amoebobacter_medium

Overview

External ontology

mediadive.medium:422 (source link unavailable) — AMOEBOBACTER MEDIUM

Applications

• Microbial cultivation

Notes

Source: DSMZ | Link: https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium422.pdf

Composition

flowchart LR
    CultureMech_001527["amoebobacter_medium"]
    ing0_Acetate["Acetate"]
    CultureMech_001527 --> ing0_Acetate
    CHEBI_30089["CHEBI:30089"]
    ing0_Acetate -.-> CHEBI_30089
    ing1_Thiosulfate["Thiosulfate"]
    CultureMech_001527 --> ing1_Thiosulfate
    CHEBI_26977["CHEBI:26977"]
    ing1_Thiosulfate -.-> CHEBI_26977
    ing2_Na2S_x_9_H2O["Na2S x 9 H2O"]
    CultureMech_001527 --> ing2_Na2S_x_9_H2O
    CHEBI_76209["CHEBI:76209"]
    ing2_Na2S_x_9_H2O -.-> CHEBI_76209

Ingredients (3)

Preparation

1. Supplement medium 28 with 2 mmol acetate and 1 mmol thiosulfate.
2. Adjust the pH with filter-sterilised 1M Na2CO3 to 7.1-7.3.
3. If needed, aliquot into sterile, N₂ gassed screw-cap tubes under N₂ gas.
4. During the first 24 h, the iron of the medium precipitates in the form of black flocks. No other sediment should arise in the otherwise clear medium.
5. Aliquot Solution A into 100 mL screw-cap bottles, filled with 46 mL each. Bubble with N2/CO2 and autoclave at 121°C for 15 min (as decribed below).
6. Prepare the following solutions (resazurin, bicarbonate and Pfennig's heterotrophic salts) and sterilize as given below.
7. Add bicarbonate solution and Pfennig's heterotrophic salts to the resazurin (complete volumina, i.e. 50 mL bicarbonate solution and 26 mL Pfennig's heterotrophic salts solution). Bubble with CO2 in an ice bath under sterile conditions.
8. Fill 50 ml of this mixture to each bottle of solution A (46 mL + 50 mL).
9. Before use, add 4 ml sulfide solution (1.5%) and 0.1 ml Vitamin B12 solution to each 100 mL bottle.
10. Feed the actively growing culture periodically with neutralized 3% solution of sodium sulfide (use 1 -3 mL/100 mL depending on strain and cultivation stage) to replenish sulfide and with other supplement solutions (see Ref. 3365).
11. The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle.

Target organisms

No target organisms are recorded.

Provenance