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Dr.T.V.Rao MD
MICROBIOLOGY FORMEDICAL GRADUATES
WHAT YOU SHOULD KNOW
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AIMS FOR LEARNING MEDICAL
MICROBIOLOGY
What is medical microbiology?
Why is it relevant?
Some important concepts.
Basic classification of organisms.
Classifying bacteria.
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WHAT IS MEDICAL MICROBIOLOGY?
The study of microorganisms
(including bacteria, viruses, fungi
and parasites) which are of
medical importance and are
capable of causing diseases inhuman beings
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THE EARLY YEARS OF MICROBIOLOGY
CONTRIBUTED BY DISCOVERY OF MICROSCOPE
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THE FIRST OBSERVATIONS
1673-1723, Antonivan Leeuwenhoekdescribed live
microorganisms thathe observed inteeth scrapings, rainwater, andpeppercorninfusions.
Figure 1.2b17-11-2012DR.T.V.RAO MD 5
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THE EARLY YEARS OF
MICROBIOLOGY How Can Microbes Be Classified?
Carolus Linnaeus (Swedish) developed taxonomic system fornaming plants and animals and grouping similar organismstogether
Leeuwenhoeks microorganisms grouped into six categories asfollows:
Fungi
Protozoa
Algae
Bacteria
Archaea
Small animals 17-11-2012DR.T.V.RAO MD 6
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WHAT IS MEDICAL MICROBIOLOGY?
THE PURPOSE OF LEARNING
What organisms cause
infection?How they cause infection.
How to treat them.
How to prevent infection.17-11-2012DR.T.V.RAO MD 7
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WHY IS IT IMPORTANT?
Infection is one of the most importantcauses of mortality and morbidity in the
population. Approximately 30% of hospital patients
are on antibiotics at any one time
1 in 10 patients acquires an infectionwhilst in hospital.
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THE HISTORICAL CONTRIBUTION IN THE
SUBJECT OF MICROBIOLOGY BY
Linnaeus Jenner
Hooke
Leeuwenhoek
Lister
Pasteur Koch
Darwin
Salk
Watson & Crick
Jacob and Monod
McClintock
Woese
Venter?
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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 organisms
genes, have provided new tools for classifyingmicroorganisms.
Proteomics is looking at the gene products
DEFINITIONS
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LEARN THE CLASSIFICATION OF
ORGANISMS All living organisms are classified into:
Kingdom
Phylum (family)
Genus
Species
Organisms that can cause disease are many and varied and include:
Viruses
Bacteria
Fungi
Parasites
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Different:
Diseases
Modes oftransmission
Treatment-e.g.
routinely use
antibiotics dont cure
vira lfungalinfections
RELEVANCE OF CLASSIFICATION
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THE GOLDEN AGE OF MICROBIOLOGY
LOUIS PASTEUR CHANGES THE FUTURE OF
MICROBIOLOGY
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FERMENTATION AND PASTEURIZATION
Pasteur demonstrated thatthese spoilage bacteria couldbe killed by heat that was not
hot enough to evaporate thealcohol in wine.
Pasteurization is the
application of a high heat for ashort time.
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THE GOLDEN AGE OF MICROBIOLOGY
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THE GOLDEN AGE OF MICROBIOLOGY
Kochs Postulates
Suspected causative agent must be found inevery case of the disease and be absent fromhealthy hosts
Agent must be isolated and grown outside thehost
When agent is introduced into a healthy,
susceptible host, the host must get the disease Same agent must be reisolated from now-
diseased experimental host
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Normal Microbiota prevent growth ofpathogens.
Normal Microbiota produce growth factors
such as folic acid and vitamin K.
Resistance is the ability of the body toward off disease.
Resistance factors include skin,
stomach acid, and antimicrobial
chemicals.
Biofilms are extremely important inmicrobial ecology
NORMAL MICROBIOTA
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NORMAL MICRO BIOTA ON THE HUMAN BODY
Table 14.117-11-2012DR.T.V.RAO MD 18
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NORMAL MICROBIOTA Animals, including humans, are usually germfree
in utero. Microorganisms begin colonization in and on thesurface of the body soon after birth.
Microorganisms that establish permanent coloniesinside or on the body without producing diseasemake up the normal microbiota.
Transient microbiota are microbes that arepresent for various periods and thendisappear.
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WE HAVE MORE MICROBES OCCUPYING OUR BODY
THAN OUR OWN CELLS
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Why bother?
