Simultaneous Emergence of Multidrug-Resistant Candida auris on 3 Continents Confirmed by Whole-Genome Sequencing and Epidemiological Analyses.

BACKGROUND: Candida auris, a multidrug-resistant yeast that causes invasive infections, was first described in 2009 in Japan and has since been reported from several countries. METHODS: To understand the global emergence and epidemiology of C. auris, we obtained isolates from 54 patients with C. auris infection from Pakistan, India, South Africa, and Venezuela during 2012-2015 and the type specimen from Japan. Patient information was available for 41 of the isolates. We conducted antifungal susceptibility testing and whole-genome sequencing (WGS). RESULTS: Available clinical information revealed that 41% of patients had diabetes mellitus, 51% had undergone recent surgery, 73% had a central venous catheter, and 41% were receiving systemic antifungal therapy when C. auris was isolated. The median time from admission to infection was 19 days (interquartile range, 9-36 days), 61% of patients had bloodstream infection, and 59% died. Using stringent break points, 93% of isolates were resistant to fluconazole, 35% to amphotericin B, and 7% to echinocandins; 41% were resistant to 2 antifungal classes and 4% were resistant to 3 classes. WGS demonstrated that isolates were grouped into unique clades by geographic region. Clades were separated by thousands of single-nucleotide polymorphisms, but within each clade isolates were clonal. Different mutations in ERG11 were associated with azole resistance in each geographic clade. CONCLUSIONS: C. auris is an emerging healthcare-associated pathogen associated with high mortality. Treatment options are limited, due to antifungal resistance. WGS analysis suggests nearly simultaneous, and recent, independent emergence of different clonal populations on 3 continents. Risk factors and transmission mechanisms need to be elucidated to guide control measures.

Tracing the Evolutionary History and Global Expansion of Candida auris Using Population Genomic Analyses.

Candida auris has emerged globally as a multidrug-resistant yeast that can spread via nosocomial transmission. An initial phylogenetic study of isolates from Japan, India, Pakistan, South Africa, and Venezuela revealed four populations (clades I, II, III, and IV) corresponding to these geographic regions. Since this description, C. auris has been reported in more than 30 additional countries. To trace this global emergence, we compared the genomes of 304 C. auris isolates from 19 countries on six continents. We found that four predominant clades persist across wide geographic locations. We observed phylogeographic mixing in most clades; clade IV, with isolates mainly from South America, demonstrated the strongest phylogeographic substructure. C. auris isolates from two clades with opposite mating types were detected contemporaneously in a single health care facility in Kenya. We estimated a Bayesian molecular clock phylogeny and dated the origin of each clade within the last 360 years; outbreak-causing clusters from clades I, III, and IV originated 36 to 38 years ago. We observed high rates of antifungal resistance in clade I, including four isolates resistant to all three major classes of antifungals. Mutations that contribute to resistance varied between the clades, with Y132F in ERG11 as the most widespread mutation associated with azole resistance and S639P in FKS1 for echinocandin resistance. Copy number variants in ERG11 predominantly appeared in clade III and were associated with fluconazole resistance. These results provide a global context for the phylogeography, population structure, and mechanisms associated with antifungal resistance in C. aurisIMPORTANCE In less than a decade, C. auris has emerged in health care settings worldwide; this species is capable of colonizing skin and causing outbreaks of invasive candidiasis. In contrast to other Candida species, C. auris is unique in its ability to spread via nosocomial transmission and its high rates of drug resistance. As part of the public health response, whole-genome sequencing has played a major role in characterizing transmission dynamics and detecting new C. auris introductions. Through a global collaboration, we assessed genome evolution of isolates of C. auris from 19 countries. Here, we described estimated timing of the expansion of each C. auris clade and of fluconazole resistance, characterized discrete phylogeographic population structure of each clade, and compared genome data to sensitivity measurements to describe how antifungal resistance mechanisms vary across the population. These efforts are critical for a sustained, robust public health response that effectively utilizes molecular epidemiology.

