# 1058a: THIOHALOSPIRA HALOPHILA MEDIUM


## Main sol. 1058a

233	g	NaCl	
1.5	g	K2HPO4	
0.5	g	NH4Cl	
1	ml	**Trace elements solution (Pfennig & Lippert,1966)**	
0.05	g	CaCl2 x 2 H2O	
0.4	g	MgCl2 x 6 H2O	
5	g	Na2S2O3 x 5 H2O	
2.5	g	NaHCO3	
1	ml	**Seven vitamins solution**	
1000	ml	Distilled water	

1. Dissolve sodium chloride, potassium hydrogenphosphate and ammonium chloride, 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. Dispense solution under air atmosphere in Erlenmeyer flasks to 10% of volume, close with screw caps and autoclave. Add trace elements, calcium chloride, magnesium chloride, thiosulfate, bicarbonate and vitamins from sterile stock solutions and adjust pH of the medium to 7.0 - 7.2, if necessary. Incubate with shaking.
2. Note: Use at least 10% (v/v) as inoculum.


## Trace elements solution (Pfennig & Lippert,1966)

5	g	EDTA	
2.2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.03	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.03	g	CuCl2 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. pH 3.0-4.0


## 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