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PubMed1996

Enhanced IgE response to Candida albicans in postoperative invasive candidiasis.

BACKGROUND: Invasive candidiasis is a life-threatening complication problem in post-operative and immunocompromized patients, e.g. those treated by intensive care. Candida is frequently cultured from the mucous membranes of hospital patients and fungal cultures offer little diagnostic help. Other diagnostic methods, such as blood cultures, serology and diagnostic imaging techniques produce results too late and, if positive, low sensitivity. OBJECTIVE: To study the value of Candida-specific antibodies, especially those of IgE class, in diagnosing invasive Candida infection. METHODS: The immunoglobulins IgE, IgG and IgM responses to antigens of Candida albicans in the sera of 14 patients with culture, biopsy and/or autopsy proven postoperative invasive candidiasis and of 11 colonized and 19 non-colonized operated patients were studied by mannan radioallergosorbent test (RAST), mannan enzyme-linked immunosorbent assay (ELISA) and immunoblotting. RESULTS: Detection of IgE antibodies to C. albicans polysaccharide (mannan) and protein antigens proved specific and sensitive in diagnostics of invasive candidiasis after major abdominal surgery. IgE rose early in the course of the infection and the method made a clear distinction between invasive infection and mucous colonization. Immunoblotting for protein antibodies was most sensitive while nitrocellulose-RAST for mannan antibodies was most specific. The combined use of immunoblotting and RAST increased the sensitivity and the specificity. Determinations of anti-Candida IgG and IgM antibodies had low sensitivity and specificity. CONCLUSION: Critically ill patients with invasive candidiasis develop IgE antibodies to Candida antigens probably because of disturbed TH1/TH2 responses. Determination of specific IgE antibodies can be used as a diagnostic aid in the early stage of invasive Candida infection.

Adult↗
PubMed1998

Itraconazole in the treatment of superficial cutaneous and mucosal Candida infections.

Superficial cutaneous and mucosal Candida infections are common and widely affect both the immunocompromised as well as the immunocompetent populations. Common infections may include Candida paronychia, cutaneous candidiasis, oral candidiasis, vulvovaginal candidiasis, and Candida onychomycosis. Although C albicans has been considered to be the most common pathogen, other Candida species have emerged as potential causes of certain infections. Currently, a variety of antifungal agents is available to treat these infections. They include fluconazole, ketoconazole, and itraconazole. These agents have been widely used to treat fungal infections, including superficial and systemic candidiasis. However, some concerns exist regarding safety associated with long-term use of ketoconazole, and emerging issues of Candida resistance to fluconazole in some patient subsets have been reported. Itraconazole has proven efficacy in treating cutaneous and mucosal Candida infections. Additionally, studies have demonstrated that itraconazole may have increased efficacy and an excellent safety profile when administered in a pulse-dose, or intermittent fashion, for superficial mycotic infections. Itraconazole is an effective agent that warrants consideration when selecting treatment for Candida infections.

Antifungal Agents↗
PubMed1995

An epidemiological study of Candida species infection in cancer patients using genetic fingerprinting and morphotyping.

During a six-week period a cluster of four cases of Candida fungaemia occurred in a mixed haematology/oncology unit of a large Dublin teaching hospital. A study was conducted to determine whether the cluster of cases was caused by a particular strain. Nine patients were studied; five who were colonized with Candida spp. and four who developed Candida fungaemia. Twenty-two clinical isolates of Candida spp. were collected and identified. Three of the patients with fungaemia yielded Candida albicans from blood cultures and C. tropicalis was isolated from the fourth patient. C. albicans isolates were serotyped, morphotyped and analysed by DNA fingerprinting of total cellular DNA using the cloned C. albicans-specific, mid-repeat sequence element 27A as a molecular probe. All C. albicans isolates were of serotype A. Eight distinguishable types were identified by both morphotyping and DNA typing from 19 C. albicans isolates recovered from seven individual patients, although there were several discrepancies. Of three patients from whom two or more isolates of C. albicans were recovered on separate occasions, two yielded recurrent isolates with different morphotype codes. However, in both cases, the recurrent isolates from individual patients yielded indistinguishable, or closely related, DNA fingerprint profiles. Both morphotyping and DNA fingerprint analysis readily distinguished the three blood culture isolates of C. albicans. We conclude that the Candida spp. infections in the unit were not due to cross-infection and were probably related to the patients' indigenous flora.

Candida↗
PubMed2004

[Evaluation of a new commercial test (Candida albicans IFA IgG) for the serodiagnosis of invasive candidiasis].

