DSMZ 212a . TREPONEMA RUMINIS MEDIUM

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

Culture medium recipe

Vol.
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Selected strain: Treponema ruminis DSM 103462

This strain uses the standard medium without modifications.

Equipment needed: Gassing station Hungate tubes Autoclave Filter Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 917 ml - -
Solution B 30 ml - -
Solution C 4 ml - -
Solution D 4 ml - -
Solution E 30 ml - -
Solution F 1 ml - -
Solution G 2 ml - -
Solution H 10 ml - -
Solution I 10 ml - -
1 Add and dissolve ingredients of solution A, adjust pH to 7.0 and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Solution B is prepared under 80% N2 and 20% CO2 gas atmosphere and autoclaved. Solutions C, D and H are autoclaved under 100% N2 gas atmosphere. Solutions E, F, G and I are prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to I are added to the sterile solution A in the sequence as indicated.
2 Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days . After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.545 4.007
MgCl2 x 6 H2O 0.33 g 0.36 1.77
NaCl 0.40 g 0.436 7.464
NH4Cl 0.40 g 0.436 8.155
CaCl2 x 2 H2O 0.05 g 0.055 0.371
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Clarified rumen fluid 50.00 ml - -
Trypticase peptone
(BD BBL)
1.00 g 1.091 -
Na2SO4 2.80 g 3.053 21.497
Vitamin K1 solution 1.00 ml - -
Volatile fatty acid mixture 3.10 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.5e-4 0.002
Distilled water 860.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Starch
(soluble)
0.4 g 100 -
Distilled water 4.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
D-Glucose 0.8 g 200 1110.12
Distilled water 4.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sugar mix 30 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate 0.5 g 250 2271.82
Distilled water 2.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Cysteine HCl x H2O 0.3 g 30 170.808
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
DL-Dithiothreitol
(DTT)
0.4 g 40 259.235
Distilled water 10.0 ml - -
Solution occurs in 263 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%)
10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
Solution occurs in 176 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
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.
Solution occurs in 12 other media (first defined in Medium 78).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin K1 0.1 ml - -
Ethanol
(95 %)
20.0 ml 950 20620.8
1 Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle.
Solution occurs in 4 other media (first defined in Medium 330).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Acetic acid 548.50 ml - -
Propionic acid 193.50 ml - -
Butyric acid 129.00 ml - -
n-Valeric acid 32.25 ml - -
iso-Butyric acid 32.25 ml - -
DL-2-Methylbutyric acid 32.25 ml - -
iso-Valeric acid 32.25 ml - -
Solution occurs in 1 other media (first defined in Medium 843).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Cellobiose 56.7 g 56.7 165.644
Sucrose 56.7 g 56.7 165.644
Maltose x H2O 60.0 g 60 175.287
Trehalose 63.3 g 63.3 184.926
D-Xylose 25.0 g 25 166.522
Distilled water 1000.0 ml - -
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 25.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes
Edit in Medium builder

Gas composition

  • 80 % N2
  • 20 % 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: 3
Treponema ruminis DSM 103462

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2SO4 2.778 19.556
Trehalose 1.885 5.507
Maltose x H2O 1.786 5.217
Sucrose 1.687 4.927
Cellobiose 1.687 4.927
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Cultivation metadata from 3 strains BacDive

pH 6 - 8 1 strain
phyla 1 3 strains
oxygen anaerobe 2 strains
oxygen obligate anaerobe 1 strain
temperature mesophilic 3 strains