# 968: ANAEROBACILLUS ARSENICISELENATIS MEDIUM ## Main sol. 968 0.1 g (NH4)2SO4 25 mg MgSO4 90 g NaCl 0.15 g K2HPO4 0.08 g KH2PO4 1 ml **Trace element solution SL-10** 1 ml **Selenite-tungstate solution** 1.25 g NaNO3 1.7 g Na-DL-lactate 0.2 g Yeast extract 0.5 ml Sodium resazurin (0.1% w/v) 10.6 g Na2CO3 4.2 g NaHCO3 0.25 g L-Cysteine HCl x H2O 0.25 g Na2S x 9 H2O 1000 ml Distilled water 1. Dissolve ingredients (except carbonate, hydrogencarbonate and reducing agents), then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add solid carbonate and bicarbonate, adjust pH to 9.8, dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before inoculation, add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 9.8, if necessary. ## Trace element solution SL-10 10 ml HCl (25%) 1.5 g FeCl2 x 4 H2O 70 mg ZnCl2 100 mg MnCl2 x 4 H2O 6 mg H3BO3 190 mg CoCl2 x 6 H2O 2 mg CuCl2 x 2 H2O 24 mg NiCl2 x 6 H2O 36 mg Na2MoO4 x 2 H2O 990 ml Distilled water 1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. ## Selenite-tungstate solution 0.5 g NaOH 3 mg Na2SeO3 x 5 H2O 4 mg Na2WO4 x 2 H2O 1000 ml Distilled water