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118 - DESULFOBACTERIUM MEDIUM
Solution A is prepared and autoclaved anaerobically under 80% N2 + 20% CO2. Solution C is filter-sterilized and gassed for 20 min with N2/CO2. Solutions B, D, E and F are filter-sterilized and gassed with N2. Solution G is autoclaved under N2. Solutions B to G are added to the sterile, cooled solution A in the sequence as indicated. Final pH of the medium should be 7.0. Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) 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.
 
Solution A
Distilled water930.0ml
Resazurin1.0mg
CaCl2 x 2 H2O0.15g
KCl0.5g
MgCl2 x 6 H2O3.0g
NaCl21.0g
NH4Cl0.3g
KH2PO40.2g
Na2SO43.0g
 
 
Solution B
Trace elements SL-10 (see medium 116)1.0ml
 
 
Solution C
Distilled water50.0ml
NaHCO32.5g
 
 
Solution D
Substrate (see below)
 
Solution D for E-001657
Distilled water10.0ml
Na-acetate x 3 H2O2.5g
 
 
Solution D for E-001658
Use a gas phase of 80% H2 + 20% CO2 (= substrate). Cells do not tolerate temperatures higher than 28°C.
 
Solution E
Vitamin solution (see medium 116)1.0ml
 
 
Solution F
Na2SeO3 x 5 H2O -solution*1.0ml
*3 mg in 1 l of 0.01 M NaOH
 
Solution G
Distilled water10.0ml
Na2S x 9 H2O0.4g
Solution A is prepared and autoclaved anaerobically under 80% N2 + 20% CO2. Solution C is filter-sterilized and gassed for 20 min with N2/CO2. Solutions B, D, E and F are filter-sterilized and gassed with N2. Solution G is autoclaved under N2. Solutions B to G are added to the sterile, cooled solution A in the sequence as indicated. Final pH of the medium should be 7.0. Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) 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.
 

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