Medium data

Search for medium no. = [1201].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1201 TREPONEMA BRYANTII MEDIUM

Rumen fluid, clarified (see below)	`300.0`	ml
CaCl₂·2H₂O	`0.12`	g
MgSO₄·7H₂O	`0.18`	g
KH₂PO₄	`0.45`	g
K₂HPO₄	`0.45`	g
NaCl	`0.9`	g
(NH₄)₂SO₄	`0.9`	g
0.05% Resazurin solution	`1.0`	ml
L-Cysteine·HCl·H₂O	`1.0`	g
Na₂CO₃	`1.5`	g
Glucose	`2.0`	g
Distilled water	`700.0`	ml

Dissolve ingredients (except cysteine, carbonate and glucose) and sparge medium with 100% CO₂ gas for 30- 45 min to make it anoxic. Add cysteine and adjust pH with KOH to 6.5. Distribute medium under 100% CO₂ gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add glucose from an anoxic stock solution prepared under 100% N₂ gas atmosphere and sterilized by filtration and carbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas atmosphere. Final pH of the medium should be 6.7 - 7.0. To prepare the clarified rumen fluid, preheat rumen content at 120C for 15 min. Use the supernatant after centrifuging at 25,000 x g for 15 min.