Maharashtra University of Health Sciences Courses

M.D. (Microbiology)

Course Syllabus

Broad Areas of Study

General Microbiology; Systematic Bacteriology, Mycology, Virology, Parasitology, Serology,Immunology, molecular diagnostics and Applied Clinical Microbiology including recent advances in Microbiology.

General Microbiology

• History and pioneers in Microbiology
• Microscopy
• Morphology of bacteria and other micro-organisms
• Nomenclature and classification of microbes
• Growth and nutrition of bacteria
• Bacterial metabolism
• Sterilization and disinfection
• Biomedical waste disposal
• Bacterial toxins
• Bacterial antagonism: Bacteriocins
• Bacterial genetics, gene cloning
• Antibacterial substances used in treatment of infections and drug resistance in bacteria
• Bacterial ecology-normal flora of human body, hospital environment, air, water and milk
• Host parasite relationship
• Quality control and Quality Assurance in Microbiology
• Laboratory Biosafety
• Health care associated infections- prevention and control

Immunology and Applied Aspects

• The normal immune system.
• Innate immunity.
• Antigens.
• Immunoglobulins
• Complement
• Antigen and antibody reactions
• Hypersensitivity
• Cell medidated immunity
• Immunodeficiency
• Autoimmunity
• Immune tolerance
• Transplantation immunity
• Tumour immunity
• Prophylaxis and immunotherapy
• Measurement of immunity
• Immunity and immunopathogenesis of specific infectious diseases
• Molecular Biology Techniques. For e.g. PCR, DNA probes

Systematic Bacteriology

• Isolation, description and identification of bacteria. The epidemiology, pathogenesis, antigenic characteristics and laboratory diagnosis of disease caused by them
• Staphylococcus and Micrococcus; Anaerobic Gram positive cocci
• Streptococcus and Lactobacillus
• Neisseria, Branhamnella and Moraxella
• Corynebacterium and other coryneform organisms
• Bacillus: the aerobic spore-bearing bacilli
• Clostridium: the spore-bearing anaerobic bacilli
• Non-sporing anaerobes
• The Enterobacteriaceae
• Vibrios, Aeromonas, Plasiomonas, Campylobacter and Spirillum,H.pylori
• Erysipelothrix and Listeria
• Pseudomonas
• Chromobacterium, Flavobacterium, Acinetobacter and Alkaligens
• Pasteurella, Francisella
• Haemophilus and Bordetella
• Brucella
• Mycobacteria
• The spirochaetes.
• Actinomyces, Nocardia and Actinobacillus
• Mycoplasmatales: Mycoplasma, Ureaplasma and Acholeplasma
• Rickettsiae
• Chlamydiae
• Emerging bacterial pathogens

Virology

• The nature of viruses
• Classification of viruses
• Morphology :virus structure
• Virus replication
• The genetics of viruses
• The pathogenicity of viruses
• Epidemiology of viral infections
• Vaccines and antiviral drugs
• Bacteriophages
• Pox viruses
• Herpes viruses
• Vesicular viruses
• Togaviridae
• Bunyaviridae
• Arenaviridae
• Marburg and Ebola viruses
• Rubella virus
• Orbi viruses
• Influenza virus
• Respiratory disease: Rhinoviruses, adenoviruses, corona viruses
• Paramyxoviridae
• Enteroviruses : Polio, Echo, Coxsackie viruses
• Other enteric viruses
• Hepatitis viruses
• Rabies virus
• Slow viruses
• Human immunodeficiency viruses
• Oncogenic viruses
• Teratogenic viruses
• Viruses of gastroenteritis
• Prion diseases
• Emerging viral infections – SARS, Avian influenza

Parasitology

• Protozoan parasites of medical importance :
  
  • Entamoeba, Giardia, Trichomonas, Leishmania, Trypanosoma, Plasmodium, Toxoplasma, Sarcocystis, Cryptosporidium, Balantidium,Isospora, Cyclospora,Microsporidium etc.
• Helminthology : All those medically important helminths belonging to Cestoda, Trematoda and Nematoda.
• Cestodes :
  
  • Diphyllobothrium, Taenia, Echinococcus, Hymenolepis, Dypyllidium, Multiceps etc.
• Trematodes :
  
  • Schistosomes, Fasciola, Gastrodiscoides, Paragonimus, Clonorchis, Opisthorchis etc.
• Nematodes :
  
  • Trichuris, Trichinella, Strongyloides, Ancylostoma, Nicator, Ascaris, Toxocara, Enterobius, Filarial worms, Dracunculus, etc.
• Ectoparasites :
  
  • Common arthropods and other vectors viz.,Mosquito, Sandfly, Ticks, Mite, Cyclops.

