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MICROBIOLOGY NOTES
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1st Year Pharmacy Technician

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By
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Dr. Rida Ali
Lecturer
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Asian Institute of Pharmacy and Health Sciences, Lahore

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 Chapter No 1
MICROBIOLOGY

The branch of biology that deals with microorganisms and their effects on other
living organisms.”

Microbiology is a Greek word

Micron means small
Biologia means studying life

HISTORY

1665 Robert Hook 1. Told the importance of Microscope

DID NOT invented it
2. Described cells based on CORK
structure

Leven Hooke 1670 1. Invented Microscope (single lens)

2. 1674 called small microorganisms as
ANIMALCLUES
3. Died in 1723

Q. Write a Note on SCOPE OF MICROBIOLOGY

Microbiology is important for the following aspects :

1. The study of micro-organism algae bacteria, protozoa, fungi, yeasts and viruses.
2. The organisms convert complex material into simple substances
3. Micro-organism affects human existence IS the foundation of microbiology.
4. The fermentation of fruit juice and the souring of milk
5. Yeasts and bacteria converted sugars to alcohols and acids.
6. Micro-organism has had a decisive role in shaping world history.
7. The growth of the pharmaceutical industry by rising standards , quality In sterile product
there should be no living detective microorganism.
8. The product should be able to pass a test for sterility
9. Medicines administered by vulnerable routes should be sterile products.
10. Disinfection and the properties of chemicals used as antiseptics, disinfectants and
preservatives
11. Sterile medicines may be manufactured by two different strategies.
a) Make the product, pack it in its final container and sterilize it by heat, radiation.
b) manufacture the product from sterile ingredients under conditions that do not permit
the entry of microorganisms.

2
Q. Write a Note on SCOPE OF MICROBIOLOGY IN FUTURE?

1. Microbiology is dynamic, exploding andrevolutionary .

2. Microbiologist discovered vaccines for the prevention and drugs for the treatment.
3. Discovery of some of the vitamins and other basic food materials
4. Knowledge of the chemistry of the cell and cell parts in heredity is a contribution of the
microbiologist.
5. The biologist may use microorganisms as source of food and oxygen for space travelers.
6. The PHYSICIAN needs knowledge of microbiology to fight disease.
7. Industrial scientists utilize microbiology in manufacturing of chemical products.
8. The GEOLOGIST frequently uses information about microorganism in his search for oil.

CHAPTER#2
BACTERIA
History
 In 1673 discovered by Leeuven van Hoek
 he Named them “Little Animal’.
 In 1773 Fredrick Muller naemed them Bacilli.
 In 1850s Casimir Bavaine named them Bacteria.
 the descendants of the earliest form of the life
Definition :
The microscopic, unicellular, prokaryotic organisms characterized by the lack of
membrane bound nucleus and membrane bound organelles .

Classification of Bacteria
(on the basis of Cellular morphology)
They are classified as :
1) Cocci
2) Bacilli
3) Spirals

Bacilli :
 As told by Muller, the rod is known as bacillus
 In various species of rod shaped bacteria
 the cylindrical may be as long 20µm or as short as o.5µm.

3
 COCCI :
 A spherically shaped bacterium
 Derived from greak kokkos, meaning berry.
 Cocci tend to be quite small being
 only 0.5µm to 1.0µm in diameter.

Diplococci : Cocci remain in pairs after reproduction.

 N. meningitidus.
 Neisseria gonorrhoeae.
Streptococci : Cocci that remain in chains called streptococci.
 Streptococcus pyogenes (strep throat)
 S.mutans (tooth decay )
 S.lactis (produce dairy products ,yogurt)

Staphylococcus:The cocci which divide randomly and form irregular grapes like
cluster of cells

 Staphylococcus aureus

Spirals
Spirals may take one of the following three forms.
1)Vibrio
2)Spirilla
3)Spirochaetae

Vibrios : They are the curved rods that resemble commas. Ex : Vibrio cholerae
(causing cholera)

Spirilla : They are helical shaped with a thick, rigid cell wall and flagella . Ex: Spirillum
volutan

Q. Write down the characteristics of CELL WALL

 Cell wall is to protect the cell and determine it shape.

 Bacterial cell wall is made up of Peptidoglyan

 Peptidoglycan is a large molecule and it contains two amino-

containing…carbohydrates

4
 N-acetyl glucosamine N-acetyl muramic acid.
These two are joined by cross bridges of amino acid.

Q. Difference between Gram +ve and Gram –ve

Gram Positive Bacteria Gram Negative Bacteria

Peptidoglycan is 25 nm Peptigoglycan is 3 nm
It has teichoic acid No teichoic acid,
About 60-90% is peptidoglycan Complex, contains
polusachharides, protiens,
lipids.
Stains Voilet by Gram staining Stains pink by Gram staining.

Q. Explain the following terms in detail ?

Chromosome:

1) The bacteria have no distinct nucleus and are hence called Prokaryotes.
2) Bacterial chromosome lie suspended in the cytoplasm.
3) It also lack protein.
4) The chromosome region is called Nucloid.

Plasmids:

1) They are extra-chromosomal rings of DNA.

2) they contain few genes (DNA)
3) they are not essential for bacterial growth,
4) carry genes for drug resistance.
5) ften called R factors
6) They are very important in genetic engineering.

Ribosomes:

1) Ribosomes are bodies of RNA and protein.

2) Involved in the synthesis of protein.

Inclusion Bodies:

1) Globules of starch, glycogen or lipids in the cytoplasm

2) They store nutrients for periods of starvation.

Volutin:

1) They are depots of phosphate.

2) They Stain deeply with dyes such as methylene blue.

5
Magnetosome:
It helps certain bacteria orient themselves to the environment toward their preferred
habitat.

Cell Membrane

1) The cell membrane boundary layer.

2) It exists inside the cell wall in plants and bacteria.

Cell Envelope

Cell Membrane + Cell Wall + Capsule = Cell Envelope.

Write a note on Capsule ?
1) Sticky, gelatinous layer of poly saccharides and proteins around the cell wall
2) Cocci and baccili contains capsule, Spiral don’t form capsule.
3) The loose layer of capsule is called Glycocalyx.
4) It contains a mass of tangled fibers of dextrin, a polysaccharide.
5) These fibers help bacteria attach to the surface of the host.

Describe The Host-Parasite Relationship?

 It is the relationship between two organisms,
the host and the parasite,
 the competition for supremacy takes place between them.
 if the host loses the competition, disease develops

Q. Explain the terms related to Host Parasite Relationship ?/

describe host parasite relationship in detail ?
Mutualism:
Relationship is beneficial to both the body and the microorganisms,

Commensalism:
Relationship is beneficial only to the microorganisms but no harm
to Host.

Parasitism :
Relationship is beneficial only to the microorganisms, and harmful
for host .
Opportunistic:

6
Certain commensals become parasites when the body’s normal
defenses are suppressed. Commensilism ------- Parasitism

Q. Define Pathogenicity?
The ability of a parasite to gain entry to the host’s tissues and bring
about a physiological or anatomical change resulting in a change of
health and thus disease.

Pathogen: An organism having pathogenicity

Q.Explain Variations in Pathogenicity ?

Parasites vary greatly in their pathogenicity.
Virulency: the degree of pathogenicity of a parasite.
Virulent: An organism that invariably causes disease is highly
virulent
Moderately Virulent : Organism that sometimes causes disease
Avirulent: They are not regarded as disease agent

How Normal Flora in the Neonates is Formed?

1) When the child passes through the birth canal.

2) When breathing begins and upon first feeding.
3) Within two to three days most of the flora have appeared.
4) During the next few weeks, contact with the mother
5) Other individuals will expose the child to microorganisms.
6) The normal flora remains throughout life,
7) Changes in response to the internal environment

7
Write a note on Normal Flora ?
Definition: A population of microorganisms that infect the body
without causing disease.

