Modified Chopped Meat Medium
- Add the following to 1 L of water
Mix meat, water and NaOH and bring to a boil with stirring
Cool to room temperature
Skim fat from surface
Filter the solution and retain both meat particles and filtrate
To filtrate, add sufficient distilled water to restore volume to 1 L.
To the filtrate add:
- 500g fat-free ground beef
- 25 ml 1N NaOH
Boil and cool medium under 80% N2, 10% H2, 10% CO2
To the solution add:
- 30 g trypticase peptone (BD 211921)
- 5 g yYeast extract
- 5 g KHPO4
- 4 ml resazurin solution (see below)
Adjust medium for final pH 7.0
Anaerobically dispense 7 ml into tubes containing meat particles (1 part meat to 5 parts fluid) under the gas phase noted above.
- 0.5 g L-Cysteine-HCl
- 10 ml Hemin solution (see below)
- 0.2 ml Vitamin K1 Solution (see below)
Vitamin K1 Solution
- Add 0.15 ml of vitamin K1 to 30 ml of 95% ethanol
- Store solution in brown bottle under refrigeration.
- Discard after one month.
- Add the following to 99 ml of distilled water
- 50 mg hemin
- 1 ml of 1N NaOH
- Autoclave to sterilize.