Role ofCandida in indirect pathogenesis of antibiotic associated diarrhoea in infants

One hundred and thirty seven isolates ofCandida species were isolated from antiobiotic associated diarrhoea cases and were examined to study the role ofCandida in the pathogenesis of diarrhoea in infants. The quantitative estimation of yeast population by simple gram stain smear revealed more than 70% of the cases had 3+ score. The isolates further screened for detection of-lactamases. Among the isolatedCandida sp,-lactamases was secreted byC. albicans, C. tropicalis, C. krusei andC. parapsilosis. Further, 46% of theCandida isolates were found to be produced 741–1110 mU/ml of-lactamases, suggesting that these enzyme would inactivate penicillin group of drugs and cause failure in the therapy directed against other diarrhoegenic bacteria.Abbreviation AAD antibiotic associated diarrhoea     

A method for taxonomic determination ofCandida albicans with DNA probes

Determination ofCandida species represents an important problem derived from the clinical implications of the species belonging to this genus. DNA probes have already been used for the epidemiology ofCandida albicans, as well as for taxonomic analysis ofCandida and other genera, although these probes are based on non-species-specific DNA sequences. In this work we carried out a 48-h assay, allowing the identification ofC. albicans from clinical isolates, using DNA probes based onC. albicans LEU2 andURA3 genes. Another probe related toC. albicans SEC18 gene was shown not to beC. albicans specific.     

Fungal diseases of the respiratory tract

The proportion ofCandida and non-Candida species in the clinical material from patients. with respiratory-tract diseases was determined.C. albicans was isolated in 102 cases. An additional 89 strains of yeasts, isolated in association with respiratory diseases, belonged to 10 non-albicans Candida spp. andCryptococcus spp. The prevailing species, which occurred in 47 cases, wasC. parapsilosis. C. tropicalis, C. glabrata, andC. guilliermondii were isolated in 12, 10, and 9 cases, respectively. Four strains ofC. krusei and three strains ofC. lusitaniae and one strain each ofC. freyschussii, C. robusta, C. zeylanoides, andCryptococcus neoformans were also isolated.     

Composition of lipids produced by some strains ofCandida species. Production of single-cell oil in a chemostat culture

Fatty acid composition of the lipids produced by four strains ofCandida species was studied. Oleic acid was the principal fatty acid. Cellular lipids ofCandida sp. andC. pulcherima were rich in palmitic acid. Lipids fromC. lipolytica contained a significant amount of palmitoleic acid, whereasC. farinosa produced oil rich in stearis and α-linolenic acid. Analysis of cellular lipids ofCandida sp. andC. pulcherima during growth on a nitrogen-limited medium showed that oils accumulated in the exponential growth phase were more unsaturated than those accumulated in the decelerating and stationary phases. In a chemostat culture,Candida sp. accumulated about 40% of lipid. The specific rate of lipid formation, at a dilution rate ofD=0.09/h, was 35 mg of lipid per g of biomass per h and the yield of lipid on glucose was 11.4%.     

Identification of Candida isolated from the cutaneous candidiasis by the combined use of confirmatory medium and slide agglutination with monofactorial antibodies

Forty strains ofCandida and one ofTorulopsis were isolated from patients with cutaneous candidiasis. The isolates comprised 29 strains ofC. albicans, 7 strains ofC. tropicalis, 2 strains ofC. guilliermondii, and one each ofC. parakrusei, C. lipolytica, andT. famata were identified by the ordinary method. Besides the common pathogenC. albicans, a few other species ofCandida may be etiologic organisms of cutaneous candidiasis. These strains were re-examined by combined use of sucrose agar slants and slide agglutination tests with IgG monofactorial antibodies as a rapid identification method, especially for determining serotypes ofC. albicans. The new method was useful and reliable for rapid identification ofC. albicans and related species. All strains ofC. albicans isolated from skin lesions proved to be standard serotypes ofC. albicans.

