Utilidad de la espectrometría de masas en el diagnóstico microbiológico de la candiduria / Utility of mass spectrometry in the microbiological diagnosis of candiduria

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Candida nivariensis as a New Emergent Agent of Vulvovaginal Candidiasis: Description of Cases and Review of Published Studies.

Candida nivariensis is a new emergent agent related to human infections in the vaginal tract and other localizations, but the phenotypic characteristics are very similar to Candida glabrata and can be misidentified and underdiagnosed. We described four cases of vulvovaginitis identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and confirmed the results with PCR amplification and sequencing of the entire ITS genomic region (ITS1, ITS2 and 5.8 rRNA). We reinforce the need for new diagnostic tools for the correct identification of yeast infections.

Infecciones por Mycobacterium simiae / Infections due to Mycobacterium simiae

Mycobacterium simiae es una micobacteria ambiental de crecimiento lento, fotocromógena, descrita por primera vez en 1965. Se asocia raramente a infecciones humanas, posiblemente por su limitada patogenicidad, principalmente a infección pulmonar en pacientes inmunocompetentes de edad avanzada con enfermedad pulmonar subyacente, e infección diseminada en pacientes jóvenes inmunodeprimidos con sida. El cultivo microbiológico es necesario para confirmar la sospecha clínica, y las técnicas de secuenciación genética son indispensables para identificar la especie. El tratamiento de las infecciones por M. simiae es complicado por su multirresistencia a los fármacos antituberculosos y por la falta de correlación de los datos de sensibilidad in vitro con la respuesta in vivo. El tratamiento adecuado aún está por definir, pero debe incluir claritromicina asociada a otros antimicrobianos, como moxifloxacino y cotrimoxazol. Es posible que las infecciones por M. simiae estén infradiagnosticadas

Mycobacterium simiae is a slow-growing photochromogenic environmental mycobacterium, first described in 1965. Rarely associated with human infections, possibly due to its limited pathogenicity, it mainly produces lung infection in immunocompetent elderly patients with underlying lung disease, and in disseminated infections in immunosuppressed young patients with AIDS. A microbiological culture is needed to confirm the clinical suspicion, and genetic sequencing techniques are essential to correctly identify the species. Treating M. simiae infections is complicated, owing to the multiple resistance to tuberculous drugs and the lack of correlation between in vitro susceptibility data and in vivo response. Proper treatment is yet to be defined, but must include clarithromycin combined with other antimicrobials such as moxifloxacin and cotrimoxazole. It is possible that M. simiae infections are undiagnosed

Evaluación de la espectrometría de masas en la identificación de levaduras de interés clínico / Evaluation of mass spectrometry for the identification of clinically interesting yeasts

INTRODUCCIÓN: La identificación de levaduras se basa en el estudio de las características morfológicas, bioquímicas y nutricionales, y en la utilización de métodos moleculares. La espectrometría de masas matrix-assisted laser desorption ionization time-of-fligh (MALDI-TOF) constituye un nuevo método de identificación de microorganismos que ha demostrado gran utilidad. Nuestro objetivo ha sido evaluar este nuevo método en la identificación de levaduras. MÉTODOS: Ensayamos un total de 600 cepas aisladas de muestras clínicas pertenecientes a 9 géneros y 43 especies. La identificación se realizó mediante secuenciación de las regiones ITS del ADN ribosómico, asimilación de compuestos de carbono (ID 32C) y espectrometría de masas en un espectrómetro Microflex (Bruker Daltonics GmbH, Alemania). RESULTADOS: Un total de 569 cepas (94,8%) fueron identificadas a nivel de especie por ID 32C, y 580 (96,7%) por MALDI-TOF. La concordancia entre ambos métodos comprendió un total de 553 cepas (92,2%), elevándose al 100% en las especies de interés clínico: Candida albicans, C. glabrata, C. parapsilosis, C. tropicalis, y casi del 100% en C. krusei. MALDI-TOF identificó especies que precisan métodos moleculares: Candida dubliniensis, C. nivariensis, C. orthopsilosis y C. metapsilosis. Observamos cierta irregularidad en la identificación de levaduras formadoras de artroconidias y de basidiomicetos. CONCLUSIÓN: La espectrometría de masas MALDI-TOF es un método rápido, rentable y económico que permite la identificación de la mayoría de las levaduras aisladas en clínica, así como la diferenciación de especies estrechamente relacionadas. Sería conveniente la inclusión de más especies en su base de datos para ampliar su rentabilidad