Skin:
viruses, fungi
bacteria staphylococci and Propionibacterium acnes.

Oral Cavity:
Neisseria, Leptotrichia,
Bacteroides, diphtherialike bacilli (diphtheroids),
fungal spores streptococci.

Respiratory Tract:
pneumococci species of Haemophilus
Mycoplasnia.

Small and Large Intestine:

Bacteroide Clostridium spores
streptococci Gram-negative rods
Enterobacter, Kiebsiella,
Proteus Pseudomonas
Escherichia coli Candida albicans( yeast)

Vagina: Lactobacillus
Blood and Urine: Sterile unless disease is present.
Stomach: Without a normal flora mainly due to the low pH .

Chapter no 3
8
 CULTURE MEDIA
Q. What is Natural Media? Give its types ?

Definition :
The media of which the exact components and quantities are not
known is called Natural Media.

Types :
Beef Broth
Nutrient Agar
Enriched Media
1)Blood Agar
2)Chocolate Agar
Selective Media
1. Eosin Methylene Blue Agar :(EMB).
2. Mannitol Salt Agar:
Differential Media:
1). Mac Conkey Agar

Q. Wrtie a complete note on Natural Media ?

It has following media’s involved:

Beef Broth:
Pasteur and Koch used beef broth for the cultivation of bacteria.

Nutrient Agar
The modern form of this liquid medium, consists of:
1) Water
2) beef extract
3) peptone( protein supplement from plant or animal)
4) Agar (polysaccharide from marine algae). It makes it media
solid so that bacteria can grow.

Enriched medium:
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It’s for fastidious bacteria which require special nutrients to grow.
Types of Enriched media are:
1) Blood Agar:
Streptococci causing strep throat and scarlet fever grow well when
whole blood is added to the nutrient medium ,is called blood agar.
2) Chocolate Agar:

 For the growth of Neisseria species,

 Blood agar is heated before solidification.
 Heating disrupts the red blood cells
 Releases the hemoglobin
 Gives charred brown appearance

Slective Media:
Contain ingredients to inhibit the growth of certain bacteria in a
mixture while permitting the growth of others
1) Mannitol Salt Agar:
Medium contains mannitol in a high salt concentration that
inhibits most other bacteria except staphylococci.
2)Eosin Methylene Blue Agar :(EMB)
Contains eosin and methylene blue, two dyes that inhibit Gram-
positive bacteria and permits E.coli and Gram –ve to grow.

Differential Medium:
Medium makes it easy to distinguish colonies of one organism from
colonies of other organisms on the same plate.

1) Mac Conkey Agar: It contains

 The dyes neutral red and crystal violet
 Carbohydrate lactose. Those bacteria that ferment the lactose
take up the dyes and form red colonies;
 Other bacteria show up as colorless colonies
 Bile salts that inhibit the growth of Gram-positive bacteria.

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This medium is selective + differential.

Q. Write a Note on Synthetic Media?

Definition :Synthetic media are chemically defined. The nature
and amount of each component are known

Components :
The glucose supplies energy .
The ammonium ions for amino acid + nucleic acid formation;
Phosphate is used in DNA and RNA synthesis;
Sulfur from magnesium sulfate is for enzyme formation;
Sodium chloride maintains a stable internal environment

Q. Difference between Bacterial and Pure culture ?

Bacterial culture: Bacteria grow on a laboratory are called a

culture.
Pure culture :Population of cells which are derived from a single
cell.

Q.WHAT ARE METHODS OF ISOLATION OF PURE CULTURE ?

1) THE STREAK PLATE TECHNIQUE
2) POUR PLATE TECHNIQUE
3) SPREAD PLATE METHOD
4) ENRICHMENT CULTURE TECHNIQUE
5) SERIAL DILUTION TECHNIQUE
6) SINGLE CELL ISOLATION TECHNIQUE
THE STREAK PLATE TECHNIQUE
It is used to spread millions of cells over the surface of a solid
medium so that cells are deposited at a distance from others. They
grow forming an isolated colony representing a Pure Culture.

POUR PLATE TECHNIQUE

1) It is used to determine the number of microbes/ml or

microbes/gram in a sample.
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 2) Not requires previously prepared plates,
3) Often used to determine bacterial contamination of foodstuffs.
PROCEDURE
1) Prepare sample.
2) Place the diluted sample in an empty sterile plate.
3) Pour in 15 ml of melted agar which has been cooled to 45°
C ,mix.
4) Let cool to solidify on a flat table top.Incubate to develop
colonies.
The range for total Colony forming per unit CFU/plate is between
30 to 300 colonies/plate
DISADVANTAGE
Colonies will be much smaller, and must be carefully counted

SPREAD PLATE METHOD

A portion of bacterial specimens is spread over the surface of a solid

medium.After appropriate incubation, growth from each culture
should be checked microscopically and culturally to verify that it a
pure culture.
LIMITATION: Small amount of specimen is spread over the agar.
ADVANTAGE: Minimal amount equipment s required,

ENRICHMENT CULTURE TECHNIQUE

It is used when bacterium to be isolated is present in relatively

small number and grows more slowly than many other species in
the mixture.

SERIAL DILUTION TECHNIQUE

12
 1) If the organism is present in a mixed culture , to obtain it in a
pure culture is done by a series of dilution in tubes of
medium,
2) When greatly diluted, the specimen contains only the one
specie
3) The purity of a culture isolated is confirmed by plating
procedure.

SINGLE CELL ISOLATION TECHNIQUE

1) Equipment, the micromanipulator, used with microscope to

pick a single organism from a hanging drop preparation.
2) Singe-cell is taken in the tube and transferred to medium for
growth.
3) The technique is reserved for use in highly specialized studies
4) It requires skilled operator.

Q. EXPLAIN GRAM STAINING :

METHOD:

1) Make a thin smear of the material let it dry.

2) Pass the slide through a flame once or twice
3) Place the slide on the rack and flood with the crystal violet
4) Was off the stain with Lougol’s iodine and leave it for one
minute.
5) Rinse in water.
6) Pour on alcohol till no more violet color comes from the slide
7) Rinse in water again
8) Stain it with counter-stain Safrainin
9) Rinse in water and allow it to dry
Result:
Gram Positive = Purple Violet
Gram Negative = Pink
Q. Explain ZIEHL-NIELSEN STAINING/ACID-FAST STAINING?
Why It’s Done?
Bacteria has ability to resist decolourization by acid and alcohol a
very strong stain is used basic fuchsin that can penetrate the waxy
13
covering of bacteria.

Method
1) Make a smear of the material
2) Allow drying at room temperature.
3) Pour strong carbolfuchsin and heat gently
4) Rinse in water
5) Pour 25% sulfuric acid
6) Leave the until the smear is pale pink in color.
7) Rinse in water and pour on alcohol for a few minutes.
8) Counter stain with malachite green ethylene blue
Result
The tubercle bacillus (T.B) resists decolorizing by acid and alcohol it
will Stain bright red

Chapter no 4
Viruses

Q.Characteristics of viruses
Virus is a Latin word meaning “Poison”
Viruses are obligate intracellular parasites

Definition :
 Viruses are non cellular
 Infectious agents
 Consist DNA or RNA, reproduce only in living cells
 Use the biosynthetic machinery of the host cell
 Direct the synthesis of virion
 Containing viral genome
 Transfer them to the other cells

Q.Write a note on Shapes of Viruses ?

Viruses may appear in several shapes.

14
1.Helix:
A tightly wound coil resembling a corkscrew or spring. Rabies and
Tobacco mosaic viruses
2.Icosahedron :
A polyhedron with20 triangular faces and 12 corners.Herpes
simplex and polio viruses
3.Complex:
A combination of helical and icosahedral symmetry is called
complex. bacteriophages (icosahedral head and a collar and tail)

Q. Explain the Sturcutre of Virus?

All viruses consist of two basic components:
 a core of nucleic acid, the genome,
 a coat of protein known as the capsid.
Genome:
1) The genome contains either DNA or RNA, but not both
2) The nucleic acid occurs as double-stranded or single-stranded
3) The genome may be folded, condensed, or coiled.

