# 81a: THIOMONAS ISLANDICA MEDIUM


## Main sol. 81a

2.3	g	K2HPO4	
2.9	g	Na2HPO4 x 2 H2O	
1	g	NH4Cl	
0.01	g	CaCl2 x 2 H2O	
0.5	g	MgSO4 x 7 H2O	
5	ml	NaVO3 x H2O (0.1% w/v)	
5	ml	**Trace element solution SL-6**	
0.5	g	NaHCO3	
0.05	g	Ferric ammonium citrate	
1	ml	**Seven vitamins solution**	
1000	ml	Distilled water	

1. Dissolve ingredients (ecxept bicarbonate, ferric ammonium citrate and vitamins), then sparge solution with 80% N2 and 20% CO2 gas mixture for at least 30 - 45 min to remove dissolved oxygen and to saturate the solution with CO2. Adjust pH to 4.5 and dispense solution under air atmosphere into serum vials suitable for anaerobic cultivation to 50% of volume. Close vials with butyl rubber septa to prevent free exchange of oxygen with the external atmosphere and autoclave. Add ferric ammonium citrate and vitamins from anoxic stock solutions sterilized by filtration and bicarbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Adjust pH of the complete medium to 5.0, if necessary.
2. After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.
3. Note: Use at least 10% (v/v) as inoculum.


## Trace element solution SL-6

0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	


## Seven vitamins solution

100	mg	Vitamin B12	
80	mg	p-Aminobenzoic acid	
20	mg	D-(+)-biotin	
200	mg	Nicotinic acid	
100	mg	Calcium pantothenate	
300	mg	Pyridoxine hydrochloride	
200	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water