INTRODUCTION: Two tests for the detection of antibodies to Candida albicans germ tubes in patients with invasive candidiasis were compared: a new commercially available test (Candida albicans IFA IgG) and the indirect immunofluorescence test generally used for this purpose. METHODS: With the use of two indirect immunofluorescence tests, retrospective study was done on 172 sera from 51 patients classified into two groups: Group I included 123 serum samples from 32 patients with invasive candidiasis, and Group II, the control, included 49 serum samples from 19 patients with no evidence of Candida infection. RESULTS: In Group I, 84% of patients presented anti-germ tube antibody titers >or= 1:160 by the Candida albicans IFA IgG test and 78.1% of patients were positive by the generally used test. There was a high correlation between the two tests (R2 =0.9512 by patients; R2 = 0.8986 by sera). When a titer value of >or= 1:160 was used as cutoff, the Candida albicans IFA IgG test showed a sensitivity of 84.4% and a specificity of 94.7%, whereas the traditional test showed a sensitivity of 78.1% and a specificity of 100%. CONCLUSIONS: The commercially available Candida albicans IFA IgG test is similar to the test generally used for the detection of antibodies to C. albicans germ tubes and provides faster and easier diagnosis of invasive candidiasis in the clinical microbiology laboratory.

Adolescent↗
PubMed2003

Examination on Candida spp. in refractory periapical granulomas.

AIM: To examine the occurrence of Candida spp. in refractory periapical granulomas. METHODOLOGY: One hundred and three surgically removed periapical granulomas were subjected to molecular analysis for the occurrence of Candida albicans. DNA was extracted from the samples using a modified phenol/chloroform/isoamyl alcohol method and was subjected to polymerase chain reaction (PCR) with OPA-03 and repetitive sequence (GACA)4 primers. The PCR products were separated in agarose gel electrophoresis, stained with ethidium bromide, visualized using UV light and the sequences were analysed. Samples indicating possible occurrence of Candida were further investigated by histological and immunohistological methods. Periodic acid-Shiff staining (PAS) was used to detect yeast cells and hyphae, and specific monoclonal antibodies to recognize high molecular mass mannoproteins present in the C. albicans cell wall. DNA extraction was controlled by running PCR using beta-actin primers (a housekeeping gene). C. albicans CCUG19915, C. tropicalis ATCC750, C. krusei ATCC6258, C. guilliermondii ATCC6260 and C. glabrata CCUG32725 served as positive controls in PCR. A tissue preparation of chronic atrophic candidosis in oral buccal mucosa served as a positive control for histological and immunohistological examinations. RESULTS: Polymerase chain reaction with beta-actin primers indicated successful DNA extraction in 68 out of 103 samples. The majority of the samples (50) were negative whereas 18 of the samples showed PCR products indicating possible occurrence of Candida spp. PAS-staining and immunohistological examination of these samples were, however, negative. Further analysis of the PCR products revealed sequences not typical for Candida spp. CONCLUSIONS: Candida spp. do not seem to occur in periapical granuloma.

Candida↗
PubMed1996

Vertical and horizontal transmission of unique Candida species to premature newborns.

The number of nosocomial bloodstream infections due to Candida species in critically ill newborns is increasing. This pathogen may be vertically transmitted from the mother or nosocomially acquired in the nursery. The goal of this study was to identify the route of transmission of unique Candida species and strains from mothers to their preterm offspring. Specimens from mothers for fungal cultures were obtained before delivery, and specimens from infants for sequential fungal cultures were obtained at defined intervals. Candida species were identified by standard methods and were typed by electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) with pulsed-field gel electrophoresis. Antifungal susceptibility testing was performed on all isolates. Fungal cultures were positive for Candida species in 12 (63%) of 19 mothers' specimens and in seven (33%) of 21 infants' specimens. EK and REAG revealed that both the mother and the infant in three (14%) of 21 mother-infant pairs were colonized with the identical strain of Candida albicans. C. albicans was most commonly transmitted vertically. Candida parapsilosis colonized other infants and could not be accounted for by a maternal reservoir.

Candida↗
PubMed1979

Identification of species of Candida, Cryptococcus, and Torulopsis by gas-liquid chromatography.

Gas-liquid chromatography was used to identify species of Candida, Cryptococcus, and Torulopsis by fatty-acid analysis of the whole-cell hydrolysate. Candida albicans characteristically revealed 2-OH C14:0 and C19:0 (chain length:number of double bonds); these were absent in other organisms. Candida curvata was characterized by a ratio of C16:1 to C16:0 of greater than 1.0. Candida guilliermondii contained C10:0, and Candida tropicalis had no C12:0, these features were used for their identification. Cryptococcus was characterized by the absence of C16:1. Torulopsis was characterized by a C16:1 to C16:0 ratio of greater than 10 accompanied by the presence of one unidentified fatty acid. These data suggested that certain Candida, Cryptococcus, and Torulopsis (the clinically important yeast-like organisms) may be identified by gas-liquid chromatography.

