temperature i.e 18-25celcius.
a. Thermophilic organisms: These organisms like high temperature 50-90celcius .

Group Temperature Types

Minimum Optimum Maximum
Pyschrophiles 0 15-20 30 Water bacteria cold storage
Mesophiles 15-20 37 43 Most pathhogens
Thermophiles 25-45 50-55 85 Soil, water & hot springs

 Nutrition: Following nutrient substances are required for growth of bacteria (i) Carbohydrate
as energy source (ii) protein as nitrogen source (iii)Minerals (iv) vitamins. Chemical changes
produced by bacteria upon nutrients to obtain energy is called metabolism. The chemical activity
of bacteria may be helpful or harmful to animals. In industry, bacteria metabolism helps in
production of ethyl alcohol, lactic acid, antibiotics, enzymes and vitamins, etc. While in disease
like tetanus, bacteria produce toxin during their metabolism, which may harmful. According to
metabolism requirements, bacteria are divided in to autotrophic bacteria.

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 Autotrophic bacteria: They obtain their nutritional requirements from inorganic sources with
enzymes. The energy for metabolism is supplied in the form of light in photosynthetic
bacteria or by oxidation of inorganic compounds in chemo synthetic bacteria .

Classification of pathogenic bacteria

The kinds of organisms found in a given environment and the rates at which
they grow can be influenced by a variety of factors, both physical and biochemical. Physical
factors include pH, temperature, oxygen concentration, moisture, hydrostatic pressure,
osmotic pressure, and radiation. Nutritional (biochemical) factors include availability of
carbon, nitrogen, sulfur, phosphorus, trace elements, and, in some cases, vitamins. Based on
the biochemical and physical factors influencing their growth, bacteria are classified as
follows:

a) Based on pH requirements
pH: The pH scale is a measure of hydrogen ion (H+) concentration. Low pH corresponds
with high concentrations of hydrogen ion, neutral pH with equal numbers of hydrogen and
hydroxyl ions (OH-), and high pHs correspond to low concentrations of hydrogen ion
I. Optimum pH
Optimum pH is that pH at which a given organism grows best. The range over
which most organisms can grow tends to vary over no more than a single pH unit
in either direction (e.g., from pH 6 to pH 8 for an organism whose pH optimum is
pH 7)
II. Acidophiles
Organisms whose optimum pH is relatively to highly acidic, which means
growing best in acidic pH
III. Neutrophiles
Organisms whose optimum pH ranges about pH 7, plus or minus approximately
1.5 pH units
IV. Alkaphiles
Organisms whose optimum pH is relatively to highly basic, which grow best in
high pH
b) Based on temperature requirement
I. Optimum temperature
Optimum temperature is the temperature at which an organism grows best.
Typically the range in temperature over which a bacterium can grow is about
30ºC.
II. Psychrophile
Cold-adapted organisms are called psychrophiles. The cut-off temperature for a
psychrophile is a 20ºC or colder. Psychrophiles may additionally be termed
obligate or facultative with obligate psychrophiles unable to grow above 20ºC, but
facultative psychrophiles are able to grow above 20ºC.
III. Mesophile
Organisms whose optimum growth temperature is found between 20ºC to 40ºC
are termed mesophiles. Human and animal pathogens, which must be able to grow
at the approximately 37ºC body temperature, are mesophiles
IV. Thermoduric
Mesophilic organisms that can endure brief exposures to relatively high
temperatures are termed thermoduric These are one category of the organisms that

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survive following inadequate heating of foods and may thereby contribute to the
spoilage of foods that have been heated (e.g pasteurization) to kill microorganisms
V. Thermophil
High-temperature-adapted organisms are called thermophiles. Examples organism
that groin hot springs.

c) Based on oxygen requirements

Organisms differ in their requirements of molecular oxygen (i.e., O2) as well as other
atmospheric gasses (e.g., carbon dioxide). Categories of organisms as per their oxygen
requirements include:
I. Obligate aerobe
Organisms that are unable to grow in the absence of oxygen or they require
oxygen for their growth. Sometimes this group of organism may be called strict
aerobes as they cannot grow without oxygen.
II. Facultative aerobe
Organisms that can grow in the absence normal level of oxygen that is otherwise
required.
III. Microaerophile
These are organisms that grow best when small amounts of oxygen are present
That is, less than atmospheric concentrations, but more than those concentrations
tolerable by obligate anaerobes
IV. Obligate anaerobe
Organisms that are strict anaerobe and require complete absence of oxygen for
their growth, which mean they cannot grow in the presence of oxygen.
d) Osmotic pressure
The concentration of dissolved substances in the environment can impact on
the growth and survival of bacterial cells.

I. Plasmolysis
Environments containing large concentrations of dissolved substances draw water
out of cells, causing shrinkage of the cytoplasm volume, a phenomenon termed
plasmolysis. Plasmolysis interferes with growth and this is why highly osmotic
environments prevent bacterial growth (e.g., brine, the high sugar concentrations
in jellies and jams, salting of meats)
II. Halophiles
Organisms that require high concentrations of dissolved salts to grow are termed
halophiles. Depending on organism, the salt concentrations required range from
those of seawater on up to those of brine
e) Based on staining
Cell shape Characteristics Genus Family
Gram negative bacteria
Cocci Aerobic Neisseria Neisseriaceae
Veilonella
Coccobacilli - Brucella Brucellaceae
Bordetella
Pasteurella, Mannheimia
Haemophilu s
Bacilli Facultative anaerobic, motile Escherichia, Shigella, Salmonella, Enterobacteriaceae
with peritrichous flagella or Proteus, Erwinia, Yersinia, Enterobacter,
non motile Serratia

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Aerobic, motile with Azotobacter Rhizobium Azotobacteraceae

peritrichous flagella or non
motile
Aerobic, motile with polar Nitrosomonas, Nitrobacter, Thiobacillus, Rhizobiaceae
flagella Pseudomonas, Acetobacter, Legionella
Facultative anaerobic with Campylobacter, Zymomonas, Aeromonas Nitrobacteraceae Pseudomona-
polar flagella daceae
Curved rods with polar Vibrio, Spirillum, Desulfovibrio Spirillaceae
flagella
GRAM POSITIVE BACTERIA
Cocci Cells in irregular clusters Staphylococcus, Micrococcus, Sarcina Micrococcaceae
Cells in chains Streptococcus, Leuconostoc Streptococcaceae
Bacilli Aerobic Spore forming Bacillus Bacillaeceae
Anaerobic Spore forming Clostridium
Lactic fermentation Lactobacillus Lactobacillacaeae
Propionic fermentation Propionibacterium Propioni-bacteriaceae
Oxidative, weakly Corynebacterium -
fermentative Listeria
Erysipelothrix
OTHER MAJOR GROUPS
Acid-fast - Mycobacterium Actinomycetales
rods Actinomyces, Nocardia,
Ray-forming Streptomyces
rods
Spiral motile Treponema Spirochetales
organisms Borrellia
Leptospira
Spirocheta
Small lack rigid wall Mycoplasma Mollicutes
pleomorphic
Small - Rickettsiae Rickettsiaceae Chlamydiaceae
intracellular Coxiella
parasites Chlamydia
Intracellular borderline with protozoa Bartonella Bartonellac
parasites

Exercise:-
1. Write in detail factors affecting the growth of microorganisms.
2. Write classification of bacteria based on Gram`s Staining.

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Chapter 6:- Sterilization and Disinfection

The destruction of microorganisms may be accomplished by physical and chemical means.
The use of physical means to destroy microorganisms is known as sterilization while the use of
chemicals is known as disinfection. However, this is not a set rule, for the ultimate action which
kills the organism may be the same when either a physical or a chemical method is employed.
An instrument is considered sterilized when it is free of living micro organisms.
A. Sterilization
Sterilization is most commonly done through heat.

1. Moist heat
a. Autoclave: Moist heat in the form of steam under regulated pressure is used for
sterilization. The equipment used for this purpose in the laboratory is called as
autoclave. Both pressure and temperature are applied for a definite period of time to
kill the micro organisms. In autoclave, most of the materials are sterilized under 15 lb
pressure at 121°C temperature for 15 min..
 Cotton, wools, various culture media, solution are sterilized with this
instrument.
b. Tyndallization: Tyndallization is a process of moist heat application some
materials like amino acids, sugar solutions, etc. are required to be heated at 90-
100°C temperature on three successive days with interval in between. During
this interval, resistant spore germinate and on subsequent exposure to heat, the
vegetative stage get destroyed. This method of successive at successive interval
is called as fractional sterilization or tyndallization.
c. Boiling: most of the vegetative forms of the microorganisms can be destroyed
through boiling, but spores cannot be destroyed with this method.
 Surgical instruments are sterilized by this method.
d. Pastuerization: it is the process by which food and food products are
protected from putrifaction and fermentation. . it involves a brief exposure to
heat at a lower temperature than that employed in normal sterilization. The
temperature of pasteurization is selected based on the thermal death time of
most resistant type of microorganisms to be destroyed by this process.
 Milk is generally pasteurized at 72 °C for 27 minutes or 63°C for 30
minutes, followed by rapid cooling.
2. Dry heat: it is done by the following methods.
a. Red hot: inoculating wire, forceps, spatula, etc are sterilized by heating them
red hot.
b. Flamming: Scalpel, needle, cultural tube, cotton wool plug, glass slide, etc are
sterilized by heating without allowing it to become red hot.
c. Hot air oven: heating is done by electricity above chosen temperature which
is maintained by thermostat. Here sterilization is done at 160°C for 1 hour.
 Materials sterilized by this method are glass wares, surgical
instruments, glass syringe, etc.
Precautions:
 All glass wares must be wrapped with a craft paper.
 All articles must be free from moisture.
 The whole system must be allowed to cool.

