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DSMZ

DSMZ 78d . MEDIUM FOR TREPONEMA PARVUM

This medium was extracted automatically and has not been curated by experts, yet.

Culture medium recipe

Vol.
×

Main sol. 78d #

ml
Equipment needed: Hungate tubes Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 78 1000 ml - -
1 To medium 78 add per liter:
Calf serum
(newborn or fetal) 		10 ml - -
Amino acid solution 5 ml - -
Vitamin solution 5 ml - -
Hemin solution 5 ml - -
Peptone mixture 5 ml - -
Ascorbic acid solution 2%(w/v) 5 ml - -
KCl
(100 g/l) 		5 ml - -
N-acetylglucosamin- and pyruvate solution 5 ml - -
Glutathione solution 14%(w/v) 10 ml - -
Sugar solution 10 ml - -
2 Prepare medium anaerobically under N2:CO2 (80:20). After inoculation of the tubes, pressurize them with hydrogen gas to 150 kP (1.5 bar).
3 All following solutions were filter sterilized

Vitamin solution #

1000
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.

Amino acid solution #

50
1 Amino acid solution per 50 ml:
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Histidin 0.6 g 12 77.342
L-Serin 0.5 g 10 95.154
L-Glutamine 0.7 g 14 95.799

Hemin solution #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Hemin 5 mg 0.1 0.153
NaOH
(1 N) 		1 ml - -
Distilled water 50 ml - -
1 Dissolve 5 mg hemin in 1 ml 1 M NaOH, fill up to 50 ml with dist. water.

Peptone mixture #

50
1 Peptone mixture per 50 ml:
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tryptone peptone
(Difco 0123-17) 		1.6 g 32 -
Casamino acids
(Difco 0230-17) 		1.6 g 32 -
Yeast extract 1.6 g 32 -

Ascorbic acid solution 2%(w/v) #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ascorbic acid 1 g 20 113.559
Distilled water 50 ml - -
1 Dissolve 1 g ascorbic acid in 40 ml dist. water, adjust pH to 7.0. fill up to 50 ml.

N-acetylglucosamin- and pyruvate solution #

50
1 N-Acetylglucosamin and pyruvate solution per 50 ml :
Compound Amount Unit Conc. [g/L] Conc. [mM]
N-Acetylglucosamine 5 g 100 452.059
Na-pyruvate 5 g 100 908.727

Glutathione solution 14%(w/v) #

50
Compound Amount Unit Conc. [g/L] Conc. [mM]
Glutathione 7 g 140 455.551
Distilled water 50 ml - -
1 Dissolve 7 g glutathione in 40 ml dist. Water, adjust pH to 7.0 and fill up to 50 ml.

Sugar solution #

50
1 Depending on organism use different modification .

Medium 78 #

1025
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 - -

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 K3 solution #

100
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.

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.

Metadata

Last modified: 22.02.22
Source: DSMZ
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
Description:

Not in use anymore!
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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: No strains found

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Ground beef 500 -
Casitone 30 -
Agar 15 44.599
K2HPO4 5 28.707
Glutathione 1.4 4.556
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