Candida↗
PubMed2003

The European Confederation of Medical Mycology (ECMM) survey of candidaemia in Italy: antifungal susceptibility patterns of 261 non-albicans Candida isolates from blood.

OBJECTIVES: To analyse the in vitro antifungal susceptibility of 261 non-albicans Candida bloodstream strains isolated during the European Confederation of Medical Mycology survey of candidaemia performed in Lombardia, Italy (September 1997-December 1999). METHODS: In vitro susceptibility to flucytosine, fluconazole, itraconazole, posaconazole and voriconazole was determined using the broth microdilution method described in the NCCLS M27-A guidelines. Etest strips were used to assess susceptibility to amphotericin B. In vitro findings were correlated with the patient's underlying condition and previous antifungal treatment. RESULTS: MICs (mg/L) at which 90% of the strains were inhibited were, respectively, 2 for flucytosine, 8 for fluconazole, 0.5 for itraconazole, 0.25 for voriconazole and 0.25 for posaconazole. Amphotericin B MIC endpoints were <0.50 mg/L in all the isolates tested. Flucytosine resistance was detected in 19 isolates (7%), mainly among Candida tropicalis strains (30%). Innate or secondary fluconazole resistance was detected in 13 strains (5%). Among the 13 patients with fluconazole-resistant Candida bloodstream infection, three were HIV positive, including one treated with fluconazole for oral candidosis; the four who were HIV negative had received the azole during the 2 weeks preceding the candidaemia. Cross-resistance among fluconazole and other azoles was a rare event. CONCLUSIONS: Resistance is still uncommon in non-albicans Candida species recovered from blood cultures. However, in fungaemias caused by C. tropicalis, Candida glabrata and Candida krusei, there is a high prevalence of resistance to fluconazole and flucytosine. Fluconazole resistance should be suspected in patients treated previously with azoles, mainly those with advanced HIV infection.

Candida↗
PubMed2003

Antifungal properties of 5-hydroxytryptamine (serotonin) against Candida species in vitro.

In this study the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of 5-hydroxytryptamine (5 HT, serotonin) against clinical isolates of Candida albicans (n = 11), Candida glabrata (n = 9), Candida tropicalis (n = 10) and Candida parapsilosis (ATCC 22019) using a broth microdilution test were investigated. In addition, it was examined whether delayed regrowth as a post-antifungal effect results following short exposure to 5 HT. 5 HT showed antifungal activity towards all isolates of Candida spp. The isolates yielded comparable MIC and MFC values of 5 HT in the range 0.91-7.34 mM and 1.83-14.68 mM, respectively. A lag in regrowth was dependent on the concentration tested. Treatment for 3 h at concentrations of 5 HT below and equipotent to the MFC resulted in a delayed regrowth of 8-12 h for isolates of Candida spp. In conclusion, these in vitro studies clearly demonstrate antifungal effects of 5 HT. Identifying the mode of action could be of great help in developing and researching new antifungal drugs.

Antifungal Agents↗
PubMedDate not supplied

In vitro susceptibility of 545 isolates of Candida spp. to four antifungal agents.

The in vitro susceptibility to amphotericin B, fluconazole, itraconazole and ketoconazole of 545 Candida strains from patients treated at the University Hospital of the Canaries was determined by means of a microdilution test. The distribution of the species was as follows: Candida albicans (342), Candida tropicalis (70), Candida glabrata (68), Candida parapsilosis (65). Of Candida albicans isolates, 8.5% and 7.6% showed resistance to itraconazole and fluconazole respectively. Of C. tropicalis isolates 34.3%, 27.1% and 2.9% were resistant to itraconazole, fluconazole and ketaconazole respectively. For C. glabrata, 10.3% and 4.4% of the isolates under study demonstrated resistance to fluconazole and itraconazole respectively. Only 4.6% and 1.5% of C. parapsilosis isolates demonstrated resistance to fluconazole and itraconazole respectively. C. tropicalis was the most resistant strain and C. parapsilosis the most sensitive. The greatest percentages of resistance in vitro were seen with the triazoles.

Amphotericin B↗
PubMed1996

[Candida in the ecological system of the orointestinal tract].

