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Clinical isolates of Candida guilliermondii include Candida fermentati.

Clinical isolates of Candida guilliermondii that were investigated by isoenzyme and randomly amplified polymorphic DNA analyses represented two distinct species. The two species were distinguished on the basis of delayed fermentation of galactose. The larger group of isolates was closely related to the anamorph C. guilliermondii ATCC 6260T (T = type strain) and its teleomorph, Yamadazyma (= Pichia) guilliermondii ATCC 46036T. The remaining group, whose members fermented galactose, was very similar to Candida fermentati CBS 2022, which had for many years been placed in synonymy with C. guilliermondii. Three additional groups were represented by individual strains; these strains included C. guilliermondii var. soya ATCC 20216, which was found to represent Yamadazyma ohmeri. The type strain of Y. guilliermondii is redefined.

Base Sequence↗

The Candida Genome Database: facilitating research on Candida albicans molecular biology.

The Candida Genome Database (CGD; http://www.candidagenome.org) is a resource for information about the Candida albicans genomic sequence and the molecular biology of its encoded gene products. CGD collects and organizes data from the biological literature concerning C. albicans, and provides tools for viewing, searching, analysing, and downloading these data. CGD also serves as an organizing centre for the C. albicans research community, providing a gene-name registry, contact information, and research community news. This article describes the information contained in CGD and how to access it, either from the perspective of a bench scientist interested in the function of one or a few genes, or from the perspective of a biologist or bioinformatician interpreting large-scale functional genomic datasets.

Alleles↗

Nature of Candida albicans-derived carbohydrate antigen recognized by a monoclonal antibody in patient sera and distribution over Candida species.

The minimal epitope of an anti-Candida albicans mannan monoclonal antibody (MAb) EB-CA1, used to detect mannanemia in patient sera, was determined, MAb EB-CA1 exhibited reactivity with oligomannosides released from the mannan acid stable domain, converted into neoglycolipids (NGLs) and coated onto ELISA plates. Reactivity occurred with mannopentaose and higher oligomers, whereas mannotriose and mannotetraose were unreactive. MAb EB-CA1 binding to mannan acid stable mannopentaose NGL displayed a dose dependent and saturable specific reactivity curve whereas there was a complete absence of binding, even at high concentrations, with NGLs constructed from the beta-1,2-linked mannopentaose derived from the mannan acid labile fraction. MAb EB-CA1 binding to acid stable mannopentaose NGL was inhibited by the homologous oligomannoside but not by mannotriose and mannotetraose. NMR analysis showed that mannotriose and mannotetraose contained exclusively alpha-1,2-linked D-mannopyranose units and that mannopentaose was a mixture of a mannopentaose alpha-1,2-linked and an isomer in which the fifth mannose was alpha-1,6-linked to the reducing unit of manno-alpha-1,2 tetraose. Western blot analysis has shown that MAb EB-CA1 epitope was expressed on a wide range of C. albicans manno-glycoconjugate as well as on manno-glycoconjugates of other pathogenic species of the genus Candida, viz. C. tropicalis, C. glabrata, C. parapsilosis and C. krusei.

Antibodies, Fungal↗

Implantation of Candida albicans and other Candida species in the oral cavity of rats.

The carriage of five Candida species in the mouths of normal and sialoadenectomised rats was determined for periods up to 30 days after inoculation into the oral cavity. In both test and control animals, Candida albicans was the species recovered in greatest quantities at all periods, followed by C. parapsilosis and C. tropicalis. In contrast, C. guilliermondii and C. krusei were isolatable only in small numbers and only from the 1st up to the 5th day; they were not present thereafter. Sialoadenectomy favoured oral colonisation only by C. albicans (P < 0.05) and did not influence the carriage of the other species.

Analysis of Variance↗

Treatment of murine Candida krusei or Candida glabrata infection with L-743,872.

L-743,872 is a broad-spectrum pneumocandin antifungal drug developed by Merck Research Co., and in the present work it was evaluated in vivo in murine models of Candida krusei and Candida glabrata infection. Mice were infected intravenously with two isolates of C. krusei and treated with fluconazole or L-743,872. Fluconazole was beneficial only in immune-competent mice infected with isolate 94-2696. At > 0.5 mg/kg of body weight/day, L-743,872 was effective against both infecting isolates in immune-competent and immune-suppressed mice. Against C. glabrata, L-743,872 was effective, at doses > or = 0.5 mg/kg, in reducing fungal cell counts in the kidneys but not in the spleen. L-743,872 has significant potential for clinical development.

Animals↗

Ecology of Candida albicans gut colonization: inhibition of Candida adhesion, colonization, and dissemination from the gastrointestinal tract by bacterial antagonism.

