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DSMZ

DSMZ 78c . CHOPPED MEAT MEDIUM FOR FRETIBACTERIUM SP.

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

Culture medium recipe

Vol.
×

Main sol. 78c #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ground beef
(fat free) 		500.0 g 586.166 -
NaOH
(1 N) 		25.0 ml - -
Casein peptone
(BD BACTO) 		30.0 g 35.17 -
Yeast extract 5.0 g 5.862 -
K2HPO4 5.0 g 5.862 33.654
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.9e-4 0.002
L-Cysteine HCl x H2O 0.5 g 0.586 3.337
Na-fumarate 6.0 g 7.034 43.953
Na-formate 6.0 g 7.034 103.43
Clarified rumen fluid , clarified 50.0 ml - -
Horse serum
(inactivated, INVITROGEN) 		50.0 ml - -
Fatty acid mixture 20.0 ml - -
Wolin's vitamin solution (10x) 1.0 ml - -
Haemin solution
(0.05% w/v) 		5.0 ml - -
Vitamin K1 solution
(0.05% w/v) 		2.0 ml - -
DL-Dithiothreitol
(DTT) 		0.3 g 0.352 2.279
Distilled water 700.0 ml - -
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 rom sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration.

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.

Fatty acid mixture #

1000
Solution occurs in 6 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Isobutyric acid 23 ml - -
DL-2-Methylbutyric acid 27 ml - -
Valeric acid 27 ml - -
Isovaleric acid 27 ml - -
Distilled water 896 ml - -
1 Adjust pH to 7.5 with concentrated NaOH.

Haemin solution #

100
Solution occurs in 15 other media (first defined in Medium 78).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Haemin 50 mg 0.5 0.767
NaOH
(1 N) 		1 ml - -
Distilled water 100 ml - -
1 Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated.

Vitamin K1 solution #

20
Solution occurs in 12 other media (first defined in Medium 78).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin K1 0.1 ml - -
Ethanol
(95 %) 		20.0 ml 950 20620.8
1 Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 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: 21.06.23
Source: DSMZ
Synonyms: CHOPPED MEAT MEDIUM FOR FERTIBACTERIUM SP.
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Ground beef 586.166 -
Casein peptone 35.17 -
Na-formate 7.034 103.43
Na-fumarate 7.034 43.953
K2HPO4 5.862 33.654
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Cultivation metadata from 1 strains BacDive

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