[Profile of antifungal susceptibility of species of Candida isolated from hemocultures in a university hospital, Maracaibo, Venezuela]. / Perfil de sensibilidad antifúngica de especies de Candida aisladas de hemocultivos en un Hospital Universitario, Maracaibo, Venezuela.

The aim of this study was to determine the in vitro susceptibility of amphotericin B, fluconazole and itraconazole, to several Candida spp recovered from blood cultures on hospitalized patients at the University Hospital of Maracaibo, Venezuela. The determination of the antifungal susceptibility was carried out according to the microdilution method in broth developed by The European Committee for Antimicrobial Susceptibility Testing (EUCAST). The profile of susceptibility of the 74 isolates showed that all the studied species were susceptible to amphotericin B, and 97.2% and 89.2% to fluconazole and itraconazole, respectively.

First report of Candida auris in America: Clinical and microbiological aspects of 18 episodes of candidemia.

OBJECTIVES: Characterization of a hospital outbreak of Candida auris candidemia that involved 18 critically ill patients in Venezuela. METHOD: Bloodstream isolates of C. auris obtained from 18 patients admitted at a medical center in Maracaibo, between March, 2012 and July, 2013 were included. Species identification was confirmed by ITS rDNA sequencing. Isolates were subsequently typed by amplified fragment length polymorphism fingerprinting (AFLP). Susceptibility testing was performed according to CLSI. Clinical data were collected from all cases by using a standard clinical form. RESULTS: A total of 13 critically ill pediatric and 5 adult patients, with a median age of 26 days, were included. All were previously exposed to antibiotics and multiple invasive medical procedures. Clinical management included prompt catheter removal and antifungal therapy. Thirteen patients (72%) survived up to 30 days after onset of candidemia. AFLP fingerprinting of all C. auris isolates suggested a clonal outbreak. The isolates were considered resistant to azoles, but susceptible to anidulafungin and 50% of isolates exhibited amphotericin B MIC values of >1 µg/ml. CONCLUSIONS: The study demonstrated that C. auris is a multiresistant yeast pathogen that can be a source of health-care associated infections in tertiary care hospitals with a high potential for nosocomial horizontal transmission.

[Phenotypic evaluation to differentiate Candida albicans from Candida dubliniensis]. / Evaluación de los caracteres fenotípicos para diferenciar Candida Albicans de Candida dubliniensis.

This study evaluated the phenotypic tests used to differentiate Candida albicans from Candida dubliniensis. A total of 55 isolates from vaginal secretions, oral cavity and hemoculture were studied. They were originally identified as C. albicans, based on their morphological and physiological characteristics. These isolates were tested for colony color development on CHROMagar Candida medium, growth at 45 degrees C on Sabouraud Dextrose agar, lipolytic activity on Tween 80 Agar medium and colony morphology and chlamydoconidia formation on Staib agar medium. Of the 55 isolates studied, seven yielded one or more phenotypic characteristics suggestive of Candida dubliniensis. These isolates were tested by PCR with specific primers for Candida dubliniensis and API ID 32. The seven isolates were confirmed as Candida albicans. All of these finding indicate that DNA based tests should be used for definitive identification of Candida dubliniensis.

Susceptibilidad a fluconazol y voriconazol por el método de disfusión de cepas de candida, aisladas de hemocultivos en Maracaibo, Venezuela / Susceptibility to fluconazole and voriconazole in candida strains isolated from blood culture by disk diffusion method Maracaibo, Venezuela

