# J439: DESULFOVIBRIO AEROTOLERANS DVO5 MEDIUM


## Main sol. J439

1	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.25	g	NH4Cl	
2	g	KH2PO4	
0.5	g	KCl	
0.5	g	Yeast extract	
4	g	Na2SO4	
1	ml	**Trace element solution**	
1	ml	**Selenite-tungstate solution**	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Mix components thoroughly and adjust pH to 7.0. Bring to a boil, and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 10 ml medium in Hungate tubes) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After cooling, anaerobically and aseptically add per liter the following solutions (autoclaved or *filter-sterilized): 

20	ml	Sodium lactate (1.0 M)	
1	ml	**Vitamin solution** (*)	
1	ml	**Thiamine solution** (*)	
1	ml	**Vitamin B12 solution 0.05 g/L** (*)	
20	ml	NaHCO3 (*, 8.0%)	

2. Prior to use, add per liter the following solution (autoclaved and stored under a N2 atmosphere): 

2	ml	Na2S x 9 H2O (5%)	


## Trace element solution

12.5	ml	HCl (25%)	
2.1	g	FeSO4 x 7 H2O	
30	mg	H3BO3	
100	mg	MnCl2 x 4 H2O	
190	mg	CoCl2 x 6 H2O	
24	mg	NiCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
144	mg	ZnSO4 x 7 H2O	
36	mg	Na2MoO4 x 2 H2O	
987	ml	Distilled water	


## Selenite-tungstate solution

0.4	g	NaOH	
6	mg	Na2SeO3 x 5 H2O	
8	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	


## Vitamin solution

4	mg	p-Aminobenzoic acid	
1	mg	Biotin	
10	mg	Nicotinic acid	
5	mg	DL-Calcium pantothenate	
10	mg	Pyridoxine hydrochloride	
100	ml	Sodium phosphate buffer (10 mM, pH 7.1)	


## Thiamine solution

10	mg	Thiamine HCl	
100	ml	Sodium phosphate buffer (25 mM, pH 3.4)	


## Vitamin B12 solution 0.05 g/L

5	mg	Vitamin B12	
100	ml	Distilled water