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Zusammenfassung Vierzig Stämme vonCandida und eins vonTorulopsis wurden aus Kranken mit kutanen Candidamykosen isoliert. Neunundzwanzig Stämme vonC. albicans, 7 vonC. tropicalis, 2 vonC. guilliermondii, und je einer vonC. parakrusei, C. lipolytica undT. famata wurden mit dem ordinären Methode identifiziert. Außer dem gewohnlichen Erreger,C. albicans, konnten auch ein Paar andere Spezies vonCandida als den Erreger betrachtet werden. Sechsunddreißig Stämme vonC. albicans undC. tropicalis wurden mit der von uns verbesserten kombinierten serologischen und biologischen Methode untersucht, besonders um den Serotypus vonC. albicans festzusetzen. Die neue Methode war gut und zuverlässig als die rapide Identification vonC. albicans und verwandten Spezies. Alle aus der Hautläsion isoliertenC. albicans waren der in Japan allgemeine Serotypus vonC. albicans.

     

Comparison of PCR-RFLP with 21-plex PCR and rDNA Sequencing for Identification of Clinical Yeast Isolates

Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which are time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR. One hundred and seventy-three yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method. The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=?C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR-RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR. The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.

     

The taxonomy of yeasts isolated fromDrosophila in the Yosemite region of California

Summary A survey was made of the yeasts occurring in the intestinal tract of wild species ofDrosophila occurring in the Yosemite Region of California. Two hundred and forty one yeasts, representing 42 species and varieties, were identified. Each isolate was obtained from a different fly. Almost half of the isolates belong toSaccharomyces. The most common species in this genus wereS. montanus (36 isolates),S. veronae (30 isolates),S. cerevisiae var.tetrasporus (22 isolates) andS. drosophilarum (13 isolates). Further species are listed in Table 1.Zygosaccharomyces fermentati Naganishi was shown to be a distinct species and not a synonym ofS. cerevisiae. In order to avoid confusion with another yeast of the same name, it has been proposed to change the nameZ. fermentati toS. montanus Naganishi. Two new species ofSaccharomyces were described,S. wickerhamii andS. kluyveri. S. mangini var.tetrasporus has been renamedS. cerevisiae var.tetrasporus. A non-cellobiose attacking strain ofS. drosophilarum has been designated tentativelyS. drosophilarum var.acellobiosa. A new species of the genusPichia was described asP. xylosa. Saccharomyces pastori andSaccharomyces pini were transferred to the genusPichia on the basis of arguments given in the preceding paper. A new species ofTrichosporon was described asTr. aculeatum on the basis of the presence of characteristic needlelike cells. Common species besides those mentioned inSaccharomyces wereHansenula angusta (19),Kloeckera apiculata (15),Kl. magna (13), andTorulopsis stellata (10). Other genera represented wereHanseniaspora, Cryptococus, Rhodotorula, Candida andOospora. Evidence was obtained that many species of imperfect genera consist of distinctly different physiological types.     

Systemic Candida infection in University Hospital 1997–1999: the distribution of Candida biotypes and antifungal susceptibility patterns

A total of 102 Candida species were isolated from blood cultures from January 1997 to October 1999. Using assimilation of carbohydrate test, 52 (51.0%) of the Candida sp. were identified as C. parapsilosis, 25.5% (26) were C. tropicalis. C. albicans made up 11.8% (12), 6.9% (7) were C. rugosa, 3.8% (4) C. glabrata and 1% (1) C. guilliermondii. No C. dubliniensis was found in the study. In vitro antifungal susceptibility tests showed that all Candida species were sensitive to nystatin, amphotericin B and ketoconazole. Although all isolates remained sensitive to fluconazole, intermediate susceptibility was found in 3 C. rugosa isolates. Antifungal agents with high frequency of resistance were econazole, clotrimazole, miconazole and 5-fluorocytosine. Candida species found to have resistance to these antifungal agents were non-C. albicans. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparative observations of ultrastructure of five species of Candida

