322. CLOSTRIDIUM MEDIUM II

Ground beef (fat free) 500.0 g

Distilled water 1000.0 ml

NaOH 1 N 25.0 ml

Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add water to a final volume of 1000.0 ml, and then add:

Casitone 30.0 g

Yeast extract 5.0 g

K₂HPO₄ 5.0 g

Resazurin solution (0.1% w/v) 0.5 ml

Glucose 4.0 g

Cellobiose 1.0 g

Maltose 1.0 g

Starch (soluble) 1.0 g

To make medium anoxic boil it, cool under 100% N₂ gas atmosphere, add 0.5 g/l cysteine hydrochloride and adjust pH to 7.0. Dispense under same gas atmosphere 7.0 ml medium into Hungate-type tubes.

 

Updated 11.08.2021