Abstract
SYNOPSIS. Paramecium multimicronucleatum has been cultured for 20 years on a medium of salts, vitamins, amino acids, fatty acids, ribosides, and stigmasterol plus a little nondialyzable fraction (NDF) of baker's yeast. Fractionations of NDF identified 2 essentials: (a) in a fraction < 100,000 daltons which contained much protein and replaceable by ovalbumin and (b) in a fraction of < 300,000 daltons; this fraction contained much polysaccharide, replaceable by glycogen, which is > 300,000 daltons. For 2 years now P. multimicronucleatum has grown well with ovalbumin and glycogen replacing NDF. Besides ovalbumin, concanavalin A satisfies the protein requirement; this lectin attaches to sugar residues in glycogen. Studies with a fluorescent dye, PGA‐1A, a stilbene derivative, provides further evidence for the polysaccharide requirement. This dye attaches to polysaccharides; when added to glycogen, and this in turn is added to a culture containing ovalbumin, fluorescent blue vacuoles appear within 2–3 h. When dye + glycogen were added to a culture without ovalbumin, no fluorescent vacuoles were found. A protein appears involved in formation of food vacuoles; this fits the pattern for endocytosis described in recent reviews. Besides glycogen, mannan gave good growth. Dextrin and amylopectin gave only fair growth through 7 serial transfers; glucose, maltose and amylose did not sustain growth. Strain 51 of P. tetratrelia, which grows well in NDF medium, grows well when NDF is replaced with ovalbumin and glycogen.
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