Biochemical Characteristics Procedures:

Starch Hydrolysis

Procedure:
1.    Inoculate a starch plate with the organism to be tested.
2.    Incubate at optimum temperature for at least 48 hours.
3.    Flood plate with iodine, observe results.
4.    Blue color indicates no hydrolyis, while a clear zone indicates hydrolysis.

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Lab Procedures

Phenol Red Fermentation Broth

Procedure:
1.    Inoculate a phenol red tube with the appropriate organism.
2.    Incubate for 48 hours, observe results
3.    Red and orange colors are negative tests for acid, yellow is a positive test for acid, a bubble is a positive test for gas formation.

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Lab Procedures

Lipid Hydrolysis

Procedure:
1.    Inoculate a Spirit Blue plate with the organism to be tested
2.    Incubate for 48 hours, observe results
3.    A deep blue color (rancid odor) indicates lipid hyrolysis by the organism, normal growth is negative for hydrolysis.

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Lab Procedures

Gelatin Hydrolysis

Procedure:
1.    Inoculate a nutrient gelatin tubes with a straight needle.
2.    Incubate for at least 48 hours, observe for liquification at 22 degrees Celsius.
3.    Liquification of the gelatin is a positive test for gelatin hydrolysis.

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Lab Procedures


Casein Hydrolysis

Procedure:
1.    Inoculate a milk agar plate with the organism to be tested.
2.    Incubate for 48 hours, observe results.
3.    Clearing of the agar around the bacterial growth indicates casein hydrolysis.

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Lab Procedures

Action on Litmus milk

Procedure:
1.    Inoculate a litmus milk tube with the organism to be tested.
2.    Incubate for 48 hours, observe results then incubate for 5 more days and observe results again.
3.    Multiple results, see results page.

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Lab Procedures

Sulfur Indole Motility Test

Procedure:
1.    Inoculate a SIM tube with the organism to be tested.
2.    Incubate for 48 hours.
3.    Add kovacs reagent to the top of the tube, pink/red color formation indicates indole formation.
4.    Black percipitate in the medium indicates hydrogen sulfide formation.
5.    Growth away from the stab line indicates motility.

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Lab Procedures


Urease Test

Procedure:
1.    Inoculate a urea tube with 3 loopfuls of slant culture.
2.    Incubate 24 hours, observe for reaction.
3.    A pink color formation indicates the breakdown of urea to ammonia and CO2

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Lab Procedures


Nitrate Reduction Test

Procedure:
1.    Inoculate a trypticase-nitrate tube with the organism to be tested.
2.    Incubate for 48 hours, test for results.
3.    Add one ml of sulfanilic acid to each tube, then add one ml of dimethyl 1-naphthylamine solution, Mix
4.    Development of a red color indicates a positive test.
5.    If no red color develops add a small amount of zinc powder
6.    If a red color develops, nitrate is present and the test is negative.  If no red color develops, nitrate has been reduced and the test is positive.

Go To Results of Nitrate Reduction Test
Lab Procedures