The relative importance of carbon dioxide, pH, anaerobiosis, and composition of medium on filamentation in Candida albicans
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Gandida albicans strain 105 from a normal human and strain 582 (from the American type Culture Collection) were used for studying the effect in vitro of pH, various amounts of carbon dioxide, nitrogen, and composition of media on filamentation in this yeast-like organism. The yeast phase of the organism was maintained on a glucose, glycine, yeast extract (GGY) medium (1%; 1%; 0.5%) at 37°C. The experiments were conducted on both solid and liquid media. All cultures were incubated at 37°C. for 48 hours. The two strains of c. albicans, although similar to one another in their yeast forms, behaved differently toward the environmental conditions used; strain 582 responded more readily to the factors inducing filament formation than did strain 105. Increasing the pH above 6.5 to 7.0, 7.5 and 8.0 induced maximum filamentation in strain 582, whereas no filaments were produced by strain 105. All the aerobic cultures on solid GGY medium showed alkalinity and were positive for ammonia at the end of the incubation period. In liquid media, no alkalinity was observed at any pH values. Presence of 75% carbon dioxide in the atmosphere increased filamentation in strain 582 to a maximum degree, and induced mycelial formation in strain 105. With 94% or 95% carbon dioxide, growth and filamentation decreased in both strains. None of the CO2 cultures showed alkalinity at the end of the incubation period. Moreover, all the CO2 cultures were negative for ammonia. Growth under nitrogen (9J%) was less than that of the aerobic cultures. However, colonies appeared larger in size. Nitrogen stimulated filamentation in strain 105 only at a pH of 8.0, whereas strain 582 formed a maximum amount of filaments at pH values of 7.0 to 8.0. All the solid cultures under nitrogen showed alkalinity, while the liquid cultures were acid at all pH values. The occurrence of deamination in a medium without glucose in both strains of C. albicans showed that this organism was able to use glycine its source of both nitrogen and carbon. However, only a sparse growth was obtained in a medium lacking glucose. Strain 105 did not form filaments in such a medium, while strain 582 did so. Since more filaments were produced by the latter strain when a fresh subculture on a GGY medium was transferred to a medium without glucose, it was concluded that possibly glucose is required for both growth and filamentation. Comparative studies of the effect of a medium containing mannose with a glucose medium showed the two sugars behaved similarly with regard to fermentation and filament induction in both strains or c. albicans. Under conditions where glucose induced filamentation (e.g., with C02 or N2), mannose also induced filamentation. The decreased growth in the presence of oleic or stearic acid in a concentration of 40 micrograms per liter was attributed to the toxic effect of the fatty acids. Moreover, it was noted that the two acids had different effects on filamentation in the two strains. Oleic acid in a solid GGY medium induced hyphal formation in strain 105 only under nitrogen; without glucose, oleic acid did not bring about filamentation under any of the atmospheric conditions tested. In liquid media, oleic acid induced filamentation for strain 105 only when glucose was omitted. With strain 562, oleic acid promoted filamentation in both liquid and solid media with or without glucose, except for solid cultures incubated under nitrogen in the absence or glucose. Stearic acid did not stimulate filamentation in strain 105 under any conditions, but did increase hypha! formation in strain 582. In the presence of stearic acid, maximum filamentation occurred in aerobic cultures wnen glucose was absent. Although maximum filamentation occurred with an increase in the pH of the medium under aerobic conditions, in the presence of 75% C02, under nitrogen or in the presence of stearic acid in a medium without glucose, yeast cells were also present, indicating that this Y to f transformation was not complete.
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