417. MEDIUM FOR HALOPHILIC ANAEROBES (MHlAnb)

KH₂PO₄ 0.2 g

MgCl₂ 0.1 g

NH₄Cl 0.5 g

KCl 0.2 g

Na₂CO₃ 9.3 g

NaHCO₃ 44.1 g

Na₂S × 9 H₂O 0.7 g

Yeast extract 0.2 g

Glucose 5.0 g

Trace element solution (see below) 1 ml

Vitamin solution (see below) 10 ml

Resazurin 0.01 g

Distilled water 990.0 ml

Trace element solution:

Nitrilotriacetic acid 1.50 g

MgSO₄ × 7 H₂O 3.0 g

MnSO₄ × H₂O 0.50 g

NaCl 1.0 g

FeSO₄ × 7 H₂O 0.10 g

CoSO₄ × 7 H₂O 0.18 g

CaCl₂ × 2 H₂O 0.10 g

ZnSO₄ × 7 H₂O 0.18 g

CuSO₄ × 5 H₂O 0.01 g

KAl(SO₄)₂ × 12 H₂O 0.02 g

H₃BO₃ 0.01 g

Na₂MoO₄ × 2 H₂O 0.01 g

NiCl₂ × 6 H₂O 0.03 g

Na₂SeO₃ × 5 H₂O 0.30 mg

Na₂WO₄ × 2 H₂O 0.40 mg

Distilled water 1000.0 ml

First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals.

Adjust final to pH 7.0 with KOH.

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca-D-pantothenate 5.0 mg

Vitamin B₁₂ 0.10 mg

p-Aminobenzoic acid 5.0 mg

(±)-Lipoic acid 5.0 mg

pH 9.05. Gas phase was N₂. Anaerobic conditions required.