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INTRODUCTION

TO
MEDICAL MICROBIOLOGY
Dr.T.V.Rao MD

Dr.T.V.Rao MD 1
What is Microbiology?
Microbes, or microorganisms are minute living
things that are usually unable to be viewed with the
naked eye.

What are some examples of microbes?


Bacteria, fungi, protozoa, algae, viruses are
examples!

Some are pathogenic


Many are beneficial
Dr.T.V.Rao MD 2
Defining Microbiology
Microbiology defined as the study
of organisms too small to be seen with
the naked eye. These organisms include
viruses, bacteria, algae, fungi, and
protozoa. Microbiologists are concerned
with characteristics and functions such as
morphology, cytology, physiology,
ecology, taxonomy, genetics, and
molecular biology. Dr.T.V.Rao MD 3
What is Microbiology

• Study of different
Microorganisms
• Can be
Bacteria
Viruses
Parasites
Fungus

Dr.T.V.Rao MD 4
What are Microorganisms

• Microbes are
products of
evolution,
Consequence of
Natural selection
operating upon
vast array of
genetically diverse
organisms
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History of Microbiology
1673-1723, Antoni
van Leeuwenhoek
(Dutch) described
live
microorganisms
that he observed in
teeth scrapings,
rain water, and
peppercorn
infusions. Dr.T.V.Rao MD 6
Anton van Leeuwenhoek 1674
- 1st person to actually see living microorganisms

荷兰人吕文虎克
(Leeuwenhoek)1632-1723

“wee animalcules”
Dr.T.V.Rao MD 7
History of Microbiology
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 Semmelweis advocated
handwashing to prevent transmission of
puerperal fever from one OB patient to
another.
Dr.T.V.Rao MD 8
The Germ Theory 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.
Dr.T.V.Rao MD 9
History of microbiology
 Anton van Leeuwenhoek (1632–1723): was the first
microbiologist and the first person to observe bacteria
using a single-lens microscope of his own design.
 Louis Pasteur (1822–1895): Pasteur developed a
process (today known as pasteurization) to kill microbes.
pasteurization is accomplished by heating liquids to 63
to 65 C for 30 minutes or to 73 to 75 C for 15 seconds.
 Robert Koch (1843–1910): was a pioneer in medical
microbiology and worked in cholera, anthrax and
tuberculosis. He was awarded a Nobel prize in 1905
(Koch's postulates) he set out criteria to test.
 Alexander Fleming (1929): Discovered penicillin.

Dr.T.V.Rao MD 10
Joseph Lister
• 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.

Dr.T.V.Rao MD 11
Course objectives
• To provide the student with the basic
knowledge of micro-organisms in
general
• To study the main characteristics of
Microbes of medical importance
• To teach aseptic techniques
• To provide an understanding of
antimicrobial agents
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Other Objectives
• To teach the basic
immunological
principles
• Immunological
methods for the
study
immunological
disorders

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Coverage of subject
• General
Microbiology
• Bacteriology
• Mycology
• Virology
• Immunology
• Parasitology
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Microbes in Our Lives
• Microorganisms
are organisms that
are too small to be
seen with the
unaided eye.
• “Germ” refers to a
rapidly growing
cell.
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Dr.T.V.Rao MD 16
Microbes make the Universe

• There are > 5 x 1030


types Microbes in
the world
• Humans have
intimate relation
with Microbes > 90%
of the cells in our
Body are Microbes
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Classification of Microorganisms
• Three domains
– Bacteria
– Archaea
– Eukarya
• Protists
• Fungi
• Plants
• Animals
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Naming and Classifying
Microorganisms
• Carolus Linnaeus (1735)
established the system of
scientific nomenclature.
• Each organism has two
names: the genus and
specific epithet.
• Are italicized or
underlined. The genus is
capitalized and the
specific epithet is lower
case.