Capsid:
1) The capsid protects the genome.
2) It also gives shape to the virus
3) It is responsible for the helical, icosahedral, or complex
symmetry.

Capsomers:
1) Capsid is subdivided into protein subunits called capsomeres
2) It gives the symmetry.
3) The number of capsomeres is characteristic for a particular
virus.
Capsid of herpes =162 capsomeres.
Capsid of adenoviruses =252 capsomeres.

Envelope:
15
 1) Many viruses are covered by flexible membrane called
Envelope.
2) It is composed of lipids and protein and is similar to the host
cell membrane
3) Envelope is generally loose-fitting structure
4) Except that it includes viral-specified components.
5) Enveloped viruses may lose their infectivity when envelope is
removed.
6) When the envelope is present, symmetry of the capsid may not
be apparent

Q What are Spikes ?

 In some viruses as influenza and measles viruses, the

envelope contains functional projections know as spikes.
 The spikes contain enzymes to assist attachment of viruses to
host cells.

Q. Describe Virion?
1) A complete assembled virus outside host cell is known as
Virion.

2) Virions lack the chemical machinery for generating energy

and synthesizing large molecules.

3) They must rely on the structures and chemical components

of their host cells to replicate themselves.

Q. Explain Viroids?
 Viroids are tiny fragments of nucleic acid
 They cause several diseases of plants
 They are involved in human and animal diseases.

Q. Write a Note on Viral Genome ?

DNA viruses:
1) The genome replication takes place in the nucleus.
2) They are entirely dependent on the host cell's DNA and RNA
synthesizing machinery.
16
 3) The viral genome must cross the cell's nuclear membrane to
access this machinery.
4) The DNA may be double stranded or single stranded.
RNA viruses
1) They are unique because their genetic information is encoded
in RNA.
2) Replication usually takes place in the cytoplasm.
3) RNA may be single-stranded or double-stranded..

Q.What is a Sense? Explain +ve and –ve senses?

 Positive-sense viral RNA is identical to viral mRNA and thus
can be immediately translated by the host cell.
 Negative-sense viral RNA is complementary to mRNA and thus
must be converted to positive-sense RNA by an RNA
polymerase before translation.

Q.What are Reverse transcribing viruses?

1) These replicate using reverse transcription

(formation of DNA from an RNA)
2) Reverse transcribing viruses containing RNA genomes use a
DNA intermediate to replicate
3) Those containing DNA genomes use an RNA intermediate
during genome

Classification
1962, André Lwoff, Robert Horne, and Paul Tournier were the first
to develop virus classification, based on the Linnaean hierarchical
systemwhich includes phylum, class, order, family, genus, and
species.

ICTV classification

Viral classification starts at the level of order and follows as thus,

Six orders have been established by the ICTV.

17
Caudovirales: They are tailed dsDNA bacteriophages,
Herpesviraleso : It contains large eukaryotic dsDNA viruses,
Mononegavirales : It includes non-segmented (-) strand ssRNA
plant and animal viruses,
Nidovirales : It is composed of (+) strand ssRNA viruses with
vertebrate hosts,
Picornavirales It contains small (+) strand ssRNA viruses that
infect a variety of plant, insect, and animal hosts, and
Tymovirales:It contains monopartite ssRNA viruses that infect
plants.

Baltimore classification
The Nobel Prize-winning biologist David Baltimore devised the
Baltimore classification system.
Baltimore classification (first defined in 1971) places viruses into
one of seven groups
1) dsDNA viruses (e.g. Adenoviruses, Herpesviruses, Poxviruses)
2) ssDNA viruses (+)sense DNA (e.g. Parvoviruses)
3) dsRNA viruses (e.g. Reoviruses)
4) (+)ssRNA viruses (+)sense RNA (e.g. Picornaviruses,
Togaviruses)
5) (−)ssRNA viruses (−)senseRNA (e.g. Orthomyxoviruses,
Rhabdoviruses)
6) ssRNA-RT viruses (+)sense RNA with DNA intermediate in life-
cycle (e.g.Retroviruses)
7) dsDNA-RT viruses (e.g. Hepadnaviruses

Chapter no 5
FUNGI
Q1. What is a thallous?
Thallous is a plant body that has no distinct roots, shoots and stems

18
Q2. Write down the names of divisions of fungi?
The four major Divisions of fungi are:

 Zygomycetes:
 Ascomycetes:
 Basidiomycetes:
 Chytridiomycetes:

Q3. Characteristics/ Features of divisions of fungi?

Zygomycetes  bread mold, Rhizopus siolanifei

Ascomycetes  Ascomycetcs/sac fungi.

Basidiomycetes  Basidiomycetes/ club fungi.

Chytridiomycetes  The simplest of the true fungi

 Cliytridiomycetes/ chytrids.

 Chytrids are derived from a protozoan

ancestor having similar flagellation.

1. Fungi are primarily terrestrial organism .Few are freshwater or marine.they are
found in the moist damp places

2. Many are pathogenic and infect plants and animals.

3. 3/4 of all vascular plants form associations between their roots and fungi called
mycorrhizae

4. Lichens are associations of fungi an either algae or cyanobacteria.

5. Fungi also are found in the upper portions of many plants. They affect plant
reproduction and palatability to herbivores
Q4. Write a note on Habitat of fungi :

Q5. Structure of Fungi

 vegetative structure ------------ thallus.

 Single-cell microscopic ------- yeasts
 Multicellular ---------------------- Molds
 Macroscopic --------------------- puffballs, and mushrooms

19
Q6. Write a note on Yeast / define yeast / write down the characteristics of yeast ?

1. Yeast is a unicellular
2. it has a single nucleus.
3. They are larger than bacteria
4. They are commonly spherical to egg shaped.
5. They have no flagella but do possess most of the other eukaryotic organelles.
6. Yeast size depends on the species
7. 3–4 µm in diameter, some yeasts can reach over 40 µm.
8. Almost 1500 species of yeast has been described.

Q7. Draw single cell yeast ?

Q8. Importance of yeasts

Fermentation:
They ferment the carbohydrates into ethanol (alcohol) and CO2 .
This process is very important in wine, beer and bread making.
Saccharomyces cerevisiae is used in baking and fermenting alcoholic beverages.

Research;
It is important in biology research and is one of the most thoroughly researched
eukaryotes.

Candida albicans:
they are opportunistic pathogens ,can cause infections in humans. Candida is found as
a in the mucus membranes of humans

Biofuel industry:
Yeasts are used to generate electricity, and produce ethanol for the biofuel industry.

20
Spoilage of Wine:
Growth of some yeast such as Zygosaccharomyces and Brettanomyces in wine can
cause wine spoilage.

Spoilage of food:
Yeasts are able to grow in foods with a low pH, (5.0 or lower) In this way they spoil
food.

Methylene Blue is used to test for the presence of live yeast

cells.

Q9. Define Hyphae?

A mold consists of long, branched, threadlike filaments of cells called hyphae that form

a mycelium ,a tangled mass or tissue like aggregation .

Q10. Differentiate between septate and non septate hyphae?

Septate hyphae The hyphae have cross walls called septa

with either a single pore or multiplepores .

Hyphae are composed of an outer cell wall

and an inner lumen, which contains the
cytosol and organelles

Non Septate hyphae They are not divide into cells by cross
walls called septa.

These hyphae are called coenocytic

hyphae.

Q11. Explain the nutrition in fungi ?

