ATCC medium: 1367 Modified OTI medium
Polypeptone Peptone (BD 211910)..........5.0 g
Heart infusion broth.....................5.0 g
Yeast Extract (BD 212750)................5.0 g
Pectin (Sigma P9135).....................0.8 g
Glucose..................................0.8 g
Starch...................................0.8 g
Sucrose..................................0.8 g
Maltose..................................0.8 g
Sodium pyruvate..........................0.8 g
Ribose...................................0.8 g
K2HPO4 ...................................2.0 g
NaCl.....................................5.0 g
MgSO4 ....................................0.1 g
L-Cysteine . HCl.........................0.68 g
Clarified rumen fluid..................500.0 ml
Agar.....................................1.6 g
Distilled water........................500.0 ml
Adjust pH to 7.0. Prepare medium under anaerobic conditions and tube in butyl rubber-stoppered tubes under oxygen-free nitrogen. Add inactivated sterile rabbit serum to give final concentration in each tube of 5%.
Add filter-sterilized thiamine pyrophosphate to each tube to give a final concentration of 0.75 mg/100 ml.
Subculture with a Pasteur pipette (20- 30 drops). Gas phase for culture system should be oxygen-free nitrogen or a mixture of 90% N2 and 10% CO2.