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B. Disinfection
Disinfectants: Disinfectants are agents which are too toxic to be applied to the tissues of
the host but which can be used in destroying contaminated inanimate objects. E.g., drains,
fecal matter, building, vehicles, cooking materials, surgical instruments, etc.
Disinfection is the process of freeing objects from harmful germs. Disinfectants are to
be used depending upon the nature of the organism. Some organisms are killed in acid
media and therefore acids have to be used. But most of the organisms especially the virus
like rabies, FMD virus are killed by alkali. Alkali containing pH 9 has antiviral properties
4 % washing soda is very effective against FMD virus. Soda lime is used for the disposal
of carcasses. Poultry sheds can be disinfected through fumigation (250 g KMnO4 in 500
ml of formalin).
C. Antiseptics
It is an agent that prevents sepsis i.e. prevents the growth of infective agents like
bacteria, virus, protozoa, etc.
The substances are substantially nontoxic for superficial application to living tissues.
Therefore, these agents can be applied externally on animals to kill or prevent the growth
of microbial population. The antiseptics belong to a variety of chemical substances e.g.
Alcohol, phenol, dyes, etc. They are used in the intact skin before surgical operation and
injection. They are applied to broken skin following wounds, burns, etc.

Sr. Compound Mode of action Use

No.
1. Phenolic compounds Denatures proteins, disrupts Antiseptic at low concentration
cell membrane and disinfectant at high
concentration
2. Alcohols, Ethanol, propanol Denatures proteins, Used on skin as antiseptic
solubilizes lipids
3. Chlorine compounds Oxidizing agent To disinfect liquid, surface
 Sodium hypochlorite Disrupts disulphide and other decontamination, instrument
 Calcium hypochlorite bonds disinfection.
 Sodium dichloro Chlorine dioxide –gas
isocyanate sterilization
 Chloramines
 Chlorine dioxide
4. Iodine, iodophors Inactivate proteins Antiseptic and surface
disinfection
5. Quaternary ammonium Disrupt cell membrane Surface decontamination
compounds
6. Aldehydes, Formaldehyde, Alkylating agent Cold sterilant - surface ,
glutaraldehyde instrument, glassware
7. Ethylene oxide Alkylating agent, disrupt Sterilization of heat/ moisture
tructure of proteins and sensitive supply, equipment,
nucleic acids instruments.
8. Heavy metals Coagulate proteins, react with Antiseptic
SH group of proteins
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PHENOL COEFFICIENT TEST

This value expresses the capacity of a disinfectant to kill bacteria when compared with
phenol. In the official test, a broth culture is diluted 1:10 with different concentrations of the test
compound. The end point is the lowest concentration that yields sterile loopful samples after
incubation for 10 minutes at 20°C. the compound is generally recommended for use at five times
this concentration. For example, a phenol coefficient may be stated to be 40, which means its
killing power is 40 times that of phenol. The three organisms used in the official test are
Salmonella typhy (gram- negative), staphylococcus aureus (gram positive) and pseudomonas
aeruginosa (gram- negative). Although of some value, the phenol coefficient dose not takes into
account such consideration as toxicity for tissues, in activation by organic matter. Corrosive
properties and other factors relating to particular situation.

Exercise:-

1. Explain physical method of sterilization.

2. Give name of gases used for gaseous sterilization.
3. Define. 1. Antiseptic 2. Disinfectant 3. Sterilization.
4. Give name of test used for evaluation of disinfectants.

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Chapter 7:- Mycology: Morphology, growth, nutrition,

reproductive and classification of fungi

General characteristics of Fungus

Mycology- The study of fungi.

Structural-functional relationships

The fungi are more evolutionarily advanced forms of microorganisms, as compared to the
prokaryotes (prions, viruses, bacteria). So, They are classified as eukaryotes. They containing
diploid number of chromosomes and nuclear membrane and have sterols in their plasma membrane.

 Fungi divided into two basic morphological forms: Yeasts and Mold/Hyphae.
Yeasts are unicellular fungi which reproduce asexually by blastoconidia formation (budding) or
fission. Hyphae are multi-cellular fungi which reproduce asexually and/or sexually.
 Dimorphism is the condition where by a fungus can exhibit either the yeast form or the
hyphal form, depending on growth conditions.
 Most fungi occur in the hyphae form as branching, threadlike tubular filaments.
 These filamentous structures either lack cross walls (coenocytic) or have cross walls (septate)
depending on the species.
 In some cases septate hyphae develop clamp connections at the septa which connect the
hyphal elements.
[1] Conidia: Aerial hyphae often produce asexual reproduction propagules termed
conidia (synonymous with spores).
[2] Macroconidia - Relatively large and complex conidia
[3] Micro conidia - smaller and simpler conidia.
 Endospores-When the conidia are enclosed in a sac (the sporangium), they are called
endospores.
 The presence/absence of conidia and their size, shape and location are major features used in
the laboratory to identify the species of fungus in clinical specimens.
 Mycelium- A mass of hyphal elements is termed the mycelium (synonymous with mold).

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CLASSIFICATION

The fungi pathogenic for animals and man are generally grouped according to one of two
methods:
A. based upon the area of body they infect
B. based upon the morphology of the infecting fungus

A. Based on the morphology of the infecting fungus

Morphologically the fungi causing infections in animals are classified in to

four groups.

 Yeast is the simplest form of fungi. They are ovoid or ellipsoid and unicelluar. On
cultivation in mycotic media produces colonies similar to bacteria. Temperature of
incubation is 37C. Eg: Crytococcus neoformans
 Yeast like fungi - these are fungi resembling yeast with presence of pseudohyphae.
Up on cultivation in mycotic media they produce colonies similar to bacteria with
fungal fringes. Optimun temperture is 37C. Eg: Candida albicans
 Moulds/filamentous fungi - this group of fungi are called true fungi because it
produces true filamentous hyphae with mycelia. Optimun temperature for the growth
is 22-25C. Eg: Aspergillus fumigatus
 Dimorphic fungi - one and the same fungi can exist in two different morphologies
based on the conditions of the growth. At 37C they grow as yeast/yeast like organisms
and at 22C--25C they grow as moulds. Eg: Sporothrix schenckii, Histoplasma
capsulatum
 Yeast cells are larger than bacterial cells. They vary in size ranging from 1-5 um in
breadth and 5-10 um or more in length.
 Yeast cells do not have flagella or organelle for locomotion.
 In moulds the thallus consists of the mycelium and the spores. The mycelium is a
complex of many filaments called hyphae. The hypha is 5-10 um in diameter. It is
made up of an outer tube like wall enclosing a cavity called the lumen. The lumen is
filled with protoplasm.
 A double layered membrane the plasmalamella surrounds the protoplasm and present
between the wall and the protoplasm. The hypha is divided into cells by cross walls.
They are formed by the centripetal invagination from the cell wall.
 The cross walls constrict the plasma membrane and grow in ward to form an
incomplete septum which has a central pore through that protoplasmic streaming
occurs. Even nuclei can migrate from one cell to another in the hypha.

B. Systemic classification of fungus:-

1. Phycomycetes/Zygomycotina : Nonseptate, asexual and sexual reproduction present,

Asexual reproduction by sporangia , bread mould (Rhizopus, Absidia,Mucor),
2. Ascomycetes: Septate mycelium, sexual reproduction by sac-like structure called ascus.
3. Basidiomycetes: Septate mycelium, sexual reproduction by basidium
4. Deuteromycetes (Fungi imperfecti): Septate mycelium, sexual reproduction not found

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GROWTH AND NUTRITION:-

 Fungi can withstand certain extreme conditions than other organisms. Moulds can
grow in medium containing high concentration of sugar that inhibits most bacteria.
Fungi also can tolerate more acidic conditions that other microbes. They are usually
aerobic organisms. Some yeasts are facultative. They can grow both under aerobic
and anaerobic conditions. The optimum temperature for saprophytic fungi is 22-30 C.
For pathogenic fungi it is 30-37C. Some fungi can even grow at 0 C and can cause
deterioration of meat and vegetables in cold storage.
 Fungi are heterotrophic. They can not use inorganic compounds like carbon dioxide
as their sole carbon source. Carbon must come from an organic source such as
glucose. Some fungi can use inorganic compounds of nitrogen but all fungi can use
organic nitrogen.
 Most fungi can grow aerobically in the usual bacteriological media at temperatures
ranging from 20 -30 C. Since many fungi grow more slowly compared to bacteria in
media which support growth of bacteria and fungi, fungi may be over grown by
bacterial contaminants in a mixed culture. Hence for isolation of fungi the medium
which favours their growth but not suited for bacterial growth is preferred. Acidic
media having high sugar concentration are tolerated by moulds but not bacteria.

Reproduction of Fungus

 Fungi reproduce by many ways.

Asexual reproduction

 In asexual reproduction or somatic or vegetative reproduction there are no sex cells or

organs and it does not involve union of nuclei. It can occur by
o fission of somatic cells or
o budding of somatic cells
o fragmentation or disjointing of hyphal cells
o spore formation
 Asexual spores are produced in large numbers.
 The different types of asexual spores are
o Sporangiospores – single cell spores formed within structures called sporangia
at the end of special hypha called sporangiophores. Motile sporangiospores are
called zoospores. They have flagella.
o Conidiospores or conidia – small single celled conidia are called
microconidia. Large multinucleated conidia are called macroconidia. Conidia
are formed at the tip or side of a hypha.
o Arthrospores or oidia – single celled spores formed by disjointing of hyphal
cells.
o Chlamydospores – thick walled single celled spores formed from cells of
vegetative hypha. They are resistant to adverse conditions.
o Blastospores – they are formed by budding.

