# 882: LEPTOSPIRILLUM (HH) MEDIUM


## Main sol. 882

950	ml	**Solution A**	
50	ml	**Solution B**	
1	ml	**Solution C**	

1. Sterilize solution A and solution C by autoclaving at 121°C for 20 min and solution B by autoclaving at 112°C for 30 min. Prior to use mix solutions A and B and add trace elements solution. The final pH of the medium should be 2.3.
2. Note: At the beginning of growth incubate statically without shaking.


## Solution A

132	mg	(NH4)2SO4	
53	mg	MgCl2 x 6 H2O	
27	mg	KH2PO4	
147	mg	CaCl2 x 2 H2O	
950	ml	Distilled water	

1. Adjust pH to 2.3 with 10 N H2SO4.


## Solution B

20	g	FeSO4 x 7 H2O	
50	ml	H2SO4 (0.25 N)	

1. The pH of this solution should be 1.2.


## Solution C

1	ml	**Trace element solution**	


## Trace element solution

76	mg	MnCl2 x 4 H2O	
68	mg	ZnCl2	
64	mg	CoCl2 x 6 H2O	
31	mg	H3BO3	
10	mg	Na2MoO4	
67	mg	CuCl2 x 2 H2O	
1000	ml	Distilled water