# 323: TREPONEMA SACCHAROPHILUM MEDIUM ## Main sol. 323 0.12 g CaCl2 x 2 H2O 0.19 g MgSO4 x 7 H2O 0.45 g KH2PO4 0.45 g K2HPO4 0.9 g NaCl 0.9 g (NH4)2SO4 2 g Yeast extract (OXOID) 2 g Trypticase peptone (BD BBL) 0.4 ml n-Butyric acid 0.4 ml iso-Butyric acid 0.2 ml DL-2-Methylbutyric acid 0.2 ml n-Valeric acid 0.2 ml iso-Valeric acid 0.5 ml Sodium resazurin (0.1% w/v) 1 g L-Cysteine HCl x H2O 2.5 g Na2CO3 2 g D-Glucose 12 g Agar (BD Bacto), for solid medium 1000 ml Distilled water 1. Dissolve ingredients (except cysteine, carbonate and glucose), adjust pH to 7.0 and sparge medium with 100% CO2 gas for 30 - 45 min to make it anoxic. Add the cysteine and carbonate, then equilibrate the medium with the CO2 gas to pH 7.0. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add glucose from an anoxic stock solution prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary. 2. For solid medium add 12.00 g/l agar (BD Bacto).