Different bacteria:
cause different diseases
are susceptible/resistant to
different antibiotics
some bacteria are common
normal flora whilst other
closely related species are
pathogens
CLASSIFYING BACTERIA
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How?
1st into broad groups based
on microscopic appearance
Then divided into speciesbased on a range of
different properties-often
biochemical reactions e.g.
some may be able tometabolise a sugar that
others cannot.
CLASSIFYING BACTERIA
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Method of differentiating bacteria.
Can be either Gram +ve or Gram
ve depending on how they
appear with the stain.
Can then be further grouped basedon shape (rod=long thin or
coccus=round).
Thus we end up with 4
combinations:
G+ rod, G+ coccus, G- rod, G-
coccus
GRAM STAIN
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BACTERIAL CELL WALL MAKES THE
BASIC DIFFERENCE
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GRAM STAIN
STAIN the slide withcrystal violet for 1-2 min.
Flood slide with Gram's
iodine for 1-2 min. Decolourise by washing
the slide briefly withacetone (2-3 seconds).
Stain with safranincounterstain for 2 min.
View under microscope
G+ve G-ve
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Gives an initial idea of the
possible identity of the
organism.
Can be done without growing
the organism (i.e. rapid
result)
Thus can be done on pus, joint
fluid, sputum, CSF1st result available on blood
cultures
GRAM STAIN
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GRAM POSITIVE COCCI
Clusters: usually
characteristic of
Staphylococcus spp., such as
S. aureus
Chain or pairs: usually characteristic
ofStreptococcus spp., such as
S. pneumoniae
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GRAM POSITIVE BACILLI Thick : usually
characteristic ofClostridium spp., suchas C. perfringens, C.
difficle,C. tetani
Thin: e.g. Listeria spp.
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GRAM NEGATIVE BACILLI
Thin rods: usuallycharacteristic ofenterobacteriaceae(coliforms), such as E. Coli
Coccobacilli: usually
characteristic ofHaemophilus spp., such asH. influenzae
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GRAM NEGATIVE BACILLI
Curved: usuallycharacteristic ofVibriospp.or Campylobacterspp.,such as V. cholerae C. jejuni
Thin needle shape: usually
characteristic ofFusobacterium spp.
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GRAM NEGATIVE COCCI
Diplococci: usually characteristic ofNeisseria spp., such as N.meningitides or N. gonorrhea.ThoughIn addition, Moraxella spp.andAcinetobacterspp.are oftendiplococcal in morphology.
Coccobacilli: usually characteristicofAcinetobacterspp., which can beeither Gram-positive or Gram-negative, and is often called Gram-variable.
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WHAT CAN YOU SEE ON THE SLIDE?
Staphylococcus aureus 100xGram
+ve
coc
ci
Gram
+ve
bacilli
Gram
ve
coc
ci
Gram
ve
bacilli
47%
0%
53%
0%
1. Gram +ve cocci
2. Gram +ve bacilli
3. Gramve cocci
4. Gramve bacilli
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WHAT CAN YOU SEE ON THE SLIDE?
Gram
+ve
coc
ci
Gram
+ve
bacilli
Gram
ve
coc
ci
Gram
ve
bacilli
16%11%
5%
68%
1. Gram +ve cocci
2. Gram +ve bacilli
3. Gramve cocci
4. Gramve bacilli
Streptococcus pneumoniae
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WHAT CAN YOU SEE ON THE SLIDE?
Pseudomonas aeruginosa
Gram
+ve
coc
ci
Gram
+ve
bacilli
Gram
ve
coc
ci
Gram
ve
bacilli
14%
36%36%
14%
1. Gram +ve cocci
2. Gram +ve bacilli
3. Gramve cocci
4. Gramve bacilli
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VIRUSES
Small (50-300nm)
Unable to replicate
independently
Invade host cells and usetheir cellular machinery to
replicate
Influenza, Chickenpox(varicella), Herpes,
Rhinovirus, HIV/AIDS
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FUNGI Complex, large organisms
Eukaryotes (as are humans!)
Divided into yeasts & moulds
Cause a range of diseasese.g.:
Thrush
Athletes foot
Invasive & allergic aspergillosis
Many diseases areopportunistic.
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PROTOZOA
Eukaryotes
Absorb or ingest
organic chemicals
May be motile via
pseudopods, cilia,
or flagella
Figure 1.1c17-11-2012DR.T.V.RAO MD 38
MULTICELLULAR ANIMAL
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MULTICELLULAR ANIMAL
PARASITES Eukaryote
Multicellular animals
Parasitic flatworms and round worms are called Helminths.
