Leptospirillum (HH) Medium

Solution A:

(NH4)2SO4 132.0 mg

MgCl2. 6H20 53.0 mg

KH2PO4 27.0 mg

CaCl2. 2H20 147.0 mg

Distilled water 950.0 ml

Adjust pH to 1.8 with 10N H2SO4

Solution B:

FeSO4. 7H2O 20.0 g

H2SO4, 0.25 N 50.0 ml

The pH of this solution should be 1.2.

Trace element solution

MnCl2. 2H2O 62.0 mg

ZnCl2 68.0 mg

CoCl2. 6H2O 64.0 mg

H3BO3 31.0 mg

Na2MoO4 10.0 mg

CuCl2. 2H2O 67.0 mg

Distilled water 1000.0 ml

Adjust pH to 1.8 with 10N H2SO4.

Autoclave the solutions separately at 112°C for 30 minutes. Prior to use mix solutions A and B and add 1 ml of Trace element solution. The final pH of the medium should be 1.8