 Sexual reproduction occurs by fusion of the compatible nuclei of two parent cells.
The process begins with the joining of two cells and fusion of protoplasts

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(plasmogamy). The two haploid nuclei fuse together to form a diploid nucleus. Then
meiosis occurs to reduce the number of chromosomes to the haploid number.
 The sex organs are called gametangia. They may form differentiated sex cells,
gametes or may contain instead one or more gamete nuclei. If the male and female
gametangia are morphologically different, the male gametangium is called
antheridium and the female gametangium is called the oogonium.
 The different methods of sexual reproduction are
o gamete copulation – fusion of naked gametes one or both of which are motile.
o gamete – gametangial copulation – gametangia come in to contact but do not
fuse. The male nucleus migrates through a pore or fertilization tube in to the
female gametangium.
o gametangial copulation – two gametangia or their protoplasts fuse and give
rise to a zygote that develops in to a resting spore.
o somatic copulation – fusion of somatic or vegetative cells.
o spermatization – union of a special male structure called spermatium with a
female receptive structure. The spermatium empties its contents in to the latter
during plasmogamy.
 Sexual spores occur less frequently and in small numbers than asexual spores. The
different sexual spores are
o Ascospores – single celled spores produced in a sac called ascus. Usually eight
ascospores in each ascus.
o Basidiospores – single celled spores are borne on a club-shaped structure
called a basidium.
o Zygospores – large thick walled spores formed when the tips of two sexually
compatible hyphae
o Oospores – formed in a special female structure called oogonium. Fertilization
of eggs or oospheres by male gametes formed in an antheridium gives rise
to oospores. One or more oospores are present in each oogonium.

Comon Fungal infection in Animals

Disease Name Causative fungus Animals affected

Ringworm Microsporum spp. All domestic animals and man
Trichophyton spp.
Aspergillosis Aspergillus spp. Cattle, horse, poultry
Thrush Candida albicans Avian spp.
Also in dogs, cats, man, pigs.
Histoplasmosis Histoplasma Man and domestic animals
Aflatoxicosis Aspergillus flavus All animals and man
Aspergillus
parasiticus

Exercise:-

1. Describe general characteristic of fungus.

2. Explain in details classification of fungus.
3. Explain in details the reproduction of fungus.

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Chapter 8:- Virology:- Classification, cultivation and replication

of viruses
VIROLOGY

Virology is the study of viruses and the diseases caused by them.

 Viruses are the minute entities that carry genetic information in one type of nucleic acid
(either DNA or RNA) and require living cells for multiplication.
 Viruses are obligatory intracellular infectious agents of sizes ranging from 20 to300
nanometer with an absolute dependence on living cells for their replication.

Nucleic acid (DNA / RNA)

Capsid Envelope

Viral structure
A virus consists of a nucleic acid genome (DNA or RNA) surrounded by a symmetric shell of
protein called capsid, which is made up of capsomeres. Nucleic acid and genome together is called
nuleocapsid. This nucleocapsid may or may not be covered by lipid containing outer layer called
envelope and accordingly the virus is called enveloped or unenveloped virus.

Structure of virus

General Characteristics and Properties of Viruses

1. Size:
Viruses are very small retaining infectivity after passing through filter with
pore size small enough to hold back the smallest bacteria. Bacteria are measured in
terms of micrometer (μm, 10-6 of a meter) whereas viruses are measured in nanometer
(nm, 10-9 of a meter). Viruses range in size from 20nm to 300nm. The picornaviruses
(e.g. Foot and Mouth-Disease virus) are the smallest viruses (20nm) while the
poxviruses are the largest viruses (300nm). Viruses cannot be seen by light
microscope because of their small size. They are seen only by the aid of electron
microscope. However, poxviruses can be seen by light microscope.

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2. Organelles:
Viruses do not possess cellular organization and do not have organelles.
3. Genome size:
The genomes of viruses are smaller than those of bacteria, ranging from about 2
kilobase pairs (kbp) to 200 kbp.
4. Viruses are completely dependent on living cells, either eukaryotes or prokaryotes for
replication and existence. Although some viruses possess their own enzymes such as
RNA-dependent RNA polymerase or reverse transcriptase, they cannot reproduce
and amplify the information in their genomes without the assistance of the cellular
machinery. They do not grow in inanimate/non-living media.
5. Viruses have their genetic information in either DNA or RNA. A virus possesses only one
species of nucleic acid either DNA or RNA but never both.
6. Viruses have a receptor-binding protein component for attaching to cells so that they can
enter such cell and take over the machinery of the cell to their own advantage in reproducing
their progeny.
7. Viruses do not multiply by binary fission but by a complex process involving protein
synthesis and nucleic acid production
8. Viruses are unaffected by antibiotics
9. Viruses induce the production of interferon by infected host cell and are sensitive to the
interferons.

Basic Components of Viruses

Viral Nucleic Acid
1. This can either be RNA or DNA
2. It contains the information necessary for directing the infected cells to synthesis virus specific
proteins
3. It may be single stranded or double stranded
4. It may be linear or circular
5. It may be positive sense or negative sense (a positive sense nucleic acid possesses the same
polarity as the mRNA and so can be translated directly into protein without first being
transcribed)
6. It may be a single piece or segmented
7. It is haploid except in retroviruses in which it is diploid
The Capsid
It is protective coat to the nuceic material made up of identical protein unit capsomere.
Capsid containing several peculiar characteristics as below,
 It offers protection for the nucleic acid against adverse conditions
 It facilitates attachment and entry of the virus into host cell
 It possesses antigens used for virus identification in serological tests
 It determines the symmetry of the virus

The Envelope
Some viruses possesses envelope which is derived from the plasma membrane of host
cells during release of the bacteria by budding. In some viral infections, the envelope is acquired
from the endoplasmic reticulum, the Golgi apparatus or the nuclear membrane. Generally,
envelopes are made up of lipids.
 In some enveloped viruses, capsomeres take the form of projections called spikes or
peplomers, protruding out through the lipid bilayer of the envelope. The spikes are
glycoprotein in nature.
 There may be a stabilizing protein membrane beneath the envelope lipid bilayer. This is
referred to as the membrane/matrix protein.

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 Enveloped viruses are usually susceptible to detergent and are rendered noninfectious
following damage to the envelope.

SYMMETRY OF VIRUS

Capsid symmetry is the arrangement between the viral genome with the capsid. Accordingly
viruses can classify in to four types of symmetry.

(a) Icosahedral/Cubical symmetry: An icosahedran is a polyhedran made up of 12

vertices, 20 equilateral triangular faces and 30 edges. The vertices have fivefold (This
means that rotation of the icosahedran by one fifth of a revolution presents a position
which is indistinguishable from the starting orientation) faces have a threefold and
edges have a twofold axis of symmetry; thus it is also known as 5:3:2 rotational
symmetry. e.g. Adeno viruses

(b) Helical symmetry In this type of symmetry the capsomeres are surrounded the
genome nucleic acid in spiral or helical manner. All helical animal viruses possess
single-stranded, negative-sense RNA genomes eg. Rabies virus
Helices are described by the number of subunits per turn of the helix (μ) and
the axial rise per subunit (p). The pitch of the helix (P) is therefore equal to: P = μ x p

(c) Complex symmetry Certain viruses have complex symmetry; their ultra structure
appears to be complex. eg. Pox viruses and Retroviruses.

Classification of Virus
Virus classification is the process of naming viruses and placing them into a taxonomic system. The
purpose of classification is to make systemic ordered arrangement of viruses that have similarity and
differences.
In earlier time the infectious agents were named based on spectrum of clinical outcomes or
the geographic location where it was found or host (animal, plant, insect, bacteria) or tissue tropism
(Neurotropic-Rabies, Enterotropic-RP, Epitheliotropic-FMD, Dermatotropic-Pox, Pantropic-Swine
fever) etc. Thus the agent that caused foot-and-mouth disease in cattle becomes foot-and-mouth
disease virus. Arboviruses include the arthropod borne viruses. Later when the methods for studying
the physical, chemical and biological properties of viruses were developed more information on
viruses became available to develop a classification system on the basis of these properties.

1. Holmes System of Virus Classification

Holmes (1948) used Carolus Linnaeus's system of binomial nomenclature to classify
viruses into 3 groups under one order, Virales. They are placed as follows:
(a) Group I: Phaginae (attacks bacteria)
(b) Group II: Phytophaginae (attacks plants)
(c) Group III: Zoophaginae (attacks animals)

2. Baltimore System of Virus Classification

In 1971, David Baltimore proposed a virus classification based on how the viral genome is
replicated and expressed. Viruses were grouped into six/seven classes based on the mode of
transcription (mRNA synthesis).
a) dsDNA viruses: e.g. Adenoviruses, Herpesviruses, Poxviruses, Papillomaviruses,
Polyomaviruses
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b) ssDNA viruses: e.g. Parvoviruses, Circoviruses

c) dsRNA viruses: e.g. Reoviruses , Birna viruses
d) ssRNA viruses (+): e.g. Picornaviridae, Togaviridae, Calciviridae, flaviviridae,
Coronaviridae, Astroviridae
e) ssRNA viruses (−): e.g. Orthomyxoviridae, Paramyxoviridae, Bunyaviridae,
Rhabdoviridae, Filoviridae
f) ssRNA-RT viruses: e.g. Retroviridae
g) dsDNA-RT viruses: e.g. Hepadnaviridae

3. ICTV System of Virus Classification

An International Committee on Nomenclature of Viruses (ICNV) was formed at the ninth
International Congress for Microbiology in 1966. This committee is now called International
Committee on Taxonomy of Viruses (ICTV). It is the international body which recognizes
and classifies the new viruses. The main properties used for classification are nature of
(a) Nucleic acid (DNA or RNA, single or double stranded, segmented or not, positive or
negative sense)
(b) Structure of virion ( Symmetry, enveloped or non enveloped)
(c) Site of replication ( nucleus or cytoplasm)
(d) The other properties used are host range, trophism, mode of transmission, surface
structures.