Microscopic stages in life cycles.
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THE ETIOLOGY OF INFECTIOUS DISEASES
Kochs postulates are criteria for establishing thatspecific microbes cause specific diseases.
Kochs postulates have the following requirements:
(a) the same pathogen must be present in every case ofthe disease;
(b) the pathogen must be isolated in pure culture;
(c) the pathogen isolated from pure culture must cause thesame disease in a healthy, susceptible laboratoryanimal;
(d) the pathogen must be reisolated from the inoculatedlaboratory animal.
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Figure 14.7
KOCHS POSTULATES
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EXCEPTIONS TO KOCHS POSTULATES
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EXCEPTIONS TO KOCHS POSTULATES
Kochs postulates are modified to establish etiologies of
diseases caused by viruses and some bacteria, which cannotbe grown on artificial media.
Some diseases, such as tetanus, have unequivocal signsand symptoms.
Some diseases, such as pneumonia and nephritis, may becaused by a variety of microbes.
Some pathogens, such as S. pyrogenes, cause several
different diseases. Certain pathogens, such as HIV, cause disease in humans
only.
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Disease-causing microorganisms arecalled pathogens.
Pathogenic microorganisms have specialproperties that allow them to invade thehuman body or produce toxins.
When a microorganism overcomes thebodys defenses, a state of diseaseresults.
Diseases and Infections
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O OG C O S S
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PATHOLOGY, INFECTION, AND DISEASE
Pathology is the scientific study of disease.
Pathology is concerned with the
etiology (cause),
pathogenesis (development),
effects of disease structural and functional changes broughtabout by disease.
Infection is the invasion and growth of pathogens in the body.
A host is an organism that shelters and supports the growth of
pathogens. Disease is an abnormal state in which part or all of the body is not
properly adjusted or is incapable of performing normal functions.
Infection disease presence of particular microorganism in part of
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IMMUNITY PROTECTS FROM EVENTS WITH
INFECTIONS
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CLASSIFYING INFECTIOUS DISEASES
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CLASSIFYING INFECTIOUS DISEASES
Every disease alters body structures and functions
A patient may exhibit
symptoms (subjective changes in body functions)
Pain or body discomfort
signs (measurable changes), which a physician uses tomake a diagnosis (identification of the disease)
Fever, swelling, paralysis
A specific group of symptoms or signs that alwaysaccompanies a specific disease is called asyndrome.
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MICROORGANISMS
Figure 1.117-11-2012DR.T.V.RAO MD 47
CLASSIFYING INFECTIOUS DISEASES
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CLASSIFYING INFECTIOUS DISEASES
Communicable diseases are transmitted directly
or indirectly from one host to another. Chicken pox, genital herpes,
A contagious disease is one that is easily
spread from one person to another. Noncommunicable diseases are caused by
microorganisms that normally grow outside the
human body and are not transmitted from one
host to another
Tetanus, Clostridium tetani
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THE MODERN AGE OF MICROBIOLOGY
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THE MODERN AGE OF MICROBIOLOGY
Microbial Genetics
Avery, MacLeod, and McCarty determined genes arecontained in molecules of DNA
Beadle and Tatum established that a genes activity is
related to protein function Translation of genetic information into protein
explained
Rates and mechanisms of genetic mutationinvestigated
Control of genetic expression by cells described
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THE MODERN AGE OF MICROBIOLOGY
Molecular Biology
Explanation of cell function at the molecular level
Genome sequencing
Pauling proposed that gene sequences could
Provide understanding of evolutionary relationships and processes
Establish taxonomic categories that reflect these relationships
Identify existence of microbes that have never been cultured
Woese determined that cells belong to bacteria, archaea, or eukaryotes
Cat-scratch fever caused by unculturable organism
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THE MODERN AGE OF
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Recombinant DNA Technology
Genes in microbes, plants, andanimals manipulated for practicalapplications
Production of human blood-clotting factor by E. colito aidhemophiliacs
Gene Therapy
Inserting a missing gene orrepairing a defective one in
humans by inserting desired geneinto host cells
THE MODERN AGE OF
MICROBIOLOGY
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DISCOVERY OF ANTIMICROBIAL AGENTS
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DISCOVERY OF ANTIMICROBIAL AGENTS
_________
Alexander Fleming (1881 1955),aScottish biologist andpharmacologist, observedbacterial staphylococci coloniesdisappearing on platescontaminated with mold.
