DSMZ 666 . SUCCINICLASTICUM MEDIUM

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

Culture medium recipe

Vol.
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Equipment needed: Autoclave Hungate tubes Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Clarified rumen fluid 400.00 ml - -
K2HPO4 0.23 g 0.23 1.321
KH2PO4 0.23 g 0.23 1.69
NaCl 0.45 g 0.45 7.7
(NH4)2SO4 0.45 g 0.45 3.405
CaCl2 x 2 H2O 0.06 g 0.06 0.408
MgSO4 x 7 H2O 0.09 g 0.09 0.365
Indigocarmine 5.00 mg 0.005 0.011
NaHCO3 6.40 g 6.4 76.184
Disodium succinate 5.00 g 5 30.855
Yeast extract 5.00 g 5 -
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 600.00 ml - -
1 Dissolve ingredients (except bicarbonate, succinate, yeast extract, cysteine and sulfide), bring medium to the boil, then cool to room temperature under 100% CO2 gas atmosphere. Add the bicarbonate and equilibrate the medium with the CO2 gas to pH 6.8. Distribute under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add succinate, yeast extract, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 6.7 - 6.8, if necessary.
2 Note: Supplementing the medium with 1.50 g/l agar stimulates growth of strains after resuscitation from ampoules.
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: 05.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.7 - 6.8
Equipment needed: Autoclave, Centrifuge, 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: 9
Strains with modifications: 6

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 6.4 76.184
Disodium succinate 5 30.855
Yeast extract 5 -
NaCl 0.45 7.7
(NH4)2SO4 0.45 3.405
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Cultivation metadata from 9 strains BacDive

phyla 1 8 strains
oxygen anaerobe 9 strains
temperature mesophilic 9 strains