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320. CLOSTRIDIUM CELLULOVORANS MEDIUM

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.00 g 0.978 4.287
2 1.00 g 0.978 18.292
3 0.50 g 0.489 6.562
4 0.50 g 0.489 1.985
5
(BD BBL)
0.50 g 0.489 -
6 0.50 g 0.489 -
7 Clarified rumen fluid , either 20.00 ml - -
8 Sludge fluid , or 20.00 ml - -
9 Trace element solution SL-10 , or 1.00 ml - -
10
(0.1% w/v)
0.50 ml 4.9e-4 0.002
11 0.15 g 0.147 0.836
12 1.00 g 0.978 9.232
13 5.00 g 4.892 14.293
14 0.15 g 0.147 0.611
15 1000.00 ml 976.719 54216.1
16 , some strains can be adapted to cellulose
(powder MN 301, MACHEREY-NAGEL)
10.00 g/l 10 -
Instructions:
  1. Dissolve ingredients (except cysteine, carbonate, cellobiose and sulfide), bring medium to the boil, then cool to room temperature under 80% N2 and 20% CO2 gas mixture and add cysteine. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Sterilize cellobiose separately by filtration under 100% N2 gas. Adjust pH of the complete medium to 7.0, if necessary.
  2. Use either #7 or #8 for medium
  3. Note: Some strains can be adapted to cellulose as substrate using 10.0 g/l cellulose powder MN 301 (MACHEREY-NAGEL).
Solution occurs in 8 other media (first defined in medium 119 )
Equipment needed: Autoclave Anaerobic workbench Centrifuge
Instructions:
  1. Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at room temperature in the dark.
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.
Solution occurs in 32 other media (first defined in medium 1310 )
Equipment needed: Centrifuge Filter Autoclave
Instructions:
  1. Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 minand 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

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Anaerobic workbench, Autoclave, Centrifuge, Filter, Hungate tubes

Associated strains

All strains for this medium: 1
Strains with modifications: 1

Gas composition

  • 100 % N2

Light

1 strain needs light for growth. Select a strain for more details.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 977.686 54269.8
Cellulose 10 -
Cellobiose 4.892 14.293
NH4Cl 0.978 18.292
Na2CO3 0.978 9.232
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
oxygen obligate anaerobe 1 strain
temperature mesophilic 1 strain

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