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Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
Biochemical Reaction - Prac. Microbiology
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Biochemical Reaction - Prac. Microbiology

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  • 1. Bacteria vary in their metabolic & enzymatic activities. Used in identification of different genera & species of bacteria. Biochemical reactions are done on bacteria grown in pure culture.
  • 2. Biochemical Reactions 1. Sugar fermentation 2. Oxidase test 3. Catalase test 4. Coagulase test 5. API Test
  • 3. 1. Sugar fermentation Sugar media are composed of: Peptone water + 1% test sugar + Andrade’s indicator + A small inverted tube (Durham’s tube) .
  • 4. Sugar fermentation test uninoculated Not fermented fermented with A only fermented with A & G
  • 5. 1. Sugar fermentation Sugar fermentation can be indicated by change colour of the medium from yellow to red. Glucose +ve Lactose +ve maltose +ve mannite +ve Sucrose - ve
  • 6. Catalase Test Principle: H2O2 Procedure: 1 2 Catalase enzyme 3 H2O2 H2 O + O2 Air bubbles
  • 7. Results: Catalase Test Positive test: rapid appearance of gas bubbles. Catalase +ve Catalase –ve
  • 8. Catalase Test
  • 9. Coagulase Test Principle: Fibrinogen Plasma Procedure: 2 Coagulase enzyme Fibrin Visible Clot 1 1 ml rabbit plasma 3 Place at 37oC, observe for formation of visible clot for up to 4 hrs.
  • 10. Coagulase Test Coagulase +ve Coagulase -ve
  • 11. Oxidase Test Principle: Tetramethyl p-phenylene diamine hydrochloride (oxidase reagent) colourless Cytochrome oxidase enzyme Indophenol (Purple colour) within 1-2 min
  • 12. Oxidase Test
  • 13. API (Analytical Profile Index)
  • 14. Incubation Interpretation acc. to given charts
  • 15. Bacterial Typing
  • 16. Classification of a certain species of bacteria into Types. Importance: Epidemiological investigation (e.g. food poisoning) Examples of Typing methods:  bacteriophage typing  bio-typing  serotyping  plasmid analysis  + others
  • 17. Bacteriophage Typing Bacterial strain on surface of media + drops of different phages homogenous bacterial growth + areas of lysis (= susceptibility)
  • 18. Questions
  • 19. 1- In sugar fermentation, Andrade's indicator is added to the sugar media to test for gas production: ( ) True ( ) False 2- In the coagulase test, coagulation of plasma occurs in positive cases: ( ) True ( ) False 3- Biochemical reactions must be done on a pure culture of the organism to be identified: ( ) True ( ) False
  • 20. MCQ 4-Catalase test is: a) negative if oxygen bubbles are released b) positive if a visible clot is formed c) done by adding a drop of H2O2 d) done to test for the presence of urease enzyme 5- Oxidase test is: a) done using catalase reagent b) positive when a deep purple color is produced c) positive after a few hours d) negative when a yellow color is produced
  • 21. Thank you

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