Within the microbial ecosystem of the orointestinal tract Candida spp. are common but not always detectable organisms. Therefore they belong to the transient flora. Candida spp. are found at various sites in healthy subjects in quantities between 10(2)-10(4)/ml or g. In different diseases an increased growth of Candida can occur depending on the presence of typical risk factors. In any case the yeasts only account for a minimal part of the bacterial flora. In this regard the contribution to the formation of metabolic products-in particular carbon dioxide, hydrogen and alcohols-is very small. Growth and development of Candida are dependent on nutrient and oxygen supply. In the colon under anaerobic conditions the flora competes for limited nutrients and fermentation is the dominant metabolic pathway in Candida. There is only minimal growth. Under aerobic conditions as can be demonstrated in experimental settings a marked growth with the formation of mycelium takes place. The generation time in vivo is much longer than under laboratory conditions and takes several hours. The detection of Candida in stool specimens can be interpreted in the first line only as a colonization. This is not equivalent with an infection, which, unfortunately, often is wrongly suggested in positive stool specimens.

Candida↗
PubMed2003

Interaction of sertraline with Candida species selectively attenuates fungal virulence in vitro.

This study investigated whether the interaction between isolates of Candida albicans (n=7), Candida parapsilosis (n=3), Candida krusei (n=2), Candida dubliniensis (n=1) and sertraline, a typical selective serotonin reuptake inhibitor, alters candidal virulence. Sertraline treatment of Candida spp. significantly (P<0.05) affected hyphal elongation, phospholipase activity, production of secreted aspartyl proteinases and fungal viability. In addition, monocyte-derived macrophages (MDMs) treated with sertraline reduced inhibition of blastoconidia germination in comparison to MDMs alone. In conclusion, our findings suggest that the interaction between sertraline and Candida spp. may also diminish the virulence properties of this fungal pathogen in vivo.

Aspartic Acid Endopeptidases↗
PubMed1987

Flow cytometric analysis of the DNA synthetic cycle of Candida species.

The total DNA per cell and DNA synthetic cycle phases were determined by flow cytometry in five Candida isolates including three species: Candida albicans 208R1, Candida tropicalis ATCC 750, and Candida parapsilosis 970, 3138, and ATCC 22019. The cells were prepared for flow cytometry by fixation in Carnoy fixative followed by staining with mithramycin. Marked but stable and reproducible inter- and intraspecific differences in total DNA per cell of stationary-phase cultures were found which did not correlate directly to diphenylamine estimates of the same parameter. This discrepancy was resolved by mathematically converting flow cytometry data into diphenylamine data. The reason for the discrepancy was found in studies of the DNA synthetic cycle of these yeasts: a large but isolate-specific variable proportion of the population is arrested in the S and G2-M phases after the culture passes from exponential to stationary phase. Histograms of exponential-growth-phase Candida isolates demonstrate that the majority of the population is in the G2-M phase of the DNA synthetic cycle. The DNA content of the C. tropicalis and C. parapsilosis isolates studied is as high as or higher than that of C. albicans. Extranuclear fluorescent particles were observed in the C. tropicalis isolate. No equivalent particles could be detected in the other four Candida isolates. The nature of the particles is unknown.

Candida↗
PubMed1996

Direct identification and recognition of yeast species from clinical material by using albicans ID and CHROMagar Candida plates.

Two chromogenic media, Albicans ID and CHROMagar Candida agar plates, were compared with a reference medium, Sabouraud-chloramphenicol agar, and standard methods for the identification of yeast species. This study involved 951 clinical specimens. The detection rates for the two chromogenic media for polymicrobial specimens were 20% higher than that for the Sabouraud-chloramphenicol agar plates. The rates of identification of Candida albicans for Albicans ID and CHROMagar Candida agar plates were, respectively, 37.0 and 6.0% after 24 h of incubation and 93.6 and 92.2% after 72 h of incubation, with specificities of 99.8 and 100%. Furthermore, CHROMagar Candida plates identified 13 of 14 Candida tropicalis and 9 of 12 Candida krusei strains after 48 h of incubation.

Agar↗
PubMed1998

Rapid identification of Candida species with species-specific DNA probes.

