A Further Analysis of the Nutrition of Paramecium.*

SUMMARY. A medium is described for the axenic culture of Paramecium multimicronucleatum which is chemically defined except for one constituent—a nondialyzable fraction (NDF) of an exhaustively dialyzed yeast extract. Using this medium it has been possible to demonstrate an absolute requirement for stigmasterol and the following B‐vitamins: folic acid, riboflavin, thiamine and pantothenic acid. There was evidence that nicotinamide and pyridoxal are also required for continued growth of the organisms. The nitrogen requirements could be met with any of the following: proteose peptone, purified casein, crystalline ovalbumin, or a mixture of amino acids. A carbon source, in addition to that supplied by the NDF or the amino acids, was essential. Sodium acetate and sodium pyruvate would satisfy this requirement.