NORMAL FLORA

NIGARISH AHSAN
PHARM-D , MPHIL . (PU)
LECTURER PHARMACEUTICS
Normal Flora
• It is population of microorganism that infest the body
without causing disease
• Relation of m.o. to human may be
• Symbiosis – organism interacts in a close and permanent
association e.g. E.Coli always found in human intestine
• Mutualism – symbiosis beneficial to body and
microorganism e.g. Lactobacillus found in Vagina
• Commensalism – symbiosis in which only one population
benefits from association e.g. E.Coli produce certain
amount of Vitamin B and Vitamin K
Development of Normal Flora
• Normal flora – Guards of First Line of Defense
• Fetus of mother is microbe free but microorganism passes
to child from birth canal
• Additional organism enters when breathing begins and
upon first feeding
• Contact with other humans also expose the child to new
microorganism
• It continue throughout the life and undergoes changes in
response to internal environment of individual
Development of Normal Flora
• Internal tissues like blood brain muscles are free of
microorganism
• Surface tissues like skin and mucous membrane are
constantly in contact with the environmental m.o.
• Many of the bacteria are either pathogenic or
opportunistic pathogens
• Bacteria of normal flora present at one anatomical site
may not be pathogenic but if travel to other site will
become virulent
Normal Flora Examples
• Streptococous mutans is involved in dental plaque
formation – opportunistic bacteria
• Streptococcus pneumoniae is present in upper respiratory
tract, but if travels to lower respiratory tract it will cause
pneumonia
• Many different strains of E.Coli are non-virulent but some
of them are become causative agent for diarroea
Composition of Normal Flora
• Makeup of human flora is influenced by genetics, age,
sex, stress, nutrition and diet of the individual
• Bacterial flora is sufficiently constant among all individuals
• Human developmental changes like weaning, eruption of
teeth, onset and cessation of ovarian function invariably
effect the normal flora of intestinal tract, oral cavity and
vagina respectively
• In rough estimate human house about 1012 bacteria on
skin, 1010 bacteria in mouth, 1014 Bacteria in GIT
Normal Flora of Skin
• Density and composition of normal flora is varied
according to anatomical site
• High moisture content area – Axilla, Groin, toes facilitates
high density of growth
• Other sites have 100 or may be 1000 bacteria per square
inch
• Most bacteria of the skin found on superficial layer of
epidermis and upper part of the hair follicles are gram
positive cocci – Staphylococcus epidermidis and
Micrococcus species – and cornybacteria such as
Propionibacterium species
Normal Flora of Skin
• Usually they are non-pathogenic, mutualistic and
commensal
• They produce fatty acid on skin surface to inhibit the
growth of fungi and yeast on skin
• Propionibacterium acne – normal inhabitant of skin, when
trapped in hair follicle starts inflammation and acne
Normal Flora of Conjunctiva
• A variety of bacteria may be cultivated from normal
conjuctivae, but the number of organism is usually small.
• Staphylococcus epidermidis, certain coryneforms,
Staphylococcus aureus, some streptococci, Haemophilius
species are present
• Neisseria gonorrhoeae and Chlamydia trachomatis are
specifically involve in infection of conjunctiva.
• The conjuctiva is kept moist and healthy by continuous
secretions from lachrymal glands .
• Blinking wipes the conjuctiva every few seconds
mechanically washing away foreign objects including
bacteria .
Normal Flora of Respiratory Tract
• A large number of bacterial species colonize the upper
respiratory tract.
• The nostrils are heavily colonized with Staphylococcus
epidermidis, certain coryneforms, and also with certain
Staphylococcus aureus,
• The pharynx (throat) is normally colonized by streptococci and
various gram negative cocci.
• Streptococcus pneumoniae, Streptococcus pyogenes,
Haemophilus influenza and Neisseria meningitides colonize
the pharynx.
• If the respiratory tract epithelium becomes damaged as in
bronchitis or viral pneumonia, the individual may become
susceptible to infection by pathogens such as H. influenzae or
S.pneumoniae descending from nasopharynx.
MICROBIOLOGY OF
WATER
Introduction
• Water is the medium of life.
• All organisms are composed
primarily of water.
• Liquid water is the medium in
which all cellular chemical
reactions occur.
• Essential for survival and growth
of organism.
• The drinking water is mostly obtained from surface
sources i.e rivers streams lakes.
• Such natural supplies are likely to be polluted with
domestic and industrial wastes.
• A greater quantity of water is required , and the used
water must be disposed of, by returning it .
• Water is a potential carrier for pathogenic
microorganisms. These pathogens can cause various
infections like typhoid, dysentery , cholera.
• It is therefore necessary to employ
• 1) treatment facilities that purify waste water prior to its
disposal .
• 2) water purification methods that provide safe drinking
water .
Water purification
Potable water:
Water that is free of
Disease producing microorganisms
Chemical substances and fit for drinking.