 Viral classification starts at the level of order, the taxon suffixes as follow:
Order (-virales): Mononegavirales, Family (-viridae): Paramyxoviridae
Subfamily (-virinae): Pneumovirinae, Genus (-virus):: Pneumovirus

DNA VIRUSES
No Genome Family Important Diseases
1 dsDNA Poxviridae Smallpox virus, Vaccinia virus, Fowl Pox,
Orf
2 Asfarviridae African swine fever

3 Iridoviridae Frog virus, Goldfish virus, Flounder virus

4 Herpesviridae Herpes simplex, varicella-zoster virus

(Chicken pox), Epstein-Barr virus, MD, ILT
5 Adenoviridae Infectious canine hepatitis, EDS-76

6 Papillomaviridae Human and Bovine papilloma viruses

7 Polyomaviridae SV 40, Mouse Polyoma virus

8 ssDNA Parvoviridae Parvovirus B19, Canine parvovirus
9 Circoviridae Porcine circovirus, Chicken anaemia virus
10 Partially Hepadnaviridae Hepatitis B virus, Duck hepatitis B virus
dsDNA
(RT)

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RNA VIRUSES

N Genome Family Important Diseases

o
1 ssRNA (Reverse- Retroviridae Leukosis, HIV, Maedi, Visna
transcribing)
2 dsRNA Reoviridae BT, African horse sickness, Rota
3 Birnaviridae IBD, Infec. Pancreatic necrosis
4 ssRNA , Paramyxoviridae Measles, Mumps, RP, PPR,CD,
Negative Sense ND
5 Nonsegmented Rhabdoviridae Rabies, Bovine ephemeral fever
6 Filoviridae Ebola, Marburg
7 Bornaviridae Borna disease
8 ssRNA , Orthomyxoviridae Influenza, Flu
Negative Sense
9 segmented Bunyaviridae CCHF, Nairobi sheep dis. Rift
valley fever
10 Arenaviridae Lymphocytic Choriomeningitis
11 ssRNA , Coronaviridae IB, Berne (Horse), Breda
Positive Sense (Calves)
12 Arteriviridae Equine Arteritis Virus, Lactate
dehydrogenase elevating virus
13 Picornaviridae Poliovirus, FMD
14 Caliciviridae Norwalk virus, Vesicular
exanthema
15 Astroviridae Astrovirus
16 Togaviridae Rubella virus
17 Flaviviridae Dengue, Hepatitis-C, Yellow
fever, CSF
18 No nucleic acid, self- Prions CJD, Kuru, Scrapie
replicating protein

Replication of virus
Stages of virus replication
Virus replication can be divided into following phases and each phase may have number of
stages:
Phase – I Initiation:
 Attachment
 Penetration
 Uncoating
This stage is characterized by introduction of genetic material of the virus into the
cell
Phase – II Replication:
 Genome synthesis
 Protein synthesis

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Phase – III Assembly, Release, Maturation:

Phase I - Initiation
1. Attachment: Virus attaches to the cell surface. Attachment is via ionic interactions.
Penetration: It is a process by which a virus enters into the cell. It is an energy
dependant reaction and occurs quickly. It occurs as fusion, endocytois or translocation
2. Uncoating: This is the general term applied to events after penetration, which allow
the virus to express its genome. For successful viral infection, nucleic acid has to be
sufficiently uncoated. The lysosomal enzymes play a major role in uncoating.

Phase II - Replication of viral nucleic acid and protein synthesis

Once uncoating has taken place, synthesis of viral nucleic acid starts. This occurs as three
different stages with differences between different families of the viruses.
1. Early transcription and translation – The proteins derived from this stage is mostly the
enzymes required for virus replication.
2. Replication of Nucleic acid
3. Late transcription and translation – The proteins produced this stage are structural
proteins.

Phase III

Assembly: This stage involves the assembly of all the components necessary for the
formation of the mature virion at a particular site in the cell. During this process, the basic
structure of the virus is formed.
The site of assembly varies for different viruses, e.g:

 Picornaviruses, Poxviruses, Reoviruses - In the cytoplasm.

 Adenoviruses, Papovaviruses, Parvoviruses - In the nucleus.
 Retroviruses - On the inner surface of the cell membrane.

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Release: For lytic viruses (most non-enveloped viruses), release is a simple process - the cell
breaks open and releases the virus.
Enveloped viruses acquire the lipid membrane as the virus buds out through the cell
membrane. Virion envelope proteins are picked up during this process as the virus is
extruded. Budding may or may not kill the cell

Maturation: At this stage of the lifecycle normally the virus becomes infectious. Usually it
involves structural changes in the particle, often resulting from specific cleavage of capsid
proteins to form the mature products, which frequently leads to a conformational change in
the capsid, or the condensation of nucleoproteins with the genome. For some viruses,
assembly and maturation are inseparable, whereas for others, maturation may occur after the
virus particle has left the cell.

Cultivation of Virus

 Viruses can grow only in living cells. The substrates used for virus cultivation are
 Animals – host or experimental
 Embryonated eggs
 Tissue cultures

Animals:
 Some viruses can not be cultivated in cell cultures or embryonated eggs. They must
be propagated in living animals.
 The host animal or experimental animals such as mice, guinea pigs or rabbits are
used. By animal inoculation we can observe the symptoms , lesions and
histopathology sections of infected tissues can be examined.
 The disadvantages are the need for containment facility,ethical issues and variation in
susceptility.

Embryonated eggs:
 One of the most economical and convenient methods for growing many animal
viruses is the chicken embryo inoculation. This method was devised in 1931.
 Different routes of inoculation are used namely yolk sac, allantoic, amniotic,
chorioallantoic and intravenous route. Chick embryos have different types of cells in
which various viruses can replicate. This method has been used in the production of
vaccines against various animal and human diseases.
 Limitation is that not all animal viruses can be grown by this method. In the case of
poultry viruses presence maternal antibody in embryonated chicken eggs may
interfere with virus multiplication.

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Tissue cultures:
 Cell cultures are the preferred method for virus cultivation at present. Convenience,
economy of maintenance compared to animals, cytopathic effect and choice of cells
for their susceptibility to particular viruses are the reasons for the preferred use of the
cell of cultures.
 There are three types
 Primary culture
 Diploid cell strains
 Continuous cell lines

Exercise:-

1. Describe general characteristic of virus.

2. Explain in details classification of virus.
3. Explain in details the replication of virus.
4. Explain in details different method of cultivation of virus.

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Chapter 9:- Source of infections, methods of transmission of

infections

SOURCES OF INFECTION
Sources of infection are animal and inanimate in nature.
1. Animal sources
 Normal flora
 Animals in incubation period -Animals in incubation period may excrete the
pathogen.
 Animals with overt disease.
 Convalescent carrier animals – In these animals shedding of the pathogen occurs
for varying periods after clinical recovery. The period may vary from weeks to
months.
 Contact carrier or subclinical infections – They acquire pathogenic organisms
from other animals with infectious disease without contracting the disease
themselves. Such animals are called as contact or subclinical carriers. The carrier
state may be temporary for a few days or lasting for months.
2. Inanimate sources (fomites)
 Contaminated utensils, feed and water troughs and vehicles

TRANSMISSION OF INFECTION / DISEASES

Disease can be transmitted by direct or indirect contact.
1. Direct contact
 Disease spread occurs by contact with the infectious agent on the infected host ie
contact with discharges from the animal. The other means is coitus.
 Vertical transmissiom from mother to offspring.
2. Indirect contact
 Organisms excreted by the infectedc animal are carried in various vehicles like
water, milk, food, litter, air or dust. Such contaminated objects are called as
fomites.
 Contaminated instruments may also spread the infection.

PORTALS OF ENTRY
 Inhalation
 Ingestion
 Inoculation or infection through the skin or mucous membrane
 Through coitus or contaminated instruments, catheters, semen in Artificial
insemination.
 Transplacenatal infection via the umbilicus
 Hospital acquired infections are called nosocomial infections.
 Physician induced infections are known as iatrogenic infections.

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Mode of transmission of disease

Transmission of disease is broadly classified into

A. Horizontal or lateral
B. Vertical transmission

Now, Horizontal transmission classified as transmission of disease from infected animals to

susceptible animal either directly or indirectly.

Transmission of
Disease

Horizontal Vertical

Infected mother of one

generation
Direct Indirect

Inherited, acquired,
Infected population Transplacental , Trans-colostral
Infected population

Vector require Offspring of next

generation
Susceptible population
Susceptible population

Direct transmission

 Transmission of disease from infected population to susceptible animals by direct contact or

discharges without involvement of vectors. Direct transmission of disease is possible through
below mentioned routes.
 By direct contact with infected animals or their discharges.

1. Physical as well as sexual contact leads to transmission of various diseases.

Example: Brucellosis, Anthrax, FMD, Vibriosis, Dourine, etc.
2. Direct Air Borne transmission:
- Droplet transmission: A tiny fluid, who suspends in the environment, is
called as droplets.

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- The Droplets, originated due to the sneezing or coughing of infected animals

may contain the pathogens, which may being inhaled by the nearby
susceptible animals, which may get infected.
- The close proximity, overcrowding and poor ventilation may serve as
predisposing factors for the transmission of diseases.
- Examples : New castle disease virus, Glander, tuberculosis
3. Mechanical transmission and infection of wounds:
- Animal bites and scratches from infected animals transmit some important
zoonotic diseases, viz. rabies, cat-scretch fever.
- Contamination of wounds by infected soils may leads to transmission of
diseases. Example: Tetanus in horses, Anthrax, cutaneous larvae migrans.

Indirect transmission

 Transmission of disease from infected animals to susceptible animals through intermediate

vector either animate or inanimate.
1. Fomite-borne transmission
- Infections are transmitted through the inanimate objects (contaminated soil,
cloths, utensils, surgical instruments, feeders, waterers, syringe and needles,
milking machines, farm equipment).
- Examples are anthrax, foot and mouth disease

Note: Some diseases like FMD, TB are transmitted by both ways (direct and indirect). So, they are included in
the both types of classification. However, for both type of arrangement, they fulfill the specific criteria for
 Air-borne
their inclusion transmission
in particular groups.

