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Composition

* Dehydrated powder, hygroscopic in nature, store in a dry place, in tightly-sealed containers below 25°C

and protect from direct Sunlight.

Instructions for Use

Dissolve 45gms in 445 ml distilled water. Gently heat to boiling with gentle swirling and dissolve the

medium completely. Sterilize by autoclaving at 15 psi (121°C) for 15 minutes. Cool to 45-50°C. Aseptically

add equal amount (445ml) of sterile 2% Haemoglobin powder (TS 021) and add 10ml of Vitamins growth

supplement (TS 022) or Yeast autolysate supplement (TS 023). Mix well. Dispense into sterile Petri plates

or sterile culture tubes.

Appearance: Light amber color, clear to slightly opalescent gel. On addition of Haemoglobin: Chocolate

brown colour appears, opaque gel

pH (at 25°C): 7.3 ± 0.2

Principle

CHOCHOLATE AGAR BASE is used for isolation and cultivation of fastidious microorganisms like

Neisseria gonorrhoeae. It can also be used for cultivation of aerobic, anaerobic and microaerophillic

microorganisms. Medium contains Proteose peptone is a nitrogen source required for the growth of wide

variety of organisms. Dextrose acts a carbon energy source. Disodium phosphate buffers the medium

whereas sodium chloride maintains the osmotic equilibrium. Agar is the solidifying agent. This medium

is supplemented with cofactor, which provides NAD to facilitate the growth

of Haemophilus influenzae, Neisseria gonorrhoeae and Neisseria meningitidis. Heated sheep blood is added to

give the medium its "chocolate" appearance. This medium is prepared, stored and dispensed under

oxygen-free conditions to prevent the formation of oxidized products prior to use.

Interpretation

Cultural characteristics observed after inoculating (103CFU/ml), on incubation at 35 ± 2°C for 48 hours

with CO2.

Ingredients Gms/Ltr.

Proteose peptone 20.0

Agar 15.0

Sodium chloride 5.00

Disodium phosphate 5.00

Dextrose 0.50

Regards

NITIN MALHOTRA

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