INTRODUCTION: The aim of the study was to analyze the incidence, management and cost associated to hematological and dermatological adverse effects (AE) in chronic hepatitis C patients on triple therapy (TT) with telaprevir (TVR) or boceprevir (BOC). METHODS: An analysis was made on the data recorded on patients who started treatment with TVR or BOC associated with peginterferon alfa and ribavirin in a 12-week follow-up period. RESULTS: Fifty-three patients were included (TVR n = 36; BOC n = 17). Thrombocytopenia (83% TVR vs. 88% BOC) followed by neutropenia (89% TVR vs. 82% BOC) were the most common AE. Dermatological AE were observed in 32% of patients. Eleven patients required treatment discontinuation (all of them received TVR), and toxicity was the main reason for discontinuation (64%). The percentage of patients who required supportive treatment for management of AE was 66%. The most used supportive treatment was erythropoietin. Eight patients required emergency health care, and 2 were hospitalized due to AE. Total cost of additional supportive resources was 32,522 Euros (625 [SD = 876] Euros/patient) (TVR 759 [SD = 1,022] Euros/patient vs. BOC 349 [SD = 327] Euros/patient; P > .05). Patients with grade iii-iv toxicity required greater supportive care with higher costs, compared to patients with grade i-ii toxicity (849 [SD = 1,143] Euros/patient vs. 387 [SD = 397] Euros/patient; P = .053). CONCLUSION: The addition of new protease inhibitors to conventional treatment leads to a higher incidence of hematological AE in our study, compared to data described in clinical trials. The elevated incidence of AE involves the use of supportive care, increasing total costs of therapy

Prevalencia de Candida orthopsilosis y Candida metapsilosis en muestras clínicas / Prevalence of Candida orthopsilosis and Candida metapsilosis in clinical samples

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Diferenciación de especies del complejo Mycobacterium fortuitum mediante espectrometría de masas / Differentiation of Mycobacterium fortuitum complex species using mass spectrometry

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[Infections due to Mycobacterium simiae]. / Infecciones por Mycobacterium simiae.

Mycobacterium simiae is a slow-growing photochromogenic environmental mycobacterium, first described in 1965. Rarely associated with human infections, possibly due to its limited pathogenicity, it mainly produces lung infection in immunocompetent elderly patients with underlying lung disease, and in disseminated infections in immunosuppressed young patients with AIDS. A microbiological culture is needed to confirm the clinical suspicion, and genetic sequencing techniques are essential to correctly identify the species. Treating M. simiae infections is complicated, owing to the multiple resistance to tuberculous drugs and the lack of correlation between in vitro susceptibility data and in vivo response. Proper treatment is yet to be defined, but must include clarithromycin combined with other antimicrobials such as moxifloxacin and cotrimoxazole. It is possible that M. simiae infections are undiagnosed.

[Evaluation of mass spectrometry for the identification of clinically interesting yeasts]. / Evaluación de la espectrometría de masas en la identificación de levaduras de interés clínico.

INTRODUCTION: Identification of yeasts is based on morphological, biochemical and nutritional characteristics, and using molecular methods. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, a new method for the identification of microorganisms, has demonstrated to be very useful. The aim of this study is to evaluate this new method in the identification of yeasts. METHODS: A total of 600 strains of yeasts isolated from clinical specimens belonging to 9 genera and 43 species were tested. Identification was made by sequencing of the ITS regions of ribosomal DNA, assimilation of carbon compounds (ID 32C), and mass spectrometry on a Microflex spectrometer (Bruker Daltonics GmbH, Germany). RESULTS: A total of 569 strains (94.8%) were identified to species level by ID 32C, and 580 (96.7%) by MALDI-TOF. Concordance between both methods was observed for 553 strains (92.2%), with 100% in clinically relevant species: C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and almost 100% in C. krusei. MALDI-TOF identified species requiring molecular methods: Candida dubliniensis, C. nivariensis, C. metapsilosis and C. orthopsilosis. Some irregularities were observed in the identification of arthroconidia yeast and basidiomycetes. CONCLUSION: MALDI-TOF is a rapid, effective and economic method, which enables the identification of most clinically important yeasts and the differentiation of closely related species. It would be desirable to include more species in its database to expand its performance.

