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78c. CHOPPED MEAT MEDIUM FOR FERTIBACTERIUM SP.

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1
(fat free)
500.0 g 580.046 -
2
(1 N)
25.0 ml 61.775 1544.49
3
(BD BACTO)
30.0 g 34.803 -
4 5.0 g 5.8 -
5 5.0 g 5.8 33.302
6
(0.1% w/v)
0.5 ml 5.8e-4 0.002
7 0.5 g 0.58 3.303
8 6.0 g 6.961 43.494
9 6.0 g 6.961 102.35
10 Clarified rumen fluid 50.0 ml - -
11
(INVITROGEN)
50.0 ml - -
12 Fatty acid mixture 20.0 ml - -
13 Vitamin solution 10.0 ml - -
14 Haemin solution 5.0 ml - -
15 Vitamin K1 solution 2.0 ml - -
16 0.3 g 0.348 2.256
17 700.0 ml 810.609 44995.6
Instructions:
  1. Use lean beef or horse meat. Remove fat and connective tissue before grinding. Mix meat, water and NaOH, then boil for 15 min with stirring. Cool to room temperature, skim fat off surface, and filter, retaining both meat particles and filtrate. To the filtrate add Casitone, yeast extract, hydrogen phosphate and resazurin, then add water to a final volume of 700 ml. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add cysteine and adjust pH to 7.0. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes containing meat particles (use ca. 7 ml liquid medium for 1 - 2 g meat particles), then autoclave at 121°C for 20 min. After autoclaving add fumarate, formate, rumen fluid, horse serum, fatty acids, vitamins, haemin, vitamin K1 and DTT from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration.
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.
Solution occurs in 10 other media (first defined in medium 119 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 23 ml - -
2 27 ml - -
3 27 ml 22.989 225.092
4 27 ml - -
5 1000 ml 904.173 50189.2
6 , required for pH adjustment
(2 N)
x - -
Instructions:
  1. Adjust pH to 7.0 - 7.3 with 2 N NaOH.
Solution occurs in 366 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.0 mg 0.002 0.008
2 2.0 mg 0.002 0.005
3 10.0 mg 0.01 0.049
4 5.0 mg 0.005 0.015
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 0.01
8 0.1 mg 1.0e-4 6.3e-5
9 5.0 mg 0.005 0.036
10 5.0 mg 0.005 0.024
11 1000.0 ml 998.207 55408.8
Solution occurs in 7 other media (first defined in medium 78 )
Equipment needed: Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2 50 mg 0.5 -
3
(1 N)
1 ml 21.3 532.539
4 100 - -
Instructions:
  1. Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated.
Solution occurs in 2 other media (first defined in medium 104 )
Equipment needed: Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2 0.1 ml - -
3
(95 %)
20.0 ml 750.5 16291
Instructions:
  1. Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle.

Metadata

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

Associated strains

All strains for this medium: 1

Gas composition

  • 100 % N2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 843.168 46802.9
Ground beef 580.046 -
NaOH 61.898 1547.58
Casitone peptone 34.803 -
Sodium formate 6.961 102.35
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Cultivation metadata from 1 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
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