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DSMZ

DSMZ 383 . DESULFOBACTERIUM MEDIUM

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

Culture medium recipe

Vol.
×
Medium for Desulfobacula phenolica DSM 3384

* Modification: The vitamin solution of medium 120 is replaced with 1.00 ml/l of seven vitamins solution of medium 503. Replace pyruvate with 0.40 g/l Na-benzoate and add 10.00 ml/l clarified rumen fluid (see medium 1310) as growth supplement. Initiation of growth requires addition of freshly prepared sodium dithionite.

Growth conditions: 7-14 days anaerobic 28 °C

Main sol. 383 #

ml
Equipment needed: Hungate tubes Autoclave Gassing station Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952.0 ml - -
Solution B 20.0 ml - -
Solution C 20.0 ml - -
Solution D 1.0 ml - -
Solution E 10.0 ml - -
Na-dithionite solution (5% w/v)* 0.5 ml - -
Clarified rumen fluid* 10.0 ml - -
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed in anoxic cultivation vials and autoclaved under the same gas atmosphere. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are prepared under 100% N2 gas and filter-sterilized. Solution E is autoclaved under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.0 – 7.2.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the beginning. For transfers use 5 - 10% inoculum. Incubate all strains in the dark.

Solution A #

952
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.151 22.186
KH2PO4 0.20 g 0.21 1.544
NH4Cl 0.30 g 0.315 5.891
NaCl 21.00 g 22.059 377.461
MgCl2 x 6 H2O 3.00 g 3.151 15.5
KCl 0.50 g 0.525 7.045
CaCl2 x 2 H2O 0.15 g 0.158 1.072
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.3e-4 0.002
Distilled water 950.00 ml - -

Solution B #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1 g 50 471.752
Distilled water 20 ml - -

Solution C #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate* 2.5 g 125 1135.91
Na-benzoate* 0.4 g - -
Distilled water 20.0 ml - -

Solution D #

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

Solution E #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.4 g 40 166.54
Distilled water 10.0 ml - -

Selenite-tungstate solution #

1000
Solution occurs in 178 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 - -

Trace element solution SL-10 #

1000
Solution occurs in 265 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.

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 - -

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

1000
Solution occurs in 7 other media (first defined in Medium 829).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 50 g 50 595.188
Na2S2O4 50 g 50 287.175
Distilled water 1000 ml - -
1 Dissolve NaHCO3 in water and make the solution anoxic by sparging with 80% N2 and 20% CO2 gas mixture. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in the dark and refrigerated. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.

Seven vitamins solution * #

1000
Solution occurs in 78 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 - -

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.

Metadata

Last modified: 08.09.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 7.0 - 7.2
Equipment needed: Autoclave, 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: 31
Strains with modifications: 27
Desulfobacula phenolica DSM 3384

+  0.5 ml Na-dithionite solution (5% w/v)
+  1.0 ml Seven vitamins solution
+  10.0 ml Clarified rumen fluid
-  Wolin's vitamin solution (10x)
  0.4 g Na-benzoate
  Na-pyruvate

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
NaCl 20.937 358.268
Na2SO4 2.991 21.058
MgCl2 x 6 H2O 2.991 14.712
Na2CO3 0.997 9.407
KCl 0.499 6.687
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Cultivation metadata from 31 strains BacDive

pH 6 - 8 8 strains
pH 4 - 6 3 strains
pH 8 - 10 2 strains
phyla 5 31 strains
nutrition chemoorganotroph 1 strain
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