Ground beef (free of fat)........500.0 g
Distilled water....................1.0 L
N 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 a boil while stirring.
Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate.
Add sufficient distilled water to filtrate to restore 1 liter original volume.
To the filtrate add:
Peptone...........................30.0 g
Yeast extract......................5.0 g
K2HPO4 .............................5.0 g
0.025% Resazurin solution..........4.0 ml
Boil, cool and add 0.5 g L-cysteine . HCl; adjust pH to 7.0. Under 97% nitrogen, 3% hydrogen, dispense 7 ml over 1 part meat particles to 4-5 parts fluid per test tube. Cap with butyl rubber stoppers under N2, H2 and autoclave in press for 15 minutes under fast exhaust.