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Edward Jenner Vaccinating a
Child

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Louis Pasteur
1922 - 95
• Contributed best in
Microbiology
• Sterilization
• Hot Air oven
• Autoclave
• Anthrax vaccine
• Rabies vaccine
• Built the Pasteur
Institute
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Louis Pasteur
• Pasteur coined the
word Vaccine
• Vacca – Cow cow
pox virus are given for
the prevention of
Small Pox
• Louis Pasteur
considered the father
of Modern
Microbiology

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Robert Koch
1843 - 1910
• A German scientist
• Formulated the
Bacteriological
techniques
• Staining Methods
• Discovered the
Mycobacterium and
Vibrio cholera

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Biological Principles illustrated by
Microbiology

Microbiology

Molecular
Biochemistry Genetics
Biology

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Microorganisms

Non-cellular organism Virus

Prokaryotes Bacterium

Eukaryotes Fungi

Others Prions Viroid


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Organisms included in the study
of Microbiology
1. Bacteria Bacteriology
2. Protozoans Protozoology
3. Algae Phycology
4. Parasites Parasitology
5. Yeasts and Molds
Fungi Mycology
6. Viruses Virology
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Man has Evolved So also the Microbes

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How to Study Medical Microbiology?

Fundamentals of Microbiology
•Biological Properties
Bacteriology •Morphology, identification,
•Antigenic structure
•Pathogenesis and Pathology
•Clinical findings
•Diagnostic Laboratory Tests
Virology •Immunity
•Treatment & Prevention
•Epidemiology & Control

Mycology
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Basic Classification of Microorganism
• Eukaryotes Prokaryotes
Large in size Small in Size
Mitochondria Present DNA not separated from
Membrane bound Nucleus cytoplasm
Eg Algae Mitochondria absent
Protozoa
Fungi Eg Bacteria
Slime Moulds
Contains all enzymes for Contains all enzymes like
production of metabolic Eukaryotes
energy

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Summary of differences between
prokaryote and eukaryote cells
Prokaryotic cells Eukaryote cells
Small cell (< 5µm) Larger cells (> 10 µm)
Always unicellular Often multicellular
No nucleus or any membrane bound organelles Always have nucleus and membranes bound
organelles.

DNA circular, without proteins DNA is linear and associated with proteins to
form chromatin.

Ribosomes are small 70S Ribosomes are large 80S


No cytoskeleton Always have cytoskeleton
Motility by rigid rotating flagellum made from Motility by flexible waving cilia or flagella
flagellin made from tubulins.

Cell division is by binary fission Cell division is by meiosis and mitosis.


Reproduction is always asexual Reproduction is sexual and asexual.
Prokaryotic Cell Structure
Prokaryotic cells are about 10 times smaller
than eukaryotic cells. A typical Escherichia coli
cell is about 1 μm wide and 2 to 3 μm long.
Structurally, prokaryotes are very simple cells
when compared with eukaryotic cells, and yet
they are able to perform the necessary
processes of life. Reproduction of prokaryotic
cells is by binary fission, the simple division of
one cell into two cells, after DNA replication and
the formation of a separating membrane and cell
wall.
Bacteria
• Prokaryotes
• Peptidoglycan cell
walls
• Binary fission
• For energy, use
organic chemicals,
inorganic chemicals,
or photosynthesis

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Bacterial Cell Wall
The structure of bacterial cell walls is quite different from
the relatively simple structure of eukaryotic cell walls,
although they serve the same functions, providing rigidity,
strength, and protection. The main constituent of most
bacterial cell walls is a complex macromolecular polymer
known as peptidoglycan (murein), consisting of many
polysaccharide chains linked together by small peptide
(protein) chains. Peptidoglycan is only found in bacteria. The
thickness of the cell wall and its exact composition vary with
the species of bacteria. The cell walls of “Gram-positive
bacteria” have a thick layer of peptidoglycan combined with
teichoic acid and lipoteichoic acid molecules. The cell walls of
“Gram-negative bacteria” have a much thinner layer of
peptidoglycan, but this layer is covered with a complex layer
of lipid macromolecules, usually referred to as bacteria
capsule.
Figure 1-9: Gram
Stain
Figure 3-1. Various forms of bacteria, including single cocci, diplococci, tetrads,
octads, streptococci, staphylococci, single bacilli, diplobacilli, streptobacilli,
branching bacilli, loosely coiled spirochetes, and tightly coiled spirochetes.
Morphologic arrangements of
bacteria.
Capsule stain. The capsule stain is an example of a negative staining technique. The
bacterial cells and the background stain, but the capsules do not. The capsules are
seen as unstained “halos” around the bacterial cells.
. Flagellar arrangement. The four basic types of flagellar arrangement on bacteria:
peritrichous, flagella all over the surface; lophotrichous, a tuft of flagella at one end;
amphitrichous, one or more flagella at each end; monotrichous, one flagellum.
Binary fission. Note that DNA replication must occur before the actual
splitting (fission) of the parent cell.
Pathogenic Prokaryotes
Bacteria

