UNIT – 1 FUNDAMENTALS OF MICROBIOLOGY

Classification of microorganisms – prokaryotic, eukaryotic, cell structure,

characteristics, importance, introduction to water, soil and air borne pathogens and
Parasites and their effects on human, animal and plant health, transmission of
pathogens, transmissible diseases – bacterial, viral, protozoan, and helminths
parasites, concentration and detection of virus. Control of microorganisms
preservation of microorganisms, DNA, RNA, replication, recombinant
DNA technology, their potential applications and intellectual property rights.

INTRODUCTION

DEFINITION – Environmental Microbiology is the scientific study of microorganisms

in the environment. This study helps us to understand, how microbes interact with the
environment and each other, including the spread of viruses and bacteria, distribution of
algae, fungi and parasitical organisms and their effects on natural ecosystem and human
health and environment.
Examples – Bacteria, Archaea, Viruses, Fungi, Algae, Protozoa, etc…

HISTORY OF ENVIRONMNTAL MICROBIOLOGY

Microbiology essentially began with the development of the microscope. It was Antonie
van Leeuwenhoek, a Dutch draper whose hobby was lens grinding and making
microscopes, was the first to provide proper descriptions and drawings included
protozoans from the guts of animals and bacteria from teeth scrapings. His records were
excellent because he produced magnifying lenses of exceptional quality. Leeuwenhoek
conveyed his findings in a series of letters to the British Royal Society during the mid-
1670s. Leeuwenhoek’s “animalcules,” remained absolutely Unique. He is known as
“Father of Microbiology”. Louis Pasteur is known as “Father of Modern
Microbiology”.

TYPES
 Air Microbiology
 Soil Microbiology
 Water Microbiology

SIGNIFICANCE
 Microbiology is actually one of the most important sub-sectors of biology.
 By analysing microorganisms closely, microbiologists play a crucial role in
combating disease, creating chemical products for agriculture, and even helping
to keep the planet healthy.
  Many work as biomedical scientists in hospitals and laboratories: testing samples
of body tissue, blood and fluids to diagnose infections, monitor treatments or
track disease outbreaks.
 The major importance of medical microbiology is that it helps in the
identification, isolation, diagnosis and treatment of pathogenic
microorganisms and also produces beneficial organisms such as yeasts and some
antibiotics.
 Biomedical research derives from many areas of life and physical sciences,
including biology.
 Microbial solutions enable farmers to drive yield and productivity in a
sustainable way. Deriving from various naturally-occurring microorganisms
such as bacteria and fungi, these solutions can protect crops from pests and
diseases and enhance plant productivity and fertility.

CLASSIFICTION OF CELLS
All living organisms have unique cellular organization, which may contain one or many
cells. The organism which possess only one cell is called Unicellular organisms
(Bacteria, Blue green algae, Protozoa, etc...). The organism which possess more than
one cell or many cells is known as Multicellular organism (Fungi, Plants, Animals,
Insects, Human beings). Any living organism may contain only one type of cell, based on
their complexity, either

ⅰ) Prokaryotic cells ⅱ) Eukaryotic cells

The terms Prokaryotic and Eukaryotic were suggested by Hans Ris in 1960’s.

ⅰ) PROKARYOTIC CELLS
Prokaryote means Before Nucleus in Greek. It is a single-celled organism that lacks
nucleus and other membrane-bound organelles. Therefore, all reactions occur within the
Cytoplasm known as Nucleoid. Prokaryotes include Bacteria and Archaea. The
photosynthetic prokaryotes include cyanobacteria that perform photosynthesis. They can
be free living or parasites. They may be spherical, rod shaped or spiral. A prokaryotic cell
lacks certain organelles like mitochondria, endoplasmic reticulum, and Golgi bodies.

CHARACTERISTICS OF PROKARYOTIC CELLS: Prokaryotic cells have different

characteristic features. The characteristics of the prokaryotic cells are mentioned below.

1. They lack a nuclear membrane.

2. Mitochondria, Golgi bodies, chloroplast, and lysosomes are absent.
3. The genetic material is present on a single chromosome.
4. The histone proteins, the important constituents of eukaryotic chromosomes, are
lacking in them.
5. The cell wall is made up of carbohydrates and amino acids.
6. The plasma membrane acts as the mitochondrial membrane carrying respiratory
enzymes.
 7. They divide asexually by binary fission. The sexual mode of reproduction involves
conjugation.

PROKARYOTIC CELL STRUCTURE

Capsule: It is an outer protective covering found in the bacterial cells, in addition to the
cell wall. It helps in retaining moisture content and helps in attachment of cells to
nutrients and surfaces.
Cell wall: It is the outermost layer of the cell which gives shape to the cell.
Cytoplasm: The cytoplasm is mainly composed of enzymes, salts, cell organelles and is a gel
like component.
Cell Membrane: This layer surrounds the cytoplasm and regulates the entry and exit of
substances in the cells.
Pili: These are hair-like outgrowths that attach to the surface of other bacterial cells.
Flagella: These are long structures in the form of a whip that help in the locomotion of a cell.
Ribosomes: These are involved in protein synthesis.
Plasmids: Plasmids are non-chromosomal DNA structures. These are not involved in
reproduction.
Nucleoid: It is the region in the cytoplasm where the genetic material is present.

PROKARYOTIC CELL DIAGRAM

The prokaryotic cell diagram given below represents a bacterial cell. It depicts the absence
of a true nucleus and the presence of a flagellum that differentiates it from a eukaryotic
cell.
 COMPONENTS OF PROKARYOTIC CELLS: The prokaryotic cells have four main
components:

Plasma Membrane- It is an outer protective covering of phospholipid molecules which

separates the cell from the surrounding environment.
Cytoplasm- It is a jelly-like substance present inside the cell. All the cell organelles are
suspended in it.
DNA- It is the genetic material of the cell. All the prokaryotes possess a circular DNA. It
directs what proteins the cell creates. It also regulates the actions of the cell.
Ribosomes- Protein synthesis occurs here.
Some prokaryotic cells possess cilia and flagella which helps in locomotion.

REPRODUCTION IN PROKARYOTES
A prokaryote reproduces in two ways:
• Asexually by binary fission
• Sexually by conjugation

Binary Fission
• The DNA of an organism replicates and the new copies attach to the cell membrane.
• The cell wall starts increasing in size and starts moving inwards.
• A cell wall is then formed between each DNA, dividing the cell into two daughter
cells.
Recombination
In this process, genes from one bacteria are transferred to the genome of other bacteria. It
takes place in three ways-conjugation, transformation, transduction.
• Conjugation is the process in which genes are transferred between two bacteria
through a protein tube structure called a pilus.
• Transformation is the mode of sexual reproduction in which the DNA from the
surroundings is taken by the bacterial cell and incorporated in its DNA.
• Transduction is the process in which the genetic material is transferred into the
bacterial cell with the help of viruses. Bacteriophages are the virus that initiates the
process.

EXAMPLES OF PROKARYOTIC CELLS

The examples of the prokaryotic cells are mentioned below:

Bacterial cells
• These are unicellular organisms found everywhere on earth from soil to the human
body.
• They have different shapes and structures.
• The cell wall is composed of peptidoglycan that provides structure to the cell wall.
• Bacteria have some unique structures such as pili, flagella and capsule.
• They also possess extra chromosomal DNA known as plasmids.
 • They have the ability to form tough, dormant structures known as endospores that
helps them to survive under unfavourable conditions. The endospores become active
when the conditions are favourable again.

Archaeal cells
• Archaea bacteria are unicellular organisms similar to bacteria in shape and size.
• They are found in extreme environments such as hot springs and other places such as
soil, marshes, and even inside humans.
• They have a cell wall and flagella. The cell wall of archaea does not contain
peptidoglycan.
• The membranes of the archaea have different lipids with a completely different
stereochemistry.
• Just like bacteria, archaea have one circular chromosome. They also possess plasmids.

ⅱ) EUKARYOTIC CELLS
Eukaryotic cells have a nucleus enclosed within the nuclear membrane and form large and
complex organisms. Protozoa, fungi, plants, and animals all have eukaryotic cells. They are
classified under the kingdom Eukaryota. They can maintain different environments in a
single cell that allows them to carry out various metabolic reactions. This helps them grow
many times larger than the prokaryotic cells.

CHARACTERISTICS OF EUKARYOTIC CELLS

The features of eukaryotic cells are as follows:

1. Eukaryotic cells have the nucleus enclosed within the nuclear membrane.
2. The cell has mitochondria.
3. Flagella and cilia are the locomotory organs in a eukaryotic cell.
4. A cell wall is the outermost layer of the eukaryotic cells.
5. The cells divide by a process called mitosis.
6. The eukaryotic cells contain a cytoskeletal structure.
7. The nucleus contains a single, linear DNA, which carries all the genetic information.

EUKARYOTIC CELL STRUCTURE - It comprises the following:

Plasma Membrane
 The plasma membrane separates the cell from the outside environment.
 It comprises specific embedded proteins, which help in the exchange of substances in
and out of the cell.
Cell Wall
 A cell wall is a rigid structure present outside the plant cell. It is, however, absent in
animal cells.
 It provides shape to the cell and helps in cell-to-cell interaction.
 It is a protective layer that protects the cell from any injury or pathogen attacks.
  It is composed of cellulose, hemicellulose, pectins, proteins, etc.
Cytoskeleton
The cytoskeleton is present inside the cytoplasm, which consists of microfilaments,
microtubules, and fibres to provide perfect shape to the cell, anchor the organelles, and
stimulate the cell movement.

