Media Biochemical Tests
Media Biochemical Tests
Media Biochemical Tests
Laboratory Objectives
Tests To Know
Case Study Tests
Indole
Methyl Red/Voges Proskauer
Citrate
H2S production in SIM
Urea hydrolysis
Motility
Lactose fermentation
Sucrose fermentation
Glucose fermentation & gas production
Staphylococcus identification tests
BAP
MSA
Mstaph broth
Coagulase
Indole Test
How to Perform Test: Inoculate Tryptone broth with inoculating loop.
Property it tests for: This test is performed to help differentiate species of
the family Enterobacteriaceae. It tests for the bacteria species’ ability to
produce indole. Bacteria use an enzyme, tryptophanase to break down the
amino acid, tryptophan, which makes by-products, of which, indole is one.
Media and Reagents Used: Tryptone broth contains tryptophan.
Kovac’s reagent—contains hydrochloric acid, dimethylaminobenzaldehyde,
and amyl alcohol—yellow in color.
Reading Results: Kovac’s reagent reacts with indole and creates a red
color at the top part of the test tube.
Indole
Methyl Red/Voges Proskauer
(MR/VP)
How to Perform Tests: Inoculate 2 glucose broths with inoculating
loop. After 48 hours of incubation, add a few drops of MR to one
tube, and VP reagents to the other tube.
Properties they test for: Both tests are used to help differentiate
species of the family Enterobacteriaceae.
MR—tests for acid end products from glucose fermentation.
VP—tests for acetoin production from glucose fermentation.
Media and Reagents Used:
Glucose Broth
Methyl Red indicator for acid
Voges Proskauer reagents—A: 5% Alpha-Naphthol, & ethanol, B: Potassium
Hydroxide, & Deionized Water.
MR/VP continued
Reading Results:
MR— a + result is red (indicating pH below 6) and a – result is yellow (indicating
no acid production)
VP—A + result is red after VP reagents are added (indicating the presence of
acetoin) and a – result is no color change.
Tolerates Salt.
Does not tolerate salt.
Coagulase
How to Perform Test: Inoculate rabbit plasma with one
single colony. Break up colony and stir until blended in plasma.
Incubate at 37 degrees C for 24 hours.
Property it tests for: This tests for the bacteria’s
ability to clot blood plasma using the enzyme
coagulase. If the organism has coagulase it will clump
rabbit plasma.
Media and Reagents: This media contains rabbit
plasma dissolved in buffer.
Coagulase Results
Reading Results:
If the organism is has coagulase it will clump the plasma.
If the organism does not have coagulase it will not clump the plasma.