MediaDive DSMZ
Content
Media Solutions Ingredients Strains Steps Gas
Search
Medium finder Solution finder Taxonomy search Isolation sources
Tools
Medium builder Unit converter Compare media Compare solutions Prediction SOON
DSMZ

DSMZ 383 . DESULFOBACTERIUM MEDIUM

What is this page?
* Medium for Desulfobacula phenolica DSM 3384

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
Equipment needed: Autoclave Filter Gassing station Hungate tubes
Final gas composition: 80 % N2 20 % CO2 read more
Final pH: 7.0 - 7.2
Select a strain (optional):

ml
Compound Amount Unit
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
Na2SO4 3.00 g
KH2PO4 0.20 g
NH4Cl 0.30 g
NaCl 21.00 g
MgCl2 x 6 H2O 3.00 g
KCl 0.50 g
CaCl2 x 2 H2O 0.15 g
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Sodium resazurin
(0.1% w/v) 		0.50 ml
Distilled water 950.00 ml

Solution B #

20
Compound Amount Unit
Na2CO3 1 g
Distilled water 20 ml

Solution C #

20
Compound Amount Unit
Na-pyruvate* 2.5 g
Na-benzoate 0.4 g
Distilled water 20.0 ml

Solution D #

1
Compound Amount Unit
Wolin's vitamin solution (10x) 1 ml
Seven vitamins solution 1 ml

Solution E #

10
Compound Amount Unit
Na2S x 9 H2O 0.4 g
Distilled water 10.0 ml

Selenite-tungstate solution #

1000
Compound Amount Unit
NaOH 0.5 g
Na2SeO3 x 5 H2O 3.0 mg
Na2WO4 x 2 H2O 4.0 mg
Distilled water 1000.0 ml

Trace element solution SL-10 #

1000
Compound Amount Unit
HCl
(25%) 		10.0 ml
FeCl2 x 4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2 x 4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2 x 6 H2O 190.0 mg
CuCl2 x 2 H2O 2.0 mg
NiCl2 x 6 H2O 24.0 mg
Na2MoO4 x 2 H2O 36.0 mg
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
Compound Amount Unit
Biotin 20 mg
Folic acid 20 mg
Pyridoxine hydrochloride 100 mg
Thiamine HCl 50 mg
Riboflavin 50 mg
Nicotinic acid 50 mg
Calcium D-(+)-pantothenate 50 mg
Vitamin B12 1 mg
p-Aminobenzoic acid 50 mg
(DL)-alpha-Lipoic acid 50 mg
Distilled water 1000 ml

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

10
Compound Amount Unit
NaOH
(1 N) 		1.0 ml
Na2S2O4 0.5 g
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.

Seven vitamins solution * #

1000
Compound Amount Unit
Vitamin B12 100 mg
p-Aminobenzoic acid 80 mg
D-(+)-biotin 20 mg
Nicotinic acid 200 mg
Calcium pantothenate 100 mg
Pyridoxine hydrochloride 300 mg
Thiamine-HCl x 2 H2O 200 mg
Distilled water 1000 ml

Clarified rumen fluid * #

1000
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.