Endoplasmic Reticulum: It is a network of small, tubular structures that divides the cell
surface into two parts: luminal and extra luminal. Endoplasmic Reticulum is of two types:
Rough Endoplasmic Reticulum contains ribosomes.
Smooth Endoplasmic Reticulum that lacks ribosomes and is therefore smooth.
Nucleus
 The nucleoplasm enclosed within the nucleus contains DNA and proteins.
 The nuclear envelop consists of two layers- the outer membrane and the inner
membrane. Both the membranes are permeable to ions, molecules, and RNA material.
 Ribosome production also takes place inside the nucleus.
Golgi Apparatus
 It is made up of flat disc-shaped structures called cisternae.
 It is absent in red blood cells of humans and sieve cells of plants.
 They are arranged parallel and concentrically near the nucleus.
 It is an important site for the formation of glycoproteins and glycolipids.
Ribosomes: These are the main site for protein synthesis and are composed of proteins and
ribonucleic acids.
Mitochondria
 These are also known as “powerhouse of cells” because they produce energy.
 It consists of an outer membrane and an inner membrane. The inner membrane is
divided into folds called cristae.
 They help in the regulation of cell metabolism.
Lysosomes: They are known as “suicidal bags” because they possess hydrolytic enzymes to
digest protein, lipids, carbohydrates, and nucleic acids.
 Plastids: These are double-membraned structures and are found only in plant cells.
These are of three types:
 Chloroplast that contains chlorophyll and is involved in photosynthesis.
 Chromoplast that contains a pigment called carotene that provides the plants yellow,
red, or orange colours.
 Leucoplasts that are colourless and store oil, fats, carbohydrates, or proteins.

EUKARYOTIC CELL DIAGRAM: Eukaryotic cell diagram mentioned below depicts

different cell organelles present in eukaryotic cells. The nucleus, endoplasmic reticulum,
cytoplasm, mitochondria, ribosomes, lysosomes are clearly mentioned in the diagram.
 EUKARYOTIC CELL CYCLE
The eukaryotic cells divide during the cell cycle. The cell passes through different stages
during the cycle. There are various checkpoints between each stage.
Quiescence (G0)
This is known as the resting phase, and the cell does not divide during this stage. The
cell cycle starts at this stage. The cells of the liver, kidney, neurons, and stomach all
reach this stage and can remain there for longer periods. Many cells do not enter this
stage and divide indefinitely throughout their lives.
Interphase
In this stage, the cells grow and take in nutrients to prepare them for the division. It
consists of three
Checkpoints
Gap 1 (G1) – Here the cell enlarges. The proteins also increase.
Synthesis (S) – DNA replication takes place in this phase.
Gap 2 (G2) – The cells enlarge further to undergo mitotic division.
Mitosis
Mitosis involves the following stages:
 Prophase
 Prometaphase
 Metaphase
 Anaphase
 Telophase
 Cytokinesis
On division, each daughter cell is an exact replica of the original cell.

EXAMPLES OF EUKARYOTIC CELLS

Eukaryotic cells are exclusively found in plants, animals, fungi, protozoa, and other complex
organisms. The examples of eukaryotic cells are mentioned below:

Plant Cells
The cell wall is made up of cellulose, which provides support to the plant. It has a large
vacuole which maintains the turgor pressure. The plant cell contains chloroplast, which
aids in the process of photosynthesis.

Fungal Cells
The cell wall is made of chitin. Some fungi have holes known as septa which allow the
organelles and cytoplasm to pass through them.

Animal Cells
These do not have cell walls. Instead, they have a cell membrane. That is why animals have
varied shapes. They have the ability to perform phagocytosis and pinocytosis.

Protozoa
Protozoans are unicellular organisms. Some protozoa have cilia for locomotion. A thin layer
called pellicle provides supports to the cell
 DIFFERENCE BETWEEN PROKARYOTIC AND EUKARYOTIC CELLS

PROKARYOTIC CELLS EUKARYOTIC CELLS

They are Unicellular (Single-celled organism) They are Multicellular (More than one cell)

well defined Nucleus is Absent (Nucleoid) Well defined Nucleus is Present

Cell wall is Present Cell wall is Absent

Membrane bound Organelles are Absent Membrane bound Organelles are Present

Endoplasmic Reticulum, Lysosome, Golgi Apparatus, Endoplasmic Reticulum, Lysosome, Golgi

Peroxisome, Centrosome, Cytoskeleton, Cilia are Absent Apparatus, Peroxisome, Centrosome(except
flowering plants), Cytoskeleton, Cilia are
Present
Plasmid is Present Plasmid is Absent

Mesosome is Present Mesosome is Absent

Ribosomes are Smaller Ribosomes are Larger

Chloroplast is Absent. Chlorophyll scattered in Chloroplast present in Plants and Algae

Cytoplasm
Cell size ranges from 0.5µm-100µm (dia) Cell size ranges from 10µm-150µm (dia)

No. of Chromosomes - Single Haploid (n) No. of Chromosomes – Paired diploid (2n)

Mode of Reproduction: Both Asexual & Sexual Mode of Reproduction: Mostly Sexual

Cell Division: Cell Division: Through Mitosis

Asexually by Binary fission
Sexually by Conjugation, transformation and
transduction
Metabolic rate is Higher Metabolic rate is Lower
Respiration via Cytoplasmic membrane Respiration via Mitochondria

Electron Transport chain found in Cell membrane ElectronTransport Chain found in

Mitochondrial membrane
DNA replication occurs in Cytoplasm DNA replication occurs in Nucleus

Generation time is Shorter (20 to 60 minutes) Generation time is Longer (12 to 24 hours)
WATER BORNE PATHOGENS
The drinking water is most often contaminated with pathogenic microbes including
bacteria, viruses, and protozoa beyond the recommended limits causing serious human
health hazards. The pathogenic microbes commonly detected in water supplies include
those related to dysentery, typhoid fever, vomiting, and cholera. Several pathogenic
microorganisms are commonly found in water as well as in effluents from wastewater
treatment plants. The three categories of pathogens encountered in the environment are:
1. Bacterial pathogens
2. Viral pathogens
3. Protozoan parasites

1. Bacterial Pathogens:

Bacteria are the most commonly identified organism, in which some of them are highly
pathogenic to plants. The most commonly found pathogenic bacteria are bacilli
(rodshaped).

These bacteria act on different parts of the plants, and localize and inhibit few functions of
the plants. Some of these pathogens (e.g., Salmonella, Shigella) are enteric bacteria. Others
(e.g., Legionella, Mycobacterium avium, Aeromonas) are indigenous aquatic bacteria.

2. Viral Pathogens:
Viruses are small particles, typically between 20 and 300 nanometers in length, containing
RNA or DNA. Viruses require a host cell to replicate.
Some of the diseases include smallpox, influenza, mumps, measles, chickenpox, ebola,
HIV, rubella, and COVID19 were caused by viral pathogens. Water and wastewater may
become contaminated by approximately 140 types of enteric viruses.
These viruses enter into the human body orally, multiply in the gastrointestinal tract, and
are excreted in large numbers in the feces of infected individuals.
These are also released into aquatic environments but are unable to multiply outside their
host cells. Their infective dose is generally lower than for bacterial pathogens.
3. Protozoan Parasites
A protozoan parasite is basically a protozoan that has adapted to invade and live in cells
and tissues of other organisms. We could say that a protozoan took lessons from a
parasite to learn how they live and survive, and then slowly started changing to become
more like them. Most of the protozoan parasites produce cysts, which are able to survive
outside their host under adverse environmental conditions. Encystment is triggered by
factors such as lack of nutrients, accumulation of toxic metabolites, or host immune
response. Under appropriate conditions, a new trophozoite is released from the cyst.
This process is called excystment . These are released into aquatic environments as
cysts or oocysts, which are quite resistant to environmental stress and to disinfection,
and do not multiply outside their hosts.

SOIL BORNE PATHOGENS: A soil borne pathogen is a disease-causing agent which lives
both in soil and in a plant host, and which will tend to infect diseased plants which are grown
in that soil. Common soil borne pathogens include
1. Fusarium
2. Pythium
3. Rhizoctonia
4. Phytophthora
5. Verticillium
6. Rhizopus 7. Thielaviopsis.

1. Fusarium:

Fusarium is a large genus of filamentous fungi, part of a group often referred to as

hyphomycetes, widely distributed in soil and associated with plants. Most species are
harmless saprobes, and are relatively abundant members of the soil microbial community.
Some species produce mycotoxins in cereal crops that can affect human and animal health
if they enter the food chain. The main toxins produced by these Fusarium species are
fumonisins and trichothecenes. Despite most species apparently being harmless (some
existing on the skin as commensal members of the skin flora), some Fusarium species and
sub specific groups are among the most important fungal pathogens of plants and animals.
2. Pythium

Pythium is a genus of parasitic oomycetes. They were formerly classified as fungi.

Most species are plant parasites, but Pythium insidiosum is an important pathogen
of animals, causing pythiosis. The feet of the fungus gnat are frequently a vector for
their transmission.

3.Rhizoctonia

Rhizoctonia is a genus of anamorphic fungi in the order Cantharellales. Species do not

produce spores, but are composed of hyphae and sclerotia (hyphal propagules) and are
asexual states of fungi in the genus Thanatephorus. Rhizoctonia
species are saprotrophic, but are also facultative plant pathogens, causing
commercially important crop diseases. They are also endomycorrhizal associates of
orchids.[1] The genus name was formerly used to accommodate many superficially
similar, but unrelated fungi.