Saprotrophs:
They get energy by decomposition of dead organic substances. They release hydrolytic
exoenzymes that digest external substrates.then absorb the soluble products. Also
called chemo-organo-heterotrophs

21
Parasites
They may be either Obligate parasites or Facultative Parasites.

Obligate parasites:
They can grow only on the host cell through special hyphal tips called haustoria.

Facultative parasites:
Besides living on their hosts they can also survive on the growth media.

Diseases:
The water mold Peronospora hyoscyami is c responsible for the "blue mold" of tobacco
Oomycetes cause late blight of potatoes and grape downy mildew.

Q12. Explain Symbiosis in fungi ?

Lichens: It is symbiotic association between a fungus and alga.

Mycorrhizae: It is symbiotic association between fungus and the roots of vascular
plants.

Q13. Explain Respiration in fungi ?

1) Fungi usually are aerobic.

2) Some yeast are facultatively anaerobic and can obtain energy by fermentation
3) Obligatory anaerobic fungi are found in the rumen of cattle.

Q14. Explain Reproduction in fungi ?

 Yeast: Budding
 Asexual Reproduction in Molds:
Spore
Conida
ragmentation
 Sexual Reproduction

CHAPTER NO 6

MICROBIOLOGY OF WATER
Q. Define water Microbiology ?
Definition:
Water microbiology is concerned with the microorganisms that live in water, or can be
22
transported from one habitat to another by water.

Q. Write down the types of water ?

a) Ground water
b) Fresh water

Q. Write down the characteristics of ground water ?

a. It originates from deep wells and subterranean springs.

b. This is free of bacteria due to filtering action of soil, deep sand and rock.
c. it may become contaminated when it flows along the channels.

Q, Write down the characteristics of fresh water ?

1. It is found in streams, lakes, and shallow wells.

2. Sources of microbial contamination of a body of water are soil and agricultural

run off, farm animals, rain water, industrial waste, discharges from sewage
treatment plants and storm water run off from urban areas.

3. Many microorganisms are found naturally in fresh and saltwater. These include
bacteria, cyanobacteria, protozoa, algae, and tiny animals such as rotifers. These
can be important in the food chain that forms the basis of life in the water.

Q, Write down the sources of contamination of fresh water ?

Sources of microbial contamination of a body of water are as follows:

1) soil and agricultural run off,

2) farm animals,
3) rain water,
4) industrial waste,
5) discharges from sewage treatment plants
6) storm water run off from urban areas.

Q. Name the zones included in fresh water microbiology?

1) littoral zone
2) benthic zone
3) limnitic zone
Q. Write down the Characteristics of littoral zone?

23
  A variety of microorganisms live in fresh water.
 The region of a water body near the shoreline ()
 It is well lighted, shallow, and warmer than other regions of
the water.
 Photosynthetic algae and bacteria that use light as energy live
here.

Q. What are the Characteristics of The Limnitic Zone?

 Further away from the shore is the.

 Photosynthetic microbes also live here.
 As the water deepens, temperatures become colder and the oxygen
concentration and light in the water decrease.
 Now, microbes that require oxygen do not thrive.
 Purple & green sulfur bacteria,that can grow without oxygen lives here

Q. What are the Characteristics of The Benthic Zone ?

 The bottom of fresh waters.

 Few microbes survive.
 Bacteria that can survive in the absence of oxygen and sunlight, such as
methane producing bacteria lives.

Q. Write down the Methods to minimize the microbial contamination?

Drinking water is usually treated to minimize the risk of microbial contamination.:

1. in pre-Christian times the water was stored in metal jugs”

2. the boiling of drinking water, as a means of protection of Water
3. Chemicals such as chlorine or chlorine derivatives has been a popular means of
killing bacteria such as Escherichia coli in water
4. use of a gas called ozone.
5. by the use of ultraviolet light
6. By filteration

CHAPTER NO 7
MICROBIOLOGY OF AIR
Q.What are the Characteristics of Air:

 The microbial flora of air is transient and variable.

24
  Air is not a medium in which micro-organisms can grow
 It is carrier of particulate matter, dust, and droplets, which may be laden
with microbes.
 No micro-organisms are indigenous to air
 The air in our environment, several miles above the earth’s surface, over
land ,over water contains various species of micro-organisms.

Q. Describe the Transmission of Air borne Micro-organisms.

Air borne microorganisms can be transmitted by

 Sneezing
 Coughing
 Talking
 dust particles
 droplets from mouth

Q. Explain the Fate of Airborne Micro-organisms ?

The fate of airborne micro-organisms depends on atmospheric conditions as;

 Humidity
 Sunlight
 Temperature
 Size of the particles bearing the microorganisms
 Nature of the micro-organisms

Q. Describe the Microbial Content of Indoor Air?

The microbial contamination of indoor depends on;

 Ventilation rates
 Crowding
 Nature and degree of activity of the individuals

Q. How the size of particle carrying micro-organisms effects them?/ effect their
spread?

1) Particles of dust raised from surfaces and droplets expelled from the
respiratory tract vary in size from micrometers to millimeters.

25
 2) Particles in the low-micrometer range can remain airborne for long periods
time
3) Larger droplets and dust particles settle rapidly as dust on surfaces.
4) This dust becomes airborne intermittently during periods activity in the
room.

Q. Micro-organisms present in In door Air?

Tubercle Bacilli: Tubercle bacilli have been isolated from the dust of Sanitoria.
Diphtheria Bacilli and Hemolytic Streptococci: They have been isolated from
floor dust near patients or carriers .

Q. Explain the microbial flora of Outdoor Air?

 Algae, protozoa, yeasts, molds and bacteria have been isolated from the air
near the surface of earth
 Mold spores constituted the largest portion of the airborne micro flora.
 The predominant mold spores were of the species Cladosporium
 Among the bacterial types were spore forming and non-spore forming gram
positive bacilli, gram positive cocci, and gram negative bacilli,

Q. What are the Bacterial Species Present in Outdoor Air?

 Micrococcus
 Sarcina
 Gram-negative rods
 Gram positive pleomorphic rods
 Aerobic spore formers

Q. what are the Fungal species present in Outdoor Air? :

 Cladosporium(Most abundant)
 Alternaria
 Pullularia
 Penicillium
 Batrytis
 Stemphylium

Q.What are Devices for Microbiological Analysis of Air ?

1) Solid impingement devices.
26
 2) Liquid impingement devices have been designed for this purpose.
3) Liquid Impingement Devices:

Q. What are the techniques and devices used for microbiological analysis of air ?

1) Setting-plate Technique:
2) Sieve and slit-type Samplers:
3) Membrane filter

Q.Write a Note on the Air Borne Diseases ?

Bacterial

 Diphtheria
 Tuberculosis
 Pneumonia
 Meningitis

Viral

 Small pox
 Measles
 Influenza
 Common cold

Fungal

 Systemic Mycosis
 Histoplasmosis
 Cryptococcosis

Q.Explain The Methods to CONTROL OF MICRO-ORGANISMS IN AIR

There methods of controlling the level air contamination;

1.Ultraviolet Radiation

 Direct Irradiation: Indirect Irradiation:

2.Chemical agents

 Triethylene glycol
 Formaldehyde Lactic Acid
27
3.Filtration

 Use of cotton plug

 Air filter

4.Laminar- Airflow system

CHAPTER NO 8
MICROBIOLOGY OF SOIL
Q.What are the characteristics of SOIL?

 The region of earth's crust where geology and biology meet is called soil.
 The characteristics of the soil environment vary with location and climate
soil differs in depth, chemical composition, physical properties and origin.

Q. What are the Soil Constituents ?

There are 5 major categories

 Mineral particles
 Organic residue
 Water
 Gases
 Biological systems.

Q. What are the Mineral Particles Present in Soil ?

 The dominant particles are Aluminum, Iron and Silicon.

 Less dominant are Calcium, Magnesium, Potassium, Sodium, Nitrogen,
Sulphur, Phosphorous, Titanium, Manganese etc.
 Particle size from small particles (0.002mm or lesser) to large pebbles and
gravel.

