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DSMZ

DSMZ 346 . DESULFALLAS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 346 #

ml
Equipment needed: Gassing station Hungate tubes Autoclave Filter Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 966.0 ml - -
Solution B 20.0 ml - -
Solution C 11.0 ml - -
Solution D 10.0 ml - -
Solution E 1.0 ml - -
Solution F 1.0 ml - -
Solution G 1.0 ml - -
Solution H 1.7 ml - -
1 Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic. Thereafter, dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Anoxic stock solutions C, D, E, F, and H are prepared under 100% N2 gas atmosphere and solution G under 80% N2 and 20% CO2 gas atmosphere. Filter sterilize solutions D, F and G. To complete the medium appropriate amounts of solutions B to H are added to the sterile solution A in the sequence as indicated. Adjust pH of the complete medium to 7.2 - 7.5, if necessary.

Solution A #

966
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.106 21.864
KH2PO4 0.20 g 0.207 1.521
NH4Cl 0.30 g 0.311 5.806
CaCl2 x 2 H2O 0.15 g 0.155 1.056
MgCl2 x 6 H2O 0.40 g 0.414 2.037
NaCl 1.00 g 1.035 17.714
Trace element solution SL-10 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.2e-4 0.002
Distilled water 965.00 ml - -

Solution B #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 1 g 50 595.188
Distilled water 20 ml - -

Solution C #

11
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ethanol 1 ml - -
Distilled water 10 ml - -

Solution D #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-benzoate 0.7 g 70 485.764
Distilled water 10.0 ml - -

Solution E #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Clarified rumen fluid 1 ml - -

Solution F #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution (10x) 1 ml - -

Solution G #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-dithionite solution (5% w/v) 1 ml - -

Solution H #

2
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 50.0 mg 25 104.088
Distilled water 1.7 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.

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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 307 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 - -

Na-dithionite solution (5% w/v) #

10
Solution occurs in 7 other media (first defined in Medium 829).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH
(1 N) 		1.0 ml - -
Na2S2O4 0.5 g 50 287.175
Distilled water 9.0 ml - -
1 Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.

Metadata

Last modified: 11.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2 - 7.5
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes
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Gas composition

  • 100 % N2

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]
Na2SO4 2.964 20.87
NaCl 0.988 16.909
Na-benzoate 0.692 4.8
MgCl2 x 6 H2O 0.395 1.944
NH4Cl 0.296 5.542
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain