ATCC medium: 1703 Chopped meat carbohydrates with rumen fluid
ATCC Medium 1490 (see below) with 0.4% glucose, 0.1% cellobiose, 0.1% maltose, 0.1% starch and 15% filtered rumen fluid.
ATCC Medium 1490:
Ground beef (fat-free)...........500.0 g
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:
Trypticase Peptone (BD 211921)....30.0 g
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 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:
Hemin.............................50.0 mg
N NaOH.............................1.0 ml
Distilled water to...............100.0 ml
Autoclave at 121C for 15 minutes