ATCC medium: 2228 Thiomicrospira pelophila medium
NaCl....................................25.0 g
(NH4)2SO4.................................1.0 g
MgSO4 . 7H2O .............................1.50 g
CaCl2 . 2H2O .............................0.42 g
Wolfe's Mineral Solution (see below).....1.0 ml
Bromothymol blue.........................4.0 mg
K2HPO4* ..................................0.50 g
Na2S2O3 . 5H2O* ...........................5.0 g
Vitamin B12 .............................15.0
mcg Distilled water..........................1.0 L
*Autoclave the K2HPO4 and Na2SO3 separately, each in 10% of the final volume.
Filter-sterilize the vitamin B12. Adjust the completed medium to pH 7.2 with sterile 0.4% (w/v) Na2CO3 solution.
Alternatively, use "aged sea water" (see medium 246), supplemented by the indicated amounts of (NH4)2SO4, bromothymol blue, K2HPO4, sodium thiosulfate, and vitamin B12.
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 pH to 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.