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ATCC medium: 1648 Modified Desulfobacterium medium
Solution A:
Na2SO4 .........................................3.0 g
KH2PO4 .........................................0.2 g
NH4Cl ..........................................0.3 g
NaCl..........................................21.0 g
MgCl2 . 6H2O ...................................3.0 g
KCl............................................0.5 g
CaCl2 . 2H2O ...................................0.15 g
Resazurin......................................1.0 mg
Distilled water..............................920.0 ml
Solution B:
Trace Element Solution SL-10 (see below).......1.0 ml
Solution C:
NaHCO3 .........................................2.5 g
Distilled water...............................50.0 ml
Solution D:
Sodium acetate . 3H2O ..........................2.5 g
Distilled water...............................10.0 ml
Solution E:
Wolfe's Vitamin solution (see below)..........10.0 ml
Solution F:
Na2SeO . 5H2O ..................................3.0 mg
0.01 M NaOH....................................1.0 L
Use 1.0 ml/L medium.
Solution G:
Na2S . 9H2O ....................................0.4 g
Distilled water...............................10.0 ml
Directions for complete medium:
Prepare and autoclave Solutions A and D anaerobically under 80% N2, 20%
CO2. Filter-sterilize and gas Solution C for 20 minutes with 80% N2, 20%
CO2. Filter-sterilize and gas Solutions B, E and F with N2. Autoclave
Solution G under N2. Add Solutions B through G to the sterile, cooled
Solution A in the sequence indicated. Final pH of the medium should be
7.0.
Addition of 10-20 mg/L sodium dithionite (e.g., from 5% (w) solution
freshly prepared under N2 and filter-sterilized) may stimulate growth of
all strains at the beginning. For transfers use 5-10% inoculum. Incubate
all strains in the dark.
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 to 1.0 L with distilled
water.
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 .