DSMZ 795a . LEBETIMONAS MEDIUM

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

Culture medium recipe

Vol.
×
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 1.25 23.369
KH2PO4 0.50 g 0.5 3.674
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 0.2 2.941
Tryptone
(BD Bacto)
1.00 g 1 14.069
Yeast extract
(OXOID)
1.00 g 1 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
NaHCO3 2.00 g 2 23.808
Wolin's vitamin solution 10.00 ml - -
Sulfur , powdered
(powdered)
10.00 g 10 311.818
Na2S x 9 H2O 0.50 g 0.5 2.082
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 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. 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. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Solution occurs in 170 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.
Solution occurs in 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -
Solution occurs in 3 other media (first defined in Medium ).
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 - -

Metadata

Last modified: 06.04.22
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 10 311.818
MgCl2 x 6 H2O 5.5 27.053
NaHCO3 2 23.808
NH4Cl 1.25 23.369
Tryptone 1 14.069
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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