PHARMACEUTICAL MICROBIOLOGY

(THEORY ) BP303T
&
PHARMACEUTICAL MICROBIOLOGY
(PRACTICAL) BP 307P
Unit I 10 Hours

Introduction, history of microbiology, its branches, scope and its importance.

Introduction to Prokaryotes and Eukaryotes

Study of ultra-structure and morphological classification of bacteria,

nutritional requirements, raw materials used for culture media and physical
parameters for growth, growth curve, isolation and preservation methods
for pure cultures, cultivation of anaerobes, quantitative measurement of
bacterial growth (total & viable count).

Study of different types of phase constrast microscopy, dark field microscopy and
electron microscopy.
 Discovery of Microbes and the Dawn of Microbiology

Microbiology is the study of living organisms of microscopic size.

The term microbiology was given by French chemist Louis Pasteur (1822-95).

Microbiology is said to have its roots in the great expansion and development of
the biological sciences that took place after 1850.

The term microbe was first used by Sedillot (1878).

Contd…

 Physics began in ancient times, mathematics even earlier, but the knowledge of
tiny living things, their biology, and their impact on human lives have only been
around since the late 19th century.

 Until about the 1880s, people still believed that life could form out of thin air
and that sickness was caused by sins or bad odors.
 Contd…
 Opinions about why diseases afflicted people differed between cultures and
parts of society and the treatments differed as well. Diseases were thought to be
caused by

• Bad smells, treated by removing or masking the offending odor

• An imbalance in the humor of the body, treated with bleeding, sweating, and vomiting

• Sins of the soul, treated with prayer and rituals

• Although the concept of contagion was known, it wasn’t attributed to tiny living creatures
but to bad odors or spirits, such as the devil.
 INTRODUCTION, HISTORY,BRANCHES, SCOPE
AND IMPORTANCE OF MICROBIOLOGY
Microbiology?
Study of Microorganisms
Branch of science that deals with the structure, function, classes and economic
importance of microorganisms.
Is one of the exciting; ever developing field of science with greater scopes as the
microbes play a major role in our day to day life.

DEFINITION?
Microorganisms are tiny creatures less than 1 millimeter in diameter which can
not be seen by the naked eye and can only be visualized under microscope .
 Contd…
Naming Microorganisms
• Names may be descriptive or honour a scientist:
Staphylococcus aureus
• Describes the clustered arrangement of the cells (staphylo-) and the golden color
of the colonies.
Escherichia coli
• Honours the discoverer, Theodor Eshcerich, and describes the bacterium’s
habitat, the large intestine or colon.

Staphylococcus aureus and Esherichia coli are found in the human body.

S. aureus is on skin and E. coli, in the large intestine.

Contd…
Branches of Microbiology

Pure Microbiology

• Taxonomic arrangement
• Integrative arrangement

Applied Microbiology
Contd…
Taxonomic Arrangement

Bacteriology
Mycology
Phycology
Virology
Protozoology
Immunology
Contd…
Integrative Arrangement

Microbial cytology
Microbial physiology
Microbial genetics
Microbial ecology
Microbial taxonomy
Cellular Microbiology
Molecular Microbiology
Contd…
Applied Microbiology

Medical Microbiology
Veterinary Microbiology
Public Health Microbiology
Industrial Microbiology
Pharmaceutical Microbiology
Agriculture Microbiology
 Plant Microbiology
 Soil Microbiology
Contd….
Applied Microbiology

• Food and Dairy Microbiology

• Environmental Microbiology
• Water/Aquatic Microbiology
• Aero-microbiology
• Microbial Biotechnology
• Vaccinology
• Chemotherapy
 Microorganisms?

OCCURRENCE OF MICROORGANISMS

they occur everywhere, even in the atmosphere, water and soil.

Almost all natural surfaces are colonized by microbes.

Some microorganisms are even adapted to live comfortably in boiling hot springs
and frozen sea ice.

Microbes are the dominant form of life in the universe.

 Contd…

More than 50 per cent of the biomass on earth consists of microorganisms.

Majority of the microorganisms are not dangerous to humans.

Microbes help to digest our food and protect our bodies from pathogens.

Additionally, they carry out essential functions such as decomposition of dead

animals and plants, nutrient cycling which enhances the soil health and crop
productivity.
 Contd…
MEMBERS OF THE MICROBIAL WORLD
Based on the structure of nucleus, fundamentally two types of cells exist.
They are
i. Prokaryotes and ii. Eukaryotes

PROKARYOTIC CELLS
Prokayote is a Greek word, pro - before and karyon - nut or kernel.
Prokaryotes are the organism with a primordial nucleus.
They have a much simpler morphology than eukaryotic cells and lack a true
membrane bound nucleus and cell organelles like mitochondria, golgi bodies,
endoplasmic reticulum, etc.
All bacteria and archaea are prokaryotic.
 Contd…
EUKARYOTIC CELLS

Eukaryote is a Greek word, eu - true and karyon - nut or kernel.

