ATCC medium: 2311 Spirochaeta americana medium
NH4Cl .................................... 1.0 g
MgCl2.6H2O ............................. 0.1 g
Yeast extract............................ 0.5 g
NaCl.................................... 30.0 g
Na2CO3 ................................... 2.76 g
NaHCO3 .................................. 24.0 g
KCl...................................... 0.2 g
K2HPO4 ................................... 0.2 g
Glucose.................................. 5.0 g
Wolfe's Vitamin Solution (see below)..... 2.0 ml
Modified Wolfe's Minerals (see below).... 1.0 ml
Na2S . 9H2O .............................. 0.4 g
Adjust the pH to 9.5. Dispense anaerobically into tubes, approximately 10 ml/tube.
Wolfe's Vitamin Solution: Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no.MD-VS). mg
Biotin....................... 2.0
Folic acid................... 2.0 mg
Pyridoxine hydrochloride.... 10.0 mg
Thiamine . HCl............... 5.0 mg
Riboflavin................... 5.0 mg
Nicotinic acid............... 5.0 mg
Calcium D-(+)-pantothenate... 5.0 mg
Vitamin B12.................. 0.1 mg
p-Aminobenzoic acid.......... 5.0 mg
Thioctic acid................ 5.0 mg
Distilled water.............. 1.0 L
Modified Wolfe's Minerals:
Na2SeO3 .................................10.0 mg
NiCl2 . 6H2O ............................ 10.0 mg
Na2WO4 . 2H2O............................ 10.0 mg
Wolfe's Mineral Solution (see below)..... 1.0 L
Wolfe's Mineral Solution: Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.) Nitrilotriacetic acid........ 1.5 g
MgSO4 . 7H2O ................. 3.0 g
MnSO4 . H2O .................. 0.5 g
NaCl......................... 1.0 g
FeSO4 . 7H2O ................. 0.1 g
CoCl2 . 6H2O ................. 0.1 g
CaCl2 ........................ 0.1 g
ZnSO4 . 7H2O ................. 0.1 g
CuSO4 . 5H2O ................. 0.01 g
AlK(SO4)2 . 12H2O............. 0.01 g
H3BO3 ........................ 0.01 g
Na2MoO4 . 2H2O................ 0.01 g
Distilled water.............. 1.0 L
Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.