Antibiotic-treated and untreated Syrian hamsters were inoculated intragastrically with Candida albicans to determine whether C. albicans could opportunistically colonize the gastrointestinal tract and disseminate to visceral organs. Antibiotic treatment decreased the total population levels of the indigenous bacterial flora and predisposed hamsters to gastrointestinal overgrowth and subsequent systemic dissemination by C. albicans in 86% of the animals. Both control hamsters not given antibiotics and antibiotic-treated animals reconventionalized with an indigenous microflora showed significantly lower gut populations of C. albicans, and C. albicans organisms were cultured from the visceral organs of 0 and 10% of the animals, respectively. Conversely, non-antibiotic-treated hamsters inoculated repeatedly with C. albicans had high numbers of C. albicans in the gut, and viable C. albicans was recovered from the visceral organs of 53% of the animals. Examination of the mucosal surfaces from test and control animals indicated further that animals which contained a complex indigenous microflora had significantly lower numbers of C. albicans associated with their gut walls than did antibiotic-treated animals. The ability of C. albicans to associate with intestinal mucosal surfaces also was tested by an in vitro adhesion assay. The results indicate that the indigenous microflora reduced the mucosal association of C. albicans by forming a dense layer of bacteria in the mucus gel, out-competing yeast cells for adhesion sites, and producing inhibitor substances (possibly volatile fatty acids, secondary bile acids, or both) that reduced C. albicans adhesion. It is suggested, therefore, that the indigenous intestinal microflora suppresses C. albicans colonization and dissemination from the gut by inhibiting Candida-mucosal association and reducing C. albicans population levels in the gut.

Adhesiveness↗

Single-copy IMH3 allele is sufficient to confer resistance to mycophenolic acid in Candida albicans and to mediate transformation of clinical Candida species.

Parasexual genetic analysis of Candida albicans utilized the dominant selectable marker that conferred resistance to mycophenolic acid. We cloned and sequenced the IMH3(r) gene from C. albicans strain 1006, which was previously identified as resistant to mycophenolic acid (MPA) (A. K. Goshorn and S. Scherer, Genetics 123:213-218, 1989). MPA is an inhibitor of IMP dehydrogenase, an enzyme necessary for the de novo biosynthesis of GMP. G. A. Kohler et al. (J. Bacteriol. 179:2331-2338, 1997) have shown that the wild-type IMH3 gene, when expressed in high copy number, will confer resistance to this antibiotic. We demonstrate that the IMH3(r) gene from strain 1006 has three amino acid changes, two of which are nonconservative, and demonstrate that at least two of the three mutations are required to confer resistance to MPA. We used this gene as a dominant selectable marker in clinical isolates of C. albicans and Candida tropicalis. We also identified the presence of autonomously replicating sequence elements that permit autonomous replication in the promoter region of this gene. Finally, we found the excision of a phi-type long terminal repeat element outside the IMH3 open reading frame of the gene in some strains. We used the IMH3(r) allele to disrupt one allele of ARG4 in two clinical isolates, WO-1 and FC18, thus demonstrating that a single ectopic integration of this dominant selectable marker is sufficient to confer resistance to MPA.

Alleles↗

CaNAT1, a heterologous dominant selectable marker for transformation of Candida albicans and other pathogenic Candida species.

A dominant selectable marker for Candida albicans and other Candida species, which confers resistance to nourseothricin, was characterized. In a heterologous promoter system and a recyclable cassette, the marker efficiently permitted deletion and complementation of C. albicans genes. Neither growth nor filamentous development was affected in strains expressing this marker.

Antifungal Agents↗

Interspecific complementation analysis by protoplast fusion of Candida tropicalis and Candida albicans adenine auxotrophs.

A protocol employing inositol starvation was used to isolate proline and adenine auxotrophs of Candida tropicalis. Interspecific hybrids between red adenine auxotrophs of C. tropicalis and Candida albicans were formed by protoplast fusion. These C. tropicalis red adenine auxotrophs were shown to fall into two complementation groups by crossing them with a known C. albicans ade1 tester strain. It is suggested that these two groups correspond to the ade1 and ade2 mutants of Saccharomyces cerevisiae and C. albicans and that these defined mutants may be useful in attempts to develop transformation systems for C. tropicalis.

Adenine↗

Candida ciferrii and Candida chiropterorum isolated from clinical specimens.

Ten clinical yeast isolates submitted to the Centers for Disease Control from diverse geographic areas were identified as Candida ciferrii and Candida chiropterorum. The association of C. ciferrii with clinical specimens, particularly its repeated isolation from a case of onychomycosis, suggests that this species may be an etiological agent of superficial yeast infections.

