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PubMed2001

Three new species of Candida from apple cider: C. anglica, C. cidri and C. pomicola.

Three new anamorphic ascomycetous yeasts are described: Candida anglica (type strain NRRL Y-27079, CBS 4262), Candida cidri (type strain NRRL Y-27078, CBS 4241), and Candida pomicola (type strain NRRL Y-27083, CBS 4242). These three species were isolated from cider produced in the United Kingdom, and their identification was determined from unique nucleotide sequences in the species-specific D1/D2 domain of large subunit (26S) ribosomal DNA. Phylogenetic analysis of D1/D2 sequences placed C. anglica near Candida fragi, C. cidri near Pichia capsulata, and C. pomicola in the Pichia holstii clade.

Beverages↗
PubMed2004

Identification of species of the genus Candida by analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers.

The PCR amplification and subsequent restriction analysis of the ribosomal region spanning the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene is applied to the identification of yeasts belonging to the genus Candida. This methodology has previously been used for the identification of some species of this genus, but in the present work this application has been applied to the identification and characterisation of a greater number of species of the genus Candida, with a special survey of species of clinical and biotechnological interest. Among the species of the genus Candida, the high variability observed, both in the length of the amplified region (ranging between 390 and 900 bp) and in their restriction patterns, allows the unequivocal identification to the species level, with the exception of the group of species that comprises C. membranifaciens, C. conglobata, C. atlantica, C. atmosphaerica, and C. oleophila, that required the sequencing of the D1/D2 domain of the 26S rRNA gene or the 5.8S-ITS region for their proper differentiation. The 5.8S-ITS restriction analysis also failed in the differentiation of species within the pairs C.aaseri/C.butyri,C.fructus/C.musae,C.santamariae var. santamariae / C. beechii and C. zeylanoides / C. krissii. In this case, the high sequence similarities obtained for their 26S D1/D2 domain and the 5.8S-ITS region indicate that each pair of species should be considered as a single species. The main purpose of this work is to generate a database for a high number of yeast species, of both biotechnological and clinical interest, and to facilitate their easy, fast, and reliable identification. The present work improves the database available online at the IATA web page (http://motor.edinfo.es/iata/) with the patterns of 75 species belonging to the genus Candida.

Animals↗
PubMed2004

Candida sergipensis, a new asexual yeast species isolated from frozen pulps of tropical fruits.

Sixteen strains of the new yeast species Candida sergipensis have been isolated from frozen pulps of the tropical fruits umbú ( Spondias tuberosa Avr. Cam.) and mangaba ( Hancornia speciosa Gom.). Candida sergipensis was one of the prevalent species in the yeast community of these substrates. The new asexual ascomycetous yeast is phylogenetically related to Candida spandovensis and Candida sorbophila, species belonging to the Wickerhamiella clade, as evidenced by the sequences of the D1/D2 domains of their large subunit ribosomal DNAs. The species C. sergipensis and C. spandovensis can be separated on the basis of growth on 50% glucose agar, xylose and succinate, negative for the first species and positive for the second. The type culture is strain UFMG-R188 (CBS 9567).

Anacardiaceae↗
PubMed2003

Evaluation of the adherence of Candida species to urinary catheters.

Catheter-associated urinary tract infections (CAUTIs) are the most common kind of nosocomial infection. Recent years have seen a significant increase in numbers of infections caused by yeasts of the genus Candida. The adherence of a microorganism to the host surface is a decisive factor in the success of colonization and the pathogenesis of infection. The objective of this work was to evaluate the adherence of species of the genus Candida to urinary catheters. In vitro adherence to the sections of latex and silicon catheters of Candida albicans and Candida parapsilosis were studied. Adherence was measured by counting the number of adhering viable cells and the results were expressed as Colonies Forming Units per mL. The results demonstrated that the latex catheter facilitated adherence more than the silicon catheter (p < 0.01). The adherence of the C. albicans was significantly greater than C. parapsilosis on latex, but it was similar on silicon.

Bacterial Adhesion↗
PubMed2006

Hyphal invasion of Candida albicans inhibits the expression of human beta-defensins in experimental oral candidiasis.

