154. DESULFOVIBRIO SULFODISMUTANS MEDIUM

Solution 1:

KH₂PO₄ 0.2 g

NH₄Cl 0.3 g

NaCl 1.0 g

MgCl₂ × 6 H₂O 0.4 g

KCl 0.5 g

CaCl₂ × 2 H₂O 0.15 g

Distilled water 920.0 ml

Solution 2:

Trace element solution SL-10 (see below) 1.0 ml

Solution 3:

NaHCO₃ 2.5 g

Distilled water 50.0 ml

Solution 4:

Na-acetate × 3 H₂O 0.3 g

Distilled water 10.0 ml

Solution 5:

D(+)-Biotin 10.0 mg

Ca-D(+)-Pantothenate 50.0 mg

Distilled water 1.0 ml

Solution 6:

Na₂S × 9 H₂O 0.4 g

Distilled water 10.0 ml

Solution 7:

Na₂S₂O₃ (0.5 M) 10.0 ml

pH 7.5-8.0 (adjust with NaOH)

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ × 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ × 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ × 6 H₂O 190.0 mg

CuCl₂ × 2 H₂O 2.0 mg

NiCl₂ × 6 H₂O 24.0 mg

Na₂MoO₄ × 2 H₂O 36.0 mg

Distilled water 990.0 ml

pH 7.1-7.4

Solution 1 is prepared and autoclaved anaerobically at under 80% N₂ + 20% CO₂ at 121°C for 15 min. Solutions 2, 4, 6 and 7 are gassed with N₂ and autoclaved separately at 121°C for 15 min. Solution 3 (gassed with N₂ + CO₂) and solution 5 (gassed with N₂) are filter-sterilized. Solutions with 2 to 7 are added to the sterile, cooled solution 1 in the sequence as indicated. When growth has started feed culture again with same amount of solution 7. After a further two days repeat feeding once more.

 

Updated 1.02.2024