Q. Explain Variations in Soil Water ?

The amount of water depends upon

 Amount of precipitation

28
  Climate conditions
 Soil composition
 Drainage
 Living population of soil.

Water is retained as:

 Free water in the spaces between soil particles.

 Absorbed to the surface of particles.
 Various organic and inorganic components of soil are dissolve in soil water
and thus are made available as nutrients for soil inhabitants.

Q. Write a note on Gases present in Soil ?

Gaseous phase of soil consists of

 Carbon dioxide
 Oxygen
 Nitrogen
 Small amount of the gases are dissolved in water
 Some are present primarily in spaces between soil particles.

Q. Explain Microbial Flora of Soil / Describe Bacteria present in Soil ?

Bacterial population is highest in both number and variety.

 Autotrophs
 Heterotrophs
 Mesophiles
 Thermophiies
 Sulphur oxidizers
 Nitrogen fixer
 Protein digesters and other kinds of bacteria are all likely to found in soil.

Q.Describe Fungi Present in Soil?

 Their numbers range from thousands to hundreds of thousands

29
  Fungi are active in decomposition of cellulose and lignin of plant tissue.
 The mold mycelium penetrates the soil and forms a net-work which
entangles soil particles and forms water stable aggregates.
 Fungi also improve the physical condition of the soil

Q. Describe Algae present in Soil?

 Algae are generally found on the surface of moist soils, where there is
sufficient light for photosynthetic reactions.
 The major types present are green algae Chlorophyceae, and the
 Their total numbers vary from several hundreds to several thousands.
 They fix nitrogen in paddy soils used for cultivation of rice.

Q. Describe Protozoa Present in Soil?

 Many types of Protozoa are found in the soil, but flagellates and amoebae
usually outnumber the ciliates.
 Their total numbers may range from a few hundred to several thousands.
 Since Protozoa do not ingest all bacteria, they maintain some equilibrium of
the bacterial flora of the soil.

Q. Write a note on the Roles of Soil Microbial flora /Biota?

1) Recycling of the energy, carbon, and nutrients in dead plant and animal
tissues into forms that are potentially useful for living plants
2) Many of these hazardous or toxic substances can be degraded by soil
microorganisms.
3) Soil microorganisms are also responsible for transformations of elements to
various forms

Q. What is Carbon Cycle ?

 In the carbon cycle, microorganisms transform plant and animal residues

into carbon dioxide and the soil organic matter known as humus.
 Humus improves the water-holding capacity of soil, supplies plant nutrients
Etc

30
Q. What is Nitrogen Cycle ?

 Soil microorganisms play key roles in the .nitrogen cycle/Nitrogen (N2)

Fixation
 The atmosphere is approximately 80% nitrogen gas (N2), a form of nitrogen
that is available to plants only when it is transformed to ammonia (NH3) by
soil bacteria .

Q. Explain Sulphur Cycle ?

 Cycling of sulphur is similar to that of nitrogen.

 Transformations between organic and elemental states and between
oxidized and reduced states of sulphur are done by especially bacteria.

CHAPTER NO 9
STERILIZATION
Q. Define sterilization and give its purposes?
Definition:
Sterilization is the process of killing or removing bacteria and all other forms of
living organism and their spores from preparation or articles.

Purpose

 To kill Micro organisms

 To preserve substances
 To purify the substances by removing bacteria , viruses etc
 To make substances sterile

Q. What are the Methods of Sterilization? ? Name the methods of sterilization

of Sterilization?

Physical Methods

 Dry heat sterilization

 Moist heat sterilization
 Sterilization by radiations

Chemical Methods

31
  Gaseous sterilization
 Sterilization by disinfectants

Mechanical Methods

 Sterilization by filtration

Q. Write a Note on Physical Methods /write a Note on Dry Heat Sterilization ?

 Substances which are destroyed by moist heat may be sterilized by Dry

heat.
 Dry heat can be used to sterilize but the heat takes longer time and temp.
 Dry heat is used on powders and heat-stable items that are affected by
steam.
 The standard setting for a hot air oven is
 2 hours at 160 C (320 0F)
 190 0C (374 0F) for 6 minutes for unwrapped objects
 12 minutes for wrapped objects

Q. What are the Substances Sterilized by Dry Heat Sterlization?

 Fixed oils , Propylene glycol

 Liquid paraffin, Petroleum
 Powders , Sterilize glass
 Surgical instruments, Surgical catgut

IMPORTANT NOTE : Volatile preparations or substances and surgical dressing

can not be sterilized by this method.

Q. What is the Mechanism of Dry Heat Sterlization ?

1) During dry heat sterilization the microorganism and bacteria spores are
killed by oxidation
2) Dry heat is less effective than moist heat higher temperature and longer
period is required.

Q. What are the Advantages of Dry Heat Sterilization ?

32
 1) Suitable for substances destroyed by moisture
2) Glass wares like flasks test tubes pipettes can be sterilized
3) Less damaging to glad and metal equipments than moist

Q. What are the Disadvantages of Dry Heat Sterilization ?

 Can not used for volatile and thermolabile substances.

 Required long heating time and high temperature
 Not suitable for surgical dressings.

Q. What are the Techniques that uses Dry Heat Sterilization ?

Flaming:

1) It is simplest method of dry heat sterilization.

2) The material to be sterilized is kept in the hot part of of the Bunsen burner
flame for few seconds and the process is repeated several times
3) This method is generally used for those articles which are to be used
immediately for example forceps ,blades, knives,needles wire loops ,metal
spatulas.

Hot Air Oven :

1) It consists of metallic chamber of aluminium or stainless steel

2) Which is electrically heated
3) Thermostatically controlled
4) They are of two types.
5) In which air is circulated by gravity convection to all parts of the chamber.
6) Mechanical convection type in which air is circulated by fan,more
satisfactory because sterilizing temperature is controlled.

Material Sterilized :

 Glass ware conical flasks

 test tubes pippets

Note : They should be plugged with non absorbent cotton wool

Incineration

1) Incineration will also burn any organism to ash.

33
 2) It is used to sanitize medical and other biohazardus waste before it is
discarded with non- hazardous waste.

Q.Write a Note on Moist heat sterilization ?

 It is the most reliable method of sterilization

 In the presence of moisture bacteria are destroyed at lower temperature .

Mechanism
Microorganisms are destroyed by denaturizing and coagulation of proteins.

Advantages

1) Microbes are killed more effectively

2) Ampoules are readily sterilized by this method
3) Bulk quantities surgical dressing and surgical instrument are effectively
sterilized

Disadvantages
Thermolabile substances and ointments can not be sterilized.

Q. What are the Methods that uses Moist Heat Sterilization ?

1) Autoclaving
2) Heating with bactericide
3) Heating with boiling water
4) Tandalization

Q.Write a Note on Autoclaving ?

Autoclaving is the process of heating in an autoclave in which saturated steam

under pressure is allowed to penetrate through the material for 20 minutes at
temperature of 121C.

Autoclave

It is an apparatus used for sterilization by steam underpressure.

34
Working

1) Autoclaves commonly use steam heated to 121 C or 134C.

2) For Sterilization,holding time of at least 15-20 minutes at 121 C or 3
minutes at 134 C is required.
3) Additional sterilizing time is required for liquid/instruments packed .
4) Autoclave treatment will kill all fungi, bacteria, viruses and also spores .
5) For prion elimination, 121-132C for 60 minute or 134C for 18 minutes.

Precautions:

 For effective sterilization steam needs to penetrate tuniformly.

 During the initial heating of the chamber, residual air must be removed.
 For autoclaving, cleaning is critical.
 Cleaning can also removed large number of organisms.
 Cleaning instruments ,cool water must be used.
 Treatment with ultrasound or pulsed air can be used to remove debris.

