Culture Media By Zahoor Ahmed B.S MLT 06
Definitions <ul><li>Medium </li></ul><ul><li>Any preparation that contains nutrients essential  for bacterial growth. </li...
Classification <ul><li>Classified on the following basis. </li></ul><ul><li>a)  Consistency </li></ul><ul><li>b)  Nutritio...
Consistency <ul><li>Sub-divided into  </li></ul><ul><li>a )   Solid  e.g   nutrient agar </li></ul><ul><li>b )   Semi-soli...
Nutritional <ul><li>Further sub-divided into </li></ul><ul><li>a )   Simple   e.g   peptone water </li></ul><ul><li>b )   ...
Functional <ul><li>Further sub-divided into </li></ul><ul><li>a )  Basic media </li></ul><ul><li>b)  Enriched media </li><...
Basic Media <ul><li>These are simple media contains only basic substances i.e Co2 ,N , &minerals. </li></ul><ul><li>Suppor...
Enriched Media <ul><li>Used for the organisms that require exact growth requirements. </li></ul><ul><li>Basic media may be...
Selective Media <ul><li>These are solid media contains substances  i.e  bile salts ,dyes etc. </li></ul><ul><li>Inhibit th...
Contin….. <ul><li>Examples: </li></ul><ul><li>MacConkey agar  for  Gram-negative  bacteria  </li></ul><ul><li>Low stein Je...
Indicator Media <ul><li>Also called differential media. </li></ul><ul><li>Used to differentiate one organisms from another...
Contin…… <ul><li>Examples </li></ul><ul><li>a)   E osin methylene blue (EMB),   which is differential for lactose and sucr...
Transport Media <ul><li>These are semi-solid media. </li></ul><ul><li>Used for the temporary storage of specimens. </li></...
Contin…… <ul><li>Ensure the survival of aerobic and anaerobic organisms. </li></ul><ul><li>Examples: </li></ul><ul><li>a) ...
Identification Media <ul><li>Help to identify bacteria isolated on primary growth. </li></ul><ul><li>Organisms are identif...
Contin…. <ul><li>Examples: </li></ul><ul><li>Peptone water </li></ul><ul><li>Urea broth </li></ul><ul><li>Klinger iron aga...
Preparation <ul><li>Wash hands & wear gloves </li></ul><ul><li>Sterilize all equipments </li></ul><ul><li>Pour D/W in glas...
Sterilizing Culture Media <ul><li>Autoclaving: </li></ul><ul><li>To remove bacterial endospores as well as  </li></ul><ul>...
Storage of Culture Media <ul><li>Nutrient broth and agar can be stored for 6 months at  12-l6°C.  </li></ul><ul><li>Agar p...
Q.C of Culture Media <ul><li>PH values. </li></ul><ul><li>Temperature </li></ul><ul><li>Sterility </li></ul><ul><li>Growth...
 
 
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Culture Medias

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