Oral epithelium reacts to microbial challenges by eliciting a defensive response that includes the production of antimicrobial peptides. This study investigated the expression of human beta-defensins-1, 2, and 3 in reconstituted human oral epithelia during experimental oral infections with six different Candida species, and a wild-type C. albicans isolate and five of its mutants. The expression of defensins was induced after 12 hours infection with the wild-type C. albicans, but this response was not seen for the noninvasive hyphal mutants nor the secreted aspartyl proteinase mutants. Furthermore, defensin expression was not detected after 48 hours in epithelia infected with either C. albicans wild-type isolate or its invasive hyphal and proteinase mutants. Most of the non-albicans Candida were capable of inducing the expression of defensins in epithelia after 24 or 48 hours of infection. These Candida-host interaction patterns suggest that the oral epithelia possess mechanisms for sensing the early invasion of C. albicans through recognition of the presence of hyphae and proteinases of Candida and respond to the insult by producing antimicrobial peptides. This hyphal-invasion-dependent inhibition of defensin expression in oral epithelium that undermines the host surveillance system represents a hitherto undescribed novel pathogenic mechanism of C. albicans.

Aspartic Acid Endopeptidases↗
PubMed2004

Low yield of ancillary diagnostic studies in neonates infected with Candida.

BACKGROUND: Fungal infection can be a significant complication for the critically ill neonate. However, the usefulness of extensive radiologic and ophthalmologic investigations in this population has not been thoroughly elucidated. OBJECTIVE: To report the incidence of organ fungal involvement diagnosed by ancillary testing (echocardiogram, ophthalmologic examination, brain imaging, and renal ultrasound (RUS)) among neonatal intensive care unit (NICU) patients with Candida infection. METHODS: This was a single center review of all NICU patients with Candida-positive cultures of blood, urine, peritoneal fluid, endotracheal tube aspirate, or cerebrospinal fluid from January 1, 1997 to June 1 2002. Data regarding the number of positive cultures, species isolated, and presence of specific risk factors and clinical symptoms were recorded for each case, as well as occurrence, timing and results of ancillary testing. RESULTS: In all, 66 patients had at least one positive culture for Candida. The majority (71%) were <1500 g at birth, and mean gestational age was 29.5+/-5.6 weeks. Echocardiograms were obtained in 54/66 (82%), and ophthalmology examinations were obtained in 36/66 (55%); none of these was consistent with fungal involvement. Brain imaging was performed in 50/66 (76%), only one of which was positive, in a patient with 16 positive blood cultures for Candida albicans. RUS were performed in 58/66 (88%) of patients, with concerning findings for fungal involvement in seven of the studies. RUS findings alone did not appear to consistently influence the length of therapy. CONCLUSIONS: Ancillary evaluations to investigate for fungal dissemination were undertaken frequently, but were of overall low yield. Although ancillary testing may be of limited additional value in centers with a low threshold for suspecting fungal infections and an aggressive approach to therapy, potentially important findings, which could impact management, may occur.

Brain↗
PubMed2000

Purification of native enolase from medically important Candida species.

The 48 kDa glycolytic enzyme, enolase, has been identified as an immunodominant antigen in Candida albicans infections. It has also been identified as an important fungal allergen. Enolase from a number of medically important Candida species has been purified using a two-step anion- and cation-exchange chromatography method that was preceded by an organic extraction. The enolases purified by this method have a high specific activity and the procedure is 40% efficient, with an average of 5 mg of enolase/g of Candida cells. The purification of native enolase from medically important Candida species will enable the immunological significance and interspecies relationships of this major fungal antigen to be investigated.

Candida↗
PubMed2001

Cytometric approach for a rapid evaluation of susceptibility of Candida strains to antifungals.