Para determinar la susceptibilidad de cepas de Candida aisladas de hemocultivos en nuestro medio, se estudiaron 78 cepas obtenidas de pacientes hospitalizados en diferentes servicios del Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM), Venezuela. Para la identificación de especies se usó el medio cromogénico Brilliance Candida Agar y Vitek-YBC. Adicionalmente, los cultivos fueron identificados por el método tradicional. La susceptibilidad fue determinada por el método de difusión con discos de fluconazol y voriconazol según la metodología M44-A2 del Clinical Laboratory Standard Institute. La frecuencia de las especies de Candida fue: C. parapsilosis 51,28%, C. tropicalis 15,38%, C. guilliermondii 11,54%, C. albicans 10,26% C. famata 6,41%, C. glabrata 3,85% y C. krusei 1,28%. La susceptibilidad fue de 96,15% y 100% para fluconazol y voriconazol, respectivamente. Tres de las 78 cepas, identificadas como C. albicans, C. guilliermondii y C. krusei fueron resistentes a fluconazol. Estos resultados sugieren que fluconazol y voriconazol pueden ser utilizados en el tratamiento de pacientes con candidemia en SAHUM, sin embargo, la vigilancia epidemiológica y la determinación de la susceptibilidad de Candida deben mantenerse

To determine the susceptibility of Candida strains isolated from blood cultures in Maracaibo, Venezuela, 78 strains obtained from hospitalized patients in different services of the autonomous Maracaibo University Hospital, Venezuela, were studied. Chromogenic Medium Brilliance Candida Agar and Vitek-YBC were used for species identification. In addition, cultures were assessed using the traditional identification method. Susceptibility was determined by the diffusion method with fluconazole and voriconazole disks, according to the Clinical and Laboratory Standard Institute, Document M44-A2. Frequency of the Candida species was: C. parapsilosis 51.28%, C. tropicalis 15.38%, C. guilliermondii 11.54%, C. albicans 10.26% C. famata 6.41%, C. glabrata 3.85% and C. krusei 1.28%. Susceptibility was 96.15% and 100% for fluconazole and voriconazole, respectively. Three isolates identified as C. albicans, C. guilliermondii and C. krusei were resistant to fluconazole. These results suggest that fluconazole and voriconazole can be useful in the treatment of patients with candidemia; however, epidemiological surveillance and susceptibility pattern determination of Candida must be maintained

Cambios en la distribución de especies de Candida aislados de hemocultivos: en pacientes del Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela / Changes in tje distribution of Candida species isolated from the hemocultures of patients in the Service Autónomo Hospital Universitario de Maracaibo, Venezuela

Se estudió la epidemiología de Candidemias en el Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM), analizándose 78 episodios de Candidemia entre Enero 2007 y Marzo 2010, procedentes de 70 pacientes de ambos sexos y edades entre 15 días y 88 años, hospitalizados en diferentes servicios de SAHUM, de cuyos hemocultivos se aisló Candida sp en el Centro de Referéncia Bacteriológica (CRB) de esa institución. Para la identificación de especies se usó el medio cromogénico Brilliance Candida Agar y Vitek-YBC en el CRB. Adicionalmente, los cultivos fueron evaluados por el método tradicional en el Laboratorio de Micología de la Universidad del Zulia. Las características de los pacientes se tomaron de fichas del CRB. La mediana de la edad de los pacientes fue 17 años. Hubo 8 aislamientos secuenciales. La frecuencia de las especies de Candida fue: C. parapsilosis 51,28%, C. tropicalis 15,38%, C. guilliermondii 11,54%, C. albicans 10,26% C. famata 6,41%, C. glabrata 3,85% y C. krusei 1,28%. El Chi cuadrado indicó que los niños en los servicios no UCI tienen mayor probabilidad de adquirir Candidemias. Se observó un cambio en la frecuencia de especies, por lo tanto se requiere una vigilancia epidemiológica permanente en SAHUM

The epidemiology of candidemia in the Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM) was studied. From January 2008 to March 2010, 78 candidemia episodes from 70 patients were analyzed. Ages ranged from 15 days to 80 years old. Patients were hospitalized in different wards at SAHUM. Candida spp was isolated in blood cultures processed at the Bacteriology Reference Center (CRB). For species identification, chromogenic Brilliance Candida Agar and Vitek-YBC were used. Additionally, the cultures were evaluated by traditional methods at the Mycology Laboratory, University of Zulia. Patients’ characteristics were obtained from the CRB register. The median patients’ age was 17 years old. Eight sequential isolates were obtained. The following Candida species were found:, C. parapsilosis 51.28%, C. tropicalis 15.38%, C. guilliermondii 11.54%, C. albicans 10.26%, C. famata 6.41%, C. glabrata 3.85% and Candida krusei 1.28%. X²showed that children in non-ICU wards were more likely to have candidemia. A change in the frequency of the species isolated was observed; for this reason, a permanent surveillance system is required

Species distribution and antifungal susceptibility profile of Candida spp. bloodstream isolates from Latin American hospitals.