Mycology

•  The morphology and reproduction of fungi and antimycotic agents
• Classification of fungi
• Contaminant and opportunistic fungi
• Fungi causing superficial mycoses
• Fungi causing subcutaneous mycoses
• Fungi causing systemic infections
• Antifungal agents

Applied Clinical Microbiology

• Epidemiology of infectious diseases
• Hospital acquired infections
• Infections of various organs and systems of the human body
• Molecular genetics as applicable to Microbiology
• Automation in Microbiology
• Rapid diagnostic techniques for microbial diseases.
• Vaccinology : principle, methods of preparation, administration of vaccines
• Outbreak investigations & disaster management
• Biological warfare

Practicals (Skills)

Bacteriology

• Must acquire:
  • Care and operation of Microscopes viz. Light, Dark ground, Phase contrast, Inverted, Fluorescent microscopes.
  • Preparation of stains viz. Gram’s, Albert’s, Ziehl- Neelson and other special stains - performing of staining and interpretation of stained smears.
  • Washing and sterilization of glassware including plugging and packing.
  • Operation of incubator, autoclave, hot air oven, inspissator, distillation plant, filters like Seitz and membrane and sterility tests.
  • Care and maintainance of common laboratory equipments like water bath, centrifuge, refrigerators, incubators etc.
  • Preparation and pouring of liquid and solid media - Nutrient agar, Blood agar, MacConkey agar, sugars, TSI agar, Robertson’s cooked meat,
  • Lowenstiein- Jensen’s, selective media.
  • Preparation of reagents – oxidase, Kovac, etc.
  • Tests for beta-lactamases including ESBLs.
  • Collection of specimens for Microbiological investigations such as blood, urine, throat swab, rectal swab, stool, pus, OT specimens.
  • Preparation, examination and interpretation of direct smears from clinical specimens,viz. Sputum for AFB – ZN & auramine O, slit smears for
  • M..leprae,-ZN stain, conjunctival smear for Chlamydiae – Giemsa/Iodine.
  • Techniques of anaerobiosis – Gaspack system,anaerobic jars-evacuation & filling with H2, CO2
  • Identification of bacteria of medical importance upto species level (except anaerobes – upto generic level)
  • Quantitative analysis of urine by pour plate method and semiquantitative analysis by standard loop test for significant bacteriuria.
  • Plating of clinical specimens on media for isolation, purification identification and quantitation.
  • Tests for motility: hanging drop, Craige’s tube, dark ground microscopy for Spirochaetes – Treponema & Leptospira.
  • In-vitro toxigenicity tests – Elek test, Nagler’s reaction
  • Special tests – Bile solubility, chick cell agglutination, sheep cell haemolysis, niacin and catalase tests for mycobacterium, satellitism, CAMP test, catalase test and slide agglutination tests, and other as applicable t identification of bacteria upto species level
  • Preparation of antibiotic discs; performance of antimicrobial susceptibility testing by Kirby-Bauer disk diffusion method; estimation of Minimum inhibitory /Bactericidal concentrations by tube/plate dilution methods. Tests for drug susceptibility of Mycobacterium tuberculosis
  • Skin tests like Mantoux, Lepromin etc.
  • Testing of disinfectants- Phenol coefficient and ‘in use’ tests.
  • Quality control of media reagents etc. and validation of sterilization procedures.
  • Aseptic practices in laboratory and safety precautions.
  • Disposal of contaminated material like cultures.
  • Bacteriology of food, water, milk, air
  • Maintenance of stock cultures.
• Desirable to acquire:
  • Care and breeding of laboratory animals viz. Mice, rats, guinea pigs and rabbits.
  • Techniques of withdrawal of blood from laboratory animals includingsheep.
  • Inoculation of infective material in animals by different routes.
  •  Animal pathogenicity /toxigenicity tests for C.diphtheriae, Cl.tetani, S. pneumoniae,  S.typhimurium, K. pnumoniae etc.
  • P erformance of autopsy on animals.
  • Isolation of plasmids and Conjugation experiments for transfer of drug resistance
  • Serum antibiotic assays eg. Gentamicin
  • Phage typing for staphylococci, S.typhi etc.
  • Bacteriocine typing eg. Pyocine, Proteocin etc.
  • Enterotoxigenicity tests like rabbit ileal loop, intragastric inoculation of mouse, Sereny’s test.
  • Mouse foot pad test for M.leprae

