# 722: OCEANIDESULFOVIBRIO MEDIUM


## Main sol. 722

3	g	Na2SO4	
0.3	g	KCl	
0.3	g	NH4Cl	
0.2	g	KH2PO4	
0.1	g	CaCl2 x 2 H2O	
50	g	NaCl	
3.3	g	MgCl2 x 6 H2O	
1.6	g	MgSO4 x 7 H2O	
0.1	g	Yeast extract	
1	ml	**Trace element solution SL-11**	
0.5	ml	Sodium resazurin (0.1% w/v)	
1.5	g	Na2CO3	
2.5	g	Na-DL-lactate	
1	ml	**Seven vitamins solution**	
0.2	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except carbonate, lactate, vitamins and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add lactate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins should be sterilized by filtration. The pH of the complete medium should be 7.0 - 7.2.


## Trace element solution SL-11

5.2	g	Na2-EDTA x 2 H2O	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. 


## 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