An electron microscopic comparison was made of five species ofCandida, namely:C. guilliermondii, C. krusei, C. parapsilosis, C. stellatoidea andC. tropicalis. The cell wall, plasma membrane and the cytoplasm with its organelles were described. The cell wall ofC. tropicalis was twice as thick as the cell wall in the other species.C. krusei appeared with distinct, rather elaborate wall sculpturing, a feature not pronounced in the other four species. A single nucleus with nucleolus appeared only in micrographs ofC. guilliermondii andC. krusei. At the same time, large central electron-luscent area (vacuole) appeared in the cells ofC. guilliermondii, C. parapsilosis andC. stellatoidea. The cytoplasm ofC. tropicalis was characterized by a granular appearance. Budding cells and pseudohyphae appeared similar to single cells in their general organelles. Such organelles in species studied were similar to these reported for other yeasts. These include: mitochondria, lipid granules, endoplasmic reticulum, ribosomes and vacuoles.Southwest Foundation for Research and Education, San Antonio, Texas.In partial fulfillment of the requirement of course work for Master of Science, Incarnate Word College, San Antonio.     

A comparative study of the extracellular and cell-wall polysaccharides of someCandida species

Candida bogoriensis, C.buffonii, C.diffluens, C.foliarum andC.javanica, produce an extracellular polysaccharide which contains galactose, glucose, mannose, fucose and rhamnose. These sugars were also found in cell-wall preparations of the same yeast species. The cell-wall preparations ofC.diffluens andC.foliarum included capsular material. The similarity in composition of the extracellular polysaccharides and components of the cell wall suggests that both are synthetized by the same enzymatic system. The fiveCandida species may be closely related.     

Species Distribution and Antifungal Susceptibility Profile of Oral Candida Isolates from HIV-infected Patients in the Antiretroviral Therapy Era

In this study, we investigated the yeasts colonization of genus Candida, including C. dubliniensis, isolated of HIV-infected patients oral cavities and we accessed in vitro susceptibility pattern of the Candida isolates to four antifungal agents. Out of 99 patients investigated, 62 (62.6%) were colonized with yeasts. C. albicans was the prevailing species (50%). C. dubliniensis isolates were not recovered in our study. We verified that 8.1% of the yeasts isolated were resistant to fluconazole, 8.1% to itraconazole and 3.2% to voriconazole. The isolates demonstrated very low voriconazole MICs, in which 79% (49/62) presented values of 0.015 μg/ml. All Candida isolates were susceptible to amphotericin B. The results reported here showed that although C. albicans continues to be present in one-half of oral Candida carriage of HIV-infected patients, Candida non-albicans species are increasing among these patients. Besides, the findings of resistant isolates endorse the role of antifungal susceptibility testing whenever antifungal treatment with azoles is planned.     

Ecology of yeasts in polluted water

The general size and composition of the extant yeast populations in 13 polluted freshwater habitats were surveyed. Subsequently the yeast populations in three of the 13 locations were quantitatively determined and compared. The three locations had (A) low pollution levels, (B) heavy industrial waste pollution, and (C) heavy domestic waste pollution.The yeast population at location A was dominated byRhodotorula andCryptococcus isolates. At station BRhodotorula andCandida were predominant.Candida isolates were in the majority at location C andRhodotorula strains were second in frequency, but were much lower in proportion of the population than at the other two habitats.These polluted waters in general had large yeast populations, ranging as high as 27,000 yeasts per 100 ml, and averaging approximately 3000 yeasts per 100 ml.The presence of human wastes was especially associated with large increases in the proportion ofCandida yeasts in the environment. The genusRhodotorula was consistently present at all locations, but the genusCryptococcus was a major component of the yeast population only in non-polluted or lightly polluted fresh water.We appreciate the assistance of the Calumet Area Surveillance Program, Federal Water Pollution Control Administration, Chicago, in the collection of water samples. This investigation was supported by U.S. Public Health Service Research Grant No. AI 04642 from the National Institute of Allergy and Infectious Diseases.     

