# 998: MEDIUM FOR ROSEOSPIRA


## Main sol. 998

0.3	g	KH2PO4	
0.5	g	NH4Cl	
0.05	g	CaCl2 x 2 H2O	
1	g	MgCl2 x 6 H2O	
0.5	g	MgSO4 x 7 H2O	
20	g	NaCl	
1	ml	**Trace element solution SL12**	
0.5	g	Yeast extract	
1000	ml	Distilled water	
1	ml	**Vitamins V7 solution (Pfennig et Trüper, 1992)**	
0.15	%(w/v)	NaHCO3	

1. Media were autoclaved and cooled under N2/CO2 (90/10, v/v). Vitamin V7 solution (1 ml/l) and NaHCO3, (0.15% w/v) were then aseptically added to the medium. Final pH was adjusted to 6.8-7.0 and the medium was dispensed into sterile 50 ml screw-capped bottles. Organic substrates (5 mM Na-acetate and/or 5 mM di-Na-succinate) were added as substrates before use. In addition, for strains DSM 15114 and AT2115, Na2S x 9 H2O (0.02% w/v) were also added to the medium prior to utilisation. Pure cultures, grown in 50-ml screw-capped bottles were stored at +4°C in the dark for preservation.


## Trace element solution SL12

1000	ml	Distilled water	
300	mg	H3BO3	
1.1	g	FeSO4 x 7 H2O	
190	mg	CoCl2 x 6 H2O	
50	mg	MnCl2 x 2 H2O	
42	mg	ZnCl2	
24	mg	NiCl2 x 6 H2O	
18	mg	Na2MoO4 x 2 H2O	
2	mg	CuCl2 x 2 H2O	

1. Sterilized by filtration.


## Vitamins V7 solution (Pfennig et Trüper, 1992)

1000	ml	Distilled water	
2	mg	Biotine	
10	mg	p-Aminobenzoate	
10	mg	Thiamine	
5	mg	Pantothenate	
50	mg	Pyridoxamine	
20	mg	Vitamine B12	
20	mg	Nicotinate	

1. Sterilized by filtration.