ATCC medium: 1357 NOS Spirochete medium
Brain Heart Infusion Broth (BD 237500)..............12.5 g
Trypticase (BD 211921)..............................10.0 g
Yeast extract........................................2.5 g
Sodium thioglycollate................................0.5 g
L-Cysteine . HCl.....................................1.0 g
L-Asparagine.........................................0.25 g
Glucose..............................................2.0 g
Agar, Noble (BD 214230), for semi-solid medium.......3.0 g
Distilled water to...................................1.0 L
Supplements: 0.2% (w/v) Thiamine pyrophosphate....................3.0 ml
Volatile Fatty Acids (see below).....................2.0 ml
10% Sodium bicarbonate..............................20.0 ml
Normal rabbit serum.................................20.0 ml
Autoclave at 121C for 15 min. Filter-sterilize all supplements.
Prepare and dispense medium anaerobically under an atmosphere of 80% N2, 10% H2, 10% CO2.
Volatile Fatty Acids:
Isobutyric acid..............0.5 ml
D,L-2-Methylbutyric acid.....0.5 ml
Isovaleric acid..............0.5 ml
Valeric acid.................0.5 ml
0.1 N KOH..................100.0 ml