The taxonomy of yeasts found in exudates of certain trees and other natural breeding sites of some species ofDrosophila

Summary A survey was made of the yeasts occurring in slime fluxes ofQuercus kellogii (black oak),Abies concolor (red fir) and in certain mushrooms and dead logs in the mountains of the Yosemite region of California. Most of the 134 isolates identified were found to be non-fermentative or poorly fermentative (fermenting glucose only and usually weakly). The isolates were placed in the following genera:Pichia (34),Debaryomyces (35), imperfect forms ofHansenula (39),Endomyces (3),Saccharomyces (2),Candida (11),Trichosporon (7),Torulopsis (1),Cryptococcus (1) andSporobolomyces (1). Four new species have been described,Pichia silvestris, Pichia quercibus, Pichia carsonii andDebaryomyces fluxorum. P. quercibus andP. carsonii are unusual representatives of the genusPichia in that they form a very primitive pseudomycelium and lack pellicles on malt extract. To accommodate such species an amended diagnosis of the genusPichia has been proposed byPhaff in an accompanying paper.Saccharomyces pastori, of which 12 isolates were obtained, has been transferred to the amended genusPichia asP. pastori (Guilliermond) nov. comb.     

Candidal urinary tract infections caused by non-<Emphasis Type="Italic">albicans Candida</Emphasis> species

The incidence of non-albicans Candida and non-Candida species isolated from the urine of patients admitted to various departments of theFaculty Hospital of the Medical Faculty of Šafárik University in Košice was examined. From a total of 94 samples of analyzed urine 58 strains ofC. albicans and 36 strains of yeasts belonging to 6 species of non-albicans Candida and non-Candida spp. were detected:C. parapsilosis (n=23), C. tropicalis (6), C. krusei (3), C. robusta (2), C. catenulata (1) andCryptococcus neoformans (1). In relation to the diagnosis, the yeasts were isolated from patients suffering from a kidneys disease, epididymitis, diabetes, neoplastic diseases, urogenital anomalies, obstructive uropathy, cystitis, prostatitis, hemolytic-uremic syndrome, and others.     

Prevalence and Antifungal Susceptibility of Yeasts Obtained from the Oral Cavity of Elderly Individuals

Infections caused by Candida yeasts are common in elderly individuals. Seventy-five isolates of Candida spp. were obtained from saliva samples of 136 institutionalized elderly individuals resident in six retirement homes of Belo Horizonte, Brazil. Forty-seven isolates (62.66%) were identified as Candida albicans, 15 (20%) as C. tropicalis, 7 (9.33%) as C. glabrata, 4 (5.33) as C. parapsilosis, and 2 (2.67%) as C. guilliermondii. Among the 136 elderly individuals studied, 49 (36%) were male and 87 (64%) were female. Ages ranged from 60 to 90 years old. Sixty-three (46.3%) of the institutionalized individuals were denture wearers and, among them, 53 (84.1%) carried Candida yeasts in the oral cavity. Forty-four subjects presented lesions in the oral mucosa and among these, 36 (82%), had positive culture for Candida spp. The samples were tested for the in vitro susceptibility to amphotericin B, itraconazole, fluconazole and 5-flucytosin, and great variations were observed in the minimum inhibitory concentrations (MIC) of these drugs according to the species.     

Interactions ofCandida albicans with bacteria and salivary molecules in oral biofilms

The yeastCandida albicans coaggregates with a variety of streptococcal species, an interaction that may promote oral colonization by yeast cells.C. albicans andCandida tropicalis are the yeasts most frequently isolated from the human oral cavity and our data demonstrate that both these species bind toStreptococcus gordonii NCTC 7869 while two otherCandida species (Candida krusei andCandida kefyr) do not. Adherence ofC. albicans was greatest when the yeast had been grown at 30° C to mid-exponential growth phase. For 21 strains ofC. albicans there was a positive correlation between the ability to adhere toS. gordonii and adherence to experimental salivary pellicle. Whole saliva either stimulated or slightly inhibited adherence ofC. albicans toS. gordonii depending on the streptococcal growth conditions. The results suggest that the major salivary adhesins and coaggregation adhesins ofC. albicans are co-expressed.     

