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J1067. FRESHWATER IRON-OXIDIZING BACTERIA MEDIUM-II

Culture medium recipe

Vol.
×
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 FeS solution 50.00 ml - -
2 Modified Wolfe's mineral solution 50.00 ml - -
3 , Noble
(BD-Difco)
1.50 g 1.333 -
4 Modified Wolfe's mineral solution 1000.00 ml - -
5 Trace minerals 10.00 ml - -
6 Trace vitamins 10.00 ml - -
7
(8%)
5.25 ml 0.806 9.595
Instructions:
  1. Bottom layer:
  2. Top layer: #4-5
  3. Autoclave both layer solutions. Shortly after autoclaving, pipette the bottom layer into each culture vessel (typically 5 ml in 60 ml glass vials, use pipette tips that have been cut to enlarge the opening). The bottom layer is allowed to cool for at least 30 min to ensure its solidification.
  4. After cooling of the top layer solution, add per liter the following solutions (filter-sterilized): #6-7
  5. Add the top layer solution in 5 to 6 times the volume of the bottom layer to the culture vessels containing the bottom layer (e.g., 25 to 30 ml in 60 ml glass vials with 5 ml of the bottom layer).
  6. Then, cap the culture vessels with silicone rubber stoppers, and replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture.
  7. Comment: The medium can be stored in culture vessels for a certain time by capping with butyl rubber stoppers, instead of silicon rubber stoppers and keeping anoxic in the vessels. Silicone rubber stopper is O2 permeable and creates a mircoaerophilic condition in the culture vessels.
Solution occurs in 88 other media (first defined in medium J151 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.50 g 1.5 7.848
2 3.00 g 3 12.171
3 0.50 g 0.5 -
4 1.00 g 1 17.111
5 0.10 g 0.1 0.36
6 0.10 g 0.1 -
7 0.10 g 0.1 0.68
8 0.10 g 0.1 -
9 0.01 g 0.01 -
10 0.01 g 0.01 0.039
11 0.01 g 0.01 0.162
12 0.01 g 0.01 0.041
13 1000.00 ml 998.207 55408.8
Instructions:
  1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.
Solution occurs in 218 other media (first defined in medium J197 )
# 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 -
4 5.0 mg 0.005 -
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 -
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 1 other media (first defined in medium J628 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 46.2 g 154 553.917
2 39.6 g 132 549.565
3 300.0 ml 998.207 55408.8
Instructions:
  1. Prewarm distilled water at 50C in a beaker with a stir bar. While rapidly starring the water, add FeSO4 x 7H2O, followed immediately by Na2S x 9H2O. A thick black precipitate will form instantly. Stir continuously for 2 to 3 min. Remove the mixture to a narrow-mouthed glass bottle (500 ml) that can be tightly stopped. The bottle is filled to the top with distilled water and capped. Allow to settle for several hours, decant and replace the overlaying water. Repeat this procedure at least five times to wash the FeS precipitate. Check pH to be close to neutral. To stock the FeS solution, fill the bottle with distilled water and cap. Remove the overlaying water when prepare the bottom layer.
Solution occurs in 3 other media (first defined in medium J628 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.00 g 1 18.695
2 0.20 g 0.2 0.811
3 0.10 g 0.1 0.68
4 0.05 g 0.05 0.367
5 1000.00 ml 998.207 55408.8

Metadata

Media status: Inofficial medium
Source: JCM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: unknown

Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 993.771 55162.5
FeSO4 x 7H2O 6.845 24.622
Na2S x 9H2O 5.867 24.425
Agar 1.333 -
NH4Cl 0.933 17.448
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