INTRODUCTION

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MEDICAL MICROBIOLOGY
Dr.T.V.Rao MD

Dr.T.V.Rao MD

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
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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.
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What is Microbiology
Study of different Microorganisms Can be Bacteria Viruses Parasites Fungus
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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.

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Anton van Leeuwenhoek 1674

- 1st person to actually see living microorganisms

Leeuwenhoek1632-1723

wee animalcules
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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.
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The Germ Theory of Disease

1860s: Joseph Lister used a chemical disinfectant to prevent surgical wound infections after looking at Pasteurs work showing microbes are in the air, can spoil food, and cause animal diseases.
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 Anton van Leeuwenhoek (16321723): was the first microbiologist and the first person to observe bacteria using a single-lens microscope of his own design. Louis Pasteur (18221895): Pasteur developed a process (today known as pasteurization) to kill microbes. pasteurization is accomplished by heating liquids to 63 to 65C for 30 minutes or to 73 to 75C for 15 seconds. Robert Koch (18431910): 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.
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History of microbiology

Joseph Lister
1860s: Joseph Lister used a chemical disinfectant to prevent surgical wound infections after looking at Pasteurs work showing microbes are in the air, can spoil food, and cause animal diseases.
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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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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 Biology

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Biochemistry

Genetics

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Microorganisms
Non-cellular organism Virus

Prokaryotes Bacterium
Eukaryotes Others

Fungi
Prions Viroid
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Organisms included in the study of Microbiology

1. Bacteria

2. Protozoans 3. Algae 4. Parasites 5. Yeasts and Molds

Fungi

Bacteriology Protozoology Phycology Parasitology

6. Viruses

Mycology Virology
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Man has Evolved So also the Microbes

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How to Study Medical Microbiology?

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

Basic Classification of Microorganism

Eukaryotes
Large in size Mitochondria Present Membrane bound Nucleus Eg Algae Protozoa Fungi Slime Moulds Contains all enzymes for production of metabolic energy

Prokaryotes
Small in Size DNA not separated from cytoplasm Mitochondria absent
Eg Bacteria Contains all enzymes like Eukaryotes

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Summary of differences between prokaryote and eukaryote cells

Prokaryotic cells Eukaryote cells

Small cell (< 5m)

Larger cells (> 10 m)

Often multicellular Always have nucleus and membranes bound organelles. DNA is linear and associated with proteins to form chromatin. Ribosomes are large 80S

Always unicellular

No nucleus or any membrane bound organelles

DNA circular, without proteins

Ribosomes are small 70S

No cytoskeleton

Always have cytoskeleton

Motility by flexible waving cilia or flagella made from tubulins. Cell division is by meiosis and mitosis. Reproduction is sexual and asexual.

Motility by rigid rotating flagellum made from flagellin

Cell division is by binary fission

Reproduction is always asexual

Prokaryotic Cell Structure

Prokaryotic cells are about 10 times smaller than eukaryotic cells. A typical Escherichia coli cell is about 1 m wide and 2 to 3 m 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.

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.

Bacterial Cell Wall

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.

Figure 3-3. Morphologic arrangements of bacteria.

Figure 1-10: 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.

Figure 1-11. 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.

Figure 1-13. 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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Bacteria
Prokaryotes Peptidoglycan cell walls Binary fission For energy, use organic chemicals, inorganic chemicals, or photosynthesis
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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 hosts 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), GerstmannStrussler-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 RNAthere 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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Kochs 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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Kochs 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 organisms 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 Bacteriology Virology Mycology
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Biological Properties Morphology, identification, Antigenic structure Pathogenesis and Pathology Clinical findings Diagnostic Laboratory Tests Immunity Treatment & Prevention Epidemiology & Control

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
Staphylococcus epidermidis

Streptococcus pneumonia
Vibrio cholera Rhodospirillium rubrum

Bacillus subtilis
Micrococcus luteus

Escherichia coli Bacillus anthrasis Salmonella enteridis Streptococcus pyogenes Steptococcus lactis Streptococcus faecalis Erlichia canis Campylobacter jujuni Helicobacter pylori 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 Plague Diarrhea Dr.T.V.Rao MD Gangrene

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Progress of Hepatitis Viruses

1947, concepts of hepatitis A and serumtransmitted 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 Note that the genus starts with a capital letter and the species is always written in italics, e.g. Plasmodium falciparum, Giardia

lamblia

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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
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Treatment and Control and Prevention

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* 1902 1905 von Behring Ross Koch Diphtheria antitoxin Malaria transmission TB bacterium

1908
1945 1952 1969 1987 1997

Metchnikoff
Fleming, Chain, Florey Waksman Delbrck, Hershey, Luria Tonegawa Prusiner
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Phagocytes
Penicillin Streptomycin Viral replication Antibody genetics Prions
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* The first Nobel Prize in Physiology or Medicine.

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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