BT221 Microbiological methods-

Unit 1-

Instrumentation- Microscopy: Principles of Microscopy- resolving power, numerical aperture, working principle and applications of light, Compound microscope, Dark field microscope, Phase contrast microscope, Fluorescence Microscope, confocal microscope, Electron Microscopes- TEM and SEM.
Analytical techniques: Working principles and applications: Centrifuge, Ultracentrifuge, Spectrophotometer, Chromatography: Paper and TLC.

Unit 2-

Sterilization techniques- Definition of terms; Physical methods of control: Principle, construction and applications of moist heat sterilization, Boiling, Pasteurization, Fractional sterilization-Tyndallization and autoclave, Dry heat sterilization-Incineration and hot air oven, Filtration – Diatomaceous earth filter, seitz filter, membrane filter and HEPA.
Radiation : Ionizing radiation-γ rays and non ionizing radiation- UVrays Chemical methods: Alcohol, aldehydes, phenols, halogen, metallic salts, Quaternary ammonium compounds and sterilizing gases as antimicrobial agents.

Unit 3-

Microbiological techniques- Culture Media: Components of media, natural and synthetic media, chemically defined media, complex media, selective, differential, indicator, enriched and enrichment media Pure culture methods: Serial dilution and plating methods (pour, spread, streak); cultivation, maintenance and preservation/stocking of pure cultures; cultivation of anaerobic bacteria
Stains and staining techniques: Principles of staining, Types of stains-simple stains, structural stains and differential stains.

Unit 4-

Antimicrobial agents and assessment of antimicrobial activity- Mode of action of antimicrobial agents: Antifungal agents: Amphotericin B, Griseofulvin Antiviral agents: Amantadine, Acyclovir, Azidothymidine Antibacterial agents: Plazomicin, Ervacycline, Omadacyclin and Imipenum,
Challenges in antimicrobial therapy; Emergence of antibiotic resistance (MDR, XDR).
Assessment of antimicrobial activity: Antibacterial-Disc and agar well diffusion techniques, Microdilution method, Zone of inhibition, MCB, Determination of IC 50, Antifungal-Determination of MFC, Time kill kinetics assay, sorbitol assay, Antiviral-CPE, virus yield reduction assay, TCID, Neutralization ASSAY, Hemagglutination inhibition.

Text Books / References:

  1. Atlas RM. (1997). Principles of Microbiology. 2nd edition. WM.T. Brown Publishers.
  2. Black JG. (2008). Microbiology: Principles and Explorations. 7th edition. Prentice Hall
  3. Madigan MT, and Martinko JM. (2014). Brock Biology of Micro-organisms. 14th edition. Parker J. Prentice Hall International, Inc.
  4. Pelczar Jr MJ, Chan ECS, and Krieg NR. (2004). Microbiology.
  5. Srivastava S and Srivastava PS. (2003). Understanding Bacteria. Kluwer Academic Publishers, Dordrecht
  6. Stanier RY, Ingraham JL, Wheelis ML and Painter PR. (2005). General Microbiology. 5th edition McMillan.
  7. Tortora GJ, Funke BR, and Case CL. (2008). Microbiology: An Introduction. 9th edition Pearson Education.
  8. Willey JM, Sherwood LM, and Woolverton CJ. (2013). Prescott’s Microbiology. 9th edition. McGraw Hill Higher Education.
  9. Cappucino J and Sherman N. (2010). Microbiology: A Laboratory Manual. 9th edition. Pearson Education Limited
  10. Microbiology- Concepts and applications by Paul A. Ketchum, Wiley Publications
  11. Fundamentals of Microbiology –Frobisher, Saunders & Toppan Publications
  12. Introductory Biotechnology-R.B Singh C.B.D. India (1990)
  13. Fundamentals of Bacteriology – Salley
  14. Frontiers in Microbial technology-P.S. Bison, CBS Publishers.
  15. Biotechnology, International Trends of perspectives A. T. Bull, G.
  16. General Microbiology –C.B. Powar