2. Vector-borne transmission

 An invertebrate host usually an arthropod is responsible for the transmission of an infectious

agent from the infected host to the susceptible host.
A. Mechanical transmission by biting flies:
- Mechanical transmission of pathogens by biting flies. In which, biting flies
are not affected with the pathogens.
- Examples are anthrax, salmonellosis
B. Multiplication of the parasite in the vector:
- Ricketssia multiply in the lice and ticks and transmitted to definitive host
during feeding of lice nad ticks.
C. A portion of lifecycle of parasite undergone in the vector
- Ex.: Trypanosomiasis.
- The sexual maturity of these protozoa attains in the flies.

Vertical transmission

 Transmission of an infectious agent from the infected mother to their offspring (from one
generation to the next generation) by hereditary, acquired or congenital at birth or infection of
the ovum / embryo or fetus itself in vivo.

 Transmission of parasite from insect to its next generation through its eggs.
- Ex.: Babesiosis.
The protozoan parasite transmits from parents to offspring through transovarian route.
Exercise:- 1. Explain source of infections.
2. Explain mode of transmission of disease.
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Chapter 10:- Important bacterial, viral and fungal disease of

animal
Johne's disease
Synonym : Paratuberculosis
Definition
 Chronic wasting illness of ruminants with prolonged course, recurrent diarrhoea, dehydration,
emaciation and death caused by Mycobaterium paratuberculosis
Aetiology
 Mycobaterium paratuberculosis - Three strains:
o Bovine strain
o Ovine strain
o Scottish strain
Incidence
 The disease is of worldwide distribution and constitutes an important economic problem in
cattle
Susceptibility
 Ruminants are usually affected. Horses and pigs may be affected very rarely
 It is less frequently encountered sheep and goats
Transmission
 Infection is ingestion. Faeces containing the organisms serve as the primary source of
infection
Pathogenesis
 Incubation period : Long and protracted even upto 2 years
 Organisms enters the body through intestinal mucosa, sets up bacteremia and settles in the
mucosa of intestine, mesenteric lymph nodes and tonsil produce a chronic granulomatous
inflammation
 Cell mediated immune response and followed by humoral immune response initiated by the
release of bacteria by dying macrophages as the disease progresses. Type III (immune
complex) reaction may contribute to the development of intestinal lesions. Type I or type III
reactions in the intestinal mucosa may result in an increased outflow of fluid and diarrhoea.
Clinical signs
 Emaciation, hide and bound with dry coat
 Normal appetite with increased thirst and diarrrhoea
 Sheep and goat – Diarrhoea is not common, emaciation observed
Gross lesions
 Carcass – Emaciation with gelatinous fat
 Terminal part of ileum – Wall thickened and oedematous
 The mucosa is folded and showed Transverse corrugation or rugae (like cerebral convolution)
 Intestinal wall stretched – Corrugations does not disappear
 Sheep and goats – corrugation – Not marked
 Uterine infection may l ead to uterus may lead tocongenital infection and abortion
Microscopic lesions
 Mucosa, submucosa and mesenteric lymph nodes infiltrated with epithelioid cells with foamy
cytoplasm
 In the submucosa, epithelioid cells may fuse to form syncytial mass
 Thickeneing of submucosa, edema, infiltration with neutrophils and eosinophils
 Giant cells may also be seen
 Nodule formation with necrosis and calcification described in sheep and goat and corrugation
is not marked in sheep and goat
 Mesenteric lymph node- swollen and juicy

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Diagnosis
 Symptoms
 Examination of bowel washings
 Examination of rectal-pinch
 Double intradermal test by Johnin in the neck (Johnin test)
 Complement Fixation Test (CFT)
 Agar Gel Immuo diffusion test (AGID)
 Enzyme Linked Immuno Sorbent Assay (ELISA)

ANTHRAX
Synonym : Splenic fever, charbon
Definition
 Acute infectious septicaemic disease commonly affecting herbivorous animals characterized
by high fever, rapid course, sudden death and greatly enlarged spleen, caused by gram
positive, capsulated spore forming rod shaped organism Bacillus anthracis
Etiology
 Gram positive capsulated rod, spore forming ' Bacillus anthracis'
Incidence
 Anthrax is worldwide in distribution. In India, it is mostly seasonal, sporadic out breaks
occurring usually in rainy season
Susceptibility
 Sheep and cattle – Most susceptible
 Buffalo, horse, mule, goat, pig, rabbit and fowls.
 Man – Contact with animal or animal products (wool, hide, bristles) - 'Wool sorters disease'
 Cutaneous anthrax-„ Malignant Carbuncle’
Transmission
 Ingestion, wound infection, flies, inhalation, vaccination
 Spread of infection through water streams, rivers, flies and wild birds etc.
Pathogenesis
 Incubation period: 1 to 14 days.
 Ingested bacilli proliferate in the tonsils and subsquently reach lymphatic glands.
 Bacilli reach the stomach are killed by gastric juice but the spores resists action and develop
into bacilli in the intestine, penetrate the wall, reach lymphoid follicles and multiply.
 Those entering through wounds of skin proliferate in the connective tissue
 The organisms are inhibited or not killed by the cells of reticuloendothelial system due to the
presence of a capsule and resist phagocytosis by neutrophils. -The capsule has fibrinolytic
property and so prevents clotting of blood. It also produces a diffusible toxin which damage
brain and other nerve cells.
Toxin
 Factor I- oedema factor
 Factor II -Protective Antigen
 Factor III - Lethal factor
 The capsule swells by absorbing fluid and so becomes gelatinous.
 The gelatinous fluid cannot be absorbed and bacteria accumulate oedematous swellings form
 The bacterial toxin injure the endothelium causes hemorrhages - Effusion of blood in organs
in which circulation is slow.
 The toxin causes nerve cell degeneration leads to apoplectic form of death
 Death within 12 to 36 hours
 Death – Asphyxia may be due to toxin acting on the respiratory centre in the brain
 Spores – Do not form in living animals
 Carcass –If opened spores form when they come in contact with environment
 “Do not open carcass ”

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Clinical signs
 The disease may occur in a peracute, acute, subacute or chronic form.
 Death may takes place without symptoms in peracute form
 Discharge of dark tarry blood from the natural orifices
 In acute type- Increase temperature, excitement, weakness, cyanosis, dyspnoea, subcutaneous
oedematous swelling
 Sheep, Horse – Subcutaneous oedema common
 Pigs – swelling on the throat and cervical lymph node enlargement is very characteristic
Gross lesions
 Postmortem examination of anthrax carcass should not be conducted
 Oedema and haemorrhages may be seen in any part of the body, particularly in serous
membrane.
 The spleen is greatly enlarged and engorged with dark unclotted blood
 Lymph nodes are swollen, oedematous and occasionally haemorrhagic
 Haemorrhages and swelling may occur in the intestinal tract, liver and kidney.
 In sheep and horse splenomegaly is not a constant lesion.
 In swine, oedema and haemorrhages are seen in the pharynx and cervical lymph nodes.
Microscopic lesions
 Large numbers of bacilli demonstrated in smears or tissue sections.
 The anthrax rods can be differentiated from pm invaders by gram's stain.
 The ends of the anthrax bacilli are truncated seen in short chains with capsule.
 Spores never noticed in anthrax
Diagnosis
 Sudden death
 Bloody discharge from the mouth, nose and anus
 PM should not be conducted, since opening of carcass may result in spreading of infection
 Diagnosis can be confirmed by demonstration of the organisms and biologicals tests.
Demonstration of casule by methylene blue -McFadyen's reaction
 Materials to be sent to laboratory include blood smear from ear, smear from oedematous
swelling of throat, blood smear from ear and dried piece of muzzle.
 Ascoli precipitation test- White precipitate form positive for anthrax
 Fluorescent antibody technique

BLACK QUARTER
Synonym: Black leg, quarter ill, Symptomatic anthrax
Definition
 Acute febrile disease of cattle (6 month to 2 years), less often of sheep, goat and swine
characterized by emphysematous, sero haemorrhagic swelling in the heavy muscles,
especially of the hind limbs caused by Clostridium chauvoei
Etiology
 Clostridium chauvoei - Gram positive, spore forming, rod shaped bacterium
Incidence
 The disease is enzootic in particular areas of India and the mortality rate may approach 100%
Transmission
 Infection is ingestion of spores
 In sheep: wound infection
Pathogenesis
 Incubation period: 1- 5 days
 Infection is by ingestion and the organisms multiply in the intestine and through blood stream
reach the muscle (thigh, shoulder)
 Necrosis of muscle with formation of gas (Necrotizing Myositis) -Gangrene
 Smell of Rancid butter(Ferments sugars by organisms)

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 Death within 24 hrs to 60 hrs. Death may be due to toxaemia

Clinical signs
 Fever, lameness, visible swelling of muscles- Tongue, diaphragm, myocardium and gluteal
muscles
 In early stages, the swelling is hot and painful to the touch but soon cold and painless and
oedema and emphysema can be felt
Gross lesions
 Crepitating swelling of the muscle(extremities)
 Rubber sponge and dark brown or black sero sanguineous fluid exudes with gas bubbles -Gas
gangrene
 Affected muscles appears black due to the formation of iron sulphide from H2S and iron of
blood
 Muscle – Centre area is dry and have odour of rancid butter
 Regional lymph nodes – Swollen and oedematous
 Large muscles – Diaphragm and tongue
 Internal organs – Heart, lung, kidneys, liver, spleen and intestine showed acute congestion
Microscopic lesions
 Muscle fibres separated and showed waxy degeneration and coagulation necrosis
 Streaks of haemorrhages - Collection of neutrophils, lymphocytes and Gram positive
organisms demonstrated in tissue sections
Diagnosis
 May be confirmed by gross lesions
 Demonstration of organism – from the fluid incised swelling
 Biological tests using guinea pigs
 Fluorescent Antibody Technique (FAT)
 Fatal course and found dead before signs of illness are seen