4. Phytophthora

Phytophthora is a genus of plant-damaging oomycetes (water molds), whose

member species are capable of causing enormous economic losses on crops
worldwide, as well as environmental damage in natural ecosystems. The cell wall
of Phytophthora is made up of cellulose.
5.Verticillium

Verticillium is a genus of fungi in the division Ascomycota, and are an anamorphic

form of the family Plectosphaerellaceae. The genus used to include diverse groups
comprising saprobes and parasites of higher plants, insects, nematodes, mollusc
eggs, and other fungi, thus the genus used to have a wide-ranging group of taxa
characterised by simple but ill-defined characters. The genus, currently thought to
contain 51 species, may be broadly divided into three ecologically based
groupsmycopathogens, entomopathogens, and plant pathogens and related
saprotrophs. However, the genus has undergone recent revision into which most
entomopathogenic and mycopathogenic isolates fall into a new group called
Lecanicillium.

6. Rhizopus

Rhizopus is a genus of common saprophytic fungi on plants and specialized parasites on

animals. They are found in a wide variety of organic substances, including "mature fruits
and vegetables", jellies, syrups, leather, bread, peanuts, and tobacco. They are
multicellular. Some Rhizopus species are opportunistic human pathogens that often cause
fatal disease called mucormycosis. This widespread genus includes at least eight species.
7. Thielaviopsis

Thielaviopsis is a small genus of fungi in the order Microascales. The genus includes
several important agricultural pathogens. The most widespread is T. basicola, the causal
agent in several root rot diseases of economically important crop species including cotton
and a variety of vegetables. In cotton, Thielaviopsis root rot, also known as black root rot
causes necrosis of the roots and stunting of the crop plants.

AIR BORNE PATHOGENS: Airborne diseases are caused by pathogenic microbes

small enough to be discharged from an infected person via coughing, sneezing, laughing
and close personal contact or aerosolization of the microbe. The discharged microbes
remain suspended in the air on dust particles, respiratory and water droplets. Few
diseases are predominantly airborne. Most diseases that spread through the air are also
contagious through larger respiratory droplet transmission. This type of infection occurs
when people are within 6 feet of each other. Common air borne pathogens causing
diseases include
1.Measles
2.TB
3.Other diseases.

1.Measles: Measles is one of the most contagious diseases, affecting up to 90% of

the people close to a person with the disease. It’s a virus that lives in the mucus of
the nose and throat and is spread through coughing and sneezing. The measles virus
survives for up to 2 hours in the air once the infected person leaves an area.

2.TB: Tuberculosis, or TB, is a bacterial disease of the lungs and throat. When a
person with TB coughs, speaks, or laughs, the TB bacteria are released into the air.
TB is not transmitted through touching, kissing, or sharing food.

3.Other Diseases: Measles and TB are airborne-exclusive diseases. There are

several other diseases that spread through respiratory droplets, which can exist
either in the air or on surfaces. These diseases include:
 • Chickenpox
• Influenza
• Pertussis (whooping cough)
• Respiratory Syncytial Virus (RSV)

PARASITES: A parasite is an organism that lives on or in a host organism and

gets its food from or at the expense of its host. There are three main classes of
parasites that can cause disease in humans:
1. Protozoa
2. Helminths
3. Ectoparasites

1.Protozoa : Protozoa are single celled organisms. They come in many different
shapes and sizes ranging from an Amoeba which can change its shape to
Paramecium with its fixed shape and complex structure. They live in a wide variety
of moist habitats including fresh water, marine environments and the soil.

2.Helminths: Helminths are worm-like parasites that survive by feeding on a living

host to gain nourishment and protection, sometimes resulting in illness of the host.
There are a variety of different helminths from the very large to the microscopic.
'Helminth' is a general term meaning worm.

3.Ectoparasites: Ectoparasites are organisms that live on the skin of a host, from which
they derive their sustenance.
EFFECTS OF PARASITES ON HUMAN: Some of the most common effects of a
parasitic infection include:
• Stomach cramps and pain.
• Nausea or vomiting.
• Dehydration.
• Weight loss.
• Swollen lymph nodes.
• Digestive problems including unexplained constipation, diarrhoea or persistent gas.
• Skin issues such as rashes, eczema, hives, and itching.
• Continuous muscle and joint pain.
EFFECTS OF PARASITES ON ANIMAL: The most common signs and effects of
parasites on animal are:
• Scooting.
• Vomiting.
• Diarrhea.
• A distended abdomen.
• Weight loss.
• Occasionally coughing.
EFFECTS OF PARASITES ON PLANT: Effects of Parasites on plant are
• Leaf rust (common leaf rust in corn)
• Stem rust (wheat stem rust)
• Sclerotinia (white mold)
• Powdery mildew
• Birds-eye spot on berries (anthracnose)
• Damping off of seedlings (phytophthora)
• Leaf spot (septoria brown spot)
• Chlorosis (yellowing of leaves)

TRANSMISSION OF PATHOGENS
PATHOGENS DEFINITION
A pathogen is defined as an organism causing disease to its host, with the severity of the
disease symptoms referred to as virulence. Pathogens are taxonomically widely diverse and
comprise viruses and bacteria as well as unicellular and multicellular eukaryotes.
Every living organism is affected by pathogens, including bacteria, which are targeted by
specialized viruses called phage.
A pathogen brings disease to its host. Another name for a pathogen is an infectious agent, as
they cause infection. As, with any organism, pathogens prioritize survival and reproduction.
The human body’s immune system acts as a defence against pathogens. The body can easily
fight off some pathogens, but others are potentially fatal.

DIFFERENT TYPES OF PATHOGENS

There are five types of pathogens. They are,

• Bacteria
• Viruses
• Fungi
• Protists Parasitic worms.

BACTERIA: Bacteria are microscopic pathogens that reproduce rapidly after entering the
body. They can release toxins that damage tissues and cause illness. Doctors typically
prescribe antibiotics to treat bacterial infections, but some bacteria are becoming resistant to
these drugs. Not all bacteria are pathogenic, though. In the body, there are many types of
harmless bacteria, and some may even support essential bodily functions.
VIRUSES: Smaller than bacteria, virus invades a host cell. It then replicates, producing
hundreds and thousands of new viruses that go on to infect more host cells. Viruses can pass
from person to person in various ways, including:

• via respiratory droplets that travel through the air

• through contact with the blood of a person with the infection

• through contact with the bodily fluids of someone with the infection
FUNGI: There are thousands of species of fungi, some of which cause disease in humans.
Common fungal skin conditions include athlete’s foot and ringworm. These conditions are
contagious and can spread through person-to-person contact. Fungal pathogens are evolving a
capacity for memory. They can use signals in the body to anticipate imminent threats to their
survival, against which they can then prepare themselves.

PROTISTS: These single cell organisms cause disease in their host. They infect other
organisms to survive and reproduce. Protist pathogens affect plants and food crops. Foods
containing protists can cause dysentery which is an infection of the intestines that causes
diarrhoea. Protist pathogens can also be parasitic and live in other organisms, such as
mosquitoes. Protists cause malaria through mosquito bites.

PARASITIC WORMS: Parasitic worms, also known as helminths, are large enough for
people to see with the naked eye, and they can live in many areas of the body. Some worms
include:

• Flatworms: These include tapeworms, which reside in the intestines.

• Thorny-head worms: This type of worm lives in the intestines.

 • Roundworms: These worms can survive in the gastrointestinal tract and
lymphatic system.

TRANSMISSION OF PATHOGENS:
Pathogens can be transmitted a few ways depending on the type. They can be spread through
skin contact, body fluids, airborne particles, contact with feces, and touching a surface
touched by an infected person. Pathogen transmitted through the environment may survive
from hours to years outside the host, depending on the organism and environment. Pathogen
may exit a host in respiratory secretions from the noise and mouth, or be shed on dead skin or
in feces, urine, saliva or tears.

ENVIRONMENTAL PATHOGENS:
They are microorganism that generally in habitat the environment and exist outside of a
human host for a large proportion of their lifecycle.

ENVIRONMENTALLY TRANSMITTED PATHOGENS:

Although humans are continually exposed to a vast array of microorganisms in the
environment, only a small proportion of these microbes are capable of interacting with the
host in such a manner that infection and disease will result.
Disease-causing microorganisms are called pathogens. Infection is the process in which the
microorganism multiplies or grows in or on the host. Infection does not necessarily result in
disease since it is possible for the organism to grow in or on the host without producing an
illness. In the case of enteric infections (i.e., diarrhoea) caused by Salmonella, only half of the
individuals infected develop clinical signs of illness. A frank pathogen is a microorganism
capable of producing disease in either normal healthy or immunocompromised persons.
Opportunistic pathogens are usually capable of causing infections only in
immunocompromised individuals (burn patients, patients taking antibiotics, those with
impaired immune systems, elderly patients with diabetes, etc.). Opportunistic pathogens are
common in the environment and may be present in the human gut or skin, but normally do
not cause disease.
To cause illness, the pathogen must usually first grow within or on the host. The time
between infection and the appearance of clinical signs and symptoms such as diarrhoea, fever
or rash, is the incubation time. This may range from as short as 6–12 h in the case of
Norwalk virus diarrhoea to 30–60 days for hepatitis A virus, which causes liver disease.
At any time during infection, the pathogen may be released into the environment by the host
through feces, urine, or respiratory secretions. Although the maximum release may occur at
the height of the disease, it may also precede the first signs of clinical illness. In the case of
hepatitis A virus, the maximum excretion in the faeces occurs before the onset of signs of
clinical illness. The concentration of organisms released into the environment varies with the
type of organism and the route of transmission. The concentration of enteric viruses during
gastroenteritis may be as high as 10 10–1012 per gram of faeces.
TRANSMISSIBLE DISEASES -BACTERIAL, VIRAL, PROTOZOAN, AND
HELMINTHS PARASITES.