Mycoplasma

Spirochetes

Chlamydiae

Rickettsia

Actinomyces

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Viruses
A viral particle consists of a nucleic acid molecule, either DNA or RNA,
enclosed in a protein coat, or capsid

Viruses lack many of the attributes of cells, including the ability to


replicate. Only when it infects a cell does a virus acquire the key
attribute of a living system: reproduction

Viruses are known to infect all cells, including microbial cells. Host-virus
interactions tend to be highly specific

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Discovery of Virus
• Iwanovski
– a Russian chemist, 1892
– Tobacco Mosaic Disease
• Beijerinck confirmed

• Walter Reed, USA


– Yellow fever virus
– Ist human virus
Tobacco mosaic disease,
caused by the tobacco
mosaic virus

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Viruses
• A virus is not a cell!
• Viruses are replicated
only when they are in a
living host cell
• Consist of DNA or RNA
core
• Core is surrounded by a
protein coat
• Coat may be enclosed in
a lipid envelope

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What are Viruses
• Viruses Dependent on
Host cells for necessary
functions and
Multiplication
• Intracellular
parasites
• Contain either
DNA or RNA
never both.
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Prion
A kind of infectious protein that can resist the digestion of proteinase

The cellular form of the prion protein (PrPc) is encoded by the host’s
chromosomal DNA

An abnormal isoform of this protein (PrPres) is the only known


component of the prion and is associated with transmissibility.

Kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-


Sträussler-Scheinker disease, fatal familial insomnia
, and Bovine spongiform encephalopathy (BSE)
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Viroid
Small, single-stranded, covalently closed circular RNA molecules
existing as highly base-paired
rod-like structures; they do not possess capsids

They range in size from 246 to 375 nucleotides in length. The


extracellular form of the viroid is naked RNA—there is no capsid of any
The RNA molecule contains no protein-encoding genes, and the viroid
kind
is therefore totally dependent on host functions for its replication
The RNAs of viroids have been shown to contain
inverted repeated base sequences at their 3' and 5' ends, a
characteristic of transposable elements and retroviruses. Thus, it is
likely that they have evolved
from transposable elements or retroviruses by the deletion of internal
sequences

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Koch’s Postulates
1 The bacterium should be constantly associated with
lesions of Disease
2 It should be possible to isolate the bacterium in pure
culture from the lesions
3 Inoculation of such pure culture into laboratory animal
should reproduce the lesions of the disease
4 It is possible to reisolate the bacterium in pure culture
from the lesions produced in the experimental animal

Additional criterion specific antibodies in the


serum of patients suffering with disease

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Koch’s postulates

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Scientific era of Antibiotics
1928: Alexander
Fleming discovered
the first antibiotic.
He observed that
Penicillium fungus
made an antibiotic,
penicillin, that killed
S. aureus.
1940s: Penicillin was
tested clinically and
mass produced.
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Discovery of Antibiotics
• Alexander Fleming (1881-1955)

Sir Alexander Fleming Ernst Boris Chain Sir Howard Walter Florey

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Microbes are used to produce
Antibiotics

• Penicillin

• Mold
– Pencillium notatum

• 1928 Alexander Fleming

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Modern Developments
• Bacteriology is the study of bacteria.
• Mycology is the study of fungi.
• Parasitology is the study of protozoa and
parasitic worms.
• Recent advances in genomics, the study of
an organism’s genes, have provided new
tools for classifying microorganisms.

