Solution A
Nitrilotriacetic acid 1.5 g, MgSO4 * 7H2O 3.0 g, MnSO4 * 2 H2O 0.5 g, NaCl 1.0 g, FeSO4
* 7 H
2O 0.1 g, CaSO4 * 7 H2O 0.18g, CaCl2 * 2H2O 0.1 g, ZnSO4 * 7H2O 0.18 g, CuSO4 *
5H
2O 0.01 g, KAl(SO4)2 * 12H2O 0.02 g, H3BO3  0.01 g, Na2MoO4 * 2 H2O 0.01 g, NiCl2 *
6 H
2O 0.025 g, Na2SeO3  0.3 mg, Distilled water 1.0 liter.
COMPOSITION
NH
4Cl 1.0 g, KH2PO4  0.33 g, K2HPO4  0.45 g, MgSO4 * 7H2O  0.1 g, Solution A 20 ml, Solution B 20 ml, Yeast extract 2.00 g, Fructose
1.0 g, Resazurin 1.0 mg NaHCO
3 10.0 g, Cysteine hydrochloride 0.5 g, Na2S * 9H2O 0.5 g, Distilled water 1.0 liter.
Acetobacterium medium
Solution B
Biotin 2.0 mg, Folic acid 20 mg, Pyridoxine-HCl 10 mg, Thiamine-HCl 5.0 mg,
Riboflavin 5.0 mg, Nicotinic acid 5.0 mg, DL-Calcium panthotenate 5.0 mg, Vitamin
B12 0.1 mg, p-Aminobenzoic acid 5.0 mg, Lipoic acid 5.0 mg, Distilled water 1.0 L
PROCEDURE

First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final ph 7.0

Dissolve ingredients except NaHCO
3, fructose, cysteine and sodium sulphide, bring to the boil for a few minutes and cool to room
temperature under N
2 + CO2 (80 + 20) gas mixture. Add NaHCO3 and equilibrate medium with the gas until a pH of approximately 7.4
is reached. Then distribute and autoclave under the same gas.

Before use adjust the pH to 8.2 by adding sterile anaerobic Na
2CO3 solution (0.25 ml of 5% Na2CO3 per 10 ml medium) and add
fructose, cysteine
and sodium sulphide from anaerobic sterile stock solutions.
(c) Costin Stoica
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