1. BACTERIAL PATHOGENS: Bacteria have been characterized and belong to the

following groups.
• Gram - negative facultatively anaerobic bacteria (e.g., Aeromonas,
Plesiomonas, Vibrio, Enterobacter, Escherichia, Klebsiella, and Shigella)
• Gram - negative aerobic bacteria (e.g., Pseudomonas, Alcaligenes,
Flavobacterium, Acinetobacter)
• Gram - positive spore - forming bacteria (e.g., Bacillus spp.);
• Non - spore - forming gram - positive bacteria (e.g., Arthrobacter
Corynebacterium, Rhodococcus).
SALMONELLA: Salmonellae are Enterobacteriaceae that are widely distributed in the
environment and include more than 2000 serotypes. They are the most predominant
pathogenic bacteria in wastewater and cause typhoid and paratyphoid fever, and
gastroenteritis. salmonellosis is primarily due to food contamination, but its transmission
through drinking water. S. typhi is the etiological agent for typhoid fever, a deadly disease
that was brought under control as a result of the development of adequate water treatment
processes (e.g., chlorination, filtration). This pathogen produces an endotoxin that causes
fever, nausea, and diarrhoea, and may be fatal if not properly treated by antibiotics.
SHIGELLA: Shigella is the causative agent of bacillary dysentery or shigellosis, an infection
of the large bowel that leads to cramps, diarrhoea, and fever. This disease produces bloody
stools as a result of inflammation (induction and release of pro - inflammatory cytokines) and
ulceration of the intestinal mucosa. There are four pathogenic species of Shigella:
• S. dysenteriae (13 serotypes),
• S. flexneri (15 serotypes), S. boydii (18 serotypes), S. sonnei (1
serotype).
Infection with bacteria producing this toxin may lead to haemolytic uremic syndrome, which
results in kidney failure. Currently, no vaccine is available for the prevention of Shigella.

VIBRIO CHOLERAE: V. cholerae is a gram - negative curved rod bacterium. This

bacterium is the causative agent of cholera. In 1854, John Snow first demonstrated that
contaminated drinking water was the cause of cholera. This pathogen releases an enterotoxin

that causes mild to profuse diarrhoea, vomiting, and a very rapid loss of fluids which may
result in death in a relatively short period of time. The infectious dose for V. cholerae is
between 10 4 and 10 6 cells. Vibrios are naturally present in many aquatic environments and
survive by attaching to solids, including zooplankton (e.g., copepods), cyanobacteria (e.g.,
Anabaena), and phytoplankton (e.g.Volvox) cells.
E.COLI: E. coli is a facultative anaerobe that colonizes the gastrointestinal tract a few hours
after birth. Several strains of E.coli many of which are harmless, are found in the
gastrointestinal tract of humans and warm - blooded animals. E. coli classified as
enterotoxigenic (ETEC), EPEC, enterohemorrhagic (EHEC), enteroinvasive (EIEC), and
enteroaggregative (EAggEC) .ETEC causes gastroenteritis with profuse watery diarrhea
accompanied with nausea, abdominal cramps, and vomiting. Approximately 2 – 8% of the E.
coli present in water was found to be EPEC E. coli , which causes traveler’s diarrhoea.Food
and water are important in the transmission of this pathogen. However, the infective dose for
this pathogen is relatively high, within the range of 10^6 to10 ^9 organisms.
YERSINIA: Yersinia enterocolitica is responsible for acute gastroenteritis with invasion of
the terminal ileum. Swine are the major animal reservoir, but many domestic and wild
animals can also serve as reservoirs for this pathogen. Foodborne (e.g., milk, tofu) outbreaks
of yersiniosis have also been documented in the United States. The role of water is uncertain,
but there are instances in which this pathogen was suspected to be the cause of waterborne
transmission of gastroenteritis. This psychrotrophic organism thrives at temperatures as low
as 4 ° C, is mostly isolated during the cold months, but is poorly correlated with traditional
bacterial indicators. This organism has been isolated from wastewater effluents, river water,
and drinking water.
CAMPYLOBACTER: This thermo tolerant bacterial pathogen (e.g., Campylobacter fetus
and Campylobacter jejuni) is known to infect humans as well as wild and domestic animals.
It is ubiquitous in domestic wastewater and in effluents from abattoirs and poultry processing
plants. Raw milk is another source of Campylobacter infections. Campylobacter is also a
common cause of acute gastroenteritis (fever, nausea, abdominal pains, bloody diarrhoea,
vomiting) and is transmitted to humans through contaminated food, mainly undercooked
poultry, unpasteurized milk, contaminated drinking water, and water from contaminated
mountain streams. Campylobacter has been detected in surface waters, potable water, and
wastewater, but no organisms have been recovered from digested sludge. Due to their
sensitivity to oxygen and inability to grow at temperatures below 30 ° C (optimum
temperature is 42 ° C) Campylobacter jejuni survives but does not grow in the environment.
LEPTOSPIRA: Leptospira is a small spirochete that can gain access to the host through
abrasions of the skin or through mucous membranes. It causes leptospirosis, which is
characterized by the dissemination of the pathogen in the patient’s blood and the subsequent
infection of the kidneys and the central nervous system. The disease can be transmitted from
animals (rodents, domestic pets, and wildlife) to humans coming into contact (e.g., bathing)
with waters polluted with animal wastes. This zoonotic disease may strike sewage workers.
This pathogen is not of major concern because it does not appear to survive well in
wastewater.
LEGIONELLA PNEUMOPHILA: This bacterial pathogen is the etiological agent of
Legionnaire’s disease .This disease is a type of acute pneumonia with a relatively high
fatality rate; it may also involve the gastrointestinal and urinary tracts as well as the nervous
system. People manifesting this syndrome have fever, headaches, and muscle aches but may
recover without any treatment. Aquatic environments and soils can act as natural reservoirs
for pathogenic species of Legionellae. Legionnaire ’s disease are associated with exposure to
L. pneumophila aerosols from cooling towers, evaporative condensers, humidifiers, shower
heads, air conditioning systems, whirlpools, mist machines in produce departments of grocery
stores, mechanical aerosolization of soil particles during gardening, or dental equipment.
B.FRAGILIS: Bacteroides species are a major part of the microorganisms in the human
colon and account for approximately 25% of all colonic isolates. This pathogen has been
found in wastewater at levels ranging from 6.2 × 10^4to 1.1 × 10^5 colony forming units/Ml.
Enterotoxin - producing strains of this anaerobic bacterium may be involved in causing
diarrhoea in humans.
2 .VIRAL PATHOGENS:

HEPATITIS A VIRUS:
(HAV) Infectious hepatitis is caused by HAV, a 27 - nm RNA enterovirus (formerly
enterovirus type 72 but now classified as a hepatovirus belonging to the family
Picornaviridae) with a relatively short incubation period (2 – 6 weeks) and displaying a fecal
– oral transmission route.
Although HAV can be replicated on primary and continuous human or animal tissue cultures,
it is hard to detect because it does not always display a detectable cytopathic effect (CPE) in
infected cells.
This disease is distributed worldwide, and the prevalence of HAV antibodies is higher among
lower socioeconomic groups and increases with the age of the infected individuals. Direct
contact transmission has been documented mainly in nurseries (especially among infants
wearing diapers), mental institutions, prisons, or military camps.
HEPATITS B VIRUS:
Serum hepatitis is caused by HBV, a 42 - nm DNA virus displaying a relatively long
incubation time (4 – 12 weeks). This virus is transmitted by contact with infected blood or by
sexual contact. The mortality rate (1 – 4%) is higher than for infectious hepatitis (< 0.5%).
HBV is responsible for approximately 60% of the 434,000 cases of liver cancer worldwide
(WHO,1996).
HEPATITS C VIRUS:
HCV is also transmitted via contact with infected blood (drug users and blood transfusion,
tattoos). One percent of adults in industrialized countries are infected with HCV compared
with about 3% in developing countries. Chronic infection with HCV may ultimately lead to
liver cancer.

HEPATITS E VIRUS:
Non - A, non - B infectious hepatitis is caused HEV. HAV causes liver damage with necrosis
and inflammation. After the onset of infection, the incubation period may last up to 6 weeks.
One of the most characteristic symptoms is jaundice.
VIRAL GASTROENTERITIS:
Gastroenteritis is probably the most frequent waterborne illness; it is caused by protozoan
parasites and by bacterial and viral pathogens (e.g., rotaviruses, noroviruses, adenoviruses,
astroviruses). Rotaviruses, belonging to the family Reoviridae, are 70 - nm particles
containing double - stranded RNA surrounded with a double - shelled capsid.
Rotaviruses are the major cause of infantile acute gastroenteritis in children less than 2 years
of age. This disease largely contributes to childhood mortality in developing countries and is
responsible for millions of childhood deaths per year in Africa, Asia, and Latin America. It is
also responsible for outbreaks among adult populations (e.g., the elderly) and is a major cause
of traveler’s diarrhoea.

Astroviruses are 27 - to 34 - nm spherical non - enveloped single - stranded RNA viruses

with a characteristic star - like appearance. Seven human serotypes have been identified to
date. Astroviruses affect mostly children and immunocompromised adults and are transmitted
by the faecal – oral route, spreading via person - to - person contacts and via contaminated
food or water. Astroviruses are the second leading cause of viral gastroenteritis in children
and adults. They are associated with outbreaks in day care centres and homes for the elderly
The mild, watery diarrhoea lasts for 3 – 4 days but can be long - lasting in
immunocompromised patients.

