Candidaemia due to Candida parapsilosis species complex at a hospital in Brazil: Clinical characteristics and antifungal susceptibility profile / Candidemia por especies del complejo Candida parapsilosis en un hospital de Brasil: características clínicas y perfil de sensibilidad a los antifúngicos

Background. Recent decades have seen a global emergence of candidaemia caused by non-Candida albicans Candida species, particularly the Candida parapsilosis complex. Aims. To evaluate the clinical features and antifungal susceptibility profiles of isolates belonging to the C. parapsilosis species complex in patients with candidaemia in a midwestern Brazilian tertiary-care teaching hospital. Methods. Yeast identification was performed using an automated Vitek 2 Compact system. PCR-RFLP was employed for species differentiation. Results. Five cases of infection by C. parapsilosis sensu stricto and two by Candida orthopsilosis were found. Of the seven cases, five were adult patients undergoing haemodialysis. The only isolate of C. parapsilosis sensu stricto resistant to fluconazole (MIC=8μg/ml) was obtained from a patient on a long-term regimen with this drug. This was the only patient who evolved to death. Conclusions. Resistance to antifungal agents poses a therapeutic challenge, especially for non-C. albicans Candida species, and requires continuous monitoring using susceptibility tests because resistance in vitro can be predictive of treatment failure. In the present study, in vitro antifungal susceptibility proved consistent with clinical outcome (AU)

Antecedentes. En las últimas décadas se ha visto un surgimiento mundial de la candidemia causada por especies de Candida no-C. albicans, en particular del complejo Candida parapsilosis. Objetivos. Evaluar las características clínicas y los perfiles de sensibilidad antifúngica en aquellos aislamientos del complejo de especies C. parapsilosis responsables de candidemia en un hospital universitario de tercer nivel en la región centro-oeste de Brasil. Métodos. La identificación se realizó en un sistema automatizado Vitek 2 compact. Se utilizó PCR-RFLP para la diferenciación de las especies. Resultados. Se encontraron cinco casos de candidemia por C. parapsilosis sensu stricto y dos por Candida orthopsilosis. Cinco eran pacientes adultos sometidos a hemodiálisis. El único aislamiento de Candida parapsilosis sensu stricto resistente a fluconazol (CIM, 8μg/ml) se obtuvo de un paciente en régimen largo de tratamiento con este antifúngico. Este fue el único paciente que murió. Conclusiones. La resistencia a los antifúngicos constituye un desafío terapéutico, en especial contra las especies de Candida no-C. albicans, que requieren la monitorización continua por medio de pruebas de sensibilidad en vista de que la resistencia in vitro puede ser predictiva de fracaso del tratamiento. En el presente estudio la sensibilidad antifúngica in vitro resultó consistente con el curso clínico (AU)

Candidaemia due to Candida parapsilosis species complex at a hospital in Brazil: Clinical characteristics and antifungal susceptibility profile.

BACKGROUND: Recent decades have seen a global emergence of candidaemia caused by non-Candida albicans Candida species, particularly the Candida parapsilosis complex. AIMS: To evaluate the clinical features and antifungal susceptibility profiles of isolates belonging to the C. parapsilosis species complex in patients with candidaemia in a midwestern Brazilian tertiary-care teaching hospital. METHODS: Yeast identification was performed using an automated Vitek 2 Compact system. PCR-RFLP was employed for species differentiation. RESULTS: Five cases of infection by C. parapsilosis sensu stricto and two by Candida orthopsilosis were found. Of the seven cases, five were adult patients undergoing haemodialysis. The only isolate of C. parapsilosis sensu stricto resistant to fluconazole (MIC=8µg/ml) was obtained from a patient on a long-term regimen with this drug. This was the only patient who evolved to death. CONCLUSIONS: Resistance to antifungal agents poses a therapeutic challenge, especially for non-C. albicans Candida species, and requires continuous monitoring using susceptibility tests because resistance in vitro can be predictive of treatment failure. In the present study, in vitro antifungal susceptibility proved consistent with clinical outcome.

Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species.

The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candida species known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei--A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. krusei demands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species

The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candidaspecies known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei - A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. kruseidemands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

Genotipo, sensibilidad a los antifúngicos y formación de biofilm de aislados de Trichosporon asahii recuperados de la orina de pacientes hospitalizados / Genotype, antifungal susceptibility, and biofilm formation of Trichosporon asahii isolated from the urine of hospitalized patients

En este estudio fueron analizadas mediante el cultivo muestras de orina de pacientes hospitalizados en la región centro-oeste de Brasil; los microorganismos aislados fueron identificados filogenéticamente como Trichosporon asahii. A través del análisis de máxima parsimonia de las secuencias de IGS1, fueron encontrados 3 genotipos que no habían sido descritos anteriormente. Las concentraciones inhibitorias mínimas frente a los 9 aislados identificados presentaron un rango de 0,06-1µg/ml en el caso de la anfotericina B, de 0,25-4µg/ml en el del fluconazol, y de 0,03-0,06µg/ml en el del itraconazol. Aproximadamente 6/9 de los aislados de T. asahii formaron biopelículas en la superficie de microplacas de poliestireno. Este trabajo documenta el aislamiento de T. asahii como agente causal de infeciones urinarias nosocomiales. Además, demuestra que la región IGS1 puede ser considerada una nueva herramienta epidemiológica para la genotipificación de los aislados de T. asahii. Los genotipos menos comunes encontrados en este estudio pueden estar relacionados con las características epidemiológicas locales

In this study, the culture analysis of urine samples from patients hospitalized in the Central-West region of Brazil was performed, and the isolated microorganisms were phylogenetically identified as Trichosporon asahii. Maximum parsimony analysis of the IGS1 sequences revealed three novel genotypes that have not been described. The minimum inhibitory concentrations of the nine isolates identified were in the range of 0.06­1µg/ml for amphotericin B, 0.25­4µg/ml for fluconazole, and 0.03­0.06µg/ml for itraconazole. Approximately 6/9 of the T. asahii isolates could form biofilms on the surface of polystyrene microplates. This study reports that the microorganisms isolated here as T. asahii are agents of nosocomial urinary tract infections. Furthermore, the IGS1 region can be considered a new epidemiological tool for genotyping T. asahii isolates. The least common genotypes reported in this study can be related to local epidemiological trends

Genotype, antifungal susceptibility, and biofilm formation of Trichosporon asahii isolated from the urine of hospitalized patients.

In this study, the culture analysis of urine samples from patients hospitalized in the Central-West region of Brazil was performed, and the isolated microorganisms were phylogenetically identified as Trichosporon asahii. Maximum parsimony analysis of the IGS1 sequences revealed three novel genotypes that have not been described. The minimum inhibitory concentrations of the nine isolates identified were in the range of 0.06-1 µg/ml for amphotericin B, 0.25-4 µg/ml for fluconazole, and 0.03-0.06 µg/ml for itraconazole. Approximately 6/9 of the T. asahii isolates could form biofilms on the surface of polystyrene microplates. This study reports that the microorganisms isolated here as T. asahii are agents of nosocomial urinary tract infections. Furthermore, the IGS1 region can be considered a new epidemiological tool for genotyping T. asahii isolates. The least common genotypes reported in this study can be related to local epidemiological trends.

Genotypic variability and antifungal susceptibility of Candida tropicalis isolated from patients with candiduria.

