培養(yǎng)基配方
Clarified rumen fluid 300.00 ml
CaCl2 x 2 H2O 0.12 g
MgSO4 x 7 H2O 0.18 g
KH2PO4 0.45 g
K2HPO4 0.45 g
NaCl 0.90 g
(NH4)2SO4 0.90 g
Sodium resazurin (0.1% w/v) 0.50 ml
L-Cysteine HCl x H2O 1.00 g
Na2CO3 1.50 g
D-Glucose 2.00 g
Distilled water 700.00 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. Therafter, 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.
For DSM 11370: Plus cellobiose (0.1%), maltose (0.1%) and soluble starch (0.1%);
anaerobic
For DSM 11371: Plus cellobiose (0.1%), maltose (0.1%) and soluble starch (0.1%);
anaerobic; grows within 12 hours, lyses quickly;
Clarified rumen fluid (from medium 1310)
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.