Q. Explain methods using moist heat for sterilization ?

Heating With A Bactericide:

1. Bactericide is added to the solutions to be sterilized which are then sealed.

2. The sealed containers are then heated at 100C for 30 minutes in water
bath.
3. Commonly used bactericide includes ( Benzalkonium chloride, Chlorocresol)

Sterilization by boiling water

1) The boiling water bath is most use full for sterilizing instruments like
syringes, knives, blades, scissors and others.
2) They are completely dipped in boiling water for 20 minutes.

Tyndallization

35
 1) Tyndallization named after john Tyndall
2) Lengthy process designed to reduce the level of activity of sporulating
bacteria
3) The process involves boiling for a period at atmospheric pressure, cooling,
incubating for a day and finally boiling again.
4) The three incubation periods are to allowed to kill heat-resistant spores
5) Tyndallization is ineffective against prions.

Q.Write a Note on/Explain Sterilization by Radiation ?

1) Sterilization by radiation is also known as cooled sterilization

2) No heat is used in this method.
3) The microorganisms are very susceptible to lethal effects of radiation.

MechanismBy radiations, alteration of chemicals takes place in

microorganisms ,which destroy the microbes, Nucleoproteins are destroyed by
radiations

Advantages

1) Used in the preservation of food and parenterals containing antibiotics.

2) Used for the sterilization of some bacterial and viral vaccines.
3) No aseptic handling is required
4) Sterilization can be done after packing.

Disadvantages

1) High cost.
2) Radiations are harmful the persons operating.
3) Radiations may lead to change in color, texture and solubility.

Methods: Methods exist to sterilize using radiation such

1) electron beams
2) X- rays
3) gamma rays
4) subatomic particles.

36
Gamma rays
Very penetrating and are commonly used for sterilization of disposable medical
equipment, such as syringes, needles, cannulas and IV sets.

Electron beam processing

 Commonly used for medical device sterilization,.

 Due to the higher dose rate, less exposure time is needed and potential
degradation to polymers is reduced.
 A limitation is that electron beams are less penetrating

X-Rays

1) X-rays are less penetration than gamma rays

2) Require longer exposure times
3) Require less shielding,
4) Generated by an x-ray machine that can be turned off for servicing and
when not in use.

Ultraviolet light

1) Useful only for sterilization of surfaces and some transparent objects.

2) Many objects that are transparent to visible light absorbed UV.
3) It also damages many plastics, such as polystyrene foam.

Q. Describe Sterilization by Chemicals/ Chemical Sterilization ?

Chemical are also used for sterilization. Chemical sterilization includes:

 Ethylene oxide
 Ozone, Chlorine bleach
 Glutaraldehyde , Formaldehyde
 Hydrogen peroxide
 Peracetic acid, Prions

Chlorine bleach ( Most Commonly Used)

1) Chlorine bleach is accepted liquid sterilizing agent.

2) House hold bleach consists of 5.25% Sodium hypochlorite.
3) It is usually diluted to 1/10 immediately before use;
37
 4) However to kill Mycobacterium tuberculosis. It should be diluted only 1/5.
5) Bleach will kill many organisms immediately, but for full sterilization it
should be allowed to react for 20 min.
6) Bleach will kill many, but not all spores.
7) It is highly corrosive and may corrode even stainless steel, surgical
instruments.

Ozone :

1) Ozone is used in industries to sterilize water, air and disinfectant for

surfaces.
2) It has the benefit of being able to oxidize most organic matter.
3) It is a toxic and unstable gas that must be produced on sight
4) It is not practical to use in may settings.

Q. What are the Mechanical methods for sterilization ?

It includes filtration.
Filtration:

1) Involves the physical removal of all cells in a liquid or gas.

2) Important for sterilization of solutions which would be denatured by heat
antibiotics, inject able drugs, amino acids, vitamins)

Q.Explain Antimicrobial agents?

 Are chemicals that kill or inhibit the growth microorganisms.

 Antimicrobial agents include chemical preservatives and antiseptics. T
 hey have both static and cidal effect .

Preservatives( Anti microbial agents)

 Static agents used to inhibit the growth of microorganisms, mostly in foods.

 If eaten they should be nontoxic and sulfur dioxide.

Q. What are the Types of antimicrobial agents used for sterilization ?

Antiseptics:
Microbial agents harmless enough to be applied to the skin and mucous
membrane; should not be taken internally.

38
Examples

 Alcohols Mercurial
 silver nitrate iodine solution
 alcohols detergents.

Disinfectants:
Agents that kill microorganisms but not spores
They are not safe for application to living tissues, used on inanimate objects such
as tables, floors, utensils etc.

Example

 hypochlorite chlorine compound dye

 copper sulfate … quaternary ammonium compounds
 Formaldehyde.. Phenolic compounds.

Q, Write down the difference between antiseptic and disinfectant ?

ANTISEPTIC
Applied to living tissues
Kill microorganisms on surface
Not taken orally
Applied on skin n mucous membrane
Example alcohol , acetone etc
DISINFECTANT
Applied to non living objects
Kill microorganisms not spores
Toxic when taken orally
Applied on non living objects
Example formaldehyde etc

Q. Write a Note on Application of sterilization processes?

Sterilization process is used in ;

1) In pharmaceutical tries for sterilization of glassware and other equipment’s

2) Preparation of injectables, ophthalmic, irrigating and sterile dosage forms
3) For the dilution of sterile medicaments.
4) In hospital practice for the preparation of sterile gauzes and dressings;
5) For sterilization of glass equipment like test tubes, pipettes, all glass
syringes etc.
6) For sterilization of operation theatre, and anything used in operation
theatre including aprons, gloves, face masks, hoods, forceps, scissors,
knives etc.
39
Q. Name the articles/medicaments which require sterilization?

Glassware
The glassware and apparatus like flasks, beakers, funnels Etc may be sterilized
either by moist or dry heat but dry heat method is preferred.

Equipment
Apparatus or devices made up of metal and surgical instruments, etc may be
sterilized by steam under pressure at a temperature of 121°C for 15 to 30 minutes

Rubber and plastics

Rubber articles such as stoppers, gloves, certain catheters etc may be sterilized in
the autoclave at 121°C for 20 minutes they must not be subjected to dry heat .

Aqueous/non-aqueous solutions and suspensions

Thermostable aqueous solutions should be sterilized by steam under pressure in
an autoclave at a temperature of 115°C to 118°C for 30 minutes

Surgical dressings and fabrics:

The materials used in surgery are sterilized by dry heat, moist heat, ethylene
oxide or ionizing radiations.

CHAPTER NO 10
FERMENTATION

Q. Define fermentation? write down its process?

Definition :
Fermentation is a unique process because an organic molecule, usually an

40
intermediary of the chemistry, accepts the electrons. It is Anaerobic respiration
because it does not use oxygen as final electron acceptor.

Process

 In fermentation of glucose by certain bacteria and viruses an intermediately

accepts the electrons and proton from NADH formed in reaction of
glycolysis.
 This regenerates the molecules for reuse as electron acceptors. NAD exists
in limited supply in the cytoplasm and must be continually regenerated so
the glycolysis may proceed.
 (When oxidative phospborylation is taking place, the NAD is regenerated by
giving up its electrons and proton to FAD.)

Q. What are the Uses/ Applications of Fermentation?

1) The food industry, fermentation results in a broad variety of useful product.

2) Vinegar, for instance, is a fermentation product of Acetobator species.
3) Swiss cheese develops its holes from fermentation gases.
4) Pickles are sour because bacteria ferment the carbohydrates in cucumbers.
41
 5) Sausage tastes like sausage because bacteria ferment the meat proteins.
6) Thus fermentation is useful not only to the microorganisms but also to
consumers enjoy the products of fermentation.

Q. What is the importance of fermentation in Pharmaceuticals and

biotechnology industry ?

