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DSMZ

DSMZ 201 . DESULFONEMA LIMICOLA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 201 #

ml
Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 930 ml - -
Solution B 1 ml - -
Solution C 30 ml - -
Solution D 10 ml - -
Solution E 1 ml - -
Solution F 1 ml - -
Solution G 20 ml - -
Solution H 10 ml - -
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to reach a pH of around 6, then distributed under same gas atmosphere into anoxic Hungate-type tubes and autoclaved.
2 Solutions B, D, E, G, and H are autoclaved separately under 100% N2 gas. Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution F is prepared under 100% N2 gas and sterilized by filtration. 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 7.6, if necessary.
3 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution freshly prepared under N2 and filter-sterilized) just before inoculation may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
4 Cultures of Desulfonema spp. show usually improved growth in media containing an artificial light sediment of aluminum phosphate. Alternatively, a viscous medium prepared with 1 - 2 g washed agar per liter can be used (Widdel et al., Arch. Microbiol. 134:286-294 (1983)): Agar (Difco) is washed three times for one hour with distilled water at room temperature. Finally, the agar is suspended (c. 20.0 g/l) in distilled water and autoclaved. 50 ml of the hot agar is added to the hot sterile medium part A (when preparing part A calculated for 1 liter, subtract the volume of the agar solution). After cooling to room temperature complete the medium by adding part B to I. For mass cultures the aluminum phosphate as artificial sediment can be used.

Solution A #

930
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.226 22.711
NaCl 15.00 g 16.129 275.993
MgCl2 x 6 H2O 2.20 g 2.366 11.636
CaCl2 x 2 H2O 0.15 g 0.161 1.097
KCl 0.50 g 0.538 7.212
KH2PO4 0.20 g 0.215 1.58
NH4Cl 0.30 g 0.323 6.031
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.4e-4 0.002
Distilled water 930.00 ml - -

Solution B #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace element solution SL-10 1 ml - -

Solution C #

30
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -

Solution D #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-acetate x 3 H2O 2.5 g 250 -
Distilled water 10.0 ml - -

Solution E #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Disodium succinate 0.1 g 100 617.094
Distilled water 1.0 ml - -

Solution F #

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

Solution G #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
Clarified rumen fluid , clarified 20 ml - -

Solution H #

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

Metadata

Last modified: 06.09.22
Source: DSMZ
Medium type: Complex medium
Final pH: 7.6
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: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 14.955 255.906
Na2SO4 2.991 21.058
Na-acetate x 3 H2O 2.493 -
MgCl2 x 6 H2O 2.193 10.789
Na2CO3 1.496 14.11
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Cultivation metadata from 0 strains BacDive

phyla 1 1 strain