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Microbes and Human Disease
• Normal micro biota prevent growth of
pathogens.
• Normal micro biota produce growth factors
such as folic acid and vitamin K.
• Resistance is the ability of the body to
ward off disease.
• Resistance factors include skin, stomach
acid, and antimicrobial chemicals.

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How to Study Medical Microbiology?

Fundamentals of Microbiology
•Biological Properties
Bacteriology •Morphology, identification,
•Antigenic structure
•Pathogenesis and Pathology
•Clinical findings
•Diagnostic Laboratory Tests
Virology •Immunity
•Treatment & Prevention
•Epidemiology & Control

Mycology
Dr.T.V.Rao MD 54
Bacteria - what comes to mind?
• Diseases
• Infections
• Epidemics
• Food Spoilage
• Only 1% of all known bacteria cause human
diseases
• About 4% of all known bacteria cause plant
diseases
• 95% of known bacteria are non-pathogens
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• Staphylococcus aureus • Escherichia coli
• Staphylococcus • Bacillus anthrasis
epidermidis • Salmonella enteridis
• Streptococcus • Streptococcus pyogenes
pneumonia • Steptococcus lactis
• Streptococcus faecalis
• Vibrio cholera
• Erlichia canis
• Rhodospirillium rubrum • Campylobacter jujuni
• Bacillus subtilis • Helicobacter pylori
• Micrococcus luteus • Enterobacter aerogenes

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Microbes Benefit Humans
1.Bacteria are primary decomposers - recycle
nutrients back into the environment (sewage
treatment plants)
2. Microbes produce various food products
– cheese, pickles, sauerkraut, green olives
– yogurt, soy sauce, vinegar, bread
– Beer, Wine, Alcohol

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Microbes are also capable of
causing many diseases
Pneumonia Whooping Cough
Botulism Typhoid Fever Measles
Cholera Scarlet Fever Mumps
Syphilis Gonorrhea Herpes 1
Chlamydia Tuberculosis Herpes 2
Meningitis Tetanus RMSV
Strep Throat Lyme Disease AIDS
Black PlagueDiarrhea Dr.T.V.Rao MD Gangrene 58
Progress of Hepatitis Viruses
• 1947, concepts of hepatitis A and serum-
transmitted hepatitis
• 1970, Dane particle was observed
(hepatitis B virus)
• 1973, hepatitis A virus
• 1978, non-A, non-B hepatitis viruses
(NANBV)
• 1989, hepatitis C virus (HCV), hepatitis E
virus (HEV)
• 1990-1994, non A-E hepatitis viruses
• 1995, hepatitis G virus (HGV)
• 1997, TT virus (TTV)
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Human Immunodeficiency Virus & AIDS
• 1981, the first cases report
about AIDS
• 1983, HIV was isolated
• 1990s, HAART (cocktail therapy)
was employed
• So far, no effective vaccine
available

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HIV – AIDS
• Luc Montaigner and
Robert Gallo
announce their
discovery of the
immunodeficiency
virus (HIV) believed
to cause AIDS.
(American Society
for Microbiology
Archives)
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Parasitology
• Parasitology is the study
of parasites .and their
interactions with their
hosts. The science of
parasitology has a long
history and has its roots
in zoology, with its
emphasis on the
identification and
classification of parasites
and of life cycles,

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Taxonomic classification of parasitic organisms

• The classification of parasites is controversial -


there is no universally accepted system
• Parasites form part of the animal kingdom which
comprises some 800,000 identified species
categorised into 33 phyla (but it is estimated
that there may be ~10m species in total)
• The parasitic organisms that are of importance
for human health are eukaryotes - they have a
well defined chromosome in a nuclear
membrane (as opposed to prokaryotes which
have no nuclear membrane, e.g. bacteria)
Taxonomic classification of parasitic
organisms
• Parasites are classified into 2 sub-
kingdoms: protozoa (unicellular) and
metazoa (multicellular)
• Protozoan (unicellular) parasites are
classified according to morphology and
means of locomotion. There are 45,000
protozoa species. Most species that cause
human disease belong to the phylum's
sarcomastigophora and apicomplexa
• Metazoa (multicellular) include the worms
(helminths) and arthropoda (posses an
external skeleton) e.g. ticks, lice
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What Are Fungi
• Considerable variation
in size.
• Internal Molecular
system
• Well defined cell wall
composed of
polysaccharides
• Gaining importance in
Immunosupressed
patients and increased
use of Antibiotics
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Zoonotic Diseases

