NCTC9336

CHOPPED MEAT MEDIUM

Ground beef (fat free) 500.0 g

1N NaOH 25.0 mL

Distilled water 1.0 L

Peptone 30.0 g

Yeast extract 5.0 g

L-cysteine·HCl 0.5 g

K2HPO4 5.0 g

0.025% Resazurin solution 4.0 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, filter and retaining both meat particles and filtrate. Add sufficient distilled water to filtrate to restore 1.0 L original volume. To the filtrate add peptone, yeast extract, K2HPO4 and resazurin solution. Boil, then cool under O2-free nitrogen and 3% hydrogen, and add 0.5 g L-cysteine·HCl. Adjust pH to 7.0. Dispense 4-5 parts fluid to 1 part meat particles per test tube. Cap with rubber stoppers under N2 and H2 and autoclave in press for 15 minutes under fast exhaust.