Candida galli as a cause of tinea unguium-molecular characterization of a rare clinical fungal entity.

We present the first clinical report of an infection caused by Candida galli, an anamorphic yeast species in the Yarrowia clade. C. galli has been described in the literature only four times, but never before it has been isolated from clinical samples. The colony morphology on Sabouraud medium and morphotype on CHROMagar Candida medium were similar to C. lipolytica as well as the carbon assimilation profile. The phenotypic differences with C. lipolytica were the non-assimilation of N-acetyl glucosamine, the absence of urease activity, growth in 10 % NaCl with 5 % glucose and in vitamin-free medium. MALDI-TOF MS could not generate reliable identification of the strain. Molecular analysis based on amplification of the ITS1-5.8S-ITS2 rDNA regions confirmed the identity as C. galli. Antifungal susceptibility test clearly demonstrated high MICs to 5-fluorocytosine, amphotericin B and fluconazole, as in the species belonging to the Yarrowia clade.

Sensibilidad a antifúngicos de especies de Scopulariopsis de origen clínico / Antifungal susceptibility of clinical isolates of Scopulariopsis species

Introducción. Scopulariopsis es un hongo saprofito del suelo. En los últimos años se ha incrementado el número y diversidad de las infecciones causadas por este hongo, incluyendo micosis superficiales e invasoras, y se ha descrito resistencia in vitro a antifúngicos, aunque existe escasa información al respecto. Nuestro objetivo fue establecer la sensibilidad in vitro de especies de origen clínico frente a un amplio número de antifúngicos. Material y métodos. Veintiocho cepas de Scopulariopsis (10 S. brevicaulis, 7 S. koningii, 3 S. acremonium, 3 S. candida, 3 S. flava, 1 S. brumptii y 1 S. fusca) fueron ensayadas mediante el método Sensititre YeastOne y el de microdilución en caldo, para determinar las concentraciones mínimas inhibitorias (CMIs) frente a anfotericina B, fluconazol, itraconazol, posaconazol, voriconazol y 5-fluorocitosina, y las concentraciones mínimas efectivas (CMEs) frente a anidulafungina, caspofungina y micafungina. Resultados. Nuestros datos confirman la alta resistencia in vitro a los antifúngicos de Scopulariopsis. Anidulafungina, caspofungina, micafungina (CMIs ≥8 mg/L), 5-fluorocitosina (CMIs ≥64 mg/L) y fluconazol (CMIs ≥128 mg/L) fueron inactivas para todas las especies. Las CMIs para anfotericina B (rango 2 a ≥8 mg/L) e itraconazol (0,5 a ≥16 mg/L) fueron altas. La mejor actividad se observó para posaconazol y voriconazol (0,5 a ≥8 mg/L). Con el método Sensititre YeastOne se obtuvieron CMIs ligeramente inferiores. Scopulariopsis candida, S. flava y S. fusca, fueron las especies más resistentes; S. acremonium y S. brevicaulis las de CMIs más bajas. Conclusiones. Las CMIs de todos los antifúngicos ensayados fueron elevadas para Scopulariopsis, lo que demuestra que las infecciones causadas por especies de Scopulariopsis podrían no responder al tratamiento. En infecciones graves, voriconazol sería una buena elección para el tratamiento, asociado a anfotericina B (AU)