Ubiquinone and urease distribution inTaphrina andSymbiotaphrina

Identifications of ubiquinone isoprenologues are presented for isolates identified with six species ofTaphrina and for isolates of the two species ofSymbiotaphrina. All had Q-10 as the major ubiquinone system. The inclusion ofT. populina andS. buchneri, the respective type species, establishes this as the value for these genera. Both species ofSymbiotaphrina were urease positive even though, according to the literature, they are unable to utilize urea as a sole nitrogen source. The urease results for theTaphrina isolates were mixed.     

Molecular Identification of <Emphasis Type="Italic">Candida orthopsilosis</Emphasis> Isolated from Blood Culture

The incidence of candidemia and invasive candidiasis have increased markedly due to the increasing number of immunocompromised patients. There are five major medically important species of Candida with their frequency of isolation in the diminishing order namely Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata and Candida krusei. In addition, there are numerous other species of Candida which differ in their genetic makeup, virulence properties, drug susceptibilities and sugar assimilation capabilities. In this report, an unusual Candida species was isolated from the blood of two leukaemic patients. Conventional culture and biochemical tests identified the Candida species as C. parapsilosis. Using fungal-specific oligonucleotide primers ITS1 and ITS4, we managed to amplify the ribosomal RNA gene and its internal transcribed spacer region from the genomic DNA of these isolates. The PCR products were then purified and subjected to automated DNA sequencing using BLAST and CLUSTAL sequence analysis identified these isolates to be Candida orthopsilosis. Candida orthopsilosis is a new species recently identified in 2005, being morphologically indistinguishable from C. parapsilosis and was previously classified as a subspecies of C. parapsilosis. This report highlights the importance of complementing traditional culture and biochemical-based identification methods with DNA-based molecular assays such as PCR as the latter is more superior in terms of its discriminatory power and speed.     

Candida-associated denture stomatitis

Candida-associated denture stomatitis was demonstrated by its cultivation in 171 out of 240 patients examined with partial or total dentures. After taking smears from lesions of the oral mucosa (tongue, cheeks, palate) and the contiguous denture surface by cotton wool swabs and inoculating them onto Sabouraud glucose agar and CHROMagarCandida, individual yeast species were identified by a germ tube. filamentous, and assimilation tests employing the commercial kit AuxaColor. SevenCandida species were identified in smears from the oral mucosa lesions and the contiguous denture surface:C. albicans (95 patients),C. tropicalis (26).C. parapsilosis (20),C. krusei (14),C. guilliermondii (12),C. lusitaniae (1) andC. freyschusii (1). Diabetes mellitus, neoplastic diseases, chemotherapy, radiotherapy, and broad-spectrum antibiotic therapy were identified as some of the large number of factors predisposing patients to stomatitis prothetica.     

Leucosporidium gen. n., the heterobasidiomycetous stage of several yeasts of the genusCandida

Investigation of yeasts from Antarctic regions revealed that certain species ofCandida have heterobasidiomycetous life cycles. Two distinct but overlapping groups of species were found: heterothallic and self-sporulating species.Candida scottii is a heterothallic species with the following life cycle: opposite mating types will conjugate and develop a dikaryotic mycelium with clamp connections. Karyogamy occurs in the teliospore which germinates and produces a promycelium. Meiosis takes place in the promycelium, followed by development of haploid sporidia to complete the life cycle. In addition,C. scottii has a self-sporulating phase. From a single cell, in the apparent absence of mating, a uninucleate mycelium is produced that lacks clamp connections. Teliospores, promycelia and sporidia develop that appear similar to those produced from dikaryotic mycelium.The self-sporulating species have life histories similar to the self-sporulating phase ofC. scottii; except that heterothallism has not been observed.Based on these life histories the new genusLeucosporidium is proposed with two heterothallic species (Leu. scottii andLeu. capsuligenum) and five self-sporulating species (Leu. antarcticum, Leu. frigidum, Leu. gelidum, Leu. nivalis andLeu. stokesii. Leu. antarcticum) andLeu. stokesii have not been described under the genusCandida.Contribution No. 1138 from the Institute of Marine Sciences, University of Miami, Fla.This study was supported by the National Science Foundation through the Office of Antarctic Programs, Grant GA-3957. The authors are grateful to Dr. E. Marelli for editing the Latin diagnosis.