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