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The presence of red (RNA) and green (DNA) fluorescent material identifying nucleic acids in the chlamydospores of Candida albicans has been well documented. Red fluorescence in chlamydospores is relatively diffused and ranges from small spots, observed in hyphal cells, to the entire protoplasmic content. Green fluorescence is rarely visible in these structures and, when present it can be observed next to the plasmalemma. The initial percentage values of the two curves related to the cell counts of red fluorescence of the suspensor cells and chlamydospores showed remarkable differences between the two structures. About 54% of the chlamydospores showed red fluorescence compared to about 28% of the suspensor cells. It seems from the results obtained in this study that much RNA was produced and/or accumulated in the chlamydospores and suspensor cells, rather than in mycelium where red fluorescence was not observed. The results obtained sustain the hypothesis that a chlamydospore should he considered a fully functional cell that is morphologically and physiologically active and independent from mycelium.

By incubating starved and unstarved yeast cells in synthetic media with a pH of 4.5 or 6.7 at 37 degrees C the effect of a 3 hours' glucose starvation on germ-tube production by Candida albicans was evaluated. In addition the endocellular content of total carbohydrates, glycogen, trehalose and proteins after and before the starvation were dosed. The most interesting result was the overcoming of the pH-regulated dimorphism, thanks to the starvation treatment. In fact the starved cultures produced germ-tubes indifferently in neutral or acid media, whereas the filamentation of the unstarved cultures was more copious in pH 6.7 medium. The endocellular content of trehalose and protein was unchanged, whereas total carbohydrates and glycogen showed a shortage after the 3 hours' glucose starvation. The possible involvements of these metabolic changes in the regulation of dimorphic transition are discussed.

Candida albicans starved cells were incubated in minimal synthetic liquid media containing different concentrations of ammonium sulphate (0.00, 0.02, 0.05, 0.10, 0.03, 0.50 g/L). Culture growth was monitored by measuring daily the optical density and by evaluating RNA and protein cellular content after 48 and 96 hours from the inoculum. The environmental availability of ammonium ion influenced the biomass production, that was maximum when its concentration was 0.10 and 0.30 g/L. In addition, an effect on cell duplication time was observed, this was particularly evident when the (NH4)2SO4 concentration was 0.10 g/L. The protein content increased in relation to the increase of ammonium ion availability, with a peak in correspondence to 0.30 g/L and a drop when the greatest concentrations were employed. RNA production was inversely proportional in respect to protein production. The optimal range of ammonium sulphate concentration for C. albicans growth was 0.10-0.30 g/L; over these concentrations there was an inhibitory effect. The rate of the protein and RNA syntheses seems to indicate the growth phase and the nitrogen nutritional conditions of the cultures, respectively.

By using the fluorescent, DNA specific stain DAPI (4',6-diamidino-2-phenylindole) some microscopic observations of Candida albicans pseudomycelium and chlamydoconidia were performed. In this manner blue fluorescent dots were noted both in yeasts, psudomycelium and chlamydoconidia, so evidencing the presence of a nucleus in C. albicans chlamydoconidia.

The growth of Cryptococcus neoformans in a minimal liquid synthetic medium with or without thiamine (10 micrograms/ml) was investigated. In these media the presence or absence of thiamine had no effect on the development of C. neoformans. To check these results, we performed a series of experiments on a solid form of the minimal synthetic medium. In this study a series of six serial transfers were carried out to starve the cells of nutrients that may have been carried over from their growth on rich media. In each of the transfers on the solid synthetic medium, C. neoformans showed a similar and scarce growth. This finding indicates that C. neoformans could be autotrophic in respect to thiamine.

A study of the vaginal yeast flora in pregnant women living in Cusco and in its region (Peru), located approximately 3000 m above sea level, is reported. We observed 300 pregnant, healthy and non-diabetic women who attended a gynaecological clinic in the Lorena, Regional or IPSS (Instituto Peruano de Seguridad Social) hospitals in Cusco. A comprehensive clinical history was obtained from each patient. It included age, work, parity, time of pregnancy, use of contraceptives or antibiotics, type of vaginal symptoms, type and amount of vaginal secretion. The yeasts were isolated from 44.3% of the cases. The positive cases were more frequently found in the following categories: 20-30 year-old patients (69.2%), ninth month of pregnancy (49.6%), first pregnancy (41.4%), no delivery (58.7%) and no abortion (66.9%). Most women complained of leukorrhea (75.9%) with mucus, scarce without odour (18.8%) or lumpy, regular, without odour (15.8%) secretion. The yeasts isolated were Candida albicans (66.2%), other Candida species (12.8%), Torulopsis glabrata (8.3%) or other Torulopsis species (2.2%), Saccharomyces cerevisiae (7.5%), Rhodotorula sp. (1.5%) and Trichosporon cutaneum (1.5%).