There are 5 major groups of commercially important fermentation:

1) Microbial cells or biomass as the product, bakers yeast, lactobacillus, etc.

2) Microbial enzymes: catalase, amylase, protease, pectinase etc
3) Microbial metabolites :
Primary metabolites – ethanol, citric acid, glutamic acid etc
Secondary metabolites: all antibiotic fermentation
4) Recombinant products: insulin, HBV, interferon, GCSF, streptokinase
5) Biotransformations: phenyl acetyl carbinol, steroid biotransformation, etc.

CHAPTER NO 11
IMMUNOLOGY

IMMUNE SYSTEM

 Immune system to be a police force of our body.

 It is a defence system of the body
 It produces resistance or responce against the foreign particles.
 Lymphocytes are the cornerstone of immune system.

42
Q. What is the difference between Immunity and Autoimmunity? OR
Q. Define Immunity and Autoimmunity?

IMMUNITY
It is a natural or acquired resistance of the body to a certain disease,pathogenic
microorganism and foreign particles produced by immune system.

Autoimmunity
Sometimes the immune system makes a mistake and attacks the body's own
tissues or organs. This is called autoimmunity.

Example is type 1 diabetes, in which the Body destroys cells in the pancreas that
produce insulin.

Q. What are the Types of Immunity ?

Immunity has two types;

1) Natural Immunity (Non-specific immunity)

2) Acquired Immunity (Specific immunity)

Q. Explain NATURAL IMMUNITY ?

43
 1) It is the natural resistance of the body against infections by a number of
mechanical and chemical stimuli.
2) It is called as nonspecific because it exists in all humans and present from
the earliest time of life.
3) Lack of such type of resistance is called as susceptibility.

Q. Explain ACQUIRED IMMUNITY ?

1) It is an acquired resistance of the body against the foreign particles or

micro-organisms.
2) It involves the formation of antibodies as a result of stimulation immune
system by foreign particles i.e. antigens.
3) They are specific and provide specific resistance

Q. What are The TYPES OF ACQUIRED IMMUNITY ?

[1] Active Immunity

[2] Passive Immunity

Q. Define Antigens and epitope( Antigen determinant ) ? What are the 2 keys
points/ properties/characteristics of antigens?

Antigen :Chemical substances capable of mobilizing the immune system and

provoking an immune response are called Antigens.

Epitope/ Antigen determinant :

Area of activity on the molecule called the antigen determinant or epitope. It
contains about six to eight amino acid molecules or monosaccharide units.

Properties:
Immunogenicity : The ability to stimulate cells of the immune system.
Reactivity: The ability to react with products of the immune system

Q.Define TOLERANCE and Specific immunologic Tolerance ?

44
 1. Tolerance is an acquired resistance to foreign particles or drugs
2. Which develops on its repeated administration over prolonged period.
Specific Immunologic Tolerance

Under normal circumstances one’s own chemicals do not stimulate an immune

response. This failure to stimulate the immune system occurs because substances
are interpreted as “self

Q.Write a Note on Types of Antigens ?

1) Autoantigens:
2) Allo-antigens:
3) Heterophiles:

Autoantigens: These are the person’s own chemical substances that stimulate an
immune response when self-tolerance breaks down.

Allo-antigens:These are the antigens existing in certain but not all members of a
species. The A, B and Rh antigens of humans .

Heterophiles: These are the antigens found in unrelated species. For instance
Erythrocytes of horses and the viruses that cause infection.

Q.What are Antibodies ?

1) A specific substance formed by the body in response to stimulation by

specific foreign antigen.
2) Antibodies are proteins composed of gamma globulins.
3) These are produced by β-lymphocytes.
4) Antibody mediated immunity s called as HUMORAL IMMUNITY.

45
Q. What are MOMOCLONAL ANTIBODIES ? Give its Applications?

1) These are the antibodies which are produced from Hybridoma cells.
2) In these antibodies variable regions of each immunoglobulin molecule are
same (monoclone).

APPLICATIONS :

It is used in.

1) Diagnostic kits to detect isolated microorganisms.

2) For specific antiserum therapy
3) For some forms of cancer therapy.
4) They are used in research for (transplantation of organ ,bone marrow)

Q. Write a Note on Types of Antibodies ?

On the basis of differences in the heavy (H) chains in the constant region.

Ig M:
It is the antibody of primary antibody response. It is the first antibody to appear in
the circulation after stimulation of B-Lymphocytes.

Ig G:
46
 1) It is the classical gamma globulin.
2) This antibody is the major circulating antibody.
3) IgG appears after 24 to 48 hours after the antigenic stimulation
4) It continues the antigen-antibody reaction begun by IgM.

IgA:

1) It is of two types. Serum Ig A and Secretory IgA.

2) Serum IgA is found in serum and is similar to IgG.
3) Secretory IgA is found in body secretions.
4) It comes from epithelial cells
5) Helps to move the antibodies into the secretions.
6) It provides resistance in the respiratory and gastrointestinal tracts.
7) It is also located in tears, saliva and closturm. (first milk by mother)

IgE: It plays a major role in allergic reactions by sensitizing cells to antigens.

IgD:

1) Both the functions and significance of the are presently unclear.

2) it is a cell surface receptor on the B-Lymphocytes together with IgM.

Q. Briefly describe the Antigen-Antibody Interactions ?

For resistance to develop, antibodies interact with antigens and antigen is altered.
The alteration may result in:

1) Death to the microorganism that possesses the antigen,

2) Inactivation of the antigen,
3) Increased susceptibility of the-antigen to other body defenses.

Q. Explain ANTIGEN-ANTIBODY REACTIONS ?/TYPES OF ANTIGEN ANTIBODY

REACTIONS : (SEROLOGICAL TESTS)
There are three main serological tests used.

 Agglutination test
 Precipitation test
 Complement fixation test

47
A) Agglutination Test:
The antigen in agglutination reactions is a cell or a particle, The addition of
homologous antibody will cause clumping or agglutination.

Agglutination tests are of following types

1) Tube agglutination test

2) Slide agglutination tests
3) Agglutinin Adsorption test

Diagnostic Applications of Agglutination Tests:

Following Agglutination Test are used for diagnosis of different diseases.

 Widal Test
 Weil-Felix Test (Agglutination adsorption Test)
 TPA Test (Treponema palladium Agglutination Test)
 COOMB'S TEST (ANTIGLOBULIN TEST)

B) PRECIPITATION TESTS:

1) In these tests, a reaction takes place between a soluble antigen and a

solution of its homologous antibody.
2) The reaction is manifested by the formation of a visible precipitate at the
interface of reactants.

There are two types of precipitation reactions.

 Ring test.  Agar-diffusion method.

DIAGNOSTIC APPLICATIONS OF PRECIPITATION TESTS:

Following tests of precipitation are important from diagnostic point of view.

 KAHN TEST (VDRL TEST)  ASCOLI TEST

C)COMPLEMENT FIXATION TEST:

It is a normal thermolabile protein constituent of blood serum that anticipate in

antigen-antibody reactions,

DIAGNOSTIC APPLICATIONS OF CFT:

 WASSERMAN TEST FOR SYPHILIS  T. PALLIDUM CFT

48
 AllERGY
Q. Define allergy and give its symptoms?

Definition :

“Allergy is a specific hypersensitivity of an individual to foreign particles Usually a

protein to which a specific individual is exposed.”

Clinical Features/ Symptoms :

 Swollen nasal mucous membranes, Complications in bronchi

 Migraine, GIT disturbances, Eczema, Dermatitis, Asthma,
 Runny nose/Stuffy nose Conjunctivitis, Tearing, Vomiting,
 Headache, Fever, Skin Rash.

