TCC medium: 1761 Modified chopped meat medium (ATCC medium 1490) plus 0.5% arginine ATCC Medium 1490 (see below) plus 0.5% arginine
ATCC Medium 1490:........... 500.0 g
Ground beef (fat-free)
Distilled water.................... 1.0 L
N NaOH............................ 25.0 ml
Mix meat, water and NaOH and bring to a boil with stirring. Cool to room temperature, skim fat from surface, and filter, retaining both meat particles and filtrate.
To filtrate, add sufficient distilled water to restore volume to 1 L.
To this filtrate, add:.... 30.0 g
Trypticase Peptone (BD 211921)
Yeast extract...................... 5.0 g
K2HPO4 ............................. 5.0 g 0
.025% Resazurin................... 4.0 ml
Agar (if necessary)............... 20.0 g
Boil and cool medium under 80% N2, 10% H2, 10% CO2, and add: L-Cysteine .
HCl................... 0.5 g
Hemin Solution (see below)........ 10.0 ml
Vitamin K1 Solution (see below).... 0.2 ml
Adjust medium for final pH of 7.0. Anaerobically dispense 7 ml into tubes containing meat particles (1 part meat to 5 parts fluid) under the same gas phase.
Vitamin K1 Solution:
Vitamin K1......................... 0.15 ml
95% Ethanol .......................30.0 ml
Store solution in brown bottle under refrigeration.
Discard after one month. Hemin.............................Solution: 50.0 mg
Hemin N NaOH............................. 1.0 ml
Distilled water to............... 100.0 ml
Dissolve hemin in NaOH and bring volume to 100 ml with distilled water. Autoclave solution at 121C for 15 minutes and cool.