• Non potable water/ polluted water:

• Water contaminated with
Domestic wastes
Industrial wastes

• Three main features to be considered to have potable water:

Source of water
Storage
Means to bring water to population
 Microbiological characteristics
• Since the composition of waste water varies . It

contains
• Fungi

• Protozoa

• Algae

• Bacteria , and

• Viruses

• Raw sewage may contain millions of bacteria per

ml
 Water born diseases
• Water can act as a vector for the transmission of bacterial, viral
and protozoan agents which cause a variety of diseases (mainly
intestinal)

• Itcan also be linked to worm invasions and viral/protozoan

diseases transmitted by insects (aquatic hosts or insect breeding
in water- indirect)

• Water is responsible for, by some estimates, approximately 80%

of all infectious disease not just waterborne diseases, but any
disease where water plays a role.
 VIRUSES PATHOGENS

Enteritis, diarrhea, and dysentery

⚫ Rotavirus
⚫ Norwalk
Liver damaging
⚫ Hepatitis A
⚫ Hepatitis E
Paralysis
⚫ Polio virus
Bacteria responsible for water diseases
• Enteritis, diarrhea, and dysentery
• Campylobacter
• Cholera
• E. coli
• Salmonella
• Shigella
• Enteric fever
• Typhoid
• Paratyphoid
• Eye, ear, and skin infections
• Miscellaneous bacteria
 Protozoa
A-Amoeba:
Entamoeba species.
B- Flagellates:
Gastrointestinal flagellates: Giardia intestinalis
Urogenital flagellates: Trichomonas vaginalis
Tissue and blood flagellates: Trypanosoma
C- Ciliophora: motile by cilia.
Example: Balantidium coli.
D- Sporozoa: intracellular infection.
Example: Plasmodium that cause Malaria
 FUNGI
•A diverse group of eukaryotes that are plant-like but that cannot
carry on photosynthesis.

• They serve as decomposers, absorbing nutrients from dead

leaves or other organic matter in soil and water.

• Fungi are found in very less quantity/numbers in water. This is

due to their competition with other heterotrophic organisms like
some bacteria and archae for food intake. their low abundance is
an indicator for their loss during competition.

• Fungi produce spores during the reproductive process.

 Algae
Algae may produce toxins in sea food which can cause illness to
human beings by using contaminated seafood.
Most important are;
a) Amnesic Shellfish Poisoning (ASP)
b) Ciguatera Fish Poisoning (CFP)
c) Diarrhetic Shellfish Poisoning (DSP)
d) Neurotoxic Shellfish Poisoning (NSP)
e) Paralytic Shellfish Poisoning (PSP)
Amnestic Shellfish Poisoning (ASP):
Causative organisms: Pseudo-nitzschia sp.
HELMINTHES

Round worm
Tape worm
Hook worm
Whip worm
Water-associated diseases can be classified
under 4 different categories: -
1. Water-borne diseases

2. Water-washed diseases

3. Water-based diseases

4. Water-related diseases
 A. Water-borne diseases
❑ They are produce by ingestion of contaminated water.

❑ They are mainly enteric diseases.

