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Culture Medias

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  • 1. Culture Media By Zahoor Ahmed B.S MLT 06
  • 2. Definitions
    • Medium
    • Any preparation that contains nutrients essential for bacterial growth.
    • Culture medium:
    • A nutrient substance (solid or liquid) that is used to grow micro-organisms .
    • Inoculation of media
    • Introduction of infected material to the medium
    • for cultivation of organisms.
  • 3. Classification
    • Classified on the following basis.
    • a) Consistency
    • b) Nutritional
    • c) Functional
  • 4. Consistency
    • Sub-divided into
    • a ) Solid e.g nutrient agar
    • b ) Semi-solid e.g peptone water.
    • c ) Liquid medium e.g nutritional broth.
  • 5. Nutritional
    • Further sub-divided into
    • a ) Simple e.g peptone water
    • b ) Complex e.g blood agar
    • c ) Synthetic e.g Davis /mingioli medium
  • 6. Functional
    • Further sub-divided into
    • a ) Basic media
    • b) Enriched media
    • c ) Selective media
    • d ) Indicator media
    • e ) Transport media
    • f ) Identification media
  • 7. Basic Media
    • These are simple media contains only basic substances i.e Co2 ,N , &minerals.
    • Support those organisms that don't require special nutrient.
    • Often used in the preparation of enriched media.
    • Also used for sub culturing of pathogens.
    • Examples:
    • Nutrient agar & Nutrient broth.
  • 8. Enriched Media
    • Used for the organisms that require exact growth requirements.
    • Basic media may be enriched with whole or lyzed blood , serum , peptones , vitamins etc.
    • Increases the no: of pathogens because it fulfill the whole nutritional requirements.
    • Examples:
    • Blood & chocolate agar .
  • 9. Selective Media
    • These are solid media contains substances i.e bile salts ,dyes etc.
    • Inhibit the growth of one organism to allow the other one i.e inhibit the growth of normal flora and allow the growth of pathogen.
  • 10. Contin…..
    • Examples:
    • MacConkey agar for Gram-negative bacteria
    • Low stein Jensen's agar for M . bacterium.
    • TCBS for vibrio.
  • 11. Indicator Media
    • Also called differential media.
    • Used to differentiate one organisms from another
    • organisms.
    • Dyes , indicator or other substances are added to differentiated organisms i.e
    • Neutral red , phenol red , eosin , or methylene blue.
  • 12. Contin……
    • Examples
    • a) E osin methylene blue (EMB), which is differential for lactose and sucrose fermentation
    • b) MacConkey (MCK), which is differential for lactose fermentation
    • c) M annitol salt agar (MSA), which is differential for mannitol fermentation
  • 13. Transport Media
    • These are semi-solid media.
    • Used for the temporary storage of specimens.
    • Maintain the viability of all organisms in the specimen.
    • Transport media typically contain only buffers and salt .
    • The lack of carbon, nitrogen, and organic growth factors prevents microbial multiplication .
  • 14. Contin……
    • Ensure the survival of aerobic and anaerobic organisms.
    • Examples:
    • a) Thioglycollate broth for strict anaerobes .
    • b) Cary-blair medium for enteric pathogen.
    • c) Amies medium for gonococci.
  • 15. Identification Media
    • Help to identify bacteria isolated on primary growth.
    • Organisms are identified by changing medium color or producing gas.
    • Organisms used to inoculate identification media must be first isolated in pure culture.
  • 16. Contin….
    • Examples:
    • Peptone water
    • Urea broth
    • Klinger iron agar.
  • 17. Preparation
    • Wash hands & wear gloves
    • Sterilize all equipments
    • Pour D/W in glassware (required amount)
    • Add powder ingredients (required amount)
    • Heat to dissolve completely
    • Autoclave
    • Dispense the medium into tubes ,bottles &plates
    • Store at required temperature
  • 18. Sterilizing Culture Media
    • Autoclaving:
    • To remove bacterial endospores as well as
    • vegetative cells.
    • Steaming at 100 c:
    • used to sterilize media containing ingredients that would be broken down or inactivated above 100 c.
    • Filtration:
    • Removing bacteria from fluids .
  • 19. Storage of Culture Media
    • Nutrient broth and agar can be stored for 6 months at 12-l6°C.
    • Agar plates should be stored at 2-8°C in sealed containers to avoid loss of moisture.
    • It is important to store all media away from light.
    • DO NOT FREEZE.
  • 20. Q.C of Culture Media
    • PH values.
    • Temperature
    • Sterility
    • Growth performance
    • Stability
  • 21.  
  • 22.