MICROBIOLOGY FOR MEDICAL GRADUATES

WHAT YOU SHOULD KNOW Dr.T.V.Rao MD

DR.T.V.RAO MD

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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 in human beings

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THE EARLY YEARS OF MICROBIOLOGY CONTRIBUTED BY DISCOVERY OF MICROSCOPE

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THE FIRST OBSERVATIONS

1673-1723, Antoni van Leeuwenhoek described live microorganisms that he observed in teeth scrapings, rain water, and peppercorn infusions.
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Figure 1.2b

THE EARLY YEARS OF MICROBIOLOGY

How Can Microbes Be Classified?

Carolus Linnaeus (Swedish) developed taxonomic system for naming plants and animals and grouping similar organisms together
Leeuwenhoeks microorganisms grouped into six categories as follows: Fungi Protozoa

Algae
Bacteria Archaea
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Small animals

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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.
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WHY IS IT IMPORTANT?
Infection is one of the most important causes 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 infection whilst in hospital.
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THE HISTORICAL CONTRIBUTION IN THE SUBJECT OF MICROBIOLOGY BY

Linnaeus Jenner Hooke Leeuwenhoek Lister Pasteur Koch
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Darwin Salk Watson & Crick Jacob and Monod McClintock Woese Venter?

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DEFINITIONS

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 classifying microorganisms.
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Proteomics is looking at the gene products

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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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RELEVANCE OF CLASSIFICATION
Different:
Diseases Modes of transmission Treatment-e.g. routinely use antibiotics dont cure vira lfungalinfections
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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 that these spoilage bacteria could be killed by heat that was not hot enough to evaporate the alcohol in wine.

Pasteurization is the application of a high heat for a short time.

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Figure 1.4 (1 of 3)

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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 in every case of the disease and be absent from healthy hosts Agent must be isolated and grown outside the host When agent is introduced into a healthy, susceptible host, the host must get the disease Same agent must be reisolated from nowdiseased experimental host
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NORMAL MICROBIOTA
Normal Microbiota prevent growth of pathogens.
Normal Microbiota 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. Biofilms are extremely important in microbial ecology

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NORMAL MICRO BIOTA ON THE HUMAN BODY

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Table 18 14.1

NORMAL MICROBIOTA
Animals, including humans, are usually germfree in utero. Microorganisms begin colonization in and on the surface of the body soon after birth.

Microorganisms that establish permanent colonies inside or on the body without producing disease make up the normal microbiota.

Transient microbiota are microbes that are present for various periods and then disappear.
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WE HAVE MORE MICROBES OCCUPYING OUR BODY THAN OUR OWN CELLS

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CLASSIFYING BACTERIA
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

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CLASSIFYING BACTERIA
How?

1st into broad groups based on microscopic appearance

Then divided into species based on a range of different properties-often biochemical reactions e.g. some may be able to metabolise a sugar that others cannot.

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GRAM STAIN
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 based on shape (rod=long thin or coccus=round). Thus we end up with 4 combinations:

G+ rod, G+ coccus, G- rod, Gcoccus

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BACTERIAL CELL WALL MAKES THE BASIC DIFFERENCE

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GRAM STAIN
STAIN the slide with crystal violet for 1-2 min. Flood slide with Gram's iodine for 1-2 min. Decolourise by washing the slide briefly with acetone (2-3 seconds). Stain with safranin counterstain for 2 min. View under microscope
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G+ve

G-ve

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GRAM STAIN
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, CSF

1st result available on blood cultures

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GRAM STAIN
Relevance of Gram reaction.

Gram +ve and gram ve organisms ae susceptible to different groups of antibiotics.

Cause different diseases Differ in their ability to survive in the environmentcleaning, infection control, outbreak management.
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GRAM POSITIVE COCCI

Clusters: usually characteristic of Staphylococcus spp., such as S. aureus

Chain or pairs: usually characteristic of Streptococcus spp., such as S. pneumoniae

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GRAM POSITIVE BACILLI

Thick : usually characteristic of Clostridium spp., such as C. perfringens, C. difficle, C. tetani

Thin: e.g. Listeria spp.

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GRAM NEGATIVE BACILLI

Thin rods: usually characteristic of enterobacteriaceae (coliforms), such as E. Coli

Coccobacilli: usually characteristic of Haemophilus spp., such as H. influenzae

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GRAM NEGATIVE BACILLI

Curved: usually characteristic of Vibrio spp.or Campylobacter spp., such as V. cholerae C. jejuni

Thin needle shape: usually characteristic of Fusobacterium spp.

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GRAM NEGATIVE COCCI

Diplococci: usually characteristic of Neisseria spp., such as N. meningitides or N. gonorrhea. Though In addition, Moraxella spp. and Acinetobacter spp.are often diplococcal in morphology.

Coccobacilli: usually characteristic of Acinetobacter spp., which can be either Gram-positive or Gramnegative, and is often called Gramvariable.

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1. 2. 3. 4.

WHAT CAN YOU SEE ON THE SLIDE?

