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DSMZ

DSMZ 795a . LEBETIMONAS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 795a #

ml
Equipment needed: Autoclave Filter Gassing station Serum bottle Water bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
Synthetic seawater (2 x conc.) 500.00 ml - -
NH4Cl 1.25 g 1.233 23.046
KH2PO4 0.50 g 0.493 3.623
NiCl2 x 6 H2O
(0.1% w/v) 		3.00 ml 0.003 0.023
Modified Wolin's mineral solution 10.00 ml - -
Na-formate 0.20 g 0.197 2.9
Tryptone
(BD Bacto) 		1.00 g 0.986 -
Yeast extract
(OXOID) 		1.00 g 0.986 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 2.00 g 1.972 23.479
Wolin's vitamin solution (10x) 1.00 ml - -
Sulfur , powdered 10.00 g 9.862 307.513
Na2S x 9 H2O 0.50 g 0.493 2.053
Distilled water 500.00 ml - -
1 Dissolve ingredients (except bicarbonate, vitamins, sulfur, and sulfide), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic and adjust pH to 5.5 with H2SO4. Dispense under the same gas atmosphere in suitable culture vessels (e.g. 20 ml of the medium in 100 ml serum bottles) and autoclave. Steam sulfur for 3 hours on each of 3 successive days (see medium 35). Aseptically mix the sterilized sulfur with the sterile medium while retaining anoxic conditions. Add vitamins from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to inoculation add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. Check pH and adjust to 5.5, if necessary.
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

Synthetic seawater (2 x conc.) #

1000
Solution occurs in 3 other media (first defined in Medium 795).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 55.40 g 55.4 947.981
SrCl2 x 6 H2O 0.03 g 0.03 0.113
MgSO4 x 7 H2O 14.00 g 14 56.801
KI 0.10 mg 1.0e-4 3.9e-4
MgCl2 x 6 H2O 11.00 g 11 54.106
Na3-citrate 20.00 mg 0.02 0.077
KCl 1.30 g 1.3 17.438
CaCl2 x 2 H2O 1.50 g 1.5 10.203
NaBr 0.20 g 0.2 1.944
H3BO3 0.06 g 0.06 0.97
Distilled water 1000.00 ml - -

Modified Wolin's mineral solution #

1000
Solution occurs in 166 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 24.07.23
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 5.5
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle, Water bath
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Gas composition

  • 80 % H2
  • 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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 9.862 307.513
MgCl2 x 6 H2O 5.424 26.68
NaHCO3 1.972 23.479
NH4Cl 1.233 23.046
Tryptone 0.986 -
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Cultivation metadata from 1 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain