433. HALODESULFOVIBRIO MEDIUM (HdvM)

Solution A:

NaCl 25.0 g

K₂HPO₄ 0.5 g

NH₄Cl 1.0 g

Na₂SO₄ 1.0 g

CaCl₂ × 2 H₂O 0.1 g

MgSO₄ × 7 H₂O 2.0 g

Na-DL-lactate 2.0 g

Yeast extract 1.0 g

Na-resazurin solution (0.1% w/v) 0.5 ml

Distilled water 980.0 ml

Solution B:

FeSO₄ × 7 H₂O 0.5 g

Distilled water 10.0 ml

Solution C:

Na-thioglycolate 0.1 g

Ascorbic acid 0.1 g

Distilled water 10.0 ml

Bring solution A to the boil, then cool to room temperature while sparging with 100% N₂ gas. Add solutions B and C, adjust pH to 7.8 with NaOH, and distribute under N₂ gas atmosphere into anoxic Hungate-type tubes. During distribution continuously swirl the medium to keep the grey precipitate suspended. Autoclave 15 min at 121°C. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.