freshwater_iron_oxidizing_bacteria_medium

Overview

External ontology

mediadive.medium:J628 (source link unavailable) — FRESHWATER IRON-OXIDIZING BACTERIA MEDIUM

Applications

• Microbial cultivation

pH

6.2

Notes

Source: JCM | Link: https://www.jcm.riken.jp/cgi-bin/jcm/jcm_grmd?GRMD=628

Composition

flowchart LR
    CultureMech_002974["freshwater_iron_oxidizing_bacteria_medium"]
    ing0_Agarose["Agarose"]
    CultureMech_002974 --> ing0_Agarose
    CHEBI_2511["CHEBI:2511"]
    ing0_Agarose -.-> CHEBI_2511
    ing1_Trace_minerals__see_Medium_No__197["Trace minerals (see Medium No. 197"]
    CultureMech_002974 --> ing1_Trace_minerals__see_Medium_No__197
    ing2_MES["MES"]
    CultureMech_002974 --> ing2_MES
    CHEBI_39010["CHEBI:39010"]
    ing2_MES -.-> CHEBI_39010
    ing3_NaHCO3["NaHCO3"]
    CultureMech_002974 --> ing3_NaHCO3
    CHEBI_32139["CHEBI:32139"]
    ing3_NaHCO3 -.-> CHEBI_32139
    ing4_FeSO4_x_7_H2O["FeSO4 x 7 H2O"]
    CultureMech_002974 --> ing4_FeSO4_x_7_H2O
    CHEBI_75836["CHEBI:75836"]
    ing4_FeSO4_x_7_H2O -.-> CHEBI_75836
    ing5_Na2S_x_9_H2O["Na2S x 9 H2O"]
    CultureMech_002974 --> ing5_Na2S_x_9_H2O
    CHEBI_76209["CHEBI:76209"]
    ing5_Na2S_x_9_H2O -.-> CHEBI_76209
    ing6_NH4Cl["NH4Cl"]
    CultureMech_002974 --> ing6_NH4Cl
    CHEBI_31206["CHEBI:31206"]
    ing6_NH4Cl -.-> CHEBI_31206
    ing7_MgSO4_x_7_H2O["MgSO4 x 7 H2O"]
    CultureMech_002974 --> ing7_MgSO4_x_7_H2O
    CHEBI_31795["CHEBI:31795"]
    ing7_MgSO4_x_7_H2O -.-> CHEBI_31795
    ing8_CaCl2_x_2_H2O["CaCl2 x 2 H2O"]
    CultureMech_002974 --> ing8_CaCl2_x_2_H2O
    CHEBI_86158["CHEBI:86158"]
    ing8_CaCl2_x_2_H2O -.-> CHEBI_86158
    ing9_KH2PO4["KH2PO4"]
    CultureMech_002974 --> ing9_KH2PO4
    CHEBI_63036["CHEBI:63036"]
    ing9_KH2PO4 -.-> CHEBI_63036

Ingredients (10)

Preparation

1. The culture medium consists of two layers to create opposing gradients of O2 and Fe(II). The top layer is the semisolid mineral salts medium and the bottom layer serves as the iron source.
2. Adjust pH to 6.2.
3. Autoclave both layer solutions. Shortly after autoclaving, pipette the bottom layer into each culture vessel (typically 0.75 ml in 10 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. Cool the top layer to 35 - 40C, add 1 ml Trace vitamins (see Medium No. 197) and adjust pH to 6.1 - 6.4 by bubbling with sterile CO2 gas. Slowly overlay the top layer onto the solidified bottom layer (3.75 ml in the 10 ml vial). Allow to solidify the top layer a minimum of 3 hr to a maximum of overnight. To inoculate, 10 �l of the inoculum is drown into a pipette tip and inject just above the bottom layer. Seal vials with butyl rubber stoppers and replace the gas phase with a N2-CO2-O2 (8:1:1, v/v) gas mixture.
4. 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.

Target organisms

No target organisms are recorded.

Provenance