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Biochemical Tests and
             Diagnostic Media




• Labs #6 (white) and #7 (green)
           white
• Overall scheme: Lab #6 (white) , last page
                             white
SIM medium: Motility test



   POSITIVE:       NEGATIVE:
Organism moves     Organism does NOT
away from stab     move away from
                   stab
SIM medium: Sulfur test



H2S-NEGATIVE:       H2S-POSITIVE:
   no black        black precipitate
  precipitate          formed
    formed
Simmon’s citrate agar slant



 negative            positive
Diagnostic Media
• Mannitol salt agar
• Eosin methylene blue (EMB) agar
• Brilliant green agar
Selective medium

   Selects for the GROWTH of some
 organisms and inhibits the growth of
            other organisms.

   i.e. Some organisms grow on a selective medium
whereas others do not grow (or do not grow well) on that
                    same medium.
Differential medium

  Differentiates between one type of
organism and another type of organism.


 Different types of organism look different from each
       other when grown on differential media.
Mannitol salt agar
Staphylococcus                                         Serratia
    aureus                                           marcescens
    grows                                           does not grow




                   Selective medium
  High salt (NaCl) concentration in medium favors organisms
  that tolerate high salt concentration, e.g. Staphylococcus.
Mannitol salt agar



      Differential medium

Differentiates among staphylococci
Mannitol salt agar



      Differential medium

Differentiates among staphylococci
Mannitol salt agar
                                                  Staphylococcus
                                                     ferments
                                                     mannitol
                                                      (yellow)




              Differential medium
Differentiates among staphylococci:
   • pathogenic Staphylococcus species ferment mannitol to

         produce acid (yellow medium)
Mannitol salt agar
                                                         Staphylococcus
Micrococcus does not                                        ferments
 ferment mannitol                                           mannitol
       (pink)                                                (yellow)




                 Differential medium
   Differentiates among staphylococci:
      • pathogenic Staphylococcus species ferment mannitol to

              produce acid (yellow medium)
      • less pathogenic Micrococcus (a staphylococcus)
Levine Eosin-Methylene Blue (EMB)
                  Agar
  Gram negatives
     (growth)




  Gram positive
(growth inhibited)


        SELECTIVE MEDIUM
Levine Eosin-Methylene Blue (EMB)
                Agar




Lactose          Nonlactose
fermenter        fermenters
            DIFFERENTIAL MEDIUM
High acid producers on EMB
             e.g. E. coli




blue-black colonies or metallic sheen
Levine Eosin-Methylene Blue (EMB)
                  Agar
 Lactose fermenter       Non-lactose fermenter
  (high acid producer:
     black colonies)




  Gram positive
(growth inhibited)
                           Lactose fermenter
                            (low acid producer:
                               pink colonies)
Brilliant Green Agar

 SELECTIVE
    •   Growth of all gram positive and
          many gram negative bacteria
          inhibited

    • Growth of many Salmonella species
       occurs

DIFFERENTIAL
Brilliant Green Agar
          DIFFERENTIAL




Differentiates lactose fermenters from
        nonlactose fermenters
Brilliant Green Agar
Lactose fermenter   Non-lactose fermenter
 E. coli                     Salmonella
Catalase Test
 Many bacteria produce the enzyme catalase.




           catalase
H2O2                  H2O       +     O2
hydrogen                              oxygen
peroxide                               (gas)
Catalase Test


                     bubbles (gas)




Catalase positive organism

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Diff mediasv

  • 1. Biochemical Tests and Diagnostic Media • Labs #6 (white) and #7 (green) white • Overall scheme: Lab #6 (white) , last page white
  • 2. SIM medium: Motility test POSITIVE: NEGATIVE: Organism moves Organism does NOT away from stab move away from stab
  • 3. SIM medium: Sulfur test H2S-NEGATIVE: H2S-POSITIVE: no black black precipitate precipitate formed formed
  • 4. Simmon’s citrate agar slant negative positive
  • 5. Diagnostic Media • Mannitol salt agar • Eosin methylene blue (EMB) agar • Brilliant green agar
  • 6. Selective medium Selects for the GROWTH of some organisms and inhibits the growth of other organisms. i.e. Some organisms grow on a selective medium whereas others do not grow (or do not grow well) on that same medium.
  • 7. Differential medium Differentiates between one type of organism and another type of organism. Different types of organism look different from each other when grown on differential media.
  • 8. Mannitol salt agar Staphylococcus Serratia aureus marcescens grows does not grow Selective medium High salt (NaCl) concentration in medium favors organisms that tolerate high salt concentration, e.g. Staphylococcus.
  • 9. Mannitol salt agar Differential medium Differentiates among staphylococci
  • 10. Mannitol salt agar Differential medium Differentiates among staphylococci
  • 11. Mannitol salt agar Staphylococcus ferments mannitol (yellow) Differential medium Differentiates among staphylococci: • pathogenic Staphylococcus species ferment mannitol to produce acid (yellow medium)
  • 12. Mannitol salt agar Staphylococcus Micrococcus does not ferments ferment mannitol mannitol (pink) (yellow) Differential medium Differentiates among staphylococci: • pathogenic Staphylococcus species ferment mannitol to produce acid (yellow medium) • less pathogenic Micrococcus (a staphylococcus)
  • 13. Levine Eosin-Methylene Blue (EMB) Agar Gram negatives (growth) Gram positive (growth inhibited) SELECTIVE MEDIUM
  • 14. Levine Eosin-Methylene Blue (EMB) Agar Lactose Nonlactose fermenter fermenters DIFFERENTIAL MEDIUM
  • 15. High acid producers on EMB e.g. E. coli blue-black colonies or metallic sheen
  • 16. Levine Eosin-Methylene Blue (EMB) Agar Lactose fermenter Non-lactose fermenter (high acid producer: black colonies) Gram positive (growth inhibited) Lactose fermenter (low acid producer: pink colonies)
  • 17. Brilliant Green Agar SELECTIVE • Growth of all gram positive and many gram negative bacteria inhibited • Growth of many Salmonella species occurs DIFFERENTIAL
  • 18. Brilliant Green Agar DIFFERENTIAL Differentiates lactose fermenters from nonlactose fermenters
  • 19. Brilliant Green Agar Lactose fermenter Non-lactose fermenter E. coli Salmonella
  • 20. Catalase Test Many bacteria produce the enzyme catalase. catalase H2O2 H2O + O2 hydrogen oxygen peroxide (gas)
  • 21. Catalase Test bubbles (gas) Catalase positive organism