TETANUS
Synonym : Lock jaw
Definition
 Acute fatal infectious disease of man and animals characterized by involuntary contraction of
voluntary muscles caused by toxins of Clostridium tetani
Aetiology
 Clostridium tetani
 Exotoxin - CI. Tetani - Gram positive sporulating rod shaped anaerobe, Spores – “Drum
Stick”
Incidence
 Tetanus occurs in all parts of the world
Susceptibility
 Hores and mules are susceptible
Transmission
 Wound infection
Pathogenesis
 Incubation period : 1 to 3 Wk
 The organisms enter the body through the nail prick, castration, docking, shearing, umbilical
wound (tetanus neonatorum) or during parturition
 Anaerobic condition allows germination of spores and release exotoxin
Three toxins
 Haemolysin – Tetanolysin – Not important
 Neurotoxin -tetanospasmin- responsible for the nervous symptoms
 Fibrinolysin- not very potent

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 Toxin gets fixed to a substance called "protagon” which act on the inhibitory synapses
interferes with action of transmitter thus producing spastic action -spasmodic contraction
 The toxins causes hyperirritability responsible for the tetanic spasms
 The toxin causing spasmodic contraction of muscles, stiffness and immobilization.
Clinical signs
 Involuntary, persistent, intense painful contraction of one or more group of muscles.
 Horse- Stiffness and moves like 'wooden horse'.
 Raised Tail, third eyelid Protrusion, and Stiffness of Jaw muscle -„ Lock Jaw’
 Ruminants – Symptoms are less severe
Gross lesion
 No characteristic lesion
 Death due to Toxaemia
Microscopic lesions
 No specific microscopic lesions. Degeneration of the neurones in the brain and spinal cord
(due to anoxaemia)
Diagnosis
 Characteristic clinical signs.
 Organisms are local but not septicaemic
 Demonstration of toxin in the serum

LEPTOSPIROSIS
Synonym : Weil‟s disease- Man, Stuttgart disease
Definition
 This is a disease of dogs and large domesticated animals, caused by several species
of Leptospira and is of zoonotic importance
Aetiology
 L. icterohaemorrhagiae - Man – weil‟s disease
 L. canicola -Dogs. L.pomona – Swine and Cattle
 Goat –L.grippotyphosa
Incidence
 Worldwide distribution and affects cattle, pig, sheep, dog and man
 Rat – Reservoir
Transmission
1. Ingestion 2. Coitus 3. abrasions 4. Intact skin 5. In utero 6. Man can be infected while
swimming in water
 Wild animal acts as a carrier
 Outside the body – organism may thrive in water for 3 months.
 Recovered animals may void organism upto 1 year
 Spirochaetes identified by dark field examination of fluid media
 Tissue sections –Organisms are black in colour in silver stains – Levaditi and warthin starry
stain
Pathogenesis
 The organism invade the blood stream and multiply producing septicaemia -Temperature rises
and last for several days
 Animals not die during this phase, the organisms settles down in liver, kidney and pregnant
uterus
 Acute form – Calves, piglets and lambs
 Sheep and goats – encephalitis occurs
 Jaundice – Intravascular haemolysis and hepatic necrosis
 Anemia , icterus, haemoglobinuria
 Leptospires – Found in Urine
 Albuminuria – Interstitial Nephritis- Focal or diffuse

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 Gravid uterus – abortion

 Uraemia – Death
Clinicalsigns
Cattle
Acute Subacute chronic
 Septicaemia milder. No Clinical
 High temp icterus symptom
 Petechiae Abortion
 on visible mucous membrane
 Jaundice, anaemia, haemorrhagic mastitis, abortion
 Death within 2 – 7 days
Gross lesions
Acute
 Icterus, anaemia, petechiae on serous membrane
 Liver – Haemorrhages –near the central veins
Miscroscopic lesions
 Liver – Cells shrink and dissociated and separated from sinusoidal epithelium
 Congestion and Haemosiderin crystals within kupffer cells and hyperplasia
Kidney
Gross lesions
 Swollen and red
Microscopic lesions
 Tubular epithelial cell degeneration followed by hydropic degeneration and necrosis
Spleen
 Haemosiderosis
 Acute haemorrhagic meningitis, necrosis of placenta
Subacute
 Interstitial nephritis,infiltration of lymphocytes and plasma cells.
 Placentitis, abortion.
 Leptospira can be found in the foetus.
Sheep and goat
 Peracute: Dullness, abortion and icterus
 Pigs: Abortion, nervous symptoms and focal interstitial Nephritis
 Horses: Periodic ophthalmia – L. pomana
 Dog:
o L. canicola, L. icterohgemorrhagiae
o Dog to dogs, Rats – Dogs
o Kidney -Nephritis
o Liver- Jaundice
Clinical signs
 Peracute: Septicaemia, fever, haemorrhages
 Acute: Fever, anaemia, incraesed ESR and icterus
Gross lesions
 Liver: No lesions may be seen
 Spleen and lymph node – Swollen
 Gastrointestinal tract: Mucosa - Swollen and dark red
Microscopic lesions
 Liver: Hyperchromatic nuclei with eosinophilic granular cytoplasm
 Kidney: Tubular epithelium – Degenerative changes and organisms seen in the tubules
 Gastrointestinal tract : Congestion and petechiae on the mucosa
 Purulent necrotic laryngitis
 Subaute and inapparent – Kidney lesion only observed
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Gross lesions
 Capsule peeled off easily
Microscopic lesions
 Focal interstitial nephritis, spirochaetes seen in the tubular lumen as clusters
 L. canicola – Nervous symptoms, purulent lymphocytic meningo encephalitis and gliosis
Diagnosis
 Symptoms
 Demonstration of organisms in the tissue sections
 Microscopic agglutination test
 Inoculation
 Dark field examination of urine

BRUCELLOSIS
Synonym
 Mediterranean fever, Gastric fever, Bruce septicaemia, Malta fever, Bang‟s disease, undulant
fever in humans.
Incidence
 1887 – Spleen of patients (Brucella mellitensis)
 1905 – The infection was traced to goat‟s milk
 1897 – Isolation and identification of organism in aborted bovine fetuses and fetal membrane
(Brucella abortus)
 1914 – Identification of Brucella suis in pigs
 1950 – Brucella ovis in rams
 1960 – Bruella canis in dogs
Etiology
 Brucella species are small (0.6 x 0.6 to 1.5 pm), non- motile, coccobacillary, Gram-negative
bacteria. As they are not decolourized by 0.5% acetic acid in the modified Ziehl-Neelsen
(MZN) staining technique, they are classed as MZN-positive.
 In MZN-stained smears of body fluids or tissues, they characteristically appear as clusters of
red coccobacilli.
 Brucella species are aerobic, capnophilic and catalase-positive. Apart from B. ovis and B.
neotomae, they are oxidase-positive.
 All Brucella species are urease-positive except B. ovis. Brucella ovis and some biotypes of B.
abortus require 5 to 10% C02 for primary isolation. Moreover, the growth of other Brucella
species is enhanced in an atmosphere of C02. Media enriched with blood or serum is required
for culturing B. abortus biotype 2 and B. ovis. Recently, brucellae have been detected in sea-
mammals.
Transmission in cattle
 By contact
 Ingestion of infective uterine discharge, aborted foetus or placenta
 Organisms are excreted in milk
 Rare through coitus
 Bulls are resistant
Pathogenesis
 Brucella organism gain entry mainly through ingestion. After ingestion, they localized into
regional lymph node. Where, they are engulfed by macrophages and remain in the phagosome
of macrophage. But they survive inside the macrophages due to their ability to prevent fusion
of lysosone with phagosomes. Due to this prevention, bacteria are not exposed to lysosomal
enzymes.
 Bacteria grows inside phagosomes of macrophages, hence antibodies are ineffective against
bacteria. They multiply inside macrophages and release by rupture and produce bacteremia.
 After bacteremia, infection become generalized and localize in reproductive organs, placenta,
fetus, mammary glands, lymph nodes, spleen, liver, joints, bones. Organisms have high
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affinity to the placenta and most especially to chorio – allantoic trophoblast due to presence of
erythritol sugar in this tissue. Their multiplication at that site leads to inflammation and
subsequently abortion.
Clinical signs
 Cattle
o Abortion in 7th and 8th months of gestation
o Persistent infection in cows exposed after puberty
 Pigs
o Transmission through coitus.
o Abortion in 2nd and 3rd months of gestation
o Orchitis in infected boars
o Localization in other tissues like skeleton more common
 Sheep and goats
o Transmission same as cattle
o Abortion in ewes
o Orchitis, epididymitis in male sheep and goats.
 Dogs
o Caused by B. canis
o Transmitted by exposure to uterine discharges or aborted fetuses or by coitus.
o Abortion in 50 days of gestation
o Male dogs – orchitis / epididymitis.
 Horses
o B. abortus + Actinomyces bovis
o “Poll evil” and “Fistulous withers”
Lesions
 Bacterial granuloma in tissues, especially in lymphoreticular system.
 No multinucleated giant cells. These ganulomas are visible grossly, or may be of microscopic
size- classical lesion of Brucellosis
Gross lesions
 Necrosis of cotyledons
 Inter – cotelydonary chorion is oedematous and filled with odourless, sticky, brownish
exudates
 Yellowish granular, necrotic areas in colytedon.
 Rest of chroion is opaque, thickened and leathery
 Induration of bovine mammary glands and supramammary lymphnodes
 Epidydymis and testicles of bulls become enlarged and hard
 Scrotal contents – suppuration & rupture
Pigs
 Tiny white yellowish nodules in all organs
Rams
 Tail of epididymis a inflammed
Bitches
 Uterine and placental lesions
 Bronchopneumonia in aborted pups
 Osteomyelitis in dogs
Horses
 Necrotizing and purulent lesions in ligamentum nuchae
 Necrotizing and purulent lesions in region of thorasic attachment of ligamentum nuchae.
Microscopic lesion
 Bovines
o Organisms in chorionic epithelial cells
o Necrosis and inflammatory exudates with macrophages and neutrophils
o Collection of epithelioid cells in endometrium.
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 Mammary gland
o Diffuse inflammation, with lymphocytes and neutrophils.
o Collection of epithelioid cells with langhan‟s giant cells.
o Later atrophy of glands and fibrosis
 Pigs
o Typical Brucella granulomas with necrosis
 Rams
o Perivascular oedema and lymphocytic infiltration
o Hyperplasia and degeneration of tubular epithelium and intertubular fibrosis
o Escape of spermatozoa from damaged tubules produced granulomatous response
 Bitches
o Hyperplasia and plasmacytosis of lymphnodes
o Orchitis
o Epididymitis
o Prostatitis
o Hyalinization of glomeruli
Diagnosis
 Symptoms and lesions
 Agglutination test to detect antibodies
 Immunological staining or molecular probes

