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78b. MEDIUM FOR TREPONEMA PARVUM

Culture medium recipe

Vol.
×
Equipment needed: Anaerobic workbench Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 Main sol. 78 1000 ml - -
2 10 ml - -
3 Amino acid solution 50 ml - -
4 Vitamin solution 5 ml - -
5 Hemin solution 5 ml - -
6 Peptone mixture 5 ml - -
7 Ascorbic acid solution 5 ml - -
8
(100 g/l)
5 ml 0.49 6.575
9 N-Acetylglucosamin and pyruvate solution 5 ml - -
10 Glutathione solution 10 ml - -
Instructions:
  1. To medium 78 add per liter: #2-12
  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 solutions were filter sterilized
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1
(Difco 0123-17)
1.6 g 32 -
2
(Difco 0230-17)
1.6 g 32 -
3 1.6 g 32 -
4 50.0 ml 998.207 55408.8
Solution occurs in 17 other media (first defined in medium 1116 )
Equipment needed: Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 50 mg 0.05 0.032
2 50 mg 0.05 0.228
3 50 mg 0.05 0.133
4 10 mg 0.01 0.049
5 20 mg 0.02 0.082
6 20 mg 0.02 0.045
7 25 mg 0.025 0.203
8 25 mg 0.025 0.205
9 50 mg 0.05 0.242
10 50 mg 0.05 0.365
11 50 mg 0.05 0.134
12 1000 ml 998.207 55408.8
Instructions:
  1. Stir for some hours, filter sterilize the solution.
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.6 g 12 77.342
2 0.5 g 10 95.154
3 0.7 g 14 95.795
4 50.0 ml 998.207 55408.8
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 5 mg 0.1 -
2 , required for pH adjustment
(1 M)
1 ml 42.6 1065.08
3 50 - -
Instructions:
  1. Dissolve 5 mg hemin in 1 ml 1 M NaOH, fill up to 50 ml with dist. water.
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1 g 20 -
2 50 ml 998.207 55408.8
Instructions:
  1. Dissolve 1 g ascorbic acid in 40 ml dist. water, adjust pH to 7.0. fill up to 50 ml.
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 5 g 100 -
2 5 g 100 908.727
3 50 ml 998.207 55408.8
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 7 g 140 -
2 50 ml 998.207 55408.8
Instructions:
  1. Dissolve 7 g glutathione in 40 ml dist. Water, adjust pH to 7.0 and fill up to 50 ml.
# Compound Amount Unit Conc. [g/L] Conc. [mM]
6 10 g 200 1332.18
7 10 g 200 1030.18
8 50 ml 998.207 55408.8
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 10 g 200 584.29
2 50 ml 998.207 55408.8
Solution occurs in 3 other media (first defined in medium 78 )
Equipment needed: Hungate tubes Filter Autoclave
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1
(fat free)
500.0 g 500 -
2 1000.0 ml 998.207 55408.8
3
(1 N)
25.0 ml 53.25 1331.35
4 30.0 g 30 -
5 5.0 g 5 -
6 5.0 g 5 28.707
7 1.0 mg 1.0e-3 0.004
11 0.5 g/l 0.5 3.172
12
optional
15.0 g/l 15 -
13
(5% w/v)
optional
1.0 g/l 1 11.904
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 water to a final volume of 1000 ml, and then add: #4-7
  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.
  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: #8 and either #9 or #10.
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 5 other media (first defined in medium 78 )
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.
Solution occurs in 4 other media (first defined in medium 78 )
Equipment needed: Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 5 g/l 5 -
2
(95 %)
x - -
3 100 ml 998.207 55408.8
Instructions:
  1. Dissolve 5 mg/ml of vitamin K3 in 95% ethanol, dilute to 0.05 mg/ml in water and filter sterilize. Store refrigerated in a brown bottle.

Metadata

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

Associated strains

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

Gas composition

  • 20 % CO2
  • 80 % N2

Light

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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 1056.93 58668.1
Ground beef 490.196 -
NaOH 52.415 1310.47
Casitone 29.412 -
Agar
optional
14.706 -
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Cultivation metadata from 1 strains BacDive

halophily moderate 1 strain
pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 1 strain
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