DSMZ 78a . CHOPPED MEAT MEDIUM FOR TREPONEMA SP.

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Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station
1 To the ready medium, add 50 ml/l amino acid solution and 5 ml/l vitamin solution. These solutions must not be prepared under anaerobic conditions.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 78 1000 ml - -
Vitamin solution 5 ml - -
Amino acid solution (filter sterilized) 50 ml - -
Equipment needed: Anaerobic workbench Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Histidine 0.6 g 12 77.345
L-Serine 0.5 g 10 95.157
L-Glutamine 0.7 g 14 95.799
Distilled water 50.0 ml - -
Solution occurs in 15 other media (first defined in Medium 461).
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 50 mg 0.05 0.037
Pantothenic acid 50 mg 0.05 0.228
Riboflavin 50 mg 0.05 0.133
Pyridoxamine hydrochloride 10 mg 0.01 0.049
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Nicotinic acid 25 mg 0.025 0.203
Nicotine amide 25 mg 0.025 0.205
alpha-lipoic acid 50 mg 0.05 0.242
p-Aminobenzoic acid 50 mg 0.05 0.365
Thiamine-HCl x 2 H2O 50 mg 0.05 0.134
Distilled water 1000 ml - -
1 Stir for some hours, filter sterilize the solution.
Solution occurs in 4 other media (first defined in Medium 78).
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ground beef
(fat free)
500.0 g 500 -
Distilled water 1000.0 ml - -
NaOH
(1 N)
25.0 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 water to a final volume of 1000 ml, and then add:
Casitone 30.0 g 30 -
Yeast extract 5.0 g 5 -
K2HPO4 5.0 g 5 28.707
Resazurin 1.0 mg 1.0e-3 0.004
2 To make medium anoxic bring it to a boil, cool under 100% N2 gas atmosphere, add 0.5 g/l L-cysteine hydrochloride and adjust pH to 7.0. Dispense under 100% N2 gas atmosphere by filling 7 ml medium into anoxic Hungate-type tubes (for strains demanding meat particles put those first into the tube (use 1 part meat particles to 4 or 5 parts fluid)). Autoclave at 121°C for 30 min. For agar slants use 15.0 g agar per 1000.0 ml medium.
L-Cysteine HCl 0.5 g 0.5 3.172
Agar , for solid medium 15.0 g 15 44.599
3 In some cases (as indicated in the catalogue) the addition of Haemin and Vitamin K1 or Vitamin K3 is necessary. Add to 1000 ml of medium after autoclaving:
Haemin solution
optional
10.0 ml - -
Vitamin K1 solution , alternative
optional
10.0 ml - -
Vitamin K3 solution , alternative
optional
10.0 ml - -
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.
Solution occurs in 7 other media (first defined in Medium ).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin K3 5 mg 0.05 -
Ethanol
(95%)
1 ml 9.5 206.208
Water 100 ml - -
1 Dissolve 5 mg vitamin K3 in 1 ml 95% ethanol, dilute to 0.05 mg/ml in water and filter sterilize. Store refrigerated in a brown bottle.
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.

Metadata

Last modified: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Anaerobic workbench, Autoclave, Filter, Gassing station, Hungate tubes
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Associated strains

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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Ground beef 473.934 -
Casitone 28.436 -
Agar 14.218 42.274
K2HPO4 4.739 27.21
Yeast extract 4.739 -
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain