Blackwell Publishing Journal Backfiles 1879-2005
SYNOPSIS. Telotrochidium henneguyi was cultured axenically. The major nutrients in medium T-3 were liver extract (1:20 National Biochemicals Co.), hydrolyzed yeast nucleic acid, glucose, and dl-β-hydroxybutyric acid; these were fortified with phosphate buffer, EDTA, and penicillin. The supplements were: 18 amino acids, 7 vitamins, 10 salts supplying trace-metals; uridylic, cytidylic, guanylic, and adenylic acids; thymidine-5-diphosphate, nicotinamide mononucleotide, and choline. Optimum conditions for axenic cultivation were obtained with phosphate buffer 2 × 10−1M, penicillin 5,000 U.S.P. units/ml, 23°C, pH 6.8. A monoxenic maintenance medium (“T-broth”) allowed prolific growth and produced trxmendous populations. It was composed of Bacillus cereus in an aqueous broth-concoction of Proteose-peptone, Cerophyl, and wheat kernels. Axenic T-3 medium supported serial subculture; yields of peritrichs were comparable to those in T-broth. Axenic yields were poorer in medium lacking the T-3 supplemenits but containing the major nutrients fortified with acid-hydrolyzed gelatin, serine, riboflavin, EDTA, CaCL, FeCI3, KCI, MgSO4·7 H2O, phosphate buffer, and penicillin. For rapid axenination, excystment was induced by vibrating encysted peritrichs ∼ 30 sec in a Vortex Jr. mixer. Freshly excysted animals were washed by centrifugation. A salt solution, not distilled water, was used for washing inoculants. Inocula consisting of 1 × 103 or more animals were obtained by conventional centrifuge methods.Extension of this investigation to construction of a chemically defined medium is discussed.
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