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DSMZ

DSMZ 212a . TREPONEMA RECTALE MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 212a #

ml
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 to 30% of their volume and autoclave. Solution B is prepared under 80% N2 and 20% CO2 gas atmosphere and autoclaved. Solutions C, D, and H are autoclaved under a 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.

Solution A #

917
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 - -

Solution B #

30
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -

Solution C #

4
Compound Amount Unit Conc. [g/L] Conc. [mM]
Starch
(soluble) 		0.4 g 100 -
Distilled water 4.0 ml - -

Solution D #

4
Compound Amount Unit Conc. [g/L] Conc. [mM]
D-Glucose 0.8 g 200 1110.12
Distilled water 4.0 ml - -

Solution E #

30
Compound Amount Unit Conc. [g/L] Conc. [mM]
Sugar mix 30 ml - -

Solution F #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -

Solution G #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate 0.5 g 250 2271.82
Distilled water 2.0 ml - -

Solution H #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Cysteine HCl x H2O 0.3 g 30 170.808
Distilled water 10.0 ml - -

Solution I #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
DL-Dithiothreitol
(DTT) 		0.4 g 40 259.235
Distilled water 10.0 ml - -

Trace element solution SL-10 #

1000
Solution occurs in 267 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.

Selenite-tungstate solution #

1000
Solution occurs in 180 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 - -

Clarified rumen fluid #

1000
Solution occurs in 28 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.

Vitamin K1 solution #

20
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.

Volatile fatty acid mixture #

1000
Solution occurs in 4 other media (first defined in Medium 330).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Acetic acid 548.50 ml 573.182 9545.09
Propionic acid 193.50 ml 192.145 2593.76
Butyric acid 129.00 ml - -
n-Valeric acid 32.25 ml 30.283 296.512
iso-Butyric acid 32.25 ml 30.638 347.719
DL-2-Methylbutyric acid 32.25 ml - -
iso-Valeric acid 32.25 ml 30.025 293.986

Sugar mix #

1000
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 - -

Seven vitamins solution #

1000
Solution occurs in 77 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: 28.03.23
Source: DSMZ
Synonyms: TREPONEMA RUMINIS MEDIUM, TREPONEMA RECTALIS MEDIUM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes
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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: 4
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2SO4 2.778 19.556
Trehalose 1.884 5.504
Maltose x H2O 1.786 5.217
Acetic acid 1.763 29.355
Cellobiose 1.688 4.93
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Cultivation metadata from 4 strains BacDive

pH 6 - 8 1 strain
phyla 1 4 strains
oxygen anaerobe 2 strains
oxygen obligate anaerobe 1 strain
temperature mesophilic 3 strains