Eukaryotes posses a membrane enclosed nucleus and cell organelles.

They are more complex morphologically and are usually larger than prokaryotes.

Algae, fungi, protozoa, higher plants and animals are eukaryotic.

Contd…
MICROBIAL GROUPS

Based on the morphological, phylogenetic and physiological characteristics,

microorganisms are divided into six distinct groups, they are as follows
1) Bacteria
2) Archaea
3) Fungi
4) Protozoa
5) Algae
6) Viruses
 Contd…
Bacteria
Prokaryotes
Peptidoglycan cell walls
Binary fission
most dominant group of microorganisms in soil, water and air.
 Some bacteria even live in environment that has extreme temperatures, pH or
salinity
Many of them play more beneficial roles in nutrient cycling, decomposition of
organic matter, production of commercial industrial products like vitamins,
antibiotics, etc.
some of them cause diseases and food spoilage.
Ex: Bacillus, Pseudomonas.
Contd…
Archaea
Prokaryotic
Lack peptidoglycan
Live in extreme environments

Include:

Methanogens
Extreme halophiles
Extreme thermophiles (Taq polymerase)

Non-pathogenic
Contd…
Fungi

Eukaryotes
Chitin cell walls
Use organic chemicals for energy
most dominant organism in the soil
Yeasts are unicellular
Moulds and mushrooms
Multicellular
Hyphae
Ex: Mucor, Rhizopus.
 Contd…
Protozoa

Unicellular Eukaryotes
Absorb or ingest organic chemicals
May be motile via pseudopods, cilia, or flagella
lack cell wall
found in many different environments
some are normal inhabitants of the intestinal tracts of animals, where they aid in
digestion of complex materials such as cellulose.
Some of them are parasitic and can cause diseases.

Ex: Amoeba, Paramecium

Contd…
Algae

Eukaryotes
Cellulose cell walls
Chlorophyll
Capable of performing photosynthesis for energy
Found most commonly in aquatic environments
Reproduce either sexually or asexually
Mostly they are used as food supplements
Mainly used in the preparation of agar

Ex: Spirulina, Gelidium

Contd…
Virus

Acellular
visualized only using electron microscopes
Consist of DNA or RNA core
Core is surrounded by a protein coat
Coat may be enclosed in a lipid envelope
Viruses are replicated only when they are in a living host cell
pathogenic to plants, animals and humans.

Ex: Cauliflower mosaic virus, Cucumber mosaic virus.

 Advantages of microbes ?
Microbes have many important and beneficial Biological functions

 Decompose organic waste

• (recycle nutrient)

 Producer in the ecosystem

• (algae, and some bacteria) capture the energy from sun light and convert it to
food, forming the basis of food chain (photosynthesis)
 Contd…
 Food industry:
Produce fermented foods such as vinegar, cheese, yogurt and bread.
Other industries:
Produce industrial chemicals such as ethyl alcohol and acetone.
Nitrogen fixation:
some bacteria can take the nitrogen from air and incorporate it into soil.
Medicine and medical research:
many antibiotic and other drugs are naturally synthesized by microbes )penicillin
made by molds).
Digestion:
Humans and animals are have microorganisms in their digestive tract that are
essential for digestion, and overall health
 History of Microbiology
The early Greeks believed that living things could originate from non living
matter (abiogenesis)

Aristotle discarded this notion, but he still held that animals could arise
spontaneously from dissimilar organisms or from soil. (spontaneous generation)

This concept was still felt as late as the 17th century, but toward the end of that
century several observations, experiments, and arguments began that eventually
refuted this concept.
 The Discovery Era

Robert Hooke, a 17th-century English scientist, was the first to use a lens to
observe the smallest unit of tissues he called “cells.”

Soon after, the Dutch amateur biologist Anton van Leeuwenhoek observed what
he called “animalcules” with the use of his homemade microscopes.

Antonie van Leeuwenhoek (1632-1723) of Delft, Holland (Netherland) was the

first person to observe and accurately describe microorganisms (bacteria and
protozoa) called ‘animalcules’ (little animals) in 1676.
 Contd…
Actually he was a Dutch linen merchant but spent much of his spare time
constructing simple microscopes composed of double convex lenses held
between two silver plates. He constructed over 250 small powerful microscopes
that could magnify around 50-300 times.

Leeuwenhoek was the first person to produce precise and correct descriptions of
bacteria and protozoa using a microscope he made himself.

Because of this extraordinary contribution to microbiology, Antonie van

Leeuwenhoek is considered as the “Father of microbiology”.