Immunology/ Serology

• Must acquire:
  • Collection of blood by venepuncture, seperation of serum and preservation of serum for short and long periods.
  • Preparation of antigens from bacteria or tissues for widal, Weil-Felix, VDRL, etc. and their standardisation.
  • Preparation of adjuvants like Freund’s adjuvant.
  • Raising of antisera in laboratory animals.
  • Performance of serological tests viz. Widal, Brucella tube agglutination, indirect haemagglutination, VDRL, Paul-Bunnel, Rose-Waaler, IFA.
  • Immunodiffusion in gels, counter immunoelectrophoresis- visualization and interpretation of bands.
  • Performance and interpretation of Enzyme linked immunosorbant assay.
  • Latex and staphylococcal co-agglutination tests.
• Desirable To Acquire:
  • Leucocyte migration inhibition test.
  • T-cell rosetting.
  • Flow Cytometry
  • Radial immunodiffusion.
  • Immunoelectrophoresis.
  • Neutrophil phagocytosis.

Mycology

• Must acquire:
  • Collection of specimens for mycology.
  • Direct examination of specimens by KOH, Gram, Kinyoun’s, Giemsa, Lactophenol cotton blue stains.
  • Examination of histopathology slides for fungal infections.
  • Isolation and identification of pathogenic yeasts and moulds and recognition of common laboratory contaminants.
  • Special techniques like Wood’s lamp examination, hair baiting, hair perforation, paraffin baiting and slide culture.
  • Maintenance of stock cultures.
  • Animal pathogenecity tests viz. Intracerebral and intraperitoneal inoculation of mice for cryptococcus.

Parasitology

• Must acquire:
  • Examination of faeces for parasitic ova and cysts etc. by direct and concentration methods (salt floatation and formol – ether methods)and complete examination for other cellular features.
  • Egg counting techniques for helminths.
  • Examination of blood for protozoa and helminths by wet mount, thin and thick stained smears.
  • Examination of other specimens for e.g. urine, C.S.F., bone marrow etc. for parasites.
  • Histopathology sections – examination and identification of parasites.
  • Performance of stains – Leishman, Giemsa, Modified Acid Fast, Toluidine Blue O.
  • Identification of common arthropods and other vectors viz. Mosquito, sand fly, ticks, mite and cyclops.
  • Collection of specimens.
  • Preservation of parasites – mounting, fixing, staining etc.
• Desirable to acquire:
  • In-vitro culture of parasites like entamoeba, leishmania, P.falciparum.
  • Maintainance of toxoplasma gondii in mice.
  • Preparation of media – NIH, NNN etc.
  • Copro-culture for larva of hook worms.
  • Antigen preparation viz. Entamoeba , Filarial , Hydatid for serological tests like IHA and skin test like Casoni’s .
  • Permanent staining techniques like iron haematoxylin

Virology

• Must acquire:
  • Preparation of glassware for tissue culture(washing, sterilization)
  • Preparation of media like Hanks, MEM.
  • Preparation of clinical specimens for isolation of viruses.
  • Serological tests-ELISA and rapid tests for HIV, RPHA for HbsAg,
  • Haemagglutination inhibition for influenza, AGD and couterimmunoelectrophoresis for detection of viral antigens or antiviral antibodies.
  • Chick embryo techniques- inoculation and harvesting.
  • Handling of mice, rats, guinea pigs, rabbits for collection of blood,pathogenicity test etc.
• Desirable to acquire:
  • Preparation of Monkey Kidney Cells (Primary) maintenance of continuous cell lines by subcutures. Preservation of cell cultures.
  • Recognition of CPE in tissue cultures.
  • Performance of haemadsorption, haemagglutination, immunofluorescence, neutralization tests for identification of viruses.