HAEMORRHAGIC SEPTICAEMIA

Synonym: Stockyards disease , Pasteurellosis, barbone

 Pasteurella are small, ovoid Gram positive rods and the ends are deeply stained than central
portion which gives them a Bipolar appearance
 Pasteurella boviseptica – H. S. incattle
 P. avi septica – Fowl cholera
 P. sui septica – Swine plague
 P. ovi septica – H. S in sheep.
 P. lepi septica – Snuffles and Septicaemia rabbits
 P. Pestis - Bubonic Plague – Man (Natural host – rat)
 P. pseudo tuberculosis – Guinea pigs, rodents and goats.
 P. haemolytica ( Mannheimia haemolytica)– Arthritis – Calves, mastitis – Cows & Sheep.
 These organisms cannot be differentiated from each other and various types are adapted to
different hosts
 All the types are classified under “Pasteurella multocida"
 P. multocida and P. haemolytica (Mannheimia haemolytica) -Cattle and buffaloes.

Incidence
 It is one of the important bacterial diseases of cattle and buffaloes in India
 Sheep and goat – Rarely affected
 Swine and Fowls – May be affected
Transmission
 Ingestion of contaminated feed, water etc.
 The organism is found in the saliva of affected animals
 Droplet infection may occur
Pathogenesis
 Organisms on entry into the system reaches blood, proliferate and spread throughout the body
due to its septicaemic nature and produces petechial and ecchymotic haemorrhjages on
serous and mucous menbranes in different organs. Hence it is called as -
“ HAEMORRHAGIC SEPTICAEMIA ”
Clinical signs
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 High temperature, dullness, dyspnoea, hot painful swelling – head, dewlap and neck with
inflammatory exudate in the subcutaneous tissues.
Gross lesions
 Petechiae on all serous and mucous membranes especially on epicardium, myocardium,
pleura, peritoneum and Gastrointestinal tract.
 Lymph glands- Swollen and haemorrhagic
 G.I tract is severely inflammed with contents mixed with blood
Microscopic lesions
 In acute and subacute cases, the predominant lesion is fibrinous bronchopneumonia
 The organism is small ovoid rods the ends of rods are more deeply stained than central
portion which gives them a bipolar appearance.
 In rabbits – Haemorrhagic tracheitis.
Diagnosis
 Examination of blood smear
 A heart blood swab may be rubbed over the scarified abdomen of a rabbit. It will die within
24 to 40 hours showing haemorrhgaes, especially haemorrhagic tracheitis is pathognomonic
lesion
Fowl cholera
 Aetiology: P. multocida (avi septica)
 Definition : Acute, chronic, generalized or local infectious disease of fowl and other species
of birds characterized by sudden onset, enteritis, sub mucous, subserous petechial
haemorrhages, enlargement of liver and spleen.
Transmission
 Transmitted by Inhalation or Ingestion
Shipping Fever
 Calves and young animals
 Transportation to great distance by rail, truck and ship
 Heavy losses are observed.
Aetiology
 Stress, viral and bacterial infections
 Heat, cold, Fatigue, trauma, insufficient feed and water, anxiety
 Virus : Myxovirus parainfluenza -3
 Infectious bovine Rhino tracheitis, entero virus, bovine adeno virus
 Bacteria: P. multocida, Streptococcus sp., Pseudomonas sp., Haemophilus sp.
Clinical signs
 Depressed, stand aloof, anorexia, dry nose, increased respiratory rate
Lesions
 Oedema and haemorrhages – Upper nasal passage, larynx, trachea, regional lymph nodes

FOOT AND MOUTH DISEASE

Synonym : Aphthous fever,Aftosa,Enzootic apthiae

Definition
 Acute highly contagious febrile disease of cloven footed animals
 Formation of vesicles in mouth and feet of cattle,sheep, goat and swine
Etiology
 Picorna virus group,
 GENUS-APHTHOUS VIRUS
 Serotypes – O, A,C, and South African type SAT1, SAT2, SAT 3 and Asia 1 of which O,A,C
and Asia 1 - prevalent in India
Incubation period
Complied by: Dr. A .I. Dadawala & Dr. F .M. Parth
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 Few hours to few days

 Sheep: Incubation period is 3 to 8 days
Susceptibility
 All cloven footed animals and it is rarely fatal except in very young animals
 The disease is also seen in elephants and camels
Transmission
 Ingestion of contaminated materials
Pathogenesis
Vrius is epitheliotropic
 Virus invades epithelial cells, multiplies resulting in vesicle firmation finally causing focal
areas of inflammation
 Invades lymph and blood results in viremia and reaches epithelium of other mucous
membrane, interdigital space, dorsum of tongue, gums etc.producing vesicles
 Causes myocardial degeneration in young calves
Clinical signs
 High temperature
 Vesicles appear on mouth
 Rupture leaving ulcer - refuses feed and water
 Vesicles on cleft of feet leading to lameness
 Also seen in lightly haired parts (Udder and teat)
 Produces smacking noise or smacking of tongue
 Drooling of saliva
Gross lesions
 Vesicles on the mucosa of lips, dorsum of tongue and palate in cattle and also in coronet,
udder and teat
 Lesions also seen – Rumen, reticulum and omasum
 Haemorrhage and diffuse oedema in mucosa of abomasum and small intestine
 Death may be due to gastroenteritis and myocardial lesions in young calves -"Tigroid heart"
 In sheep: Oral lesions appear on the dental pad only
Microscopic lesions
 Vacuoles appear in the cells of epithelium with eosinophilic cytoplasm and pyknotic nuclei.
These small vesicles coalesce to form larger ones and the epithelium gets detached
 Hyaline degeneration and necrosis of myocardial fibres and also in sketetal muscles -
"Tigroid heart appearance"
 Secondary bacterial infections produces osteomyelitis and suppurative arthritis
Secondary Effects
 Mastitis
 Anaemia
 Disturbances in sexual function
 Diabetes
 Over growth of hair
 Cardiac cicatrices
 Panting
 Dyspnoea
 Delayed growth of young animals
Important sequelae of FMD is infertility and decreased production probably due to
endocrine damage especially the thyroid gland
Diagnosis
 Clinical signs and lesions
 Complement fixation test
 Virus isolation and polymerase chain reaction

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PESTE- DES-PETITS- RUMINANTS (PPR, goat plague)

Synonyms
 Plague of small ruminants (goats, sheep), Erosive stomatitis, Enteritis of goats
 Stomatitis – pneumoenteritis complex

Definition
 It is an acute, highly contagious disease of goats and sheep characterized by fever, anorexia,
lymphopaenia, erosive stomatitis, diarrhoea, oculo-nasal discharge and respiratory distress
Etiology
 Morbilli virus
Incidence
 First reported in 1942 in Africa
 In India, first reported in sheep flocks during 1989 in the Villupuram district of Tamil Nadu
Suceptibility
 Disease is more severe in goats than sheep; fatal in young animals
Transmission
 Close contact with infected animal -Direct contact
 Contaminated fomites
 Inhalation / conjunctival or oral routes
 Large amount of virus is present in excretions and secretions
Pathogenesis
 Virus penetrates the retro-pharyngeal mucosa sets up a viraemia
 Damages the alimentary, respiratory and lymphoid system
 Infected cells undergo degeneration and necrosis
 In respiratory mucosa and lungs, proliferation of cells occurs
 Death in young goats occur because of diarrhoea and dehydration in severe infection
 Concurrent infection with other diseases aggravate the condition
 Lymphoid necrosis is very marked
 Hyperplasia of cells and inclusion bodies
Clinical signs

 Acute or subacute form

Acute form
Clinical signs similar to Rinder pest in cattle
 High fever
 Dullness
 Sneezing
 Serous discharges from eyes and nostrils which turns mucopurulent later
 Leucopenia
 Necrotic lesions in mouth, oral mucosa forming diphtheretic plaques
 Halitosis
 Diarrhoea within 3-4 years after onset of fever (mucoid or blood tinged)
 Dyspnoea and coughing
 Death within one week of onset of illness
 Pregnant animals abort
Subacute form