Gram +ve cocci Gram +ve bacilli Gram ve cocci Gram ve bacilli

53% 47%

0%
i ci lli cc co cc i co ba

0%

+v e

+v e

v e

ra m

ra m

ra m

ra m

v e

Staphylococcus aureus 100x

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ba ci

lli

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WHAT CAN YOU SEE ON THE SLIDE?

1. 2. 3. 4. Gram +ve cocci Gram +ve bacilli Gram ve cocci Gram ve bacilli
68%

Streptococcus pneumoniae

16% 5%
i ci lli i

11%

cc

co cc

+v e

+v e

v e

ra m

ra m

ra m

ra m

v e

ba ci

co

ba

lli

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WHAT CAN YOU SEE ON THE SLIDE?

1. 2. 3. 4. Gram +ve cocci Gram +ve bacilli Gram ve cocci Gram ve bacilli
36% 36%

14% 14%

Pseudomonas aeruginosa
i ci lli cc co cc +v e +v e v e ra m ra m ra m ra m v e G ba ci co ba lli i

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VIRUSES
Small (50-300nm)
Unable to replicate independently

Invade host cells and use their cellular machinery to replicate

Influenza, Chickenpox (varicella), Herpes, Rhinovirus, HIV/AIDS Often difficult to treat
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FUNGI
Complex, large organisms Eukaryotes (as are humans!) Divided into yeasts & moulds Cause a range of diseases e.g.:
Thrush Athletes foot Invasive & allergic aspergillosis

Many diseases are opportunistic.

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PROTOZOA
Eukaryotes
Absorb or ingest organic chemicals May be motile via pseudopods, cilia, or flagella
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Figure 1.1c

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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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Figure 12.28a

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THE ETIOLOGY OF INFECTIOUS DISEASES

Kochs postulates are criteria for establishing that specific microbes cause specific diseases.
Kochs postulates have the following requirements:

(a) the same pathogen must be present in every case of the disease;
(b) the pathogen must be isolated in pure culture; (c) the pathogen isolated from pure culture must cause the same disease in a healthy, susceptible laboratory animal; (d) the pathogen must be reisolated from the inoculated laboratory animal.
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KOCHS POSTULATES

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Figure 14.7

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EXCEPTIONS TO KOCHS POSTULATES

Kochs postulates are modified to establish etiologies of diseases caused by viruses and some bacteria, which cannot be grown on artificial media. Some diseases, such as tetanus, have unequivocal signs and symptoms. Some diseases, such as pneumonia and nephritis, may be caused 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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Diseases and Infections

Disease-causing microorganisms are called pathogens. Pathogenic microorganisms have special properties that allow them to invade the human body or produce toxins. When a microorganism overcomes the bodys defenses, a state of disease results.
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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 brought about 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 the body where is not usually found. DR.T.V.RAO MD 17-11-2012 44

IMMUNITY PROTECTS FROM EVENTS WITH INFECTIONS

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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 to make a diagnosis (identification of the disease)
Fever, swelling, paralysis

A specific group of symptoms or signs that always accompanies a specific disease is called a syndrome.
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MICROORGANISMS

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Figure 1.1

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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 are contained 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 mutation investigated 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 MICROBIOLOGY

Recombinant DNA Technology Genes in microbes, plants, and animals manipulated for practical applications Production of human bloodclotting factor by E. coli to aid hemophiliacs Gene Therapy Inserting a missing gene or repairing a defective one in humans by inserting desired gene into host cells

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DISCOVERY OF ANTIMICROBIAL AGENTS

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Alexander Fleming (1881 1955), a Scottish biologist and pharmacologist, observed bacterial staphylococci colonies disappearing on plates contaminated with mold. Fleming extracted the compound from the mold responsible for destruction of the bacterial colonies. The product of the mold was named penicillin, after the Penicillium mold from which it was derived. Nobel Prize in Physiology of Medicine in 1945.
DR.T.V.RAO MD From the Virtual Microbiology Classroom on ScienceProfOnline.com Images: Penicillium mold, PHIL #8396; Staphylococcus aureus on 17-11-2012 53 antibiotic test plate, PHIL #2641; Poster attached to a mailbox offering advice to World War II servicemen, 1944, NIH

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 of chemicals 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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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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THE BIRTH OF MODERN CHEMOTHERAPY

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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 Immunology is the study of immunity. Vaccines and interferons are being investigated to prevent and cure viral diseases.

MODERN DEVELOPMENTS IN MICROBIOLOGY

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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Figure 1.4 (3 of 3)

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MODERN BIOTECHNOLOGY AND GENETIC ENGINEERING

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 can produce a variety of proteins including vaccines and enzymes.
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SELECTED NOBEL PRIZES IN PHYSIOLOGY OR MEDICINE

1901* 1902 1905 1908 1945 1952 1969 1987 1997 von Behring Ross Koch Metchnikoff Fleming, Chain, Florey Waksman Delbrck, Hershey, Luria Tonegawa Prusiner Diphtheria antitoxin Malaria transmission TB bacterium Phagocytes Penicillin Streptomycin Viral replication Antibody genetics Prions

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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 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
* The first Nobel Prize in Physiology or Medicine. DR.T.V.RAO MD
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SELECTED NOVEL PRIZES IN PHYSIOLOGY OR MEDICINE

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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 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 email

doctortvrao@gmail.com

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