DSMZ 140 . RUMINICLOSTRIDIUM MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

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Selected strain: Clostridium polysaccharolyticum DSM 1801

This strain uses the standard medium without modifications.

Equipment needed: Hungate tubes Filter Autoclave Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.5 3.674
NaCl 1.00 g 1 17.112
(NH4)2SO4 0.50 g 0.5 3.784
MgSO4 x 7 H2O 0.10 g 0.1 0.406
CaCl2 x 2 H2O 0.10 g 0.1 0.68
K2HPO4 0.50 g 0.5 2.871
Clarified rumen fluid 300.00 ml - -
Sludge fluid , alternative
optional
300.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Na2CO3 2.50 g 2.5 23.588
Cellobiose 5.00 g 5 14.607
L-Cysteine HCl x H2O 0.25 g 0.25 1.423
Na2S x 9 H2O 0.25 g 0.25 1.041
Distilled water 700.00 ml - -
1 Dissolve ingredients (except carbonate, cellobiose and reducing agents), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Use either clarified rumen fluid or sludge fluid as supplement. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose, sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Cellobiose has to be sterilized by filtration. Adjust pH of complete medium to 6.8, if necessary.
Solution occurs in 8 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 4 g 4 -
Sludge 1000 ml - -
1 Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours. Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at 8-12°C in the dark.
Solution occurs in 27 other media (first defined in Medium 1310).
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.

Metadata

Last modified: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.8
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes
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Gas composition

  • 100 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 2
Clostridium polysaccharolyticum DSM 1801

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Cellobiose 5 14.607
Na2CO3 2.5 23.588
Yeast extract 1.2 -
NaCl 1 17.112
(NH4)2SO4 0.5 3.784
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Cultivation metadata from 2 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 2 strains
oxygen anaerobe 2 strains
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