Introduction. Scopulariopsis is a common soil saprophyte. In the last years the infections caused by Scopulariopsis species have increased, included superficial and invasive mycoses. This fungi has been reported resistant in vitro to some antifungal agents, although there is little information about this. The aim of the study was to establish in vitro antifungal susceptibility of clinical isolates of Scopulariopsis species against to broad-spectrum antifungal agents. Methods. A total of 28 Scopulariopsis strains (10 S. brevicaulis, 7 S. koningii, 3 S. acremonium, 3 S. candida, 3 S. flava, 1 S. brumptii and 1 S. fusca) were tested using Sensititre YeastOne and broth microdilution methods to determine the minimum inhibitory concentrations (MICs) to amphotericin B, fluconazole, itraconazole, posaconazole, voriconazole and 5-fluorocytosine, and minimun effective concentration (MECs) to anidulafungin, caspofungin and micafungin. Results. Our data confirm the high in vitro resistance of Scopulariopsis to antifungal agents. Anidulafungin, caspofungin, micafungin (MICs ≥8 mg/L), 5-fluorocytosine (MICs ≥64 mg/L), and fluconazole (MICs ≥128 mg/L) were inactive in vitro in all species. MICs of amphotericin B (range 2 to ≥8 mg/L) and itraconazole (0.5 to ≥16 mg/L) were high. The best antifungal activity was observed for posaconazole and voriconazole (0.5 to ≥8 mg/L). With Sensititre YeastOne method MICs obtained slightly lower. Scopulariopsis candida, S. flava and S. fusca were the most resistant species, while S. acremonium and S. brevicaulis showed the lowest MICs. Conclusions. MICs of all tested antifungal agents for Scopulariopsis were very high. Infections caused by Scopulariopsis species may not respond to antifungal treatment. Voriconazole is the drug of choice for treatment. We consider it appropriate to add amphotericin B in serious infections (AU)

Evaluation of the Sensititre Yeast One microdilution method for susceptibility testing of Candida species to anidulafungin, caspofungin, and micafungin

Introducción. Las equinocandinas representan un nuevo grupo de antifúngicos con gran actividad frente a especies de Candida. El propósito de este estudio fue evaluar el método Sensititre Yeast One para determinar la actividad in vitro de anidulafungina, micafungina y caspofungina frente a especies de Candida aisladas de muestras clínicas. Métodos. Un total de 131 cepas de Candida identificadas como: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides y 1 C. lipolytica, fueron ensayadas mediante el método colorimétrico de microdilución Sensititre Yeast One. Se consideraron sensibles las cepas inhibidas por concentraciones <=2 mg/L de anidulafungina, caspofungina o micafungina. Resultados. El 80,1% de las cepas fueron inhibidas por concentraciones <=0,25 mg/L de anidulafungina y micafungina. La actividad de caspofungina fue ligeramente inferior (78,6% de las cepas inhibidas por concentraciones <=0,25 mg/L). El 96,9% de las cepas resultaron sensibles frente a las tres equinocandinas. Dos cepas de C. parapsilosis (9,5%), 1 de C. guilliermondii y 2 de C. famata no mostraron sensibilidad a una o más equinocandinas. Conclusiones. En nuestra serie, anidulafungina, micafungina y caspofungina fueron efectivas frente a C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae y C. lipolytica. El 96,9% de las cepas fueron sensibles a las tres equinocandinas. Se puede afirmar que las equinocandinas tienen una excelente actividad frente a las especies de Candida más frecuentes en infecciones humanas, excepto Candida parapsilosis(AU)

Introduction. Echinocandins represent a new antifungal group with potent activity against Candida species. The purpose of our study was to evaluate the utility of the Sensititre Yeast One method to determine the in vitro activity of anidulafungin, micafungin, and caspofungin against Candida species isolated from clinical specimens. Methods. A total of 131 Candida strains were tested using Sensititre Yeast One colorimetric microdilution method. They belonged to the following species: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides, and 1 C. lipolytica. For being considered susceptible the strains had to be inhibited by concentrations <=2 mg/L of anidulafungin, caspofungin or micafungin. Results. The 80.1% of the strains tested were inhibited by concentrations <=0.25 mg/L of anidulafungin and micafungin. The activity of caspofungin was slightly lower (78.6% of strains inhibited by concentrations <=0.25 mg/L). The 96.9% of strains turned out susceptible to concentrations <=2 mg/L against the three echinocandins. Two strains of C. parapsilosis (9.5%), one of C. guilliermondii, and two of C. famata showed non-susceptible to one or more echinocandins. Conclusions. In our series, anidulafungin, micafungin, and caspofungin were effective against C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae and C. lipolytica. The 96.9% of strains were susceptible to all three echinocandins. Thus, echinocandins are proved to exhibit excellent activity to the Candida species most frequently involved in human infections, except Candida parapsilosis(AU)

Sensibilidad a fluconazol de levaduras de interés clínico: nuevos puntos de corte / Susceptibility to fluconazole of clinical interest yeasts: new breakpoints