Q. Define Hypersensitivity and describe types of Hypersensitivity

Definitions:

“It is an exaggerated are in inappropriate reaction of immune system which are

harmful to body”

Types of Hyper sensitivity reactions

Hyper sensitivity reaction is classified by Gell and Coombs in to 4 major types

1) Anaphylactic hypersensitivity ( immediate hypersensitivity )

2) Cytotoxic Hypersensitivity
3) Complex mediated Hypersensitivity
4) Cell mediated Hypersensitivity

Q. Explain TYPE I- ANAPHYLACTIC HYPERSENSITIVITY ?

1) This is a typical allergic response mediated by IgE type of antibodies in

response to specific antigen called allergen.
2) This reaction may be life threatening
3) Can be accompanied by Anaphylaxis, in which the mediators induce severe
contraction of body's smooth muscles.

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Common Allergens:

 Plant pollens,
 Serum proteins,
 Drugs like penicillin etc.

Q. Explain TYPE II, CYTOTOXIC HYPERSENSITIVITY ?

1) These reactions occur when a specific antibody (Typically IgG or IgM)

causes destruction of host cells.
2) Cells Involved are WBCs, RBCs, and platelets.

EXAMPLES

 Blood transfusion reaction.

 Hemolytic disease of newborn (Erythroblastosis fetalis).
 Autoimmune disorders: (body produces antibodies against its own cells)
 Thrombocytopenia (low platelets)
 Agranulocytosis (low WBC’s)
 Myasthenia gravis (Serious Muscle weakness)

Q. Explain TYPE III, IMMUNE COMPLEX HYPERSENSITIVITY ?

the Deposition of immune-complex (Antigen-antibody complex) at various body

locations,mainly blood vessels, kidneys, joints, lungs & skin".

Examples:

 Serum sickness
 Arthus reaction
 Rheumatoid arthritis

Q. Explain TYPE IV, CELL MEDIATED HYPERSENSITIVITY ?

1) It is an exaggeration of CMI (T-lymphocytes) and develops beginning 18-24

hours following contact with antigen & peaks in 2-3 days
2) Various substances can elicit this type of response. Include metals,
cosmetics, microbes & plant products.

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TYPES: Two major forms of this allergy are

 Infection allergy
 Contact dermatitis

CHAPTER NO 12
VACCINES
Define Vaccine and Write down its Characteristics/History/Facts of Vaccines?
DEFINITION

"Vaccine is a suspension of living or killed pathogenic microorganism modified to

make it non pathogenic and administration of which induce immune response in
the recipient sufficient to prevent susceptible disease."

Characteristics/History/Facts of Vaccines:

1) A vaccine is a preparation which is used to improve immunity to a disease.

2) Term derives from Edward Jenners use of cowpox ("vacca" means cow in
Latin),
3) Vaccines are based on the concept originating in China
4) Process of distributing and administrating vaccines is referred vaccination.
5) Vaccines can be prophylactic or therapeutic.
6) Vaccines may be dead or inactivated organisms or purified products
derived from them.

Q. How the IMMUNITY is Developed? Write down the Aims of Vaccination ?

1) The immune system recognizes vaccine agents as foreign, destroys them,

and 'remembers them.
2) When the virulent version of an agent comes along the body recognize the
protein coat on the virus, and thus prepared to respond by.
 Neutralizing the target agent before it can enter cells,
 By recognizing and destroying infected cells before that agent can multiply
to vast numbers.

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AIM OF VACCINATION?

The aim of vaccination in the individual is to induce a prime state such that on
contact with the relevant infection a more rapid and effective secondary response
could be mounted leading to the prevention of disease thus the primary aim is to
eliminate the disease.

Q. Write down the characteristics of IDEAL VACCINE?

Some important requirements (features) of an ideal vaccine are:

1) Non- pathogenicity: Non pathogenic not cause original disease.

2) Immunogenicity: Strongly immunogenic to produce antibodies.
3) Efficacy: Greater than 90%.
4) Effective: When given orally.
5) Produce long lived immunity: ( hope full life long )
6) Low cost.
7) Induce a wide range of appropriate responses.
8) Compatible with co administrated vaccine.
9) Stable genetically and thermally.

Q. Explain TYPES OF VACCINES

LIVE ATTENUATED VACCINES

There are conventionally used live viral bacterial vaccines, consist of mutants of
wild type microbes which are limited in their ability to infect body e.g. BCG
(bacterial vaccines),
Example:
Measles, polio, yellow fever small pox (viral vaccines). They are prepared by same
method as killed vaccines except the process of sterilization.

2)KILLED VACCINES.
These consist of killed or inactivated microorganisms are used where living
vaccines are not available
Example :
rabies, influenza, polio (viral vaccines), cholera, pertusis, plague, typhoid
(bacterial vaccines).
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HETEROLOGOUS VACCINES.
The antigenic site is available on some other organism inspite of using actual
molecule, we can use this heterosite to induce some immune - response

SUB-UNIT VACCINES (2nd GENERATION VACCINES).

These are vaccines which contain one or more pure or semi pure part or subunits
of microorganisms
Example :
pilli capsule, coat etc. e.g. Haemophilus meningitis vaccine

MARKER VACCINES
These are vaccines which can be used in conjunction with a diagnostic test to
differentiate a vaccinated animal from a carrier animal

Example: Used for Herpes viruses of pigs & cattle's.

DNA VACCINES (3rd GENERATION VACCINES).

These are the vaccines that contain microbial fraction produced by genetic
engineering. These are also called polynucleotide or genetic vaccines.

VECTORED VACCINES.
The vector completed with inserted gene itself act as vaccine.Viruses used as
vector are adenovirus, Herpes virus.

ONE-SHOT VACCINE.
It will be the vaccine of future consisting of viral or bacteria vector containing
genes for all required vaccines.

PEPTIDE VACCINES.
These are the subunit vaccines prepared by chemical synthesis of short
immunogenic peptides"

GENE-DELETED VACCINES.These are genetically engineered vaccines which

involve the removal or mutation of virulence gene of the pathogen"

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ANTI-IDIOTYPE VACCINES.

 It is a vaccine in which antibody molecules are used to prepare antigen such

that these antibody molecules themselves are copies of antigen.
 The antibody, which recognizes the antigen is called idiotype.
 Which is use to raise second antibodies (antiideotype antibodies) e.g. this
technique is successfully used against streptococci.

ANTISERA
(IMMUNE SERA, SEROTHERAPY)

Q. What is Antisera ? write down its types?

1) These are preparations containing antibodies introduced into the body of

patient to provide passive immunity.
2) They provide immediate and effective supply of antibodies.
3) Antisera are used both prophylactically and therapeutically.

Types:

 Non specific Antisera e.g. Normal human imrnunoglobulins.

 Specific Antiseia e.g. antitoxins, antibacterial and Antiviral sera.

Q.Write a Note on NON-SPECIFIC ANTISERA ?

(HUMAN NORMAL IMMUNOGLOBULINS)

1) Y - Globulin injection contains almost all globulins of human plasma

together with smaller amounts of other plasma proteins.
2) Y - Globulin consists of three distinct components IgA, IgG, IgM.
3) It is given I/m and is useful in number of viral and life threatening bacteria!
Infections diseases.
4) Since they are prepared from humans, so are well tolerated.

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Q, Write a Note on SPECIFIC ANTISERA ANTITOXINS?

These are antibodies to toxins of microorganisms which combine with toxins to

neutralize its toxicity."

EXAMPLES

1) Diphtheria antitoxin
2) Gas gangrene antitoxins
3) Tetanus antitoxins
4) Staphylococcus antitoxins.

Q.Write a note on ANTI-BACTERIAL SERA ?

1) These provide passive immunity against diseases caused by endotoxin

producing bacteria.
2) They are prepared in the same way as antitoxins except.
3) I/V routes are used for injection.
4) Methods of refining the sera are different because antibacterial antibodies
are associated with y-globulin fractions.

EXAMPLE

Leptospira antiserum (BPC)

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