Introduction to Pharmaceutical microbiology

Introduction to Microbiology
By Someshwar Mankar
25/06/2020Mr.S.D.Mankar
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 Definition:-
It is the study of living organisms of microscopic size
which include bacteria, fungi, algae, protozoa and
viruses.
This term is introduced by a French chemist i.e Louis
Pasteur
He is known as father of microbiology
Micro-organisms can only be seen by magnifying their
image with microscope
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 Branches of Microbiology:-
Pure science
Bacteriology:- study of bacteria
Microbiology :- study of fungi
Protozoology :- study of protozoa
Phycology :- study of algae
Parasitology:- study of parasites
Taxonomy :- study of classification
Genetics: Study of variations and Hereditary
Immunology : Study of mechanism in development of
resistance by body to infection 25/06/2020Mr.S.D.Mankar
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 Applied Science:
a) Medical microbiology:
Study of causative agents of infectious disease
b) Pharmaceutical microbiology:
Study of M. O. which is responsible for production of
antibiotics, enzymes, vitamins, vaccines and other
pharmaceutical products.
c) Industrial microbiology:
Study of M. O. which is responsible for production of
vitamins, amino acids, alcoholic beverages and other
substances. 25/06/2020Mr.S.D.Mankar
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d) Microbial Biotechnology:
Study of scientific manipulation of living m. o. based on
genetic and molecular level
e) Food microbiology:
Study of m. o. relation to food bioprocessing, food
spoilage, food born diseases and preservation of food
product
f) Soil microbiology:
Study of microbial flora and interaction among soil m. o
g) Agricultural microbiology:
Study of m.o and relationship of m.o and crops
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h)Aquatic microbiology:
Study of m.o and their activities into fresh and marine
water
i)Air microbiology:
Study which deal with role of aerospora in
contamination and spoilage of food and
determination of plant and animal through air
j) Epidemiology:
Study which deal with consult with monitoring,
and spread of disease in community.
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CLASSIFICATION OF MICROORGANISMS:
Taxonomy:
It is the science that deal with logical arrangement of
living thing into categories
1) Aristotle classified living thing into plant and animals
2) karls linaus classified living thing into kingdom Plantae
and animalea
He also devised binomial (genus and species) scheme of
nomenclature
In 18th century the German zoologist E.H Haeckel
proposed 3rd kingdom named as the protista
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Include unicellular m.o typically they are neither plant
nor animal
Protista include bacteria, fungi, algae, protozoa but
virus not classified as protista bacteria is classified as a
lower protista and fungi, algae, protozoa classified as
a higher protista
In 1940 it was discovered in some cell e.g typical
bacteria, the genetic material was not enclosed by
nuclear membrane
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In other cells i.e (algae, fungi, protozoa) the nucleus
was enclosed in membrane this resulted into division
of organism
Prokaryotes
Eucaryotes
Bacteria are prokaryotic microbes (absence of nuclear
membrane)
Fungi, algae, protozoa, plant and animal cell are
eukaryotic microbes (presence of nuclear membrane)
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Application and scope of microbiology:
1) Production of antibiotic:
2/3rd of antibiotics are produced from m.o.
Pharmaceutical / industrial microbiology concerned
with isolation of antibiotics producing m.o. from
environment. E.g: soil, water, Mang antibiotic isolated
from natural m.o by process of fermentation
e.g: penicillin from penicillium species
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2) Production of enzyme ,vaccine , biosurfactants ,
alcoholic and other pharmaceutical product: many
microbial cell converts raw material or substrate into
valuable organic compound such as butanol, ethanol etc
microbial cell produce intracellular and extra cellular
enzyme those are imp for pharmaceutical fermentation
e.g: Amalyse and protease
Different types of biosurfactants are synthesized by no. of
m. o. e.g: Acinotobacter
3) Diagnosis of disease and treatment:
Different test are used to detect infectious m. o.
e.g: ELISA test and Widal test:
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Antimicrobial suitability testing is mainly usd for
selection of antibiotics for treatment of microbial
infections.