Introducción. Recientemente, Pfaller y colaboradores (Drug Resist Updat 2010; 13:180-95), han propuesto nuevos puntos de corte para determinar la sensibilidad in vitro a fluconazol de Candida albicans, C. parapsilosis y C. tropicalis. El objetivo de este trabajo ha sido establecer las variaciones de sensibilidad de estas especies al aplicar estos puntos de corte, en relación con los del Clinical and Laboratory Standards Institute (CLSI). Métodos. Analizamos 112 cepas de Candida: 49 C. albicans, 40 C. parapsilosis y 23 C. tropicalis. La sensibilidad a fluconazol se ensayó por el método Sensititre YeastOne. Los puntos de corte para categorizar la concentración mínima inhibitoria (CMI) fueron los del CLSI y los propuestos por Pfaller y colaboradores. Resultados. Según los criterios del CLSI, todas las cepas fueron sensibles a fluconazol. Las CMI50 y CMI90 fueron 0,5 mg/L y 2 mg/L para C. albicans y C. parapsilosis, 0,5 mg/L y 1 mg/L para C. tropicalis. Con los nuevos criterios, 109 (97%) cepas fueron sensibles. Solamente se apreciaron variaciones en C. albicans (6% sensibles dosis dependientes). Conclusiones. Al aplicar los puntos de corte recomendados por Pfaller y colaboradores, y los del EUCAST, el número de cepas sensibles a fluconazol disminuye en relación con los criterios del CLSI, especialmente de C. albicans(AU)

Introduction. Recently, Pfaller et al (Drug Resist Update 2010; 13:180-95), have proposed new breakpoints for determining the in vitro susceptibility to fluconazole of Candida albicans, C. parapsilosis and C. tropicalis. The aim of this study was to establish the variations in sensitivity of these species applying these breakpoints, in relation to those of the Clinical and Laboratory Standards Institute (CLSI). Methods. We analyzed 112 strains of Candida: 49 C. albicans, 40 C. parapsilosis and 23 C. tropicalis. Susceptibility to fluconazole was performed by the method Sensititre YeastOne. The breakpoints used to determine the minimum inhibitory concentration (MIC) were identified by CLSI and the ones proposed by Pfaller et al. Results. According to the CLSI criteria, all isolates were susceptible to fluconazole. MIC50 and MIC90 were 0.5 mg/L and 2 mg/L for C. albicans and C. parapsilosis, 0.5 mg/L and 1 mg/L for C. tropicalis. With the new criteria, 109 (97%) strains were susceptible. Variations were seen in C. albicans, with 3 strains (6%) susceptible dose-dependent. Conclusions. When applying the breakpoints recommended by Pfaller et al, and EUCAST, the number of fluconazole-susceptible strains decreased according to the CLSI criteria, especially C. albicans(AU)

Infecciones por micobacterias de crecimiento rápido / Infections due to rapidly growing mycobacteria

Las micobacterias de crecimiento rápido (MCR) son ubicuas en la naturaleza y están distribuidas ampliamente en el agua, suelo y animales. Durante las últimas 3 décadas se ha observado un notable incremento de las infecciones causadas por MCR, tanto localizadas como diseminadas, así como de los brotes nosocomiales por contaminación de equipos médicos. El diagnóstico microbiológico de las infecciones por MCR incluye la observación directa al microscopio y el cultivo. La identificación taxonómica se realiza mediante técnicas fenotípicas, bioquímicas, cromatográficas y de biología molecular. El tratamiento difiere del de la tuberculosis y otras micobacteriosis, debido a la variable sensibilidad in vitro de las especies de este grupo. Las MCR son resistentes a los fármacos antituberculosos convencionales, pero pueden ser sensibles a antimicrobianos de amplio espectro. En este trabajo comentamos aspectos relevantes de las infecciones por MCR, incluyendo su biología, epidemiología, patología, diagnóstico microbiológico, identificación taxonómica, sensibilidad a los antimicrobianos y tratamiento (AU)

Rapidly growing mycobacteria (RGM) are ubiquitous in nature and widely distributed in water, soil and animals. During the past three decades we have observed a notable increment of infections caused by RGM, both localized and disseminated, as well as nosocomial outbreaks of contaminated medical equipment. The microbiological diagnosis of RGM infections includes direct microscopic observation and culture. The taxonomic (..) (AU)

[Infections due to rapidly growing mycobacteria]. / Infecciones por micobacterias de crecimiento rápido.