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How Humans Respond to Infections
Study of Immunology

• In spite of Infection
we survive with our
ability to protect
with a system
inherent in our Body
• Called the Immune
response comprises
the Medical
Immunology

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Pathogenesis
Immunity

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Immunity Protects the Living by
Complex Mechanisms

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Why we should
Medical Microbiology
• We study the
Microbes which
infects and causes
Diseases
• We study their
Diagnosis
Prevention
Treatment
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Modern Developments in
Microbiology
• Immunology is the study
of immunity. Vaccines and
interferons are being
investigated to prevent
and cure viral diseases.
• The use of immunology to
identify some bacteria
according to serotypes
(variants within a species)
was proposed by Rebecca
Lancefield in 1933.

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Must learn

• Natural History of the Disease


• Etiology
• Pathogenesis
• Laboratory Diagnosis
• Treatment and Control and
Prevention Dr.T.V.Rao MD 72
We must be familiar with Knowledge
On ….

• Names of the Microbes


• Names of the diseases
• Mode of transmission
• Pathogenic Microbes
• Commensal Organisms
• Identify wether Bacteria, Virus, Parasite or
Fungi
• Treating and Preventing

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The Birth of Modern Chemotherapy

• Treatment with chemicals is chemotherapy.


• Chemotherapeutic agents used to treat infectious
disease can be synthetic drugs or antibiotics.
• Antibiotics are chemicals produced by bacteria and
fungi that inhibit or kill other microbes.
• Quinine from tree bark was long used to treat malaria.
• 1910: Paul Ehrlich developed a synthetic arsenic drug,
salvarsan, to treat syphilis.
• 1930s: Sulfonamides were synthesized.

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Commonly Used Antibiotics
• Penicillin
• Cephalosporins,
• Tetracycline's
• Quinolones
• Vancomycin
• Chloramphenicol
• Drugs for Tuberculosis eg Streptomycin

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Vaccines Produce Immunity
and Prevents Several Infections

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Commonly used Vaccines
• Small pox eradicated
• BCG,
• MMR
• Polio oral Vaccine
• Triple Antigen
• Hepatitis B Vaccine
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What Skills You should
Develop

Able to identify the Infective Conditions


Timely Diagnosis
Choosing appropriate tests
Selection of Antibiotics
Implement measures to prevent diseases
in patients and Society
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Protect Yourself from
Infections
• Certain infections
can infect you
• Eg HIV, Hepatitis B
infections,Tubercul
osis,Many
respiratory
infections

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Working In the Hospital

• Hospitals are not safe


• Follow Universal
precaution protect
yourself as our patients
can be source of
Infection if you don't
handle the matters with
scientific knowledge.

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Medical Microbiology advanced Beyond our
Imagination
Can we handle it ???

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Major Selected Nobel Prizes in Physiology or
Medicine
1901* von Behring Diphtheria antitoxin
1902 Ross Malaria transmission
1905 Koch TB bacterium
1908 Metchnikoff Phagocytes
1945 Fleming, Chain, Florey Penicillin
1952 Waksman Streptomycin
1969 Delbrück, Hershey, Luria Viral replication
1987 Tonegawa Antibody genetics
1997 Prusiner Prions
* The first Nobel Prize in Physiology or Medicine.

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Students requirement for the course

• Timetable
• Literature – books, etc
• Practical manual
• Laboratory coat
• Attendance and active participation
• Seek advice timely
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• The Programme created by
Dr.T.V.Rao MD for Medical students
in the Developing world
• Email
• doctortvrao@gmail.com

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