BACKGROUND: Candida tropicalis is an emerging major human pathogen in nosocomial infections, and it is considered the second or third species of Candida most isolated from urine cultures. AIMS: The study aimed at characterizing genotypically C. tropicalis strains from patients with candiduria in a university hospital, and assessed the antifungal susceptibility profile. METHODS: The study was conducted with hospitalized patients who developed urinary tract infection from C. tropicalis from June 2010 to June 2011 at the Grande Dourados University Hospital of the Federal University, Dourados, MS, Brazil. Susceptibility to the antifungal agents amphotericin B and fluconazole was determined by broth microdilution. The genotypic variability of isolates of C. tropicalis was analyzed by microsatellite markers and RAPD-PCR. RESULTS: Only one isolate was resistant to amphotericin B (MIC→16µg/ml); the others were susceptible to fluconazole and amphotericin B. The genotypic variability by RAPD-PCR resulted in distinct profiles for RAPD markers. A total of 10 alleles were observed for the microsatellite loci, URA3 and CT14, which were grouped differently, and four associations were observed for locus URA3 and eight for locus CT14. CONCLUSIONS: C. tropicalis isolates from urine were susceptible to the antifungal agents tested. The genotyping techniques make possible proving the similarity and genetic diversity among isolates of C. tropicalis involved in nosocomial infections. This knowledge is important for the control and prevention of nosocomial infections caused by this yeast species.

Antifungal susceptibility and distribution of Candida spp. isolates from the University Hospital in the municipality of Dourados, State of Mato Grosso do Sul, Brazil.

INTRODUCTION: Hospital infections caused by Candida spp. are a leading cause of morbidity and mortality in hospitalized patients, particularly those that are critically ill or immunocompromised. In this study, the distribution of Candida species in isolates from the University Hospital of the Federal University at Grande Dourados and their in vitro susceptibility to antifungal drugs were analyzed. METHODS: Yeasts were phenotypically identified using classical methodologies. Antifungal susceptibility tests to amphotericin B and fluconazole were performed using the broth microdilution technique. RESULTS: A total of 50 Candida isolates were obtained from hospitalized patients during the study period. We analyzed yeast isolates from urine (n=31; 62%), blood (n=12; 24%), and tracheal secretions (n=7; 14%). The following Candida species were identified: C. tropicalis (n=21; 42%), C. albicans (n=18; 36%), C. glabrata (n=10; 20%), and C. krusei (n=1; 2%). Antifungal susceptibility tests demonstrated that C. albicans was susceptible to both antifungal agents. However, 31.2% of the non-C. albicans Candida isolates displayed dose-dependent susceptibility to fluconazole, and 3.1% were resistant to amphotericin B. CONCLUSIONS: In contrast to previous reports, our results indicated that C. tropicalis was the most commonly isolated yeast species among the hospital patients. The predominance of non-C. albicans Candida infections confirms the importance of species-level identification for implementing appropriate antifungal therapies.

Antifungal susceptibility and distribution of Candida spp. isolates from the University Hospital in the municipality of Dourados, State of Mato Grosso do Sul, Brazil

Introduction Hospital infections caused by Candida spp. are a leading cause of morbidity and mortality in hospitalized patients, particularly those that are critically ill or immunocompromised. In this study, the distribution of Candida species in isolates from the University Hospital of the Federal University at Grande Dourados and their in vitro susceptibility to antifungal drugs were analyzed. Methods Yeasts were phenotypically identified using classical methodologies. Antifungal susceptibility tests to amphotericin B and fluconazole were performed using the broth microdilution technique. Results A total of 50 Candida isolates were obtained from hospitalized patients during the study period. We analyzed yeast isolates from urine (n=31; 62%), blood (n=12; 24%), and tracheal secretions (n=7; 14%). The following Candida species were identified: C. tropicalis (n=21; 42%), C. albicans (n=18; 36%), C. glabrata (n=10; 20%), and C. krusei (n=1; 2%). Antifungal susceptibility tests demonstrated that C. albicans was susceptible to both antifungal agents. However, 31.2% of the non-C. albicans Candida isolates displayed dose-dependent susceptibility to fluconazole, and 3.1% were resistant to amphotericin B. Conclusions In contrast to previous reports, our results indicated that C. tropicalis was the most commonly isolated yeast species among the hospital patients. The predominance of non-C. albicans Candida infections confirms the importance of species-level identification for implementing appropriate antifungal therapies. .