Antonie van Leeuwenhoek is also considered to be the “Father of bacteriology

and protozoology (protistology)”.
 Transition Period
 When microorganisms were known to exist, most scientists believed that such
simple life forms could surely arise through spontaneous generation. That is to
say life was thought to spring spontaneously from mud and lakes or anywhere
with sufficient nutrients.
This concept was so compelling that it persisted until late into the 19th century.

So now there are two hypothesis:

The hypothesis that living organisms arise from non-living matter is called
spontaneous generation. According to spontaneous generation, a “vital force’
Forms life.

The Alternative hypothesis, that the living organisms arise from preexisting life, is
called biogenesis.
 Contd…
• The main aspects were to solve the controversy over a spontaneous generation
which includes experimentations mainly of Francesco Redi, John Needham,
Lazzaro Spallanzani, and Nicolas Appert, etc, and to know the disease
transmission which mainly includes the work of Ignaz Semmelweis and John
Snow.
• Francesco Redi (1626-1697) (Italian)

Performed an experiment to disprove spontaneous generation.

He showed that the maggots would not arise from decaying meat, when it is
covered.
He set up an experiment to disprove spontaneous generation of maggots. Filled
2 jars with decaying meat, sealed one and left the other open (only the open jar
developed maggots).
 Contd…

Another experiment was set up in which a jar was covered with a fine mesh
instead of being sealed so that fresh air could enter the jar again, ( only the open
jar developed maggots)

Proved that the flies had to come in contact with the meat and lay their eggs on
it, maggots did not arise spontaneously.

Can you think of an experiment that could disprove spontaneous

generation?
Contd….
 Redi filled six jars with decaying meat.
 Contd…
John Needham (1713-1781):

He was probably the greatest supporter of the theory of spontaneous

generation.

He proposed that tiny organisms the animalcules arose spontaneously on his
mutton gravy.

He covered the flasks with cork as done by Redi and even heated some flasks.

Still the microbes appeared on mutton broth.

 Contd….
• Lazzaro Spallanzani (1729-1799):

 He was an Italian Naturalist who attempted to refute Needham’s experiment.

He boiled beef broth for longer period, removed the air from the flask and then sealed
the container.

 Followed incubation no growth was observed by him in these flasks.

He showed that the heated nutrients could still grow animalcules when exposed to air
by simply making a small crack in the neck.


 Contd…
• Nicolas Appert

 followed the idea of Spallanzani’s work.

 He was a French wine maker who showed that soups and liquids can be
preserved by heating them extensively in thick champagine bottles.

• Ignaz Semmelweis and John Snow

 were the two persons who showed a growing awareness of the mode of disease
transmission.
 Contd…
Schulze (1815-1873) and Theodor Schwan (1810-1882)

Two German scholars

air was the source of microbes and sought to prove this by passing air through
hot glass tubes or strong chemicals into boiled infusions in flasks.
 free from the microbes.

• George Schroeder and Theodor Von Dusch (1854)

 first to introduce the idea of using cotton plugs for plugging microbial culture
tubes.
 The Golden Age (1857-1914)

Beginning with Pasteur’s work, discoveries included the relationship between

microbes and disease, immunity, and antimicrobial drugs

1861: Louis Pasteur “Father of Modern Microbiology / Father of Bacteriology”

 demonstrated that microorganisms are present in the air.
 set up experiments involving a goose-necked flask.
 Boiled broth in a flask with a straight neck and left it exposed to air, organisms grew.
 With goose-necked flask, nothing grew.
 S-shape of this second flask trapped dust particles from the air, preventing them
from reaching the broth.
By showing that he could allow air to get into the flask but not the particles in the air,
Pasteur proved that it was the organisms in the dust that were growing in the broth.
Next experiment, Pasteur’s S-shaped flask kept microbes out but let air in.
These experiments form the basis of aseptic technique…..
 Contd…
 Pasteur, thus in 1858 finally resolved the controversy of spontaneous generation
versus biogenesis and proved that microorganisms are not spontaneously
generated from inanimate matter but arise from other microorganisms.

 Pasteur showed that microbes are responsible for fermentation.

 Fermentation is the conversion of sugar to alcohol to make beer and wine.
 Microbial growth is also responsible for spoilage of food.
 Bacteria that use alcohol and produce acetic acid spoil wine by turning it to
vinegar (acetic acid).
 Pasteur demonstrated that these spoilage bacteria could be killed by heat
(62.8°C/(145°F)) that was not hot enough to evaporate the alcohol in wine.
 This application of a high heat for a short time (30 mins) is called Pasteurization.
Contd…
Contribution of Louis Pasteur
•
 coined the term “ Microbiology”

 disproved the Spontaneous generation Theory

 demonstrated that anthrax was caused by bacteria.

 invented the process of pasteurization and fermentation.

 development of effective vaccine (rabies and anthrax)

 Contd…
• John Tyndall (1820 – 1893): English physicist

 Final blow to spontaneous generation in 1877.