OBJECTIVE: To achieve a fast and reliable determination of the susceptibility of Candida strains to amphotericin B (Am B), fluconazole (Flu) and 5-fluorocytosine (5-FC), using cytometric methods as an alternative to the classical dilution method. METHODS: Twenty-three clinical isolates of Candida with different susceptibility patterns were treated for 1 h with two concentrations each of Am B (2 and 8 mg/L), Flu (8 and 64 mg/L) and 5-FC (4 and 32 mg/L), followed by staining with three different fluorochromes, under conditions previously defined through an optimisation study. These were 1 mg/L propidium iodide (PI)/10(6) cells for 30 min at 30 degrees C (a marker that only penetrates cells with severe lesions of the membrane); 0.5 microM FUN-1/10(6) cells for 30 min at 30 degrees C (a fluorescent probe which after entering the yeast cell is converted, by metabolically active yeasts, from a diffuse cytosolic pool with a yellow-green fluorescence into red cylindrical intravacuolar structures) and 0.25 microM of JC-1/10(6) cells for 15 min at 37 degrees C (a monomer that changes reversibly from green to red the J-aggregates, with the increased membrane potential). About 50 000 yeast cells were analysed by flow cytometry (FCM), at FL3 (red, 620 nm) for PI and FL2 (yellow-green, 575 nm) for FUN-1 and the ratio of FL3 to FL1 was determined (red, 620 nm/green, 525 nm) for JC-1; 200 cells of each suspension were also analysed by epifluorescence microscopy (EPM). Viability studies were performed in parallel to count the number of colony forming units. RESULTS: Susceptible (S) strains exposed to Am B and stained with JC-1 showed a dose-dependent decrease in the mitochondrial potential, i.e. a decreased ratio between red/green fluorescence by FCM and a decrease in J-aggregates by EPM. Neither FUN-1 nor PI was useful in the study of Am B activity. Susceptibility to Flu and 5-FC could be detected with FUN-1 staining: metabolic changes were detected by an increase in yellow-green intensity of fluorescence by FCM or a decrease of cylindrical intravacuolar structure formation by EPM, although no decrease in total viability was registered. Staining with JC-1 could predict resistance to both drugs, but did not allow distinction between sensitive dose-dependent strains (S-DD) or intermediate (I) resistance to Flu or 5-FC, respectively, from S strains. PI did not stain Candida cells treated with Flu or 5-FC under our experimental conditions. CONCLUSION: Susceptibility patterns of Candida strains to Am B can be determined by using JC-1, and to Flu and 5-FC by using FUN-1. PI was not a useful probe with which to study the effect of such antifungals under the conditions described here.

Antifungal Agents↗
PubMed2004

Candida yeasts in chronic periodontitis tissues and subgingival microbial biofilms in vivo.

The frequency of Candida infection in periodontal tissues of chronic periodontitis (CP) patients and the extent of candidal penetration into gingival tissues was studied. Tissue specimens collected from 25 CP patients during periodontal flap operations of initial periodontal therapy were examined by immunohistochemistry using Candida albicans-specific antibodies. Sections were also stained with periodic acid-Schiff (PAS) and subgingival plaque samples from 17 patients were cultured. Immunoreactivity for Candida was present in four of the 25 CP specimens (16%). Only one yeast-positive specimen was found when PAS-staining was used (4%) and two yeast-positive specimens were found with plaque culture (8%). Hyphal formation seemed essential and candidal hyphae were found to extend into the periodontal connective tissue. The degree and type of inflammation adjacent to the hyphae varied from a site and a patient to another. The sensitivity of specific antibodies was superior to PAS-stain or plaque culture in detection of Candida in tissues.

Biofilms↗
PubMed2000

Effect of serum concentration on Candida biofilm formation on acrylic surfaces.

The biofilm formation of the oral fungal pathogen Candida on denture acrylic strips coated with saliva, serum and, saliva-serum pellicle were examined in vitro using Candida albicans (four isolates), Candida glabrata (three isolates) and Candida tropicalis (three isolates). The degree of biofilm activity varied depending upon both the isolate and the pellicle. Significantly increased biofilm activity on the pellicle (particularly serum)-coated strips was observed with three isolates of C. albicans and another of C. glabrata on protein-coated acrylics, with increasing concentration of serum in the pellicle. Similar trends were observed with one isolate of C. albicans and C. glabrata, although the effects of pellicles were not significant. In contrast, with all three isolates of C. tropicalis and a single isolate of C. glabrata, although the biofilm activity on the protein-free control strips was significantly higher than that of saliva-coated strips, the increase in activity of pellicle-admixed biofilm depended upon the serum concentration. Candidal biofilm formation on acrylic surfaces is essentially promoted with increasing concentration of serum in the pellicle. This suggests that inflammation in the oral environment would facilitate fungal colonization on denture acrylic.