Enteric Adenoviruses Human enteric adenoviruses are emerging pathogens that belong to
subgroup F adenoviruses, which comprises two serotypes, types 40 and 41, among 51 human
adenoviruses serotypes.
Recent viral pathogen:
Coronaviruses:
A notorious coronavirus is the severe acute respiratory syndrome (SARS) virus, which caused
several deaths in Asia in 2003. In addition to respiratory problems, this emerging pathogen
appears to cause gastrointestinal symptoms. It is detected in large concentrations in faeces of
infected individuals.
PROTOZOAN AND HELMINTHS PARASITES:
Most of the protozoan parasites produce cysts or oocysts that are able to survive outside their
host under adverse environmental conditions. Encystment is triggered by factors such as lack
of nutrients, accumulation of toxic metabolites, or host immune response. Under appropriate
conditions, a new trophozoite is released from the cyst. It is estimated that about 63% of the
Chinese population (approximately 707 million people) is infected with one or more helminth
parasites, particularly with Ascaris lumbricoides Trichuris trichiura , and hookworms
Ancylostoma duodenale and Necator americanus .
Most of these infections are acquired by the foodborne route. The ova (eggs) constitute the
infective stage of parasitic helminths; they are excreted in feces and spread via wastewater,
soil, or food. The most important helminth parasites are given below.
Organism Disease (main site affected)

Nematodes(roundworms) Ascariasis-intestinal obstruction in children

Ascaris lumbricoides (small intestine)
Trichuris trichiura Whipworm-(trichuriasis) (intestine)
Hookworms

Necator americanus hookworm disease (GI tract) hookworm

Ancylostomas duodenale disease (GI tract)

Cestodes (tapeworms)
Taenia saginata Beef tapeworm- abdominal discomfort,
hunger pains chronic indigestion (GI tract)

Tanenia solium Pork tapeworm (GI tract)

Trematodes(flukes) Schistosoma Schistosomiasis (complication in

mansoni liver[cirrhosis], bladder and large intestine)

CONCENTRATION AND DETECTION OF VIRUS

VIRUS DETECTION AND ENUMERATION: There are several approaches to virus
detection and enumeration:

• Animal Inoculation This was the traditional method for detecting viruses before the
advent of tissue cultures. Newborn mice are infected with the virus and are observed for
symptoms of disease. Animal inoculation is essential for the detection of enteroviruses such
as Coxsackieviruses A.

• Tissue Cultures Viruses are quantified by measuring their effect on established host
cell lines, which, under appropriate nutritional conditions, grow and form a monolayer on the
inner surface of glass or plastic bottles.
There are two main types of host cell lines:
(1) Primary cell lines-These cells are removed directly from the host tissues and can be
sub cultured for only a limited number of times; and
(2) Continuous cell lines- Animal cells, after serial subculturing, acquire characteristics
that are different from the original cell line, allowing them to be sub cultured indefinitely;
they are derived from normal or cancerous tissues. Cell lines traditionally used in water
virology laboratories include HEp -2, HeLa, VERO, or Buffalo Green Monkey (BGM) cells.
The BGM cell line is the most popular and perhaps the most sensitive of all cell lines used for
the detection of enteroviruses. Brief survey of microbial groups 41 Many viruses
(enteroviruses, reoviruses, adenoviruses) infect host cells and display a cytopathic effect. The
presence of the latter needs to be confirmed by other tests, including immunological
procedures, monoclonal antibodies (MAbs), or nucleic acid probes. Other viruses (e.g.,
Norwalk type virus) cannot yet be detected by tissue cultures.

• Plaque Assay A viral suspension is placed on the surface of a cell monolayer and,
after adsorption of viruses to the host cells, an overlay of soft agar or carboxymethylcellulose
is poured on the surface of the monolayer. Virus replication leads to localized areas of cell
destruction called plaques . The results are expressed in numbers of plaque - forming units
(PFU). Bacteriophages are also assayed by a plaque assay method based on similar
principles. They form plaques that are zones of lysis of the host bacterial lawn.

• Serial Dilution Endpoint Aliquots of serial dilutions of a viral suspension are

inoculated into cultured host cells and, after incubation, viral cytopathic effect (CPE) is
recorded. The titer or endpoint is the highest viral dilution (i.e., smallest amount of viruses)
capable of producing CPE in 50 % of cultures and is referred to as TCID 50 (tissue culture
infectious dose).
• Most Probable Number ( MPN ) Virus titration is carried out in tubes or 96 - well
microplates, using three dilutions of the viral suspension. Virus - positive tubes or wells are
recorded and the MPN is computed from MPN tables.

RAPID DETECTION METHODS:

Immunoelectron Microscopy
Viruses are incubated with specific antibodies and examined by electron microscopy for the
presence of virus particles aggregated by the antibody. This is a useful technique for
examining viruses such as the Norwalk -type agent.
Immunofluorescence
A fluorescent dye- labeled antibody is combined with the viral antigen, and the complex
formed is observed with a fluorescence microscope. This approach enables the detection of
rotaviruses as fluorescent foci in MA - 104 or CaCo-2 cultured cells. This method can be
accelerated by using flow cytometry. Immunomagnetic separation has been used to detect
rotavirus and HAV in environmental samples.
Enzyme Linked Immunosorbent Assay (ELISA)

A specific antibody is fixed on a solid support and the antigen (virus) is added to form an
antigen – antibody complex. An enzyme – labelled specific antibody is then added to the
fixed antigen. The presence of the virus is detected by the formation of a coloured product
upon addition of the enzyme substrate. This enzymatic reaction can be conveniently
quantified with a spectrophotometer.
Radioimmunoassay (RIA)
This assay is also based on the binding of an antigen by a specific antibody. The antigen is
quantified by labelling the antibody with a radioisotope (e.g., 125 I) and measuring the
radioactivity bound to the antigen – antibody complex. When viruses growing in host cells 42
THE MICROBIAL WORLD are treated with a 125 I - labelled antibody, the radioactive foci
can be enumerated following contact with a special film. This test, the radioimmune focus
assay (RIFA), is used for the detection of HAV.

Molecular - Based Methods

Molecular - based methodology has helped in the detection of viruses that show no growth or
grow marginally in tissue cultures (e.g., Norwalk - like viruses, rotaviruses). Gene probes are
pieces of nucleic acid that help identify unknown microorganisms by hybridizing (i.e.,
binding) to the homologous organism’s nucleic acid. For easy detection, Viral enumeration
by plaque assay:
(a) bacterial phage;
(b) animal virus (poliovirus type 1).
Brief survey of microbial groups 43 the probes can be labelled with radioactive isotopes such
as 32 P, fluorescent compounds, or enzymes such as alkaline phosphatase, peroxidase, or β -
galactosidase. Nucleic acid probes have been used for the detection of viruses (e.g.,
polioviruses, HAV) in environmental samples (water, sediments, shellfish). RNA probes
were used for the detection of HAV in shellfish concentrates. Unfortunately, these probes are
not sensitive and detect a minimum of 10^6 HAV particles.
Amplification of the target viral sequences by PCR has also been considered. Presently, a
popular method for detecting viruses (enteroviruses, adenoviruses, rotaviruses, astroviruses,
Norwalk-like viruses) in environmental samples is the RT- PCR method. A new method for
rapidly detecting HAV in environmental samples is the use of a combined cell culture -
molecular beacon assay. A molecular beacon is a single - stranded oligonucleotide probe that
is labelled with a fluorophore and a quencher at the 5 ′ and 3 ′ ends, respectively. The beacon
is introduced into permeabilized and fixed host cells that have been infected with HAV, and
the fluorescent host cells are visualized 6 h after infection instead of 1 week.

CONTROL OF MICROORGANISM
Control of microorganisms is essential in order to prevent the transmission of diseases and
infection, stop decomposition and spoilage, and prevent unwanted microbial
contamination. Microorganisms are controlled by means of physical agents and chemical
agents.

• Physical agents include such methods of control as

• High or low temperature, desiccation,
• Osmotic pressure, Radiation, and
• Filtration.
Control by chemical agents refers to the use of disinfectants, antiseptics, antibiotics, and
chemotherapeutic antimicrobial chemicals.

Basic terms used in discussing the control of microorganisms include:

1. Sterilization: The process of destroying all living organisms and viruses. A sterile
object is one free of all life forms, including bacterial endospores, as well as viruses.
 2. Disinfection: It is the elimination of microorganisms from inanimate objects or
surfaces.
3. Decontamination: The treatment of an object or inanimate surface to make it safe to
handle.\ 3. Disinfectant- A disinfectant is an agents used to disinfect inanimate objects
but generally to toxic to use on human tissues.
4. Antiseptic: An antiseptic is an agent that kills or inhibits growth of microbes but is
safe to use on human tissue.
5. Sanitizer: A sanitizer is an agent that reduces, but may not eliminate, microbial
numbers to a safe level.
6. Cidal: An agent that is cidal in action will kill microorganisms and viruses.
7. Static: An agent that is static in action will inhibit the growth of microorganisms.
Keep in mind that when evaluating or choosing a method of controlling
microorganisms, you must consider the following factors which may influence
antimicrobial activity
1. The concentration and kind of a chemical agent used.
2. The intensity and nature of a physical agent used.
3. The length of exposure to the agent.
4. The temperature at which the agent is used.

