Module5b biochemicalactivitiesforthelab-120620065407-phpapp02
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This document discusses various biochemical tests used to identify bacteria. It provides details on tests like indole, methyl red, Voges-Proskauer, and citrate utilization (IMVIC) tests. For each test, it lists the principle, required reagents and media, and positive and negative results. Students are assigned to research additional biochemical tests, tabulating the working principle, reagents, media, and results for each. They are also instructed on proper media preparation and inoculation techniques to perform the tests in class and report their results.
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Module 5b biochemical activities for the lab
This document summarizes various biochemical tests used to identify bacteria, including:
- Fermentation tests using Durham tubes to detect gas and acid production from carbohydrates.
- The IMVIC series of tests (Indole, Methyl Red, Voges-Proskauer, Citrate) which identify metabolic pathways and substrate utilization through color changes.
- Other common tests like catalase, coagulase, urease, and oxidase which detect enzymatic activity through production of gas or color changes.
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This document summarizes various biochemical tests used to identify bacteria, including:
- Fermentation tests using Durham tubes to detect gas and acid production from carbohydrates.
- The IMVIC series of tests (Indole, Methyl Red, Voges-Proskauer, Citrate) which identify metabolic pathways and substrate utilization through color changes.
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This document discusses various biochemical tests used to identify bacteria, including:
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CATALASE TEST
CATALASE TEST-Principle:-
This test demonstrates presence of catalase enzyme. This enzyme catalyses the release of O2 from H2O2.
catalase
2H2O2 H2O + increase O2
Reagents:- 1) 3% H2O2.
2) 24 hrs cultured organisms
Procedure:-
With sterile wooden stick transfer culture organisms to test tube containing 3% H2O2 and observe for production of effervescence.
It can also be tested directly on growth plate.
Positive Control: Staphylococci.
Negative Control: Streptococci.
False positive reactions:
If culture medium contains catalase enzyme e.g., blood agar, chocolate agar.
If iron wire loop is used
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[1] Fermentation tests detect acid and gas production from carbohydrates like glucose, lactose, and sucrose. The MR-VP tests identify bacteria that produce acids or alcohols from glucose fermentation. The Methyl Red test checks for acid production and the Voges-Proskauer test detects alcohol production.
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This document discusses several physiological tests used to identify microbes based on their biochemical properties and reactions. It describes tests such as sugar fermentation, indole production, methyl red, Voges-Proskauer, citrate utilization, nitrate reduction, and more. Each test examines how microbes metabolize certain substrates like carbohydrates or amino acids and analyzes physical responses like color changes or gas production to determine whether microbes are positive or negative for desired biochemical pathways and properties.
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Module5b biochemicalactivitiesforthelab-120620065407-phpapp02
- 1. Principle, Reagents & Media Procedures & End results BIOCHEMICAL ACTIVITIES
- 3. BIOCHEMICAL ACTIVITIES OF BACTERIA: Biochemical properties could be used as tools of identification demonstrate certain metabolic activity of microbes Example : 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
- 4. 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
- 5. 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
- 6. IMVIC - VOGUES PROSKAUER • production of acetyl-methyl carbinol from glucose •media: MR-VP broth •reagent: 40% KOH •Positive (+) result = production of an eosin like color
- 7. IMVIC - CITRATE UTILIZATION TEST Principle: utilizes citrate as a source of Carbon Media :Simmon Citrate (SCA) (+) blue color (--) green color
- 8. OTHER COMMON BIOCHEMICAL TESTS: Coagulase test Catalase test Urease test Hemolysis test Greig’s Test Oxidase Test Hydrogen Sulfide Test
- 9. ASSIGNMENT: Find the other biochemical Tests & tabulate them based on: Working principle Reagents & Media Positive & Negative Results
- 10. 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
- 11. 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 )
- 12. 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