❑ Classical H2O -borne diseases are mostly:

❑ Protozoal infection; Amoebiasis, Giardia and Cryptosporidium etc.

❑ Bacterial infection; cholera and typhoid fever (rare),

Campylobacteriosis, bacillary dysentery, leptospirosis (rare),
dysentry, E.coli infections.
❑ Viral infections; infectious hepatitis, poliomyelitis, SARS
(Severe Acute Respiratory Syndrome).
2. Water-washed diseases
• Diseases caused by poor personal hygiene.

• Obviously more common in tropical, 3rd world countries where

maybe water scarcity is present.

• Intestinal and non-intestinal infections (Eye & skin diseases)

• Intestinal: Shigella (dysentery); typhoid; cholera; scabies, Yaws

(Yaws is caused by T. Pallidum subspecies pertenue, affects
skin, bone and cartilage), leprosy, conjunctivitis, other skin
infections and ulcers. Campylobacter; Giardia;
Cryptosporidium; viruses.
3. Water-based diseases
• Diseases caused by pathogens that have a complex life-cycle
which involves an intermediate aquatic host.

• All of these diseases are caused by worms, e.g.

✔ Schistosomiasis caused by the Schistosoma worm which
uses aquatic snails as an intermediate host,

✔ Dracunculiasis cause by Guinea worm which uses a small

crustacean as an intermediate host
4. Water-related diseases
• Diseases caused by pathogens carried by insects that live near
H2O and act as mechanical vectors.
• Difficult to control and diseases are severe.
Examples:
• Yellow fever (viral disease) is transmitted by the mosquito Aedes spp.

• Dengue (viral) transmitted by the mosquito Aedes aegypti (breeds in

water);

• Malaria is caused by a protozoan (Plasmodium spp.) and is transmitted

by a mosquito (Anopheles spp.)

• Trypanosomiasis (Gambian sleeping sickness) is also caused by a

protozoan transmitted by the riverine Tsetse fly (Glossina spp.)

HOW TO PREVENT….?

Always wash your hand before handling food especially

and after using toilet

Wash your fruits and vegetable before use as it contain

harmful chemicals and microbes.

Don’t drink water from river, lakes and ponds.

Try to adopt healthy and good hygiene as possible

Factors affecting microbial flora
• Nutrients
• Temperature and pH
• Oxygen and other gases
• Waste (inorganic and organic constituents)
• Hydrostatic pressure
• Light
• Salinity
• Turbidity
Water Purification
• Sedimentation
• Filtration
• Chlorination
• Additional treatment
• SEDIMENTATION:
• Leaves , particles of sand, and other materials from soil
are removed in large reservoirs or setting tanks.
• Some chemicals ( Alum , iron sulphate etc) are added to
coagulate suspended particles, in the form of flocs.
• Flocs cling to organic particles and microbes, while falling
through water, drag a major portion to bottom.
• FILETRATION:
• A slow sand filter:
• It utilizes fine particles of sand
• It is efficient for small scale operation
• Schmutzdecke / dirty layer ( a layer of microorganisms
developed within sand ) it acts as supplementary filter .
• This process may purify 3 million gallons of water / acre
per day.
• A rapid sand filter:
• it contains coarse particles of gravel.
• Commonly used method in municipal water systems.
• Both slow and rapid filters remove approximately 99% of
microorganisms from water