5. The number of microorganisms present.

6. The organism itself.
7. The nature of the material bearing.

PHYSICAL AGENTS IN MICROBIAL CONTROL

TEMPERATURE:
Microorganisms have a minimum, an optimum, and a maximum temperature for growth.
Temperatures below the minimum usually have a static action on microorganisms. They
inhibit microbial growth by slowing down metabolism but do not necessarily kill the
organism. Temperatures above the maximum usually have a cidal action, since they denature
microbial enzymes and other proteins. Temperature is a very common and effective way of
controlling microorganisms. High Temperature Vegetative microorganisms can generally be
killed at temperatures from 50°C to 70°C with moist heat. Bacterial endospores, however, are
very resistant to heat and extended exposure to much higher temperature is necessary for their
destruction. High temperature may be applied as either moist heat or dry heat.

a. Moist heat
Moist heat is generally more effective than dry heat for killing microorganisms because of
its ability to penetrate microbial cells. Moist heat kills microorganisms by denaturing their
proteins (causes proteins and enzymes to lose their three-dimensional functional shape). It
also may melt lipids in cytoplasmic membranes.
>Autoclaving: Autoclaving employs steam under pressure. Water normally boils at
100°C; however, when put under pressure, water boils at a higher temperature. During
autoclaving, the materials to be sterilized are placed under 15 pounds per square inch of
pressure in a pressure-cooker type of apparatus. When placed under 15 pounds of
pressure, the boiling point of water is raised to 121°C, a temperature sufficient to kill
bacterial endospores. The time the material is left in the autoclave varies with the nature
and amount of material being sterilized. Given sufficient time (generally 15-45 minutes),
autoclaving is cidal for both vegetative organisms and endospores, and is the most
common method of sterilization for materials not damaged by heat. Boiling water (100°C)
will generally kill vegetative cells after about 10 minutes of exposure. However, certain
viruses, such as the hepatitis viruses, may survive exposure to boiling water for up to 30
minutes, and endospores of certain Clostridium and Bacillus species may survive even
hours of boiling. b. Dry heat
Dry heat kills microorganisms through a process of protein oxidation rather than protein
coagulation. Examples of dry heat include:

>Hot air sterilization: Microbiological ovens employ very high dry temperatures: 171°C
for 1 hour; 160°C for 2 hours or longer; or 121°C for 16 hours or longer depending on the
volume. They are generally used only for sterilizing glassware, metal instruments, and
other inert materials like oils and powders that are not damaged by excessive temperature.

>Incineration: Incinerators are used to destroy disposable or expendable materials by

burning. We also sterilize our inoculating loops by incineration.

>Pasteurization: Pasteurization is the mild heating of milk and other materials to kill
particular spoilage organisms or pathogens. It does not, however, kill all organisms.
Milk is usually pasteurized by heating to 71.6°C for at least 15 seconds in the flash
method or 62.9°C for 30 minutes in the holding method. Low temperature inhibits
microbial growth by slowing down microbial metabolism. Examples include refrigeration
and freezing. Refrigeration at 5°C slows the growth of microorganisms and keeps food
fresh for a few days. Freezing at -10°C stops microbial growth, but generally does not kill
microorganisms, and keeps food fresh for several months.

c. Dessication

Desiccation, or drying, generally has a static effect on microorganisms. Lack of water

inhibits the action of microbial enzymes. Dehydrated and freeze-dried foods, for example,
do not require refrigeration because the absence of water inhibits microbial growth.

d. Osmotic pressure

Microorganisms, in their natural environments, are constantly faced with alterations in

osmotic pressure. Water tends to flow through semipermeable membranes, such as the
cytoplasmic membrane of microorganisms, towards the side with a higher concentration
of dissolved materials (solute). In other words, water moves from greater water (lower
solute) concentration to lesser water (greater solute) concentration.

When the concentration of dissolved materials or solute is higher inside the cell than it is
outside, the cell is said to be in a hypotonic environment and water will flow into the
cell. The rigid cell walls of bacteria and fungi, however, prevent bursting or plasmoptysis.

If the concentration of solute is the same both inside and outside the cell, the cell is said to
be in an isotonic environment. Water flows equally in and out of the cell. Hypotonic and
isotonic environments are not usually harmful to microorganisms. However, if the
concentration of dissolved materials or solute is higher outside of the cell than inside, then
the cell is in a hypertonic environment. Under this condition, water flows out of the cell,
resulting in shrinkage of the cytoplasmic membrane or plasmolysis. Under such
conditions, the cell becomes dehydrated and its growth is inhibited.

The canning of jams or preserves with a high sugar concentration inhibits bacterial growth
through hypertonicity. The same effect is obtained by salt-curing meats or placing foods
in a salt brine. This static action of osmotic pressure thus prevents bacterial decomposition
of the food. Moulds, on the other hand, are more tolerant of hypertonicity. Foods, such as
those mentioned microorganism.

e. Radiation
>Ultraviolet Radiation: The ultraviolet portion of the light spectrum includes all
radiations with wavelengths from 100 nm to 400 nm. It has low wave-length and low
energy. The microbicidal activity of ultraviolet (UV) light depends on the length of
exposure: the longer the exposure the greater the cidal activity. It also depends on the
wavelength of UV used. The most cidal wavelengths of UV light lie in the 260 nm - 270
nm range where it is absorbed by nucleic acid. In terms of its mode of action, UV light is
absorbed by microbial DNA and causes adjacent thymine bases on the same DNA strand
to covalently bond together, forming what are called thymine-thymine dimers. As the
DNA replicates, nucleotides do not complementary base pair with the thymine dimers and
this terminates the replication of that DNA strand. However, most of the damage from UV
radiation actually comes from the cell trying to repair the damage to the DNA by a
process called SOS repair. In very heavily damaged DNA containing large numbers of
thymine dimers, a process called SOS repair is activated as kind of a last ditch effort to
repair the DNA. In this process, a gene product of the SOS system binds to DNA
polymerase allowing it to synthesize new DNA across the damaged DNA. However, this
altered DNA polymerase loses its proofreading ability resulting in the synthesis of DNA
that itself now contains many misincorporated bases. In other words, UV radiation causes
mutation and can lead to faulty protein synthesis. With sufficient mutation, bacterial
metabolism is blocked and the organism dies. Agents such as UV radiation that cause high
rates of mutation are called mutagens. The effect of this inproper base pairing may be
reversed to some extent by exposing the bacteria to strong visible light immediately after
exposure to the UV light. The visible light activates an enzyme that breaks the bond that
joins the thymine bases, thus enabling correct complementary base pairing to again take
place. This process is called photoreactivation. UV lights are frequently used to reduce the
microbial populations in hospital operating rooms and sinks, aseptic filling rooms of
pharmaceutical companies, in microbiological hoods, and in the processing equipment
used by the food and dairy industries. An important consideration when using UV light is
that it has very poor penetrating power. Only microorganisms on the surface of a material
that are exposed directly to the radiation are susceptible to destruction. UV light can also
damage the eyes, cause burns, and cause mutation in cells of the skin.
>Ionizing Radiation: Ionizing radiation, such as X-rays and gamma rays, has much more
energy and penetrating power than ultraviolet radiation. It ionizes water and other
molecules to form radicals (molecular fragments with unpaired electrons) that can disrupt
DNA molecules and proteins. It is often used to sterilize pharmaceuticals and disposable
medical supplies such as syringes, surgical 5 gloves, catheters, sutures, and petri plates. It
can also be used to retard spoilage in seafoods, meats, poultry, and fruits. F.
FILTRATION Microbiological membrane filters provide a useful way of sterilizing
materials such as vaccines, antibiotic solutions, animal sera, enzyme solutions, vitamin
solutions, and other solutions that may be damaged or denatured by high temperatures or
chemical agents. The filters contain pores small enough to prevent the passage of
microbes but large enough to allow the organismfree fluid to pass through. The liquid is
then collected in a sterile flask. Filters with a pore diameter from 25 nm to 0.45 µm are
usually used in this procedure. Filters can also be used to remove microorganisms from
water and air for microbiological testing.

CHEMICAL AGENTS IN MICROBIAL CONTROL

 Chemical agents: chemical means to destroy or remove contaminants.
 Three Levels of Chemical Decontamination
>High-level germicides kill endospores, and if properly used, are sterilants
>Intermediate germicides kill fungal (not bacterial) spores, resistant pathogens such
as tubercle bacillus, and viruses (used for non-invasive equipment).
>Low-level germicides kill vegetative bacteria and fungal cells, and some viruses
(used for materials that may touch the skin, not mucous membrane).
 Dilutions
Chemical (solute) is added to water (solution), aqueous.
Noted at (solute):(solution) --> For 1 part Lysol there are 100 parts
water(1:100).
 Penetration
Smooth, solid objects are easier to disinfect due to rough objects can collect dirt or
contaminants in crevices that the disinfectant can't penetrate.
 Germicidal Categories According to Chemical Group:
1. Halogens: non-metallic elements that commonly occur in minerals, seawater, and
salts, Fluorine, bromine, chlorine, and iodine.
2. Fluorine and bromine difficult and dangerous to handle.
3. Microbicidal (sporicidal with longer exposure time) 4. Chlorine (CL2), hypochlorite
(Clorox), chloramines.
PRESERVATION OF MICROORGANISMS:
 Short term
 Long term

SHORT -TERM PRESERVATION METHODS:

A. Direct transfer to subculture
1. Simplest method for maintaining a short-term viability of microorganism,
saved for more then1 week.
2. Maintenance medium- support the survival of microorganism but minimize
its metabolic processes and slow its rate of growth, distilled water, tryptic
soy broth and nutrients broth with or without cryo-preservatives.
3. Storage done at lower temperature 5-8℃

B. Immersion in oil
1. An alternative to capping tubes -add a layer of mineral oil to the top of the
specimen with specific gravity of 0.865 to 0.890 and heated to 170℃ for 1
to 2 hr in an oven: sterilization.

2. Many bacteria and fungi can be stored for periods of upto 2 -3 years by this
method and transfers are not needed as frequently.

C. Freezing at -20℃
1. Refrigeration or freezing in ordinary freezers at -20℃ may be used to
preserve microorganism for periods longer than those that can be
accomplished by repeated transfers.
2. Viability may be maintained for as long as 1 to 2yrs for specific
microorganism but overall damage cause by the ice crystal formation and
electrolyte fluctuation results in poor long-term survival. D. Drying:

1. Moulds and some spore -forming bacteria may be dried and stored for
prolonged periods.
2. This method is used for quality control microorganism.

