Medium data

Search for medium no. = [1149].

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

1149 METHANOMETHYLOPHILUS ALVI MEDIUM

KH₂PO₄	`0.5`	g
MgSO₄·7H₂O	`0.4`	g
NaCl	`5.0`	g
NH₄Cl	`1.0`	g
CaCl₂·2H₂O	`0.05`	g
Sodium acetate·3H₂O	`2.7`	g
Yeast extract	`2.0`	g
Bacto peptone (BD-Difco)	`2.0`	g
Casamino acids	`2.0`	g
Trypticase peptone	`2.0`	g
Trace element solution (see Medium No. 439)	`1.0`	ml
L-Cysteine·HCl·H₂O	`0.5`	g
Selenite-tungstate solution (see Medium No. 431)	`1.0`	ml
Resazurin	`1.0`	mg
Distilled water	`980.0`	ml

Mix components and adjust pH to 7.0. Bring to a boil and cool down under a H₂-CO₂ (4:1, v/v) gas stream. Distribute the medium into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, aseptically and anaerobically add per liter the following solutions (autoclaved or *filter-sterilized):

8% NaHCO₃ solution*	`20.0`	ml
50% (v/v) Methanol	`6.3`	ml
5% Na₂S·9H₂O solution	`8.0`	ml

Pressurize inoculated bottles to 100 kPa H₂-CO₂ (4:1, v/v).
Comment:
For cultivation of JCM 31474, it needs to add an cultural filtrate of Eggerthella sp., Eggerthella lenta or rumen fluid in the medium. One example of the preparation of the bacterial cultural filtrate is as follows: cultivate Eggerthella lenta JCM 9979 in the same medium and filtrate the culture when it grows significantly (generally it takes two to three days). It can be autoclaved to make sterile certainly. Then, add 10 to 30% volume of the culture filtrate in the growth medium.