Module 5b biochemical activities for the lab
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  • 1. Principle, Reagents & MediaProcedures & End resultsBIOCHEMICAL ACTIVITIES
  • 2. BIOCHEMICAL ACTIVITIES OF BACTERIA:Biochemical properties could be used as tools of identification  demonstrate certain metabolic activity of microbesExample : Fermentation – durham tubes Carbohydrates, amino acids, organic acids  1. gases – Carbone dioxide, methane, hydrogen  2. Acid production – use of indicators to detect  Either acid or gas or both can be collected
  • 3. IMVIC - INDOLE TEST Principle : product of hydrolysis of an amino acid tryptophan Media – SIM (Sulfide Indole Motility) Reagent - (Ehrlich’s) Para-dimethyl- aminobenzaldehyde Positive result :(+) pink or red color Negative Result : (-) absence of pink or red color
  • 4. IMVIC - METHYL RED TEST  Methyl red is a pH indicator  Principle : production of sufficient acidity from glucose  Media- MR-VP broth  (+) red - acid  (-) yellow - alkaline
  • 5. IMVIC - VOGUES PROSKAUER• production of acetyl-methyl carbinolfrom glucose•media: MR-VP broth•reagent: 40% KOH•Positive (+) result = production ofan eosin like color
  • 6. IMVIC - CITRATE UTILIZATION TEST  Principle: utilizes citrate as a source of Carbon  Media :Simmon Citrate (SCA)  (+) blue color  (--) green color
  • 7. OTHER COMMON BIOCHEMICAL TESTS: Coagulase test Catalase test Urease test Hemolysis test Greig’s Test Oxidase Test Hydrogen Sulfide Test
  • 8. ASSIGNMENT: Find the other biochemical Tests & tabulate them based on:  Working principle  Reagents & Media  Positive & Negative Results
  • 9. WHAT TO DO? Place all your glassware in a plastic bag Autoclave 15 psi, 121 C, 15 minutes Wash , clean and dry Wrap and sterilize in the oven, 150 C, 45 minutes
  • 10. MEDIA PREPARATION Prepare 200 ml each ( for the whole class)  SIM  MRVP  SCA Dispense into 4 test tubes 1 tube SIM  2 tubes MRVP  1 tube SCA ( Slant )
  • 11.  Inoculate with bacteria ( get from me ) Incubate in incubator 48 hours Add the reagents ( from previous slides ) Check results Make a one page write – up Submit