LONG -TERM PRESERVATION METHODS:

A. ultra- low temperature freezing: Maintained at temperatures of -70℃ or lower for
prolonged periods using ultra low temperature electric freezers and liquid nitrogen
storage units.
 STORAGE VIALS: Plastic (polypropylene) or glass (borosilicate) tubes able to
withstand very low temperatures and maintain a seal for their contents.

 CRYOPROTECTIVE AGENTS: To protect microorganism from the damage

during the freezing process, storage, thawing are added to the culture suspension.
B. Freezing method:
Preparation of microorganism for freezing:
 Cultures are allowed to mature to the late growth or stationary phase before being
harvested.
 Broth specimens are centrifuged (low revolution) to create a pellet of microorganisms
and resuspended in 2to 5 ml of broth with appropriate concentration of cryoprotectant
additive.
 Agar- scarping
 Volume of the aliquots to be frozen is typically 0.2 to 0.5 ml.

Methodology:
 American type culture collection (ATTC) recommends slow, controlled- rate freezing
at the rate of 1℃ per min until the vials cool to a temperature of at least -30℃
followed by more rapid cooling until the final storage temperature is achieved.
 When organism are stored in liquid nitrogen, however it is still recommended that
vials be placed initially in a -60℃ freezer for 1 hr and then transferred into the liquid
nitrogen (-196℃). C. Thawing:

 Damage to microorganism occurs as they are unwanted from the frozenstate.

 critical temperature appears to be between -40 and -5℃ (improves recovery rates)
 once a vial is thawed, it should be opened and organism should be transferred to an
appropriate growth medium immediately.

DNA
DNA is known as Deoxyribonucleic Acid. It is an organic compound that has a unique
molecular structure. It is found in all prokaryotic cells and eukaryotic cells.

“DNA is a group of molecules that is responsible for carrying and transmitting the
hereditary materials or the genetic instructions from parents to off-springs.”
This is also true for viruses as most of these entities have either RNA or DNA as their
genetic material. For instance, some viruses may have RNA as their genetic material,
while others have DNA as the genetic material. The Human Immunodeficiency Virus
(HIV) contains RNA, which is then converted into DNA after attaching itself to the host
cell. Apart from being responsible for the inheritance of genetic information in all living
beings, DNA also plays a crucial role in the production of proteins. Nuclear DNA is the
DNA contained within the nucleus of every cell in a eukaryotic organism. It codes for the
majority of the organism’s genomes while the mitochondrial DNA and plastid DNA
handles the rest. The DNA present in the mitochondria of the cell is termed as
mitochondrial DNA. It is inherited from the mother to the child. In humans, there are
approximately 16,000 base pairs of mitochondrial DNA. Similarly, plastids have their
own DNA and they play an essential role in photosynthesis.
DNA TYPES: There are three different DNA types:
A-DNA: It is a right-handed double helix similar to the B-DNA form. Dehydrated DNA
takes an A form that protects the DNA during extreme condition such as desiccation.
Protein binding also removes the solvent from DNA and the DNA takes an A form.
B-DNA: This is the most common DNA conformation and is a right-handed helix. Majority
of DNA has a B type conformation under normal physiological conditions.
Z-DNA: Z-DNA is a left-handed DNA where the double helix winds to the left in a zig-
zag pattern. It was discovered by Andres Wang and Alexander Rich. It is found ahead of
the start site of a gene and hence, is believed to play some role in gene regulation.

DNA STRUCTURE:
The DNA structure can be thought of like a twisted ladder. This structure is described as a
double-helix, as illustrated in the figure above. It is a nucleic acid, and all nucleic acids are
made up of nucleotides. The DNA molecule is composed of units called nucleotides, and
each nucleotide is composed of three different components, such as sugar, phosphate
groups and nitrogen bases.
The basic building blocks of DNA are nucleotides, which are composed of a sugar group,
a phosphate group, and a nitrogen base. The sugar and phosphate groups link the
nucleotides together to form each strand of DNA.
Adenine (A), Thymine (T), Guanine (G) and Cytosine (C) are four types of nitrogen bases.
These 4 Nitrogenous bases pair together in the following way: A with T, and C with G.
These base pairs are essential for the DNA’s double helix structure, which resembles a
twisted ladder. Among the three components of DNA structure, sugar is the one which
forms the backbone of the DNA molecule. It is also called deoxyribose. The nitrogenous
bases of the opposite strands form hydrogen bonds, forming a ladder-like structure. The
DNA molecule consists of 4 nitrogen bases, which ultimately forms the structure of a
nucleotide. The A and G are purines and the C and T are pyrimidines. The two strands of
DNA run in opposite directions.
These strands are held together by the hydrogen bond that is present between the two
complementary bases. The strands are helically twisted, where each strand forms a
righthanded coil and ten nucleotides make up a single turn. The pitch of each helix is 3.4
nm. Hence, the distance between two consecutive base pairs (i.e., hydrogen-bonded bases
of the opposite strands) is 0.34 nm. The DNA coils up, forming chromosomes and each
chromosome has a single molecule of DNA in it. Overall, human beings have around
twentythree pairs of chromosomes in the nucleus of cells. DNA also plays an essential
role in the process of cell division.
DNA Function
DNA is the genetic material which carries all the hereditary information. Genes are the
small segments of DNA, consisting mostly of 250 – 2 million base pairs. A gene code for
a polypeptide molecule, where three nitrogenous bases sequence stands for one amino
acid.

Polypeptide chains are further folded in secondary, tertiary and quaternary structure to
form different proteins. As every organism contains many genes in their DNA, different
types of proteins can be formed. Proteins are the main functional and structural molecules
in most of the organisms. Apart from storing genetic information, DNA is involved in:
Replication process: Transferring the genetic information from one cell to its daughters
and from one generation to the next and equal distribution of DNA during the cell division
Mutations: The changes which occur in the DNA sequences

• Transcription

• Cellular Metabolism

• DNA Fingerprinting

• Gene Therapy

DNA REPLICATION:
DNA replication is an important process that occurs during cell division. It is also known as
semi-conservative replication, during which DNA makes a copy of itself.
DNA replication takes place in three stages
Initiation
The replication of DNA begins at a point known as the origin of replication. The two DNA
strands are separated by the DNA helicase. This forms the replication fork.

Elongation
DNA polymerase III reads the nucleotides on the template strand and makes a new strand
by adding complementary nucleotides one after the other. For eg., if it reads an Adenine
on the template strand, it will add a Thymine on the complementary strand.While adding
nucleotides to the lagging strand, gaps are formed between the strands. These gaps are
known as Okazaki fragments. These gaps or nicks are sealed by ligase.

Termination
The termination sequence present opposite to the origin of replication terminates the
replication process. The TUS protein (terminus utilization substance) binds to terminator
sequence and halts DNA polymerase movement. It induces termination.

RNA
RNA is a ribonucleic acid that helps in the synthesis of proteins in our body. This nucleic
acid is responsible for the production of new cells in the human body. It is usually
obtained from the DNA molecule. RNA resembles the same as that of DNA, the only
difference being that it has a single strand unlike the DNA which has two strands and it
consists of an only single ribose sugar molecule in it. Hence is the name Ribonucleic acid.
RNA is also referred to as an enzyme as it helps in the process of chemical reactions in the
body.
BASIC STRUCTURE OF RNA: The basic structure of RNA is shown in the figure below-

The ribonucleic acid has all the components same to that of the DNA with only 2 main
differences within it. RNA has the same nitrogen bases called the adenine, Guanine,
Cytosine as that of the DNA except for the Thymine which is replaced by the uracil.
Adenine and uracil are considered as the major building blocks of RNA and both of them
form base-pair with the help of 2 hydrogen bonds. RNA resembles a hairpin structure and
like the nucleotides in DNA, nucleotides are formed in this ribonucleic material (RNA).
Nucleosides are nothing but the phosphate groups which sometimes also helps in the
production of nucleotides in the DNA.
FUNCTION OF RNA:
The ribonucleic acid – RNA, which are mainly composed of nucleic acids, are involved in
a variety of functions within the cell and are found in all living organisms including
bacteria, viruses, plants, and animals. These nucleic acid functions as a structural
molecule in cell organelles and are also involved in the catalysis of biochemical reactions.
The different types of RNA are involved in various cellular process. The primary
functions of RNA:
Facilitate the translation of DNA into proteins
Functions as an adapter molecule in protein synthesis
Serves as a messenger between the DNA and the ribosomes.
They are the carrier of genetic information in all living cells
Promotes the ribosomes to choose the right amino acid which is required in building up of
new proteins in the body.

RNA TYPES:
There are various types of RNA, out which most well-known and most commonly studied
in the human body are: t-RNA – Transfer RNA

The transfer RNA is held responsible for choosing the correct protein or the amino acid
required by the body in-turn helping the ribosomes. It is located at the endpoints of each
amino acid. This is also called as soluble RNA and it forms a link between the messenger
RNA and the amino acid. r-RNA - Ribosomal RNA
The rRNA is the component of the ribosome and are located within the in the cytoplasm
of a cell, where ribosomes are found. In all living cells, the ribosomal RNA plays a
fundamental role in the synthesis and translation of mRNA into proteins. The rRNA is
mainly composed of cellular RNA and are the most predominant RNA within the cells of
all living beings.
m-RNA – Messenger RNA.
This type of RNA functions by transferring the genetic material into the ribosomes and
pass the instructions about the type of proteins, required by the body cells. Based on the
functions, these types of RNA is called the messenger RNA. Therefore, the mRNA plays
a vital role in the process of transcription or during the protein synthesis process.
RNA REPLICATION / TRANSCRIPTION

“Transcription is the first step of gene expression that involves the formation of RNA
molecule from DNA.”
It is one of the first processes in gene expression. The genetic information flows from
DNA to protein and this flow of information takes place in a sequential process of
transcription and translation. Only one strand of DNA is copied during the process of
transcription known as the template strand and the RNA formed is called the mRNA. The
main motive of transcription is to make a copy of RNA from the DNA sequence. The
RNA transcript carries the information used to encode a protein.

