# 888: AQUASPIRILLUM MEDIUM


## Main sol. 888

1	g	(NH4)2SO4	
1	g	MgSO4 x 7 H2O	
30	mg	CaCl2 x 6 H2O	
10	mg	Na2HPO4	
1	g	Sodium succinate	
1.5	g	Casamino acids (DIFCO)	
1	ml	**Trace elements solution (Pfennig & Lippert,1966)**	
0.5	g	Agar (DIFCO)	
1000	ml	Distilled water	

1. pH 7.5
2. Add after autoclaving sterile solutions of:

1	ml	Na2S2O3 x 5 H2O (10% solution)	
1	ml	**Vitamine mixture**	


## 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


## Vitamine mixture

100	ml	**Acidic thiamine / Ca pantothenate solution**	
100	ml	**Basic vitamin solution**	
100	ml	**Neutral folic acid / riboflavin solution**	
100	ml	**Neutral B12 solution 0.005 g/L**	

1. Sterilize by filtration.
2. Mix the four solutions in the ratio 1:1:1:1.


## Acidic thiamine / Ca pantothenate solution

5	mg	Thiamine	
5	mg	Calcium pantothenate	
100	ml	Distilled water (acidified to pH 3.0 with HCl)	


## Basic vitamin solution

2	mg	Biotin	
5	mg	PABA (p-aminobenzoic acid)	
5	mg	Nicotinic acid	
10	mg	Pyridoxine	
100	ml	NaOH (0.1N)	


## Neutral folic acid / riboflavin solution

2	mg	Folic acid	
5	mg	Riboflavin	
100	ml	Distilled water	


## Neutral B12 solution 0.005 g/L

0.5	mg	B12	
100	ml	Distilled water