Rapid identification of Candida species has become more important because of an increase in infections caused by species other than Candida albicans, including species innately resistant to azole antifungal drugs. We previously developed a PCR assay with an enzyme immunoassay (EIA) format to detect amplicons from the five most common Candida species by using universal fungal primers and species-specific probes directed to the ITS2 region of the gene for rRNA. We designed probes to detect seven additional Candida species (C. guilliermondii, C. kefyr, C. lambica, C. lusitaniae, C. pelliculosa, C. rugosa, and C. zeylanoides) included in the API 20C sugar assimilation panel, five probes for species not identified by API 20C (C. haemulonii, C. norvegica, C. norvegensis, C. utilis, and C. viswanathii), and a probe for the newly described species C. dubliniensis, creating a panel of 18 Candida species probes. The PCR-EIA correctly identified multiple strains of each species tested, including five identified as C. albicans by the currently available API 20C database but determined to be C. dubliniensis by genotypic and nonroutine phenotypic characteristics. Species identification time was reduced from a mean of 3.5 days by conventional identification methods to 7 h by the PCR-EIA. This method is simple, rapid, and feasible for identifying Candida species in clinical laboratories that utilize molecular identification techniques and provides a novel method to differentiate the new species, C. dubliniensis, from C. albicans.

Base Sequence↗
PubMed1998

Stable phenotypic resistance of Candida species to amphotericin B conferred by preexposure to subinhibitory levels of azoles.

The fungicidal activity of amphotericin B (AmB) was quantitated for several Candida species. Candida albicans and C. tropicalis were consistently susceptible to AmB, with less than 1% survivors after 6 h of exposure to AmB. C. parapsilosis and variants of C. lusitaniae and C. guilliermondii were the most resistant, demonstrating 50 to 90% survivors in this time period and as high as 1% survival after a 24-h exposure time. All Candida species were killed (<1% survivors) after 24 h of exposure to AmB. In contrast, overnight exposure to either fluconazole or itraconazole resulted in pronounced increases in resistance to subsequent exposures to AmB. Most dramatically, C. albicans was able to grow in AmB cultures after azole preexposure. Several other Candida species did not grow in AmB but showed little or no reduction in viability after up to 24 h in AmB. Depending on the growth conditions, Candida cells preexposed to azoles may retain AmB resistance for days after the azoles have been removed. If this in vitro antagonism applies to the clinical setting, treatment of patients with certain antifungal combinations may not be beneficial. The ability of some Candida isolates to survive transient exposures to AmB was not reflected in the in vitro susceptibility changes as measured by standard MIC assays. This finding should be considered in studies attempting to correlate patient outcome with in vitro susceptibilities of clinical fungal isolates. Patients who fail to respond to AmB may be infected with isolates that are classified as susceptible by standard in vitro assays but that may be resistant to transient antifungal exposures which may be more relevant in the clinical setting.

Amphotericin B↗
PubMed1993

Oral Candida in HIV infection and AIDS: new perspectives/new approaches.

Oral candidosis has become an increasingly important problem in HIV-infected individuals. At present, the small body of published literature on the characterization of the Candida strains and species found in HIV+ patients is full of confusion and contradictions. Some of these difficulties are the result of the methodological shortcomings of a number of the techniques that have been used. Examples of the problems that may be encountered on primary isolation and subculture are described and the drawbacks associated with the systems used to date for phenotyping Candida are quoted. While molecular characterization techniques would appear to offer a reliable and objective alternative, they too have their strengths and weaknesses. An attempt is made to summarize the progress that has been made recently in the detection and identification of Candida albicans and also the non-albicans species from HIV-infected individuals. What emerges is that the commensal Candida species that inhabit the oral cavities of HIV+ patients are subjected to a number of significant pressures that probably promote the selection of organisms with unusual phenotypes and genotypes. These Candida are more difficult to characterize and behave differently compared to their counterparts in HIV- individuals. It is clear that uncovering the factors that are important for the selection of treatment regimens and will be predictive of outcome will not be easy. Candida organisms are neither as benign nor as simple as once thought.

AIDS-Related Opportunistic Infections↗
PubMed1999

[The in vitro properties of a new hydroxypyridone antimycotic rilopirox, with special reference to its anti-Candida activity].

In vitro properties of a new hydroxypyridone antimycotic rilopirox (RIL) with special reference to its anti-Candida activities were studied in comparison with the three reference drugs, ciclopirox olamine (CPO), oxiconazole nitrate (OCZ) and isoconazole nitrate (ICZ), using several strains of Candida albicans and Candida glabrata as the test organisms. RIL was potently fungicidal for growing cultures of these Candida strains, whereas all the three reference drugs were slightly fungicidal or fungistatic. Unlike OCZ and ICZ whose anti-Candida activity was decreased by lowering pH or adding serum to culture media, the activity of RIL was scarecely affected by change in pH or serum addition. However, RIL became less potent in the presence of Fe3+ at concentrations of 10(-5) mmol/ml or above. These findings suggest that RIL will be useful as a topical anti-Candida agent.

Antifungal Agents↗