1310 .
BUTYRIVIBRIO M2 MEDIUM
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Equipment needed:
Autoclave
Centrifuge
Gassing station
Hungate tubes
Final gas composition:
100 % CO2
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Final pH:
6.5 - 6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Casitone
(BD Bacto) |
10.00 | g | ||
|
|
Yeast extract
(BD Bacto) |
2.50 | g | ||
|
|
D-Glucose | 2.00 | g | ||
|
|
Cellobiose | 2.00 | g | ||
|
|
Maltose | 2.00 | g | ||
|
|
Na-DL-lactate | 5.00 | g | ||
|
|
Clarified rumen fluid | 200.00 | ml | ||
|
|
K2HPO4 | 0.05 | g | ||
|
|
KH2PO4 | 0.05 | g | ||
|
|
(NH4)2SO4 | 0.10 | g | ||
|
|
NaCl | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.01 | g | ||
|
|
MgSO4 x 7 H2O | 0.01 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na2CO3 | 4.00 | g | ||
|
|
L-Cysteine HCl x H2O | 0.30 | g | ||
|
|
Na2S x 9 H2O | 0.30 | g | ||
|
|
Distilled water | 800.00 | ml | ||
| 1 Dissolve ingredients (except carbonate, cysteine and sulfide), boil medium for 1 min, then cool to room temperature under 100% CO2 gas atmosphere. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Add cysteine and sulfide form sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of the complete medium to 6.5 - 6.8, if necessary. | |||||
Clarified rumen fluid #
1000
| 1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C. |