 Conducted experiments in an aseptically designed box to prove that dust indeed carried the
germs.

Demonstrated that if no dust was present, sterile broth remained free of microbial growth for
indefinite period even if it was directly exposed to air.

 Discovered highly resistant bacterial structure, later known as endospore, in the infusion of
hay.

 Prolonged boiling or intermittent heating was necessary to kill these spores, to make the
 Contd…
The Germ Theory of Disease

1835: Agostino Bassi showed a silkworm disease was caused by a fungus.

1865: Pasteur believed that another silkworm disease was caused by a
protozoan.
1840s: Ignaz Semmelwise Advocated washing hands to stop the spread of
disease.
1860s: Joseph Lister used a chemical disinfectant to prevent surgical wound
infections after looking at Pasteur’s work showing microbes are in the air, can
spoil food, and cause animal diseases.
1876: Robert Koch provided proof that a bacterium causes anthrax and provided
the experimental steps, Koch’s postulates, used to prove that a specific microbe
causes a specific disease.

 Contd…

Robert Koch (1893-1910)

german physician

• Koch's Postulates
are used to prove
the cause of an
infectious disease.
 Contd…
• Koch's Postulates
are a sequence of
experimental steps
to relate a specific
microbe to a specific disease.
 Contd…
Robert Koch (1893-1910) German physician

 Was working on finding the causes of some very nasty animal diseases (first anthrax,
and then tuberculosis).

 He gave the first direct demonstration of the role of bacteria in causing disease.

 Isolated anthrax bacillus (Bacillus anthracis, the cause of anthrax) in 1876.

 1882 he discovered Mycobacterium tuberculosis.

 He proposed Koch postulate which were published in 1884 and are the corner stone of
the germ theory of diseases and are still in use today to prove the etiology (specific
 Contd…
• Koch’s four postulates are:

The organism causing the disease can be found in sick individuals but not in
healthy ones.
The organism can be isolated and grown in pure culture.
The organism must cause the disease when it is introduced into a healthy animal.
The organism must be recovered from the infected animal and shown to be the
same as the organism that was introduced.
The combined efforts of many scientists and most importantly Louis Pasteur and
Robert Koch established the Germ theory of disease. The idea that invisible
microorganisms are the cause of disease is called germ theory.
 Development in Medicine and Surgery

• Lord Joseph Lister (1827-1912): English surgeon

Notable contribution to the antiseptic treatment for the prevention and cure of
wound infections.
In 1867, developed a system of antiseptic surgery designed to prevent
microorganisms from entering wounds by the application of phenol on surgical
dressings and at times it was sprayed over the surgical areas.
Devised a method to destroy microorganisms in the operation theatre by
spraying a fine mist of carbolic acid into the air
first to introduce aseptic techniques for control of microbes by the use of
physical and chemical agents which are still in use today.
 Joseph Lister is known as the Father of Antiseptic surgery.
 Contd…

Edward Jenner (1749-1823) English physician

First to prevent small pox
Discovered the techniques of vaccination

Sir Alexander Fleming

 Discovered the “Wonder Drug” Penicillin from Penicillium notatum that destroy
several pathogenic bacterias
 Contd…
Paul Ehrlich
He was the first to report the acid-fast nature of tubercle bacillus.
He developed techniques to stain tissues and blood cells.
He proposed a toxin-antitoxin interaction called an Ehrlich phenomenon and also
introduced methods of standardizing toxin and antitoxin
He proposed the ‘side-chain theory for antibody production’.
He discovered ‘salvarsan’, an arsenical compound (magic bullet) for treatment of
syphilis
Paul Ehrlich is known as the father of chemotherapy.
The bacteria ‘Ehrlichia’was named after him.
 Scope and importance of
Microbiology
There is vast scope in the field of microbiology due to the advancement in the
field of science and technology.

The scope in this field is immense due to the involvement of microbiology in

many fields like medicine, pharmacy, diary, industry, clinical research, water
industry, agriculture, chemical technology and nanotechnology.

The study of microbiology contributes greatly to the understanding of life

through enhancements and intervention of microorganisms. There is an increase
in demand for microbiologists globally.
 Contd…
• Genetics: Mainly involves engineered microbes to make hormones, vaccine,
antibiotics and many other useful products for human being.
• Agriculture: The influence of microbes on agriculture; the prevention of the
diseases that mainly damage the useful crops.
• Food science: It involves the prevention of spoilage of food and food borne
diseases and the uses of microbes to produce cheese, yoghurt, pickles and beer.
• Immunology: The study of immune system which protect the body from
pathogens.
• Medicine: deals with the identification of plans and measures to cure diseases of
human and animals which are infectious to them.
• Industry: it involves use of microbes to produce antibiotics, steroids, alcohol,
vitamins and amino acids etc.