• CLORINATION:
• Chlorine gas ( an active oxidizing agent) is added to
water.
• Chlorine is continuously added until a residue is present.
• Standard permissible residue is 0.2 to 1.0 parts of chlorine
/ million ( PPM)
• Under these conditions , most remaining microorganisms
die within 30 minutes.
• Additional Treatments :
• Sometimes water is softened by removing Mg and Ca and
other salts.
• Water may also be flourinated which help to prevent from
tooth decay
• How can microorganisms be made useful during
water treatment:
• Microorganisms can be used to decompose contaminants
in fresh water.
• This kind of water treatment is called biological water
treatment .
• During biological water treatment microorganisms
breakdown organic matter , nitrates and phosphates .
SOIL MICROBIOLOGY
 Soil Microbiology:
⚫ A study of the microorganisms in soil, their functions,
and the effect of their activities on the character of the
soil and the growth and health of plant life.
⚫ It is believed that between two and four billion years
ago, the first ancient bacteria and microorganisms came
about in Earth's primitive seas. These bacteria could fix
nitrogen, in time multiplied and as a result released
oxygen into the atmosphere.
• This release of oxygen led to more advanced microorganisms.

• Microorganisms in soil are important because they affect the

structure and fertility of different soils by showing phenomenon
of mutualism and symbiosis.
Soil:
• It is the outer, loose material of earth’s surface which is
distinctly different from the underlying bedrock and
the region which support plant life.
• Soil is the region which supports the plant life by
providing mechanical support and nutrients required
for growth.
Layers of Soil:

• Organic matter: Litter layer of plant residues in relatively

undecomposed form.
• Surface soil: Layer of mineral soil with most organic
matter accumulation and soil life.
• Subsoil: This layer accumulates iron, clay.

• Parent rock: Layer of large unbroken rocks. This layer

may accumulate the more soluble compounds.
• Bedrock: It denotes the layer of partially weathered
bedrock at the base of the soil profile.
Components of Soil:

• Organic matter.
• Mineral matter.
• Soil air.
• Soil water.
• Soil microorganisms.
 Classification of Soil Microorganism

Soil microorganisms can be classified as:

• Bacteria.
• Actinomycetes.
• Fungi.
• Algae.
• Protozoa.
 Bacteria
• Bacteria are the smallest organisms in the soil and are the
only soil microorganisms that are prokaryotic.

• Bacteria are the most abundant microorganisms in the soil,

and serve many important purposes, one of those being
nitrogen fixation among other biochemical processes.

Bacteria dot the surface of strands of fungal hyphae.

Biochemical processes
• One of the most distinguished features of bacteria as a whole is
their biochemical versatility.

• Bacterial genera, Pseudomonas can metabolize a wide range of

chemicals and fertilizers.

• Another genera known as Nitrobacter can only derive its energy

by turning nitrite into nitrate, which results in a gain of oxygen
and is known also as oxidation.
• Pseudomonas (SOIL BACTERIA)
•NITROBACTER(SOIL BACTERIA)

•First exploration
of Nitrobacter diversity in soils by a
PCR
Nitrogen fixation
• Bacteria are responsible for the process of nitrogen
fixation, which is the conversion of atmospheric
nitrogen into nitrogen-containing compounds (like
ammonia) which can be used by plants to uptake.
• Autotrophic bacteria, or bacteria that derives its
energy making its own food by oxidation, like
the Nitrobacters species are responsible for nitrogen
fixation.
• These bacteria are very important because almost
every plant and organism require nitrogen in some
way, and would have no way of obtaining it if not for
nitrogen-fixing bacteria.
Nitrogen Cycle:
•The nitrogen cycle is the biogeochemical
cycle by which nitrogen is converted into
various chemical forms as it circulates among
the atmosphere, terrestrial, and marine
ecosystems.
•Nitrogen is present in the environment in a
wide variety of chemical forms including
organic nitrogen,
• Ammonium, Nitrite ,Nitrate , nitrous oxide ,
Nitric oxide or inorganic nitrogen gas .
• Atmospheric nitrogen must be processed, or "fixed", in a
usable form to be taken up by plants.
• This cycle consists of following steps.
• Nitrogen Fixation
• Assimilation.
• Ammonification
• Nitrification
• Denitrification
• 1) NITROGEN FIXATION:
• Nitrogen fixation is done by free-living or symbiotic bacteria
known as diazotrophs.
• These bacteria have the nitrogenase enzyme that combines
gaseous nitrogen with hydrogen to produce ammonia, which is
converted by the bacteria into other organic compounds.