4)Treatment of Industrial waste material:
Microbial species are used for decomposition of
industrial waste material and organic components.
e.g: actinomycetes, fungi
Anaerobic bacteria decomposes of organic matter by
yielding simple organic molecule such as acid, alcohol,
glycerol and amines.
Cynide is degraded by Streptomyces lavendulae.
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5) Plant growth promotions:
Many Microbial cell are present in soils play an
important role in soil fertility, herbicidal resistance,
insect resistance, change in protein, oil content,
enhance the quality of plant product.
e.g: Rhizobium species, Azotobacter species
Many nitrogen fixing soil m.o are usefull for providing
nitrogen for growth of plants, germination of seeds,
flavoring and fruiting of medicinal plants.
Rhizobacter bacteria may release various plant growth
promoting substances as secondary metabolites.
(PGPR) e.g: Pseudomonas
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6)Sterile product preparations:
It deals with the preparations of sterile rooms, aseptic
techniques, and detection of microbes by sampling and
testing of different sterile preparations.
7) Sterilization:
Different pharmaceutical products are sterilized by killing of
m.o.
The process of killing of m.o is known as sterilization.
All the sterilization process may be validated by using
indicator.
A biological indicator preparation of m.o usually bacterial
spores.
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8) Steroid Biotransformation
Important steroid can be produced by microbiological
transformation of naturally occurring steroid.
Microbial conversion of steroids enzyme m.o
streptomyces.
9) Identification of M. O:
 Two vital function of microbiology and biotechnology
are enumeration and identification of m.o found in
product and Mfg environment.
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10) Testing of Pharmaceuticals:
Raw material and finished product
Microbial assay of antibiotics
Evalution of disinfectant
Antimicrobial, preservative efficiency testing
Endotoxin testing
Test for support of the sterility assurance system
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History of Microbiology

Contribution of anatomy van Leeuwenhoek: - (1632- 1720)
 He was a first to observe & accurately to describe the
shape of human RBCs as well as little agent of disease i.e.
animalcule’s.
He observed the motility of bacteria.
In 1683 he described different shapes or morphological
forms of bacteria.
Antonie van Leeuwenhoek (1632–1723) was one of the first
people to observe microorganisms, using a microscope of
his own design, and made one of the most important
contributions to biology.
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He had hobby of glass grinding and preparation of
lenses and this led him to assemble 250 single
microscope.
He describe the inhibitory effect of acetic acid on M.O.
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Contribution of Louis Pasteur: - (1822-1895)
The credit for providing foundation to bacteriology as a
science goes to Louis pasture
1) Demonstration:-
Louis pasture first demonstrated that air control micro-
organism.
He pass large quantity of air through a tube which contain
a plug of guncotton to serve as filter. The guncotton was
then removed & dissolved in mixture of alcohol-ether
sediment was examine microscopically.
He found that this sediment contains not only organic
matter but also a large number of small micro-organism.
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 He performed a series of experiment to prove that
although micro-organism were present in the air they
were not spontaneously produce.
Pasture took boiled meat infusion in several swan neck
flack. The flack opening were freely open to the air but
curve so that gravity would cause any air born dust
particles to deposited in the lower part of the neck.
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The flask were heated to sterilize the broth & then
incubated. No growth occurred even through the
contents of the flack were exposed to air.
Pasture pointed out that no growth took place because
dust & germs had been trapped on the wall of the
curved necks but if the necks were broken off so that
dust fell directly down into the flack, microbial growth
commenced immediately.
In 1861, pasture finally resolved the controversy of
spontaneous generation vs. biogenesis & proved that
micro-organism are spontaneously generated from in
animated matter.
