JCM 1067 . FRESHWATER IRON-OXIDIZING BACTERIA MEDIUM-II

Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeS solution 50.00 ml - -
Modified Wolfe's mineral solution 50.00 ml - -
Agar , Noble
(BD-Difco)
1.50 g 1.333 3.964
1 Top layer:
Modified Wolfe's mineral solution 1000.00 ml - -
Trace minerals 10.00 ml - -
2 After cooling of the top layer solution, add per liter the following solutions (filter-sterilized):
Trace vitamins 10.00 ml - -
NaHCO3
(8%)
5.25 ml - -
3 Bottom layer:
4 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.
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 1 other media (first defined in Medium J628).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 46.2 g 154 553.927
Na2S x 9 H2O 39.6 g 132 549.583
Distilled water 300.0 ml - -
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]
NH4Cl 1.00 g 1 18.695
MgSO4 x 7 H2O 0.20 g 0.2 0.811
CaCl2 x 2 H2O 0.10 g 0.1 0.68
KH2PO4 0.05 g 0.05 0.367
Distilled water 1000.00 ml - -
Solution occurs in 97 other media (first defined in Medium J151).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x n H2O 0.50 g 0.5 -
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.10 g 0.1 0.645
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 0.01 g 0.01 0.039
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
Distilled water 1000.00 ml - -
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 220 other media (first defined in Medium J197).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium pantothenate 5.0 mg 0.005 0.01
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 03.04.22
Source: JCM
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Autoclave, Filter
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 6.845 24.622
Na2S x 9 H2O 5.867 24.426
Agar 1.333 3.964
NH4Cl 0.933 17.449
MgSO4 x 7 H2O 0.213 0.866
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Cultivation metadata from 1 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 1 1 strain
nutrition lithotroph 1 strain
oxygen microaerophile 1 strain
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