GELATIN HYDROLYSIS TEST Purpose :

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Presentation transcript:

GELATIN HYDROLYSIS TEST Purpose : Determination of the ability of the organisms to digest (hydrolyse) gelatin, as a sole source of protein by activity of the gelatinase enzyme. Materials : 1. Aeromonas hydrophila on BA. 2. Escherichia coli on BA.

GELATIN HYDROLYSIS TEST Media & Reagents : 1. Gelatin agar plates. 2. Acid mercuric chloride solution Method : 1. Inoculate by cross streaking of 1 & 2 on each of gelatin plates. 2. Incubate at 370C for 24 hrs.

GELATIN HYDROLYSIS TEST Interpretation : Before reading, flood the cultures with acid mercuric chloride, leaving for 5-10 minutes. Positive : clear zone around colony. Negative : no clear zone around colony.

GELATIN HYDROLYSIS TEST Positive : clear zone around colony. Negative : no clear zone around colony.

STARCH HYDROLYSIS TEST Purpose : To determine the ability of the organism to produce amylase enzyme which digests or hydrolyses starch. Materials : 1. Bacillus subtilis on BA. 2. Escherichia coli on BA.

STARCH HYDROLYSIS TEST Media & Reagents : 1. Two starch agar plates. 2. Lugol’s iodine solution. Method : 1. Inoculate 1 & 2 on each of starch agar plate. 2. Incubate at 370C for 24 hrs.

STARCH HYDROLYSIS TEST Interpretation : Flood the cultures with Lugol’s solution, and remain for 5-10 minutes. Positive : clear zone around colonies. Negative : no clear zone around colonies. Remark : Some orgamisms, produce weak reaction, clear zone present under colonies.

STARCH HYDROLYSIS TEST Positive : clear zone around colonies. Negative : no clear zone around colonies.

CASEIN DIGESTION TEST Purpose : Materials : To determine the ability of organisms to produce caseinase (proteinase) enzyme which digest casein. Materials : 1. Aeromonas hydrophila on BA. 2. Escherichia coli on BA.

CASEIN DIGESTION TEST Media & Reagents : Method : Two casein agar plates. Method : 1. Spot inoculation of 1 & 2 on each of casein agar plates. 2. Incubate at 370C for 24 hrs.

CASEIN DIGESTION TEST Interpretation : Positive : clear zone around colonies. Negative : no clear zone around colonies.

Lecithinase & Lipase activities Purpose : LECITHOVITELLIN REACTION (LV) Lecithinase & Lipase activities Purpose : To determine the ability of the organisms to produce lecithinase or/and lipase. The enzymes are demonstrated by cultivation on egg yolk agar.

Lecithinase & Lipase activities LV REACTION Lecithinase & Lipase activities Materials : 1. Bacillus cereus on BA. 2. Aeromonas hydrophila on BA. 3. Bacillus subtilis on BA. Media & Reagents : Two egg yolk agar plates.

LV REACTION Lecithinase & Lipase activities Method : 1. Spot inoculate 1, 2 & 3 on each of L-V agar plate. 2. Incubate at 370C for 24 hrs. Reaction : Zone of opalescent around colony indicated lecithinase activity against lecithovitellin in egg yolk and precipitation around colony indicated lipase activity.