Acrylic Resins↗
PubMed2000

The frequency of Candida parapsilosis in onychomycosis. An epidemiological survey in Israel.

Candida albicans is regarded as the major pathogen in yeast-induced onychomycosis. Based on our impression of an increasing prevalence of Candida parapsilosis in this disease, we examined the data of two mycology laboratories in the same geographic location, from 1994 to 1996 in one (centre A) and for 1995 (6 months) in the other (centre B). A total of 954 and 230 toenails and 621 and 190 fingernails, respectively, underwent KOH microscopy and culture studies in each centre. Positive findings were noted in 45 and 65% of the toenails and 44 and 72% of the fingernails, respectively. In the toenails, Candida spp. were found in 22 and 15%, respectively, and in the fingernails, in 77 and 63%, respectively. The most frequent Candida species was C. parapsilosis (39.5% in toenails, 36.7% in fingernails), followed by C. albicans (19.5% in toenails, and 34.4% in fingernails). These results demonstrate a higher frequency of isolation of C. parapsilosis compared with C. albicans in onychomycosis. This might have important therapeutic implications.

Candida↗
PubMed2002

Bacterial and Candida adhesion to intact and denatured collagen in vitro.

Several recent reports imply the possibility of cariogenicity and periodontal disease linked to denture plaque containing Candida albicans. Adhesion of oral bacteria and Candida species to the extracellular matrix, such as type I collagen, fibronectin and denatured type I collagen, was examined by using adenosine triphosphate (ATP) analysis. The adhesion of C. albicans to intact and denatured type I collagen was significantly greater than those of oral bacteria and other species of Candida. This result suggests that C. albicans possesses the ability to adhere specifically to extracellular matrix, as compared with other Candida species or oral bacteria.

Bacterial Adhesion↗
PubMed2003

Oropharyngeal carriage of Candida species in HIV-infected patients in India.

The present investigation represents the first study of oropharyngeal carriage of Candida and other yeasts in HIV-infected patients in India. One hundred and fifty HIV-positive patients were investigated by culturing their swish samples on plates of CHROMagar Candida. Ninety-eight patients (65.3%) were positive for Candida and four (2.7%) were positive for other yeasts. Among them, the first Indian C. dubliniensis isolate has been recovered. Molecular typing of selected C. albicans isolates by AP-PCR revealed two major genotypes based on the banding patterns. The susceptibilities of 30 Candida isolates to five antifungal agents including the new triazole voriconazole were determined in a micro-dilution test, according to the NCCLS protocol M 27. All the 22 C. albicans isolates were susceptible to five antimycotic agents (flucytosine, amphotericin B, fluconazole, voriconazole and itraconazole) except one isolate (VPCI-122), which was resistant to flucytosine (MIC > or = 64 mg l-1). The azole-resistant isolates reported here endorse the role of antifungal susceptibility testing whenever antifungal treatment with azoles is planned.

AIDS-Related Opportunistic Infections↗
PubMed2003

Efficacy of a single intravenous dose of amphotericin B for Candida urinary tract infections: further favorable experience.

Studies in experimental animals and humans have shown that Amphotericin B (AmB) persists in urine for days to weeks after a single IV dose in levels that should inhibit candidal organisms and thereby obviate the need for frequent dosing. Including data from four previously described patients, we have now treated a total of 11 patients (12 episodes) with Candida urinary tract infections with single-dose AmB (six, Candida albicans; two, C. tropicalis; four, other nonalbicans Candida). The duration of candiduria prior to entry ranged from 18 to 180 days. Predisposing conditions included renal transplantation (1), diabetes mellitus (8), genitourinary stones (1) or anomalies (4), catheterization (2), and antibacterial therapy (11). A single patient was intolerant of AmB. Out of 11 evaluable candiduric episodes, eight resolved. Failure occurred in one patient with a chronic indwelling bladder catheter and in the allograft recipient. The data suggest that the sustained urinary excretion of AmB may permit successful single- or paucidose therapy of Candida urinary tract infections in some patients with a minimum of toxicity.