A new minimal synthetic medium, with low amount of glucose, without aminoacids, vitamins and neutral pH, which induces germ-tubes production in Candida albicans, is reported in this work. The results indicate a perfect agreement between the germ-tube test performed with the standard method in human or animal serum and this test performed in minimal synthetic medium. In this medium the germ-tube test for the presumptive identification of Candida albicans can be performed with the same formality, time and reproducibility as those in human or animal serum. This constitutes an interesting finding because it is easy to prepare, to store and is highly reproducible.

Candida albicans strain B 311-10 with and without starvation was cultivated in the minimal synthetic medium of Shepherd et al., modified without biotin, amino acids, low glucose concentration and with decreasing amounts of (NH4)2SO4, to determine the optimal growth requirement for this strain. All the experiments were carried out under sterile conditions at 25 degrees C in a thermostat with initial O.D.s (675 nm) of 0.500 and 0.100. Cell growth was generally monitored everyday for six days with a spectrophotometer by determining the absorbance of the cultures at 675 nm. All the experiments were repeated three times and a statistical analysis of the data with a probability of 99% and 1% of error was performed to confirm the validity of the results. Best growth was obtained with starved cells at an initial O.D. of 0.100 and with a 0.1 g/L concentration of (NH4)2SO4. At this concentration, the growth of C. albicans B 311-10 was best between the first and the fourth day with the maximum at the third day. With (NH4)2SO4 concentrations of 0.05 and 0.5 g/L, cell growth was the same.

A total of 90 cases of suspected dermatophytoses was observed over a period of 9 months in the city of Cusco (Peru) and in its environs. The age groups most commonly infected were 1-10 and 11-20 year old. The most common tineae diagnosed were tinea capitis (13.3%) and tinea unguium (11.1%). Only four species of dermatophytes were isolated: Microsporum canis (52.4%), Trichophyton mentagrophytes (35.7%), Tr. rubrum (9.5%) and Epidermophyton floccosum (2.4%). Dry weather, low temperature and high altitude do not seem to affect proliferation and infections by dermatophytes.

The growth of some opportunistic yeasts (Candida albicans, Cryptococcus neoformans, Torulopsis glabrata, Rhodotorula rubra, Geotrichum candidum, Trichosporon cutaneum, Saccharomyces cerevisiae) was essayed in a solid minimal synthetic medium without vitamins. From the preliminary results some species (Candida albicans, Cryptococcus neoformans, Rhodotorula rubra, Geotrichum candidum and Saccharomyces cerevisiae) seem to be autotrophic regarding the growth factor considered indispensable for their metabolism and for this reason normally added to the common synthetic media employed.

Different physical and chemical methods were used to detach the chlamydoconidia of Candida albicans from its mycelium. The action of concentrated H2SO4 acid for a 4-min period on cultures lysed both the mycelium and the outer but not the inner wall layer of the chlamydoconidia. The sulfuric acid procedure is recommended as the best method to obtain mycelium free chlamydoconidia because of its simplicity, rapidity and low cost.

Cells of Candida albicans, after 24 hours of growth in YM, were starved, alternatively, in citrate buffer, physiological solution, MMS deprived of glucose or ammonium sulphate. The eventual growth was monitored by determining the absorbance at 675 nm. Simultaneously, the cell morphology was also controlled. In a second series of experiments, the C. albicans cells taken from YM were starved for 72 hours in one of the mediums as stated above, and then reinoculated in MMS liquid without, alternatively, glucose or ammonium sulphate. Again the eventual growth was monitored as in the above method. The achieved results indicate the presence of a reserve of nitrogen, which can be utilized when a source of C is given to the cell. We therefore discuss the apparent lack of glucidic reserve and we propose a method for the consumption of nitrogen reserve. The aim of the work is to define how to obtain cells that contain the smallest amount possible of endogenous reserve.