• 2) ASSIMILATION:
• Plants take nitrogen from the soil by absorption through their
roots as amino acids, nitrate ions, nitrite ions, or ammonium
ions. Most nitrogen obtained by terrestrial animals can be
traced back to the eating of plants at some stage of the food
chain.
• 3) AMMONIFICATION
• When a plant or animal dies or an animal expels waste, the
initial form of nitrogen is organic. Bacteria or fungi convert the
organic nitrogen within the remains back into ammonium , a
process called ammonification.

• 4) NITRIFICATION:
• The conversion of ammonium to nitrate is performed primarily
by soil-living bacteria and other nitrifying bacteria. In the
primary stage of nitrification, the oxidation of ammonium is
performed by bacteria such as the Nitrosomonas species,
which converts ammonia to nitrites .
• Other bacterial species such as Nitrobacter, are responsible
for the oxidation of the nitrites into nitrates.
• It is important for the ammonia to be converted to nitrates or
nitrites because ammonia gas is toxic to plants.
• 5) DENITRIFICATION:
• Denitrification is the reduction of nitrates back into
nitrogen gas (N2), completing the nitrogen cycle.
• This process is performed by bacterial species such as
Pseudomonas and Clostridium in anaerobic conditions.

• The denitrifying bacteria use nitrates in the soil to carry

out respiration and consequently produce nitrogen gas,
which is inert and unavailable to plants.
 ACTINOMYCETES
• Actinomycetes(actinobacteria) are soil micro
organisms. Actinobacteria are a group
of Gram-positive bacteria.
• Actinobacteria include some of the most common soil
life, playing an important role in decomposition of
organic materials, such as cellulose and chitin, and
thereby playing a vital part in organic matter turnover.
• This replenishes the supply of
nutrients in the soil and is imp.
part of humus formation.

•Scanning electron micrograph of Actinomyces israelii.

• Some types of Actinobacteria are responsible for the
peculiar odor emanating from the soil after rain
(Petrichor).
• They are sensitive to acidity and waterlogged soil
conditions.
• The population of actinomycetes increases with depth of
soil even up to horizon ‘C’ of a soil profiler.
 Soil-related bacterial infections:
• Tetanus (caused by the toxin-producing, anaerobic, spore-bearing,
Gram-positive bacteria Clostridium tetani).

• Botulism (caused by the toxin-producing, anaerobic, spore-bearing,

Gram-positive bacteria Clostridium botulinum).

• Wound infections

• Gastroenteritis (Clostridium perfringens is present in soil, from which it

is ingested into GIT. It is associated with a variety of human diseases
including classic food poisoning. Evidence suggests that most cases
of gastrointestinal disease caused by C. perfringens have their
source from food contaminated by other humans or by animal feces
rather than directly from the soil).
• Other Organisms involved in Gastoentritis include:
Bacillus cereus, Campylobacter jejuni, E. coli & Listeria
monocytogenes

• Anthrax (caused by the Gram-positive, spore-forming rod

Bacillus anthracis)
Fungi
• Next to bacteria, fungi are abundant in soil population
compared to other microorganisms.

Fungi are important in the soil as:

• Food sources for other, larger organisms.
• Pathogens.
• Beneficial symbiotic relationships with plants or other
organisms.
• Help to reduce crop residues.
• Biochemically process nutrients to improve the soil
they inhabit.
Pathogenic fungi:
• The growth and distribution of bacteria and actinomycetes also
influence fungi.
• The quality as well as quantity of organic matter in the soil has a
direct correlation to the growth of fungi, because most fungi
consume the organic matter for nutrition.
• Fungi also grows well in dry, arid soils because fungi are
aerobic, or dependent on oxygen, and the higher the moisture
content in the soil, the less oxygen is present for fungi.
• Coccidioides, a dimorphic (mycelium/spherule) fungus has been associated
with alkaline, highly salinic, sandy soils and extremes of temperature.