Adult↗
PubMed2002

Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance.

Non-albicans Candida (NAC) species cause 35-65% of all candidaemias in the general patient population. They occur more frequently in cancer patients, mainly in those with haematological malignancies and bone marrow transplant (BMT) recipients (40-70%), but are less common among intensive care unit (ITU) and surgical patients (35-55%), children (1-35%) or HIV-positive patients (0-33%). The proportion of NAC species among Candida species is increasing: over the two decades to 1990, NAC represented 10-40% of all candidaemias. In contrast, in 1991-1998, they represented 35-65% of all candidaemias. The most common NAC species are C. parapsilosis (20-40% of all Candida species), C. tropicalis (10-30%), C. krusei (10-35%) and C. glabrata (5-40%). Although these four are the most common, at least two other species are emerging: C. lusitaniae causing 2-8% of infections, and C. guilliermondii causing 1-5%. Other NAC species, such as C. rugosa, C. kefyr, C. stellatoidea, C. norvegensis and C. famata are rare, accounting for less than 1% of fungaemias in man. In terms of virulence and pathogenicity, some NAC species appear to be of lower virulence in animal models, yet behave with equal or greater virulence in man, when comparison is made with C. albicans. Mortality due to NAC species is similar to C. albicans, ranging from 15% to 35%. However, there are differences in both overall and attributable mortality among species: the lowest mortality is associated with C. parapsilosis, the highest with C. tropicalis and C. glabrata (40-70%). Other NAC species including C. krusei are associated with similar overall mortality to C. albicans (20-40%). Mortality in NAC species appears to be highest in ITU and surgical patients, and somewhat lower in cancer patients, children and HIV-positive patients. There is no difference between overall and attributable mortality, with the exception of C. glabrata which tends to infect immunocompromised individuals. While the crude mortality is low, attributable mortality (fungaemia-associated mortality) is higher than with C. albicans. There are several specific risk factors for particular NAC species: C. parapsilosis is related to foreign body insertion, neonates and hyperalimentation; C. krusei to azole prophylaxis and along with C. tropicalis to neutropenia and BMT; C. glabrata to azole prophylaxis, surgery and urinary or vascular catheters; C. lusitaniae and C. guilliermondii to previous polyene (amphotericin B or nystatin) use; and C. rugosa to burns. Antifungal susceptibility varies significantly in contrast to C. albicans: some NAC species are inherently or secondarily resistant to fluconazole; for example, 75% of C. krusei isolates, 35% of C. glabrata, 10-25% of C. tropicalis and C. lusitaniae. Amphotericin B resistance is also seen in a small proportion: 5-20% of C. lusitaniae and C. rugosa, 10-15% of C. krusei and 5-10% of C. guilliermondii. Other NAC species are akin to C. albicans-susceptible to both azoles and polyenes (C. parapsilosis, the majority of C. guilliermondii strains and C. tropicalis). Therefore, 'species directed' therapy should be administered for fungaemia according to the species identified-amphotericin B for C. krusei and C. glabrata, fluconazole for other species, including polyene-resistant or tolerant Candida species (C. lusitaniae, C. guilliermondii). In vitro susceptibility testing should be performed for most species of NAC in addition to removal of any foreign body to optimize management.

Adult↗
PubMed2003

Candida tropicalis fungemia in a tertiary care hospital.