 More common in sheep

 Mucopurulent discharge from eyes and nostrils
 Low grade fever
 Intermittent diarrhea
 Recovery after 10 – 14 days
 Lasting immunity in recovered animals
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Gross lesions
 Erosion, necrosis, ulceration on oral mucosa, pharynx, upper oesophagus; abomasum, small
intestine
 Haemorrhage and ulcers in ileo – caecal junction, colon and rectum forming “Zebra stripes”
 Retropharyngeal and mesenteric lymph nodes are enlarged and haemorrhagic
 Spleen enlarged
 Mucopurulent exudate from nasal opening to larynx
 Hyperaemia of trachea and bronchi
 Congestion and oedema of lungs, pneumonia
 With secondary bacterial complications, fibrinous bronchopneumonia and pleuritis is
common
Microscopic lesions
 Syncytia formation in stratified squamous epithelium of upper respiratory tract
 Degeneration and necrosis of infected cells
 Intracytoplasmic inclusion bodies in epithelial cells of upper respiratory tract or intestine
 Proliferative rhino tracheitis, bronchitis, bronchiolitis
 Intracytoplasmic and intranuclear eosinophilic inclusion bodies in the respiratory epithelial
cells / syncytia
Diagnosis
 Differential diagnosis from Rinderpest – Pneumonia is not seen in Rinderpest
 Isolation and identification of the virus
 AGID (agar gel immuno diffusion test) or CIE (counter-immmuno electrophoresis) for
demonstration of antigen in lymphnodes and other tissues
 Immuno capture sandwich ELISA
 RT – PCR
RABIES

Synonym: LYSSA, HYDROPHOBIA ,RAGE

Definition
 Acute viral infection in man and other warm blood animals characterized by abnormal
behaviour, nervous disturbances, ascending paralysis, impairment of consciousness and death.
Aetiology
 RNA Virus – Rhabdoviridae
Incidence
 Rabies has been recorded in various parts of the world.
 In Asia, Africa, Latin America and the middle east , rabies is endemic
 Australia, New Zealand, Hong Kong, Singapore, Britain, Hawaii – free from Rabies
Street virus is that found in natural cases and passage through rabbits increases its virulence
and is called fixed virus
STREET VIRUS FIXED VIRUS
Variable long incubation period Incubation period is fixed and
short
Following intracerebral inoculation, death of rabbit occurs in 14 Death of rabbit occurs in 7
– 20 days days
Negri bodies present Absent
Salivary glands affected Not affected

Suscepibility

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 Extremely susceptible: Dog, fox, wolf, jackal, cat, mongoose, bat.

 Moderately susceptible: Cattle, goat, sheep, horses,
 Cattle and equine – Dead end host
 Vampire bats acts as a vector.
Incubation period :
Depends on the site of the bite. The nearer the bite to the head, the shorter the
incubation period. It may vary from 2 weeks to 9 months.
Transmision
 By bite of an affected animal
 Since the virus is present in the saliva, infection by flies is facilitated
 Licking of the wound by the rabid dog may also result in infection
 Haematogenous spread
 Infection may be spread to the foetus in utero from an affected bitch
Pathogenesis
 After the bite, virus deposited in the wound through a break in the skin
 Local replication of virus in epithelial cells and myocytes
 Cross neuromuscular spindles
 Move centripetally through the nerves
 Reach Central Nervous System
 After reaching the brain, it parasitizes the ganglion cells, grows and then spreads through
various nerves that emanate from the centrifugal fashion
 Virus has got great affinity for salivary glands and s o virus spreading along the nerves that
supply the salivary glands, the facial (vii) and glossopharyngeal (ix) nerves and from there it
is excreted through saliva
 In CNS: Virus damage the nerve cell and vascular endothelium
 Irritation of nerve cells
 Increased excitability (Furious form)
 Neurons of medulla ( Fever, polyuria, glycosuria)
 Neuronal degeneration
 Paralysis of muscles of deglutination unable to swallow / Dribbling of saliva
 Paralysis of jaw muscles , jaws cannot close and so hang down -Dropped jaw
 Paralysis of respiratory muscles – Asphyxia and death
Clinial signs
 Furious form
o Excited, animal goes into rages ( violent anger), indiscriminate biting, barking at
imaginary objects, chews inanimate objects, red eye – Vacant look, champing of
jaws, dribbling of saliva, change of voice.
 Dumb form (Paralytic form)
o Sick, sluggish, progressive weakness, vacant look, does not obey orders, does not
recognize family members, try to hide in corners and under the cot
o Death is 3 to 4 days after onset of symptoms (or) atleast within 10 days.
o Horse: Colic
o Cattle: Bellowing
o Cat -Furious form only
o Rabbit: Dumb form only
o Bull and rams may show sexual urge
Gross lesions
 Limited only to CNS
 Hyperaemia, oedema with petechiae of meninges
Microscopic lesions
 Non suppurative encephalitis
 Perivascular cuffing by lymphocytes
 Microglial cell proliferation and form small nodules – "BABES NODULE "
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 Satellitosis around ganglion cells

 Degeneration of ganglion cells – Neuronophagia seen in hippocampus, brain stem Similar
changes in cerebrospinal and sympathetic ganglia
 Ganglioneuritis of paravertebral ganglion
 Acute catarrh of respiratory and digestive tract
 Hyperemia of kidney, spleen, liver and salivary gland
 Negri bodies: Intracytoplasmic inclusion bodies found in Neurons
 In dogs – Hippocampus impressioin smear
 Cattle – purkinje cell of cerebellum impression smear
 Aggregate of virus
 Round/oval/acidophilic (magenta red) with basophilic granules
Clinical signs
 Under observation of rabies
 Impression smear of Hippocampus stained by seller‟s (or) modified Mann‟s method or
Williams modification of von Gieson method
 Habel‟s mouse Inoculation test: Make an emulsion of piece of ammon‟s horn and dilute 20
times with sterile water. This may be incubated with an antibiotic for ½ hour. 0.03 ml of this
emulsion is injected intracerebrally into 6 mice. On the 5th, 6th and 7th days one mouse is
sacrificed and the brain is examined for negri bodies. Negri bodies is usually seen on the
5thand 6th days
 A biological test, injecting the brain tissue into rabbits intracranially can also done for
conformation
 Corneal or saliva test (FAT)
 Fluorescent Antibody Test FAT)
 Immunoperoxidase Test (IPT)
 Complement Fixation test (CFT)
IMPORTANT FUNGAL DISEASES OF ANIMALS

Sr. Disease Host Characteristics of disease Causative fungus Methods of laboratory diagnosis
No (Animal
affected)

1 Ringworm Cattle, Ring shaped lesions on the skin Microsporum, 1. DME of skin scrapping and
Horse, Trichophyton, affected hair for demonstration
Dogs, Epidermophyton of ectothrix and endothrix.
Cats, 2. Isolation of causative fungi from
Buffaloes skin scrapings and hair on SDA
containing chloramphenicol and
cycloheximide. Incubation at
25C for 2-3 weeks.
Identification by colony
characters and microscopic
examination (for typical
macroconidia).

2 Brooder Chicks Small necrotic foci in lungs Aspergillus 1) Isolation from lung on SDA.
pneumonia fumigatus Incubation at 25C for 3-6 days.
Identification by colony
characters and microscopic
examination.
2) On post-mortem necrotic foci
in lungs.
3) HPE of lung secretions for
demonstration of septic
hyphae.
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3 Bovine Cows, Pulmonary infection followed Aspergillus sp. (70% a. Gross examinations of
mycotic Buffaloes by abortion cases). cotyledons, necrotic
abortion Mucor sp. (30% cotyledons.
cases). b. Impression smears from
Rhizopus sp. and cotyledons for presence of
Candida sp. (very hyphae
rarely) c. Isolation of causative fungi
from cotyledons, placeta and
fetal stomach content.
d. HPE of placental tissue for
presence of hyphae.
4 Rhinosporidi- Cattle, Chronic granulomatous tumours or Rhinosporidium DME of nasal exudate for
osis Horses polyps in nasal mucosa seeberi sporangium and sporangiospores.

5 Mycotic Cows, Mostly after intramammary Sacchromyces, 1) DME of milk for presence of
mastitis buffaloes antibiotic therapy, Udder Cryptococcus yeast.
becomes hard and reduction of neoformans, 2) Isolation of causative fungi
milk yield. Aspergillus, from milk on SDA. Incubation
Candida. at 25 C for mould and at 37 C
for yeast.
6 Cryptococcal Dogs, Incoordination, circling and Cryptococcus  DME of cerebrospinal fluid for
meningitis Cats blindness due to involvement of neoformans, presence of capsulated yeast
CNS (CSF centrifugation
sediment + 1% Negrosin)
7 Thrush Poulty Circular, whitish, raised, Candida albicans a) Isolation from crop lesions on
ulcerative lesions in the mucosa SDA. Incubation at 37 C for 3-
of crop giving appearance of 4 days for yeast.
“Turkish Towel”. b) Subculturing of growth from
SDA on cornmeal agar.
Incubation at 25 C for
demonstration of
pseudomycelium,
chlamydospores and
blatospores.
8 Histoplasmosis Dogs RE system is affected. Lesions Histoplasma A. Blood smears stain by Giemsa
are found in lungs. Enlargement capsulatum method for presence of yeast
of liver, spleen and lymph cells within mononuclear cells.
nodes. B. Isolation from affected organs
on SDA. Incubation at 37 C
and at 25 C for observing
dimorphism.
C. HPE of liver section for
demonstration of fungus in
macrophages.
9 Epizootic Horse Nodules and suppurating Histoplasma 1. Pus smears for demonstration
Lymphangitis lesions with thick yellow pus on farciminosum of yeast cells.
shoulders, neck and limbs. 2. Isolation from lesions on SDA.
Incubation at 37 C and at 25 C
for observing dimorphism.

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References:-
1. Veterinary Microbiology And Microbial Disease by Quinn, P.J., Markey, J.K., Carter,

M.E., Donnelly, W.J. and Leonard, F.C. (2002). . Pub: Wiley – Blackwell.

2. Microbiology by Prescott, Herley and Klein Fourth Edition.

3. Microbiology - Pelczar,JR, Chan and Krieg

4. Practical Medical Microbiology - Collee, Dugid, Frazer and Marnion

5. Veterinary Virology - Murphy, Gibbs, Horzineck and Studert

6. Clinical Veterinary Microbiology - Quinn, Carter, Markey and Carter.

7. Essentials of Veterinary Microbiology - Carter, Chengappa and Roberts

8. e-source - http://www.elearnvet.net.

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