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 Pasture also showed that micro-organism are not
evenly distributed in the atmosphere & that there
number varies from place to place for this experiment ,
he took a large number of sealed flack containing
boiled & cooled infusions & opened a few at a time for
a short period at various places & resealed
Out of the 20 flacks which he opened & resealed on a
dusty road & showed spoilage while out of the 20 that
he opened on the top of a mountain only 5 showed
spoilage & out of 20 that he opened near a glacer, only
one showed spoilage.
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After incubation, only those flacks carrying micro-
organism from the air showed growth & spoilage.
 From these experiment, pasture concluded that the air
contained micro-organism & there number of varied
from place to place.
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Pasture made an intensive study of the bear & wine
fermentation. He found that wine spoilage was caused
by the growth of undesirable contaminating micro-
organism.
After some experiment, he showed that wine did not
undergo spoilage if it was held for a few min. at 50 to
600c. This gave rise to the new process of preserving
wine, fruit juice, milk etc. & was called pasteurization.
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Pasture also found that many other micro-organism
including yeast could grow either in presence or
absence of food, oxygen.
He designated life in the presence of oxygen as
‘aerobic’ in the absence of oxygen as ‘anaerobic’ & in
presence or absence of oxygen as ‘facultative
anaerobic’.
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5) Pasture opened the field of sterilization by stating
the boiling rendered the fluid sterile. Pasture
established the practice of heating fluid material to
1200c under pressure of sterilization (autoclaved).
He also introduced the particle of sterilizing glassware
by dry heat at 1700c.
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6) In 1880, pasture isolated the bacterium responsible for
chicken cholera & grow it in pure culture.
But on the day of public demonstration he failed to repeat his
experimental results. The chicken get survived after injecting the
isolated bacterial culture & this result surprised him.
On subsequent experiment his mead conclusion that on long
preservation or sub culturing the virulence power of the
pathogenic bacteria was destroyed.
Such microbial culture with decreased virulence was referred as
attenuated culture. Such attenuated culture cannot caused
infection but induced immunity in host.
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7) Pasture further introduced the vaccine for anthrax
&rabies.
He know that the causative agent of rabies attacked the
brain & spinal cord.
He took spinal cord from rabbits which died from the
disease & suspended these cord in dry sterile air.
By these process rabies micro-organism in the nervous
tissue lost their virulence & emulsion of the spinal cord
was used as vaccine.
It was found to be effective in dose.
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Robert Koch 7 Anthrax:-
The first direct detection of the role of bacteria in
causing was provided by the Robert Koch (1843-1910)
He was a German physician, first isolated bacillus
anthracis.
 His method of preparing, fixing & staining, film
preparation of bacteria using staining film.
Preparation of bacteria using aniline dyes, opened a
new area in the bacteriological techniques.
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In 1881, he described his method of preparing culture
on solid media & it made possible for the isolation of
pure strains of bacteria from single colonies.
In 1881, solid media using potato & gelatin were in
practice for isolation of fungi
In 1882, he discovered mycobacterium tuberculosis &
in 1883 the vibrio cholera
The most notable constitution of Koch was the
establishment of the causal relationship between a
micro-organism & a specific disease by applying a set
of criteria referred to as a Koch’s postulates
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The postulate are:-
1) The micro-organism must be presently in every case
of the disease
2) The micro-organism must be isolated from the
disease animal & grown in pure culture.
3) The disease must be reproduced when a pure culture
of the micro-organism is inoculated into a susceptible
host
4) The same micro-organism must be isolated from the
experimentally infected host.
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Paul Ehrlich:-
In 1904 found that the dye trypan red was active
against the trypanosome that causes African sleeping
sickness. This dye with antimicrobial activity was
referred to as’ magic bullet’.
In 1910 Ehrlich in collaboration with sakahirohata,
Japanese physician the drug salvarsan as a treatment
for syphilis caused by treponema pallidum.
In 1912, he discovered neosalvarsan. This gave him the
title “the father of chemotherapy”.
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