RNA POLYMERASE:
The RNA polymerase is the main enzyme involved in transcription. It uses single-strand
DNA to synthesize a complementary RNA strand. The DNA-dependent RNA polymerase
binds to the promoter and catalyses the polymerization in the 5’ to 3’ direction on the
template strand. Once it reaches the terminator sequence, the process terminates and the
newly synthesised RNA strand is released.
Transcription Unit is a stretch of a DNA transcribed into an RNA molecule. Its function is
to encode at least one gene. Suppose if gene encodes protein than mRNA is produced by
transcription. A protein encoded by the DNA transcription unit may comprise a coding
sequence. Compared to DNA replication, transcription has a lower copying fidelity.

STAGES OF TRANSCRIPTION:
Transcription proceeds in enzymatically catalysed steps i.e.
Initiation.
Elongation
Termination
INITIATION:
RNA polymerase attaches to the DNA molecule and moves along the DNA strand until it
recognises a promoter sequence. These are known as the transcription start sites. The
DNA double helix then unwinds and all the bases on each of the DNA strands are
exposed. This acts as a template for a new mRNA strand.
ELONGATION: Ribo-nucleoids are added to the template strand that enables the growth of
mRNA growth.
TERMINATION: RNA polymerase encounters a terminator sequence and the transcription
stops. RNA polymerase then releases the DNA template.

RNA PROCESSING
The transcribed RNA is known as the pre-mRNA. It is processed further to convert it into
mature RNA. RNA processing include:
 Capping
 Polyadenylation
 Splicing
CAPPING: A methylated guanine cap is added to protect the mRNA. It involves,
Addition of methylated guanine. It occurs at 5′ end of mRNA transcript. It protects the
mRNA from degradation
POLYADENYLATION: The poly-A tail also protects the mRNA from degradation. It
involves, The endonucleases cleave the mRNA at a specific sequence. The enzyme poly A
polymerase facilitates the addition of several adenine nucleotides.

SPLICING: The non-coding sequences, i.e., the introns are removed by spliceosome
excision. The coding sequences or the exons join together by ligation. Thus several
proteins can be made from a single pre-mRNA. A mature mRNA is obtained at the end of
transcription.
RECOMBINANT DNA TECHNOLOGY
The technology used for producing artificial DNA through the combination of different
genetic materials (DNA) from different sources is referred to as Recombinant DNA
Technology. Recombinant DNA technology is popularly known as genetic engineering.
The recombinant DNA technology emerged with the discovery of restriction enzymes in
the year 1968 by Swiss microbiologist Werner Arber, Inserting the desired gene into the
genome of the host is not as easy as it sounds. It involves the selection of the desired gene
for administration into the host followed by a selection of the perfect vector with which
the gene has to be integrated and recombinant DNA formed. Thus the recombinant DNA
has to be introduced into the host. And at last, it has to be maintained in the host and
carried forward to the offspring.

TOOLS OF RECOMBINANT DNA TECHNOLOGY

The enzymes which include the restriction enzymes help to cut, the polymerases- help to
synthesize and the ligases- help to bind. The restriction enzymes used in recombinant
DNA technology play a major role in determining the location at which the desired gene is
inserted into the vector genome. They are two types, namely Endonucleases and
Exonucleases. The Endonucleases cut within the DNA strand whereas the Exonucleases
remove the nucleotides from the ends of the strands. The restriction endonucleases are
sequence-specific which are usually palindrome sequences and cut the DNA at specific
points. They scrutinize the length of DNA and make the cut at the specific site called the
restriction site. This gives rise to sticky ends in the sequence. The desired genes and the
vectors are cut by the same restriction enzymes to obtain the complementary sticky notes,
thus making the work of the ligases easy to bind the desired gene to the vector.

The vectors – help in carrying and integrating the desired gene. These form a very
important part of the tools of recombinant DNA technology as they are the ultimate
vehicles that carry forward the desired gene into the host organism. Plasmids and
bacteriophages are the most common vectors in recombinant DNA technology that are
used as they have a very high copy number. The vectors are made up of an origin of
replication- This is aS sequence of nucleotide from where the replication starts, a
selectable marker – constitute genes which show resistance to certain antibiotics like
ampicillin; and cloning sites – the sites recognized by the restriction enzymes where
desired DNAs are inserted.

Host organism – into which the recombinant DNA is introduced. The host is the ultimate
tool of recombinant DNA technology which takes in the vector engineered with the
desired DNA with the help of the enzymes. There are a number of ways in which these
recombinant DNAs are inserted into the host, namely – microinjection, biolistics or gene
gun, alternate cooling and heating, use of calcium ions, etc.

PROCESS OF RECOMBINANT DNA TECHNOLOGY:

The complete process of recombinant DNA technology includes multiple steps, maintained
in a specific sequence to generate the desired product.

Step-1: Isolation of Genetic Material.

The first and the initial step in Recombinant DNA technology is to isolate the desired DNA
in its pure form i.e., free from other macromolecules.

Step-2: Cutting the gene at the recognition sites.

The restriction enzymes play a major role in determining the location at which the desired
gene is inserted into the vector genome. These reactions are called restriction enzyme
digestions.
Step-3: Amplifying the gene copies through Polymerase chain reaction (PCR).
It is a process to amplify a single copy of DNA into thousands to millions of copies once the
proper gene of interest has been cut using the restriction enzymes.

Step-4: Ligation of DNA Molecules.

In this step of Ligation, joining of the two pieces – a cut fragment of DNA and the vector
together with the help of the enzyme DNA ligase.

Step-5: Insertion of Recombinant DNA into Host.

In this step, the recombinant DNA is introduced into a recipient host cell. This process is
termed as Transformation. Once after the insertion of the recombinant DNA into the
hostcell, it gets multiplied and is expressed in the form of the manufactured protein under
optimal conditions.
APPLICATION OF RECOMBINANT DNA TECHNOLOGY:
• DNA technology is also used to detect the presence of HIV in a person.

• Gene Therapy – It is used as an attempt to correct the gene defects which give rise to
heredity diseases.

• Clinical diagnosis – ELISA is an example where the application of recombinant

• Recombinant DNA technology is widely used in Agriculture to produce

geneticallymodified organisms such as Flavr Savr tomatoes, golden rice rich in
proteins, Btcotton to protect the plant against ball worms and lot more.

• In the field of medicines, Recombinant DNA technology is used for the production of
Insulin.

INTELLUCTUAL PROPERTY RIGHTS

Science of microbiology, having its origins in the early human attempt to modify
biological organisms for its needs have reached critical heights from the days of brewing
to the current research in genomics, stem cells etc. The commercial application of
microbiology in the agricultural industry has shown tremendous expansion in the last few
years. Along with these developments, the ethical and legal issues have also arisen. A
creation of human brain which is termed as intellect can also be a property. Intellectual
Property thus refers to inventions, industrial designs, literary and artistic works, symbols
etc. used in commerce. Protection of intellectual properties may be by patents, copyrights,
trademarks etc. The versatile functions performed by microorganisms have rendered their
exploitation and commercial use in both industrial & agricultural sectors. Thus,
microorganisms and microbial products have become an important and lucrative
component of global business. But, monetary value of microorganisms and their products
concurrently bring the issue of “Rights” for their use and commercial exploitation. Patent
is the strongest form of intellectual property right (IPR)of TRIPS Agreement allows
member states to deny patents for “plants and animals, other than microorganisms, and
essentially biological processes for the production of plants or animals other than non-
biological and microbiological processes.”

As a result, TRIPS makes it obligatory for all its signatories to extend patents for
microorganisms, non-biological, and microbiological processes. In compliance with TRIPs,
the Patents Act 1970, as amended in June 2002, gives patent rights for new microorganisms.
Naturally occurring microorganisms are not patentable in India but genetically modified
microbes or microbe-based products can be protected by patent rights. Earlier, inventions
pertaining to microorganisms and other biological materials were subjected to product patent
in India, unlike many developed countries where microorganisms themselves are patentable.
But with effect from 20th May, 2003, India started granting patents in respect of inventions
related to microorganisms.

The grant of patent with respect to microorganisms is regulated by the requirements for the
deposition of microorganisms under the Budapest Treaty of and accessibility of that
microorganism from the depositories. According to the provision (ii) to section 10(d) the
microorganism if not being described fully and particularly and is not available to public, the
said microorganism is to be deposited before the International Depositary Authority (IDA)
under the Budapest Treaty 2002. Since 2001, India has become a member of Budapest Treaty
on deposition of microorganisms and consequently two microbial repositories in India viz.
Microbial Type Culture Collection (MTCC) and Microbial Culture Collection (MCC) have
acquired the status of IDA.

REFERENCES
[1] Bhatia S.C., “Hand book of Environmental Microbiology”, Part 1 and 2, Atlantic
publisher, 2008.
[2] Gabriel Bitton, Wastewater Microbiology, 2 nd Edition & 4th Edition, AJohn Wiley &
Sons, Inc., Publication.
[3] Ian L. Pepper, Charles P. Gerba, Terry J. Gentry, 3rd Edition, “Environmental
Microbiology”, Academic press, 2000.
[4] Nduka Okafor, Environmental Microbiology of Aquatic and waste systems. Springer
publishers, 2011, ISBN978-94-007-1459-5.