# 380a: TELMATOSPIRILLUM (PEAT) MEDIUM


## Main sol. 380a

0.68	g	KH2PO4	
0.12	g	NaNO3	
0.37	g	L(+)-Tartaric acid	
0.37	g	Succinic acid	
0.05	g	Na-acetate	
5	ml	**Modified Wolin's mineral solution**	
2	ml	**Fe(III) quinate solution 0.01 M**	
0.5	ml	Sodium resazurin (0.1% w/v)	
1	g	D-Xylose	
1	ml	**Wolin's vitamin solution (10x)**	
0.05	g	Na-thioglycolate	
1000	ml	Distilled water	

1. Dissolve ingredients (except xylose, vitamins and thioglycolate) and adjust pH to 5.5 - 6.0. Sparge medium with 100% N2 gas for 30 -45 min to make it anoxic and dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume. Seal vials with gas tight butyl rubber closures. Autoclave at 121°C for 15 min. Before inoculation add xylose, vitamins and thioglycolate from anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration. Then add sterile air (with hypodermic syringe through the rubber closure) to 1% (v/v) O2 concentration in the gas phase.


## Modified Wolin's mineral solution

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
0.4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.


## Fe(III) quinate solution 0.01 M

4.5	g	FeCl3 x 6 H2O	
1.9	g	Quinic acid	
1000	ml	Distilled water	

1. Sterilize by filtration under 100% N2 gas atmosphere.


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water