ATCC Medium: 2781 Modified Chopped Meat Carbohydrate Medium
Complete Medium
Trypticase……………………………………………….30.0 g
Yeast Extract……………………………………………5.0 g
K2HPO4…………………………………………………5.0 g
Glucose………………………………………………….4.0 g
Cellobiose……………………………………………….1.0 g
Maltose…………………………………………………..1.0 g
Starch……………………………………………………1.0 g
Cysteine HCl…………………………………………….0.5 g
Resazurin Solution (0.1% solution)……..……………1.0 ml
Hemin Solution (see below)……………………………10 ml
Vitamin K1 Solution
(see below)…………………...…..0.2 ml
Agar (if required)………………………………………..15.0 g
Chopped Meat
Broth (See Below)…………………….1000 ml
Adjust pH to 7.0. Boil, cool and dispense anaerobically under N2:H2:CO2 (80:10:10) gas. Dispense in 7 ml volumes for test tubes. Cap with rubber stoppers and autoclave at 121?C. If meat particles are required, use one part meat per 4-5 parts broth.
Chopped Meat Broth Ground Beef (fat-free)……………………………………500.0 g
DI Water…………………………………………………...1000 ml
NaOH (1N)…………………………………………………25.0 ml
Mix ingredients well. Bring to boil for 20 minutes with stirring, simmer for 3 hours. Allow to cool to room temperature and skim fat. Alternatively, you may cool in a refrigerator and remove hardened fat from surface. Filter through cheese cloth and retain both meat particles and the filtrate. Bacteriology should specify whether they want the meat particles in the final medium or not. To the filtrate, add sufficient water to restore volume to 1.0 L.
Vitamin K1 Solution
Vitamin K1…………………………………………………0.15 ml
Ethanol…………………………………………………….30.0 ml
Filter sterilize and store in dark bottle. Refrigerate
Discard after 30 days.
Hemin Solution
Hemin………………………………………………………50.0 mg
NaOH (1N)……………………………………………….....1.0 ml
DI Water…………………………………………………….100.0 ml
Filter sterilize and store in dark bottle, refrigerate. Discard after 30 days.