• Blastomyces dermatitidis occurs in soil and near-soil environments.

• Pulmonary blastomycosis has a wide differential diagnosis and may be

asymptomatic or present as mild, moderate, or severe acute pneumonia.

Other infections due to fungi residing in soil relateed environment include;

• Penicillium marneffei, may cause a fatal systemic mycosis in patients

infected with HIV.

• Infections caused by Aspergillus species (excludes allergic brochopulmonary

aspergillosis) are usually in immunocompromised patients .
Algae
• Algae are present in most of the soils where moisture and
sunlight are available. Their number in soil usually ranges from
100 to 10,000 per gram of soil.
• Blue-green algae and grass-green algae are more abundant in
soil.
• The green-grass algae and diatoms are dominant in the soils of
temperate region while blue-green algae predominate in tropical
soils.
• Blue green algae are unicellular, photoautotrophic prokaryotes.
They are common in neutral to alkaline soils.
• Some blue green algae posses specialized cells known
as "Heterocyst" which is the sites of nitrogen fixation.
Protozoa
• Their population in soil ranges from l0,000 to 1,00,000 per
gram of soil and are abundant in surface soil.
• They can withstand adverse soil conditions as they are
characterized by "cyst stage" in their life cycle.
• The soil protozoa belonging to the class ciliate / ciliophora are
characterized by the presence of cilia around their body, which
helps in locomotion.
• Protozoa are abundant in the upper layer (15 cm) of
soil. Organic manures protozoa. Soil moisture,
aeration, temperature and PH are the important factors
affecting soil protozoa.

THE
LIVING SOIL: PROTOZOA
 Scope and Importance of Soil
Microbiology
• Living organisms both plant and animal types
constitute an important component of soil.

• Though these organisms form only a fraction (less than

one percent) of the total soil mass, but they play
important role in supporting plant communities on the
earth surface.

• While studying the scope and importance of soil

microbiology, soil-plant-animal ecosystem as such
must be taken into account.
• The scope and importance of soil microbiology, can be
understood in better way by studying aspects like:

1. Soil as a living system

2. Soil microbes and plant growth
3. Soil microorganisms and soil structure
4. Organic matter decomposition
5. Humus formation
6. Biogeochemical cycling of elements
7. Soil microorganisms as bio-control
agents
8. Soil microbes and seed germination
9. Biological Nitrogen fixation
10. Degradation of pesticides in soil
MICROBIOLOGY OF AIR
• The microbial flora of air is transient and variable

• Air is not a medium in which microorganisms can grow but

a carrier of particulate matter, dust, and droplets, which
may be laden with microbes.
• Airborne microorganisms:

• Airborne particles are a major cause of respiratory ailments of

humans, causing allergies, asthma, and pathogenic infections of the
respiratory tract. Airborne fungal spores are also important agents of
plant disease, and the means for dissemination of many common
saprotrophic (saprophytic) fungi.

• During a sneeze, millions of tiny droplets of water and mucus are

expelled at about 200 miles per hour (100 metres per second). The
droplets initially are about 10-100 micrometres diameter, but they dry
rapidly to droplet nuclei of 1-4 micrometres, containing virus
particles or bacteria. This is a major means of transmission of several
diseases of humans.
• Transmission of airborne microorganisms:
• Organisms are sprayed by:
❑ Coughing
❑ Sneezing

• Air microorganisms are carried by:

