# 408: PSYCHRILYOBACTER SP. MEDIUM ## Main sol. 408 10 g Peptone (BD Bacto) 3 g Yeast extract (OXOID) 10 g Casamino acids (BD Bacto) 20 g L-Glutamic acid 2 g NH4Cl 2 g K2HPO4 x 3 H2O 25 g NaCl 10 ml **Modified Wolin's mineral solution** 0.5 ml Sodium resazurin (0.1% w/v) 0.5 g L-Cysteine HCl x H2O 1000 ml Distilled water 1. Dissolve ingredients (except cysteine), bring medium to the boil, then cool to room temperature under 100% N2 gas atmosphere. Add and dissolve cysteine, adjust pH to 7.0 and dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. ## Modified Wolin's mineral solution 1.5 g Nitrilotriacetic acid 3 g MgSO4 x 7 H2O 0.5 g MnSO4 x H2O 1 g NaCl 0.1 g FeSO4 x 7 H2O 0.18 g CoSO4 x 7 H2O 0.1 g CaCl2 x 2 H2O 0.18 g ZnSO4 x 7 H2O 0.01 g CuSO4 x 5 H2O 0.02 g AlK(SO4)2 x 12 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.03 g NiCl2 x 6 H2O 0.3 mg Na2SeO3 x 5 H2O 0.4 mg Na2WO4 x 2 H2O 1000 ml Distilled water 1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.