DSMZ 159 . TREPONEMA MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Clarified rumen fluid 300.00 ml - -
CaCl2 x 2 H2O 0.12 g 0.12 0.816
MgSO4 x 7 H2O 0.18 g 0.18 0.73
KH2PO4 0.45 g 0.45 3.307
K2HPO4 0.45 g 0.45 2.584
NaCl 0.90 g 0.9 15.4
(NH4)2SO4 0.90 g 0.9 6.811
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
L-Cysteine HCl x H2O 1.00 g 1 5.694
Na2CO3 1.50 g 1.5 14.153
D-Glucose 2.00 g 2 11.101
Distilled water 700.00 ml - -
1 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.
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: 26.10.22
Source: DSMZ
Synonyms: TREPONEMA BRYANTII MEDIUM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.7 - 7.0
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: 4
Strains with modifications: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
D-Glucose 2 11.101
Na2CO3 1.5 14.153
L-Cysteine HCl x H2O 1 5.694
NaCl 0.9 15.4
(NH4)2SO4 0.9 6.811
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Cultivation metadata from 4 strains BacDive

phyla 3 4 strains
oxygen anaerobe 4 strains
temperature mesophilic 4 strains