Adult↗

Enzyme immunoassays for invasive Candida infections: reactivity of somatic antigens of Candida albicans.

The main problem encountered with serodiagnostic tests for Candida infections is their failure to differentiate between invasive and superficial candidosis. Recent immunoblotting studies suggested that the use of selective somatic proteins of Candida albicans as antigens might be a promising approach toward developing a new generation of serodiagnostic assays. In this study major cytoplasmic protein antigens with molecular weights of 47,000 (47K), 46,000 (46K), 45,000 (45K), and 29,000 (29K) were identified as potential marker antigens for antibody detection in invasive candidosis. Continuous-flow isoelectric focusing was employed to enrich the proteins in two fractions, one of them containing the 47K and 29K proteins and the other one containing predominantly the 47K and 45K major proteins. These antigens and a whole somatic antigen extract were used to establish enzyme immunoassays (EIAs) for antibody detection. Whereas all tests were able to discriminate between patients with invasive candidosis (n = 27) and normal healthy volunteers (n = 167), as proved by graphic marker analysis, the selective antigen EIAs were highly superior to the whole somatic antigen EIA and two serological standard assays (indirect immunofluorescence assay and indirect hemagglutination assay) when a panel of sera from patients with superficial candidosis (n = 34) was used as a negative control group. The use of the 47K-29K antigen fraction allowed the best differentiation between invasive and noninvasive candidosis. The corresponding immunoglobulin G class-specific EIA had a sensitivity of 81.5% and a specificity of 97% for both negative control groups as well.

Antibodies, Fungal↗

Prevalence of Candida species, including Candida dubliniensis, in Singapore.

We present data on the prevalence of Candida species isolated from inpatients in three Singapore hospitals and from vaginal samples collected at community clinics. Strikingly, approximately 20% of the isolates from blood and vaginal samples were fluconazole-resistant species. By analyzing species-specific gene sequence signatures, we identified Candida dubliniensis from both groups of patients.

Candida↗

Casein agar: a useful medium for differentiating Candida dubliniensis from Candida albicans.

Production of chlamydospores on casein agar at 24 degrees C for 48 h provides a simple means for differentiating Candida dubliniensis from Candida albicans based on chlamydospore production. Of 109 C. dubliniensis isolates tested on this medium, 106 (97.2%) produced abundant chlamydospores and three produced few chlamydospores. In contrast, of the 120 C. albicans isolates tested, 111 (92.5%) failed to produce any chlamydospores, whereas the remaining nine isolates produced few chlamydospores. These findings indicate that abundant chlamydospore production on casein agar is a useful test for discriminating between C. dubliniensis and C. albicans.

Bacterial Typing Techniques↗

Detection of the Candida antigen mannan in cerebrospinal fluid specimens from patients suspected of having Candida meningitis.

Cerebrospinal fluid samples from five patients from which Candida cells were cultured were tested for the presence of mannan. Samples from four patients categorized as having proven candidosis reacted positively. Samples from the remaining patient and from patients with other central nervous system infections were negative. Detection of mannan may be valuable in the diagnosis of Candida meningitis.

Aged↗

Experimental and clinical studies of 5-fluorocytosine activity in Candida ocular infections. I. In vitro activity of 5-fluorocytosine on Candida species isolated from ocular infections.

In vitro sensitivity to 5-fluorocytosine (5-FC) of 35 Candida strains (14 C. albicans, 10 C. tropicalis, 5 C. parapsilosis, 2 C. guilliermondii, 2 C. krusei, 2 C. pseudotropicalis), isolated from the eyes of patients suffering from various external ocular inflammations who had not been treated with 5-FC, was tested and compared to that of standard Candida species. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of 5-FC for the strains isolated from ocular infections ranged from 0.06 to 1.95 mug/ml and from 0.24 to 7.81 mug/ml, respectively, indicating that no primary resistant strains to 5-FC were found in this series.

Animals↗

Anti Candida activity of induced transferrin in mice immunized with inactivated Candida albicans.

Mice immunized with formalin-killed Candida albicans were resistant to challenge by a lethal amount of viable C. albicans. The growth-inhibitory activity to C. albicans was detected in sera from the immunized mice, and was inhibited by the addition of anti-transferrin antibody or ferric sulfate. Both the amount of transferrin and the unsaturated iron-binding capacity (UIBC) in the serum were significantly increased, indicating that apo-transferrin increased in the immunized mice. Moreover, the intraperitoneal administration of apo-transferrin enhanced the protection from the Candida infection in vivo.

Animals↗