BIOL 351 Lab Practical - Selective Media and Differential Tests

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Eugon Agar Deeps

Purpose: Aerotolerance

- Prepared with Tryptic Soy Agar using 10mL instead of the customary 7mL to ensure that the bottom portion is anaerobic.

Anaerobic Jar

Purpose: A means of cultivating anaerobic and microaerophilic bacteria.

GasPak system
- Envelop containing sodium borohydride and sodium bicarbonate
- Paper indicator strip saturated with Methylene blue. Methylene blue is colorless when reduced and blue when oxidized.
- Palladium, which will act as a catalyst to convert H2 and O2 to water.

Mannitol Salts Agar

Purpose: Isolation and differentiation of Staphylococcus aureus.

The Agar:
Phenol red - pH indicator. Yellow below 6.8, red at 7.4 to 8.4, and pink at 8.4 and above.
Mannitol - substrate for fermentation (makes medium differential).
Sodium chloride - High concentration of salt (makes medium selective). Kills most bacteria, but Staphylococci can thrive here.

Mannitol Salts Agar Demo

Staphylococcus aureus (+): growth and color change to yellow
Escherichia coli, Staphylococcus epidermidis, Klebsiella oxytoca, sporosarcina ureae (-): no color change

MacConkey Agar

Purpose:

MacConkey Agar Demo

Escherichia coli and Klebsiella oxytoca (+): growth and red colored
Staphylococcus aureus, Staphylococcus epidermidis, Sporosarcina ureae (-): no growth, not really any color

Methyl Red and Voges-Proskauer Test

Purpose:

Methyl Red and Voges-Proskauer Demo

Methyl Red:
Escherichia coli (+)
Enterobacter aerogenes (-)

Voges-Proskauer:
Enterobacter aerogenes (+)
Escherichia coli (-)

Catalase Test

Purpose:

Catalase Test Demo

-

Oxidase Test

Purpose:

Oxidase Test Demo

-

Nitrate Reduction Test

Purpose:

Nitrate Reduction Test Demo

Micrococcus luteus
Escherichia coli
Soil: bubble present

Citrate Test

Purpose:

Citrate Test Demo

Enterobacter aerogenes (+)
Escherichia coli (-)

Starch Hydrolysis

Purpose:

Starch Hydrolysis Demo

Bacillus subtilis (+)
Escherichia coli (-)

Urea Hydrolysis

Purpose:

Urea Hydrolysis Demo

Proteus vulgaris (+)
Escherichia coli (-)

Indole Test

Purpose:

Indole Test Demo

Escherichia coli (+)
Enterobacter aerogenes (-)

Litmus Milk Medium

Purpose:

Litmus Milk Test Demo

Acid - litmus color change (light blue to pink)

Alkaline - litmus color change (light blue to darker blue): Alcaligenes faecalis

Litmus reduction - light blue to white

Curd - change from fluid to solid: Escherichia coli

Proteolysis - decrease in turbidity of milk (clearer, amber-colored fluid, watery): Proteus vulgaris

Bacitracin Susceptibility Test

Purpose:

Bacitracin Susceptibility Demo

Micrococcus luteus (+): susceptible
Staphylococcus epidermidis (-): resistant

Blood Agar

Purpose:

Blood Agar Demo

Staphylococcus aureus (+): beta
Staphylococcus epidermidis (-): gamma

Coagulase Test

Purpose:

Coagulase Test Demo

Staphylococcus aureus (+)
Staphylococcus epidermidis (-)

Motility Test

Purpose:

Motility Test Demo

Proteus vulgaris (+)
Staphylococcus epidermidis (-)

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