Transfer of spores, bacteria and yeast into toluene containing phospholipids and low amounts of water: preservation of the bacterial respiratory chain.

A method that allows the transfer of spores, bacteria and yeasts into a ternary system composed of toluene, phospholipids and low amounts of water is described. Initially an emulsion is formed by sonication of cells suspended in water in presence of toluene and phospholipids. The emulsion formed was subsequently clarified by blowing N2 on its surface and transparency was achieved when the water content of the system was reduced to 1-3 microliter/ml of organic solvent. The cells in ternary systems exhibit a reduction of cell volume, but the general structure is preserved. About 1/10,000 bacilli or yeast were viable, but spores were viable after 30 days in the ternary system. Yeast cells transferred to the ternary system, and back to an all water media failed to show oxygen uptake. In contrast bacilli that remained in the ternary system for 2 days respired to 80% of their maximal capacity when returned to an aqueous media.