Fleming extracted thecompound from the moldresponsible for destruction ofthe bacterial colonies.
The product of the mold wasnamed penicillin, after the
Penicilliummold from which itwas derived.
Nobel Prize in Physiology ofMedicine in 1945.
Images: Penicilliummold, PHIL #8396; Staphylococcus aureusonantibiotic test plate, PHIL #2641; Poster attached to a mailboxoffering advice to World War II servicemen, 1944, NIHFrom the Virtual Microbiology Classroom on ScienceProfOnline.com
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THE MODERN AGE OF
http://phil.cdc.gov/phil/home.asphttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:PenicillinPSAedit.jpghttp://en.wikipedia.org/wiki/File:PenicillinPSAedit.jpghttp://www.scienceprofonline.com/virtual-micro-main.htmlhttp://www.scienceprofonline.com/http://en.wikipedia.org/wiki/File:PenicillinPSAedit.jpghttp://www.scienceprofonline.com/http://www.scienceprofonline.com/virtual-micro-main.htmlhttp://en.wikipedia.org/wiki/File:PenicillinPSAedit.jpghttp://en.wikipedia.org/wiki/File:PenicillinPSAedit.jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_(AB_Test).jpghttp://phil.cdc.gov/phil/home.asp7/30/2019 Medical Microbiology for Graduates
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THE MODERN AGE OF
MICROBIOLOGY
How Do We Defend Against Disease? Serology
The study of blood serum
Von Behring and Kitasato existence in the blood ofchemicals and cells that fight infection
Immunology
The study of the bodys defense against specific pathogens
Chemotherapy
Fleming discovered penicillin
Domagk discovered sulfa drugs
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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 fungithat 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.
THE BIRTH OF MODERN
CHEMOTHERAPY
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1928: Alexander
Fleming discovered the
first antibiotic.
He observed thatPenicillium fungus
made an antibiotic,
penicillin, that killed S.
aureus.
1940s: Penicillin was
tested clinically and
mass produced.
THE BIRTH OF MODERN CHEMOTHERAPY
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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 aspecies) was proposed by
Rebecca Lancefield in 1933.
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Biotechnology, the use of
microbes to produce foods
and chemicals, is centuries
old.
Genetic engineering is a
new technique for
biotechnology. Through
genetic engineering,
bacteria and fungi canproduce a variety of
proteins including vaccines
and enzymes.
MODERN BIOTECHNOLOGY AND GENETIC
ENGINEERING
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SELECTED NOBEL PRIZES IN PHYSIOLOGY
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1901* von Behring Diphtheria antitoxin1902 Ross Malaria transmission
1905 Koch TB bacterium
1908 Metchnikoff Phagocytes
1945 Fleming, Chain, Florey Penicillin1952 Waksman Streptomycin
1969 Delbrck, Hershey, Luria Viral replication
1987 Tonegawa Antibody genetics
1997 Prusiner Prions
SELECTED NOBEL PRIZES IN PHYSIOLOGY
OR MEDICINE
.
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SELECTED NOVEL PRIZES IN PHYSIOLOGY OR
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* The first Nobel Prize in Physiology or Medicine.
SELECTED NOVEL PRIZES IN PHYSIOLOGY OR
MEDICINE1901* von Behring Diphtheria antitoxin
1902 Ross Malaria transmission1905 Koch TB bacterium
1908 Metchnikoff Phagocytes
1945 Fleming, Chain, Florey Penicillin
1952 Waksman Streptomycin
1969 Delbrck, Hershey, Luria Viral replication
1987 Tonegawa Antibody genetics
1997Prusiner Prions
2003Agre, Mackirron water and ion channels
2005 Marshall, Warren Helicobacter and ulcers
2008 Hausen Papilloma and viruses
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UNIVERSAL PRECAUTIONS SET
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UNIVERSAL PRECAUTIONS SET
UP BY CDC
Use gloves, gowns, masks and goggles
Minimize risk of needle sticks
Disinfections procedure Preventative treatment after exposure
Reduce risk
Treat all patients the same
HBV greater risk than HIV
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DEAR STUDENTS NEVER FORGET TO WASH HANDS
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AFTER HANDLING PATIENTS OR INFECTED
MATERIAL
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VISIT ME FOR MORE ARTICLES OF INTEREST ON
MICROBIOLOGY, INFECTIOUS DISEASES
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Programme Created by Dr.T.V.Rao MD
for Undergraduate Medical and
Paramedical Students for orientation in
Learning Medical Microbiology
doctortvrao@gmail.com
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