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IMViC Biochemical Test

(WIDAYU MUTIYA R. / 24020115140122)

Indole Production Test: The test bacteria were stab inoculated into the
medium and incubated at 370C for 48 hours. The indole production was tested
by adding Kovac‘s Reagent and then observed for the result. A positive result
had a red layer at the top. A negative result had a yellow or brown layer (Fig.
8).

Composition of the medium, containing (w/v)


Ingredients Gms/ Litre
Peptone 30.0g/l
Beef Extract 3.0g/l
Ferrous ammonium sulfate 0.2g/l
Sodium thiosulfate 0.025g/l
Agar 3.0 g/l
Distilled water 1000ml
pH 7.3 + 0.2

Methyl Red-Voges Proskauer Test: The test bacteria was inoculated in the
medium and incubated at 370C for 48 hours. Five drops of Methyl Red
indicator was added to the tube and change in color was observed. A positive
result had a distinct red layer at the top of the broth. A negative result had a
yellow layer (Fig. 11a).

The medium was inoculated with the test bacteria and incubated at 370C for
48 hours then VP reagents (Barritt's reagent A and Barritt reagent B) were
added to the tubes and then observed for the result. Pink or red indicated a
positive reaction. No change in color or copper color indicated a negative
reaction (Fig. 11b).

Composition of glucose phosphate medium, containing (w/v)


Ingredients Gms/ Litre
Glucose 5.0 g/l
Peptone 7.0 g/l
K2HPO4 5.0g/l
Distilled water 1000ml
pH 7.0 + 0.2
Citrate Utilization Test: The plates of citrate medium were inoculated with
the test bacteria and incubated at 370C for 48 hours and then observed for the
results. A positive reaction was indicated with a Prussian blue color and
growth of bacteria. A negative test had no growth of bacteria and remained
green (Fig. 4a and 4b).

Composition of medium, containing (w/v)


Ingredients Gms/ Litre
Magnesium sulfate (heptahydrate) 0.2 g/l
Ammonium dihydrogen phosphate 1.0 g/l
Dipotassium phosphate 1.0 g /l
Sodium citrate (dehydrate) 2.0 g/l
Sodium chloride 5.0 g/l
Agar 15.0 g/l
Bromothymol blue 0.08 g/l
Deionized water 1,000 ml
pH 6.8+ 0.2

Motility test: The agar tubes containing the media were stab inoculated with
the test bacteria and incubated at 370C for 48hours and then observed for
the growth. Diffusion of colony indicated positive test (Fig. 10).

Composition of the medium, containing (W/V)

Ingredients Gms/ Litre


Peptone 30.0g/l
Beef Extract 3.0g/l
Ferrous ammonium sulfate 0.2g/l
Sodium thiosulfate 0.025g/l
Agar 3.0 g/l
Distilled water 1000ml
pH 7.4 + 0.2