Candida tropicalis is a frequent cause of fungemia in hospitals in Latin America. Candida albicans (33%) was the most frequently isolated species, followed by Candida parapsilosis (27%), and Candida tropicalis (24%) in tertiary care hospital in Brazil. We identified and retrospectively reviewed 27 cases of C. tropicalis fungemia that occurred at Hospital de Clinicas de Porto Alegre from 1996 to 1999. The mean age of the patients was 32 years (range 6 months to 88 years). Eight patients (29.6%) had hematological malignancy, and four (14.8%) had solid tumors. All the patients were taking broad-spectrum antibiotics, including vancomycin for at least 7 days. Antibiotics were given through a central venous catheter for the majority of the patients (77.7%). Relevant risk factors for candidemia in our patients included neutropenia (59.2%), and use of corticosteroids (37.0%) or cytotoxic drugs (40.7%). The onset of fever was the most frequent clinical manifestation (92.5%) of fungemia. Most of the patients (81.4%) were treated with amphotericin B or fluconazole. Overall mortality was 48.1%, and 7 (53.4%) of 13 deaths occurred within 10 days of the detection of candidemia. Results of the in vitro susceptibility testing of nine isolates of C. tropicalis from seven patients did not show resistance to fluconazole and amphotericin B.C. tropicalis presents as an important cause of fungemia in oncological and nononcological patients with central venous catheters taking broad-spectrum antibiotics. Although there was no evidence of resistance of C. tropicalis to amphotericin B and fluconazole, patients treated with antifungal agents presented with a high mortality rate in the hospital setting.

APACHE↗
PubMed2003

Reassessing the presence of Candida albicans in denture-related stomatitis.

OBJECTIVE: The aim of this study was to reevaluate the link between Candida albicans and denture-related stomatitis according to a modified Newton classification, which reflects the classic types of inflammation as well as the extent to which the tissue is affected. STUDY DESIGN: Two groups of denture wearers were evaluated for denture-related stomatitis. The presence and number of yeasts on the dentures, the identification of the Candida species present, and the amount of plaque coverage were determined. Putative risk factors were included in the study. Relations between these variables and stomatitis were analyzed statistically. RESULTS: According to Newton's classification, the presence of yeast on the denture was not linked to whether subjects had stomatitis. But with our classification, higher prevalence of yeast carriers, yeast colony number, and plaque coverage were found on the dentures of individuals with the most extensive inflammation, regardless of Newton type. Among risk factors evaluated, wearing dentures at night and smoking were associated with the most extensive inflammation. We also demonstrated that the presence of C albicans as well as the cohabitation of different Candida species was more frequent in denture-related stomatitis. The differences were statistically significant. CONCLUSIONS: Statistical analysis of microbiologic data from different denture-related stomatitis categories according to our modified classification showed that the presence of yeast on dentures was significantly associated with the extent of the inflammation, rather than with the Newton type. Our findings suggest that the inflammatory process of stomatitis favors the colonization of Candida. These results could have new implications for diagnosis and management of the condition.

Analysis of Variance↗
PubMed2002

Yeast flavohemoglobin from Candida norvegensis. Its structural, spectral, and stability properties.

Flavohemoglobin was isolated directly from the yeast Candida norvegensis and studied on its structural, spectral, and stability properties. In Candida flavohemoglobin, the 155 N-terminal residues make a heme-containing domain, while the remaining 234 C-terminal residues serve as a FAD-containing reductase domain. A pair of His-95 and Gln-63 was assigned to the proximal and distal residues, respectively. In purification procedure FAD was partially dissociated on a Butyl-Toyopearl column, so that FAD-lacking flavohemoglobin was also obtainable. In this ferric species, the Soret and charge-transfer bands were all characteristic of a penta-coordinate form. Compared with the recombinant heme domain expressed in Escherichia coli, we have measured the autoxidation rate over a wide pH range. The resulting pH dependence curves were then analyzed in terms of a nucleophilic displacement mechanism. As a result, the heme domain was found to be extremely susceptible to autoxidation, its rate being more than 100 times higher than that of sperm whale MbO2. However, this inherently high oxidation rate was dramatically suppressed in Candida flavohemoglobin to an extent almost comparable to the stability of mammalian myoglobins. These new findings lead us to conclude that Candida flavohemoglobin, differently from bacterial flavohemoglobins, can serve as an oxygen storage protein in aerobic conditions.

Amino Acid Sequence↗