ATCC medium: 1296 Spirochaeta zuelzerae medium
Solution 1:
KH2PO4 .......................0.75 g
NaH2PO4 . H2O.................0.25 g
L-Cysteine . HCl.............0.5 g
Distilled water............480.0 ml
Adjust pH to 7.2 with 5.0% (w) KOH.
Solution 2:
Resazurin................................1.0 mg
Yeast extract............................0.2 g
NH4Cl ....................................1.0 g
MgSO4 . 7H2O .............................0.5 g
CaCl2 ....................................0.02 g
FeCl3 . 6H2O (250 mg/L) .................10.0 ml
Trace Element Solution (see below).......2.0 ml
Distilled water........................480.0 ml
Solution 3:
NaHCO3 .......................5%
Solution 4:
Glucose.....................10%
Trace Element Solution:
H3BO3 .......................56.0 mg
ZnSO4 . 7H2O ................44.0 mg
CoCl2 . 6H2O ................20.0 mg
CuSO4 . 5H2O .................2.0 mg
MnCl2 ........................2.0 mg
Na2MoO4 . 2H2O...............75.0 mg
Distilled water............100.0 ml
Autoclave Solutions 1 and 2 separately. Filter-sterilize Solutions 3 and
4 with 3 under pressure. Add 480.0 ml of Solution 1 to 480.0 ml of
Solution 2 under 80% N2, 20% CO2. While gassing, add 20.0 ml of NaHCO3
solution and 20.0 ml of glucose solution. Dispense anaerobically and
aseptically; cap with butyl rubber stoppers. Adjust pH to 7.2.