DSMZ 201 . DESULFONEMA LIMICOLA MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Gassing station Hungate tubes Autoclave Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 920 ml - -
Solution B 1 ml - -
Solution C 30 ml - -
Solution D 10 ml - -
Solution E 1 ml - -
Solution F 10 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.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.261 22.957
NaCl 15.00 g 16.304 278.993
MgCl2 x 6 H2O 2.20 g 2.391 11.762
CaCl2 x 2 H2O 0.15 g 0.163 1.109
KCl 0.50 g 0.543 7.29
KH2PO4 0.20 g 0.217 1.597
NH4Cl 0.30 g 0.326 6.096
Sodium resazurin
(0.1% w/v)
0.50 ml 5.4e-4 -
Distilled water 920.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace element solution SL-10 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-acetate x 3 H2O 2.5 g 250 -
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Disodium succinate 0.1 g 100 -
Distilled water 1.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Clarified rumen fluid , clarified 20 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.4 g 40 166.54
Distilled water 10.0 ml - -
Solution occurs in 263 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.
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 - -
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: 19.08.22
Source: DSMZ
Medium type: Complex medium
Final pH: 7.6
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
Edit in Medium builder

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 14.97 256.161
Na2SO4 2.994 21.079
Na-acetate x 3 H2O 2.495 -
MgCl2 x 6 H2O 2.196 10.8
Na2CO3 1.497 14.124
Show more

Cultivation metadata from 0 strains BacDive

phyla 1 1 strain