Ground beef (free of fat) 500.0 g

DW 1.0 L

1N NaOH 25.0 ml

Use lean beef or horse meat. Remove fat and connective tissue before grinding mix meat, water and NaOH, and bring to boilwhile stirring. Cool to room temperature,skim fat of surface, and filter retaining both meat particles and filtrate. Add sufficient distilled water to filtrate to restore 1.0 L. original volume. to the filtrate add:

Peptone 30.0 g

Yeast extract 5.0 g

K2HPO4 5.0 g

0.025% resazurin sol. 4ml

Boil, cool, and add 0.5 g. L-cysteine HCl-H2O. Adjust pH to 7.0 under O2-free nitrogen and 3% hydrogen, dispense 7 ml. Over 1 part meat particles to 4-5 parts fluid per test tube. Cap with rubber stoppers under N2 and H2 and autoclave in press for 15 minutes under fast exhaust.