482. THERMODESULFOVIBRIO MEDIUM (THDVM)

Na₂SO₄ 4.00 g

Na₂HPO₄ 0.20 g

NH₄Cl 0.25 g

MgCl₂ x 6 H₂O 0.40 g

KCl 0.50 g

CaCl₂ x 2 H₂O 0.15 g

Trace element solution (see below) 10.00 ml

Yeast extract 1.0 g

Na-DL-lactate 2.50 g

Na-resazurin solution (0.1% w/v) 0.50 ml

NaHCO₃ 1.30 g

Vitamin solution (see below) 10.00 ml

Na-thioglycolate 0.20 g

L-Ascorbic acid 0.20 g

Distilled water 1000.00 ml

Trace element solution:

Nitrilotriacetic acid 1.50 g

MgSO₄ x 7 H₂O 3.00 g

MnSO₄ x H₂O 0.50 g

NaCl 1.00 g

FeSO₄ x 7 H₂O 0.10 g

CoSO₄ x 7 H₂O 0.18 g

CaCl₂ x 2 H₂O 0.10 g

ZnSO₄ x 7 H₂O 0.18 g

CuSO₄ x 5 H₂O 0.01 g

AlK(SO₄)₂ x 12 H₂O 0.02 g

H₃BO₃ 0.01 g

Na₂MoO₄ x 2 H₂O 0.01 g

NiCl₂ x 6 H₂O 0.03 g

Na₂SeO₃ x 5 H₂O 0.30 mg

Na₂WO₄ x 2 H₂O 0.40 mg

Distilled water 1000.00 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:

Vitamin B₁₂ 100.00 mg

p-Aminobenzoic acid 80.00 mg

D-(+)-biotin 20.00 mg

Nicotinic acid 200.00 mg

Calcium pantothenate 100.00 mg

Pyridoxine hydrochloride 300.00 mg

Thiamine-HCl x 2 H₂O 200.00 mg

Distilled water 1000.00 ml

Dissolve ingredients except bicarbonate, vitamins, thioglycolate and ascorbic acid, then sparge medium with 100% N₂ gas for 30 – 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins, thioglycolate and ascorbic acid from anoxic stock solutions prepared under 100% N₂ gas and sterilized by filtration and bicarbonate from a sterile anoxic stock solution prepared under 80% N₂ and 20% CO₂ gas mixture. Prior to use adjust pH of complete medium to 8.0, if necessary.

Updated: 07.01.2024