ATCC medium: 1779 Clostridium litorale medium
ATCC Medium 1770 (see below) with NaCl reduced to 10 g/L and MgSO4 . 7H2O reduced to 500 mg/L.
ATCC Medium 1770: Solution 1:
NaCl..........................................60.0 g
MgSO4 . 7H2O ...................................5.0 g
NH4Cl ..........................................1.0 g
CaCl2 . 2H2O ..................................25.0 mg
Trace Elements Solution SL-10 (see below)......1.0 ml
Wolfe's Vitamin Solution (see below)..........10.0 ml
Na2SeO3 . 5H2O.................................15.0 mcg
Resazurin......................................1.0 mg
Betaine........................................5.86 g
Yeast extract..................................1.0 g
L-Alanine......................................2.2 g
Distilled water..............................900.0 ml
Solution 2:
NaHCO3 .......................5.0 g
Distilled water.............80.0 ml
Bubble under N2 for 20 minutes; add to Solution 1.
Solution 3:
K2HPO4 .....................358.0 mg
KH2PO4 .....................223.0 mg
Distilled water.............10.0 ml
Autoclave under N2.
Solution 4:
Na2S . 9H2O ................300.0 mg
Distilled water.............10.0 ml
Autoclave under N2. Boil and cool Solution 1 under 80% N2, 20% CO2. Add Solution 2. Adjust to pH 8.3 +/- 0.1, tube and autoclave under 80% N2, 20% CO2 gas phase.
Add Solutions 3 and 4 aseptically to cooled, sterile medium. 2 Trace Elements Solution SL-10:
HCl (25%).........................10.0 ml
FeCl2 . 4H2O .......................1.5 g
ZnCl2 .............................70.0 mg
MnCl2 . 4H2O .....................100.0 mg
H3BO3 ..............................6.0 mg
CoCl2 . 6H2O .....................190.0 mg
CuCl2 . 2H2O .......................2.0 mg
NiCl2 . 6H2O ......................24.0 mg
Na2MoO4 . 2H2O.....................36.0 mg
Distilled water..................990.0 ml
Dissolve FeCl2 in the HCl, dilute with water, add and dissolve the other salts, adjust pH to 6.0 with NaOH, and fill up to 1.0 L. Wolfe's Vitamin Solution: Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS). Biotin.............................2.0 mg
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