DSMZ 1455 . ALTERNATIVE RUMEN BACTERIA MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.23 g 0.23 1.321
KH2PO4 0.23 g 0.23 1.69
NaCl 0.45 g 0.45 7.7
(NH4)2SO4 0.45 g 0.45 3.405
CaCl2 x 2 H2O 0.06 g 0.06 0.408
MgSO4 x 7 H2O 0.09 g 0.09 0.365
Brain heart infusion
(BD BBL)
6.00 g 6 -
Yeast extract 5.00 g 5 -
Volatile fatty acid mixture 3.10 ml - -
Haemin solution
(0.05% w/v)
2.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
NaHCO3 6.40 g 6.4 76.184
Disodium succinate 5.00 g 5 30.855
Wolin's vitamin solution 10.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.3 1.708
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, succinate, vitamins, cysteine and sulfide, then sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the bicarbonate and equilibrate the medium with the CO2 gas to reach a pH between 6.5 and 7.0. Dispense medium under 100% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add succinate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Prior to use adjust pH of complete medium to 6.6 - 6.8, if necessary.
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 15 other media (first defined in Medium 78).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Haemin 50 mg 0.5 0.767
NaOH
(1 N)
1 ml - -
Distilled water 100 ml - -
1 Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated.
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 - -

Metadata

Last modified: 22.02.22
Source: DSMZ
Medium type: Complex medium
Final pH: 6.6 - 6.8
Equipment needed: Autoclave, 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: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 6.4 76.184
Brain heart infusion 6 -
Disodium succinate 5 30.855
Yeast extract 5 -
NaCl 0.45 7.7
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