Rapidly growing mycobacteria (RGM) are ubiquitous in nature and widely distributed in water, soil and animals. During the past three decades we have observed a notable increment of infections caused by RGM, both localized and disseminated, as well as nosocomial outbreaks of contaminated medical equipment. The microbiological diagnosis of RGM infections includes direct microscopic observation and culture. The taxonomic identification is performed by phenotypic, biochemical, chromatographic and molecular biology techniques. The treatment differs from that of other mycobacteriosis like tuberculosis, owing to the variable in vitro susceptibility of the species of this group. The RGM are resistant to conventional antituberculous drugs, but can be susceptible to broad spectrum antimicrobial agents. In this study we comment on the significant aspects of human infections by RGM, including their biology, epidemiology, pathology, microbiological diagnosis, taxonomic identification, antimicrobial susceptibility and treatment.

Evaluation of the Sensititre Yeast One microdilution method for susceptibility testing of Candida species to anidulafungin, caspofungin, and micafungin.

INTRODUCTION: Echinocandins represent a new antifungal group with potent activity against Candida species. The purpose of our study was to evaluate the utility of the Sensititre Yeast One method to determine the in vitro activity of anidulafungin, micafungin, and caspofungin against Candida species isolated from clinical specimens. METHODS: A total of 131 Candida strains were tested using Sensititre Yeast One colorimetric microdilution method. They belonged to the following species: 42 C. albicans, 36 C. glabrata, 21 C. parapsilosis, 12 C. tropicalis, 10 C. krusei, 3 C. guilliermondii, 2 C. famata, 3 C. kefyr, 1 C. lusitaniae, 1 C. zeylanoides, and 1 C. lipolytica. For being considered susceptible the strains had to be inhibited by concentrations ≤2 mg/L of anidulafungin, caspofungin or micafungin. RESULTS: The 80.1% of the strains tested were inhibited by concentrations ≤0.25 mg/L of anidulafungin and micafungin. The activity of caspofungin was slightly lower (78.6% of strains inhibited by concentrations ≤0.25 mg/L). The 96.9% of strains turned out susceptible to concentrations ≤2 mg/L against the three echinocandins. Two strains of C. parapsilosis (9.5%), one of C. guilliermondii, and two of C. famata showed non-susceptible to one or more echinocandins. CONCLUSIONS: In our series, anidulafungin, micafungin, and caspofungin were effective against C. albicans, C. glabrata, C. tropicalis, C. krusei, C. kefyr, C. lusitaniae and C. lipolytica. The 96.9% of strains were susceptible to all three echinocandins. Thus, echinocandins are proved to exhibit excellent activity to the Candida species most frequently involved in human infections, except Candida parapsilosis.

[Susceptibility to fluconazole of clinical interest yeasts: new breakpoints]. / Sensibilidad a fluconazol de levaduras de interés clínico: nuevos puntos de corte.

INTRODUCTION: Recently, Pfaller et al (Drug Resist Update 2010; 13:180-95), have proposed new breakpoints for determining the in vitro susceptibility to fluconazole of Candida albicans, C. parapsilosis and C. tropicalis. The aim of this study was to establish the variations in sensitivity of these species applying these breakpoints, in relation to those of the Clinical and Laboratory Standards Institute (CLSI). METHODS: We analyzed 112 strains of Candida: 49 C. albicans, 40 C. parapsilosis and 23 C. tropicalis. Susceptibility to fluconazole was performed by the method Sensititre YeastOne. The breakpoints used to determine the minimum inhibitory concentration (MIC) were identified by CLSI and the ones proposed by Pfaller et al. RESULTS: According to the CLSI criteria, all isolates were susceptible to fluconazole. MIC50 and MIC90 were 0.5 mg/L and 2 mg/L for C. albicans and C. parapsilosis, 0.5 mg/L and 1 mg/L for C. tropicalis. With the new criteria, 109 (97%) strains were susceptible. Variations were seen in C. albicans, with 3 strains (6%) susceptible dose-dependent. CONCLUSIONS: When applying the breakpoints recommended by Pfaller et al, and EUCAST, the number of fluconazole-susceptible strains decreased according to the CLSI criteria, especially C. albicans.