❑ Dust particles
❑ Droplet nuclei
• Fate of Airborne microorganisms:
• The ultimate fate of airborne microorganisms depends
upon various factors:
❑ Humidity
❑ Sunlight
❑ Temperature
❑ Size of particles bearing microorganisms
❑ Nature of microorganisms i.e. the degree of susceptibility
or resistance of particular species to new physical
environment.
❑ Indoor air
Example: Tubercle bacilli
• Outdoor Air
• Algae
• Protozoa
• Yeasts
• Molds
❑ Mold spore are predominant,

e.g cladosporium
❑ Bacterial species are,

Spore forming, Non spore forming

• Bacterial species
• Micrococcus
• Sarcina
• Gram negative rods
• Gram positive rods
• Aerobic spore forming bacteria
• Fungal species
• Cladosporium
• Alternaria
• Pullularia
• Penicillium
• Batrytis
• Stemphylium
Some important diseases of humans transmitted from person to person by inhaled
airborne particles

Virus diseases (virus type in brackets) Bacterial diseases (bacterial name in brackets)

Chickenpox (Varicella) Whooping cough (Bordetella pertussis)

Flu (Influenza) Meningitis (Neisseria species)

Measles (Rubeola) Diphtheria (Corynebacterium diphtheriae)

Pneumonia (Mycoplasma pneumoniae,

German measles (Rubella)
Streptococcus species)

Mumps (Mumps) Tuberculosis (Mycobacterium tuberculosis)

Smallpox (Variola)
Several other diseases, below, are acquired by inhaling particles from environmental sources, not
directly from an infected person.
Disease Source

Dried, powdery droppings from infected birds (parrots,

Psittacosis (Chlamydia psittaci)
pigeons, etc.)

Droplets from air-conditioning systems, water storage

Legionnaire's disease (Legionella pneumophila)
tanks, etc., where the bacterium grows.

Acute allergic alveolitis (various fungal & Fungal or actinomycete spores from decomposing
actinomycete spores) organic matter (composts, grain stores, hay, etc.)

Aspergillosis (Aspergillus fumigatus, A. flavus, A. Fungal spores inhaled from decomposing organic
niger) matter

Spores of the fungus, in old, weathered bat or bird

Histoplasmosis (Histoplasma capsulatum)
droppings

Spores in air-blown dust in desert regions (Central,

Coccidioidomycosis (Coccidioides immitis) South and North America) where the fungus grows in
Control of microorganism in Air:
There are several effective methods of controlling the level
of air contamination which are as follows:

ULTRAVIOLET RADIATION:
• This method has great potential value for reducing the
microbial flora of air.
• It is done by following ways :
1. Direct irradiation.
Used in unoccupied rooms.
• e.g. aseptic filling rooms for pharmaceutical preparations
• In sterile rooms ultraviolet lights are left ON when the
room is not in use.
2. Indirect Irradiation.
e.g. occupied rooms, offices, wards, hospital nurseries ,
school room e.t.c.
CHEMICAL AGENTS:
• Chemical substances vaporized into air of room are
effective in reducing the microbial flora.
• Chemicals are dispersed as aerosol and show its anti
microbial action.
• Characteristics of chemical agent as germicide:
• It should be highly bactericidal.
• It should be effective at normal room temperature and
humidity.
• It should not stain , discolor otherwise damage objects.
• Examples:
• Triethylene glycol
• Lactic acid
• Formaldehyde
• Resorcinol e.t.c
• FILTRATION:
• Use of cotton plug

• Air filters
LAMINAR AIR FLOW
• A laminar flow cabinet or laminar flow closet designed to
prevent contamination biological samples, or any particle
sensitive materials , air is passed through a HEPA (High
Efficiency Particulates Air) filter which removes all
airborne contamination to maintain sterile conditions.
• A laminar flow hood consists of a filter pad, a fan and a
HEPA (High Efficiency Particulates Air) filter
• The fan sucks the air through the filter pad where dust is
trapped
• After that the prefiltered air has to pass the HEPA filter
where contaminating fungi, bacteria, dust etc are removed
• Sterile air flows into the working area where you can do all
your work without risk of contamination.