307. FERVIDOBACTERIUM MEDIUM

NH₄Cl 0.9 g

NaCl 0.9 g

MgCl₂ × 6 H₂O 0.4 g

KH₂PO₄ 0.75 g

K₂HPO₄ 1.5 g

Trace element solution (see below) 9.0 ml

FeSO₄ × 7 H₂O solution (0.1% w/v in 0.1 N H₂SO₄) 3.0 ml

Yeast extract 3.0 g

Trypticase peptone 10.0 g

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

Vitamin solution (see below) 5.0 ml

D-Glucose 5.0 g

Na₂S × 9 H₂O 1.0 g

Distilled water 1000.0 ml

Trace element solution:

Nitrilotriacetic acid (NTA) 12.8 g

FeCl₂ × 4 H₂O 0.2 g

MnCl₂ × 4 H₂O 0.1 g

CoCl₂ × 6 H₂O 0.17 g

CaCl₂ × 2 H₂O 0.1 g

ZnCl₂ 0.1 g

CuCl₂ × 2 H₂O 0.02 g

H₃BO₃ 0.01 g

Na₂MoO₄ × 2 H₂O 0.01 g

NiCl₂ × 6 H2O 0.03 g

NaCl 1.0 g

Na₂SeO₃ × 5 H₂O 0.03 g

Distilled water 1000.0 ml

First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.0 ml.

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl × 2 H₂O 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

D-Ca-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Dissolve ingredients (except vitamins, glucose and sulfide), sparge medium with 100% N₂ gas for 30–45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N₂ gas. Adjust pH of complete medium to 7.2-7.4.

 

Updated 1.02.2024