From March 1999 to March 2000, we conducted a prospective multicenter study of candidemia involving five tertiary care hospitals from four countries in Latin America. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to the National Committee for Clinical Laboratory Standards microbroth assay method. During a 12 month-period we were able to collect a total of 103 bloodstream isolates of Candida spp. C. albicans was the most frequently isolated species accounting for 42% of all isolates. Non-albicans Candida species strains accounted for 58% of all episodes of candidemia and were mostly represented by C. tropicalis (24.2%) and C. parapsilosis (21.3%). It is noteworthy that we were able to identify two cases of C. lusitaniae from different institutions. In our casuistic, non-albicans Candida species isolates related to candidemic episodes were susceptible to fluconazole. Continuously surveillance programs are needed in order to identify possible changes in the species distribution and antifungal susceptibility patterns of yeasts that may occurs after increasing the use of azoles in Latin American hospitals.

Candidemia en el Hospital Universitario de Maracaibo estado Zulia Venezuela 2000-2002 / Candidemia at the University Hospital of Maracaibo Zulia state Venezuela 2000-2002

El objetivo de este estudio fue realizar la caracterización fenotípica y conocer la distribución de las especies de Candida aisladas en 92 pacientes con candidemia (niños y adultos) internados en el Hospital Universitario de Maracaibo (Servicios UCI y No UCI), Edo. Zulia, Venezuela, de Marzo de 2000 a Septiembre de 2002. La identificación de las levaduras aisladas de los hemocultivos fue realizada mediante el uso de CHROMagar Candida y métodos tradicionales; en algunos casos se aplicó API 20C e ID32. Los resultados indican la presencia de C. albicans (33,7 por ciento) y Candida no albicans (66,3 por ciento): Candida tropicalis (30,4 por ciento), Candida parapsilosis (21,7 por ciento), Candida guilliermondii (5,4 por ciento), Candida pelliculosa (5,4 por ciento), Candida famata (2,2 por ciento) y Candida glabrata (1,1 por ciento). Entre los ni¤os (n= 70) internados en la UCI y servicios no UCI, predominaron aislamientos de C. parapsilosis y C. albicans respectivamente. Por otra parte, Candida tropicalis fue la levadura más frecuentemente aislada en adultos en los servicios evaluados. El predominio de Candida no albicans observado en este estudio, ha sido reportado en América Latina.

Species distribution and antifungal susceptibility profile of Candida spp. bloodstream isolates from Latin American hospitals

From March 1999 to March 2000, we conducted a prospective multicenter study of candidemia involving five tertiary care hospitals from four countries in Latin America. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to the National Committee for Clinical Laboratory Standards microbroth assay method. During a 12 month-period we were able to collect a total of 103 bloodstream isolates of Candida spp. C. albicans was the most frequently isolated species accounting for 42 percent of all isolates. Non-albicans Candida species strains accounted for 58 percent of all episodes of candidemia and were mostly represented by C. tropicalis (24.2 percent) and C. parapsilosis (21.3 percent). It is noteworthy that we were able to identify two cases of C. lusitaniae from different institutions. In our casuistic, non-albicans Candida species isolates related to candidemic episodes were susceptible to fluconazole. Continuously surveillance programs are needed in order to identify possible changes in the species distribution and antifungal susceptibility patterns of yeasts that may occurs after increasing the use of azoles in Latin American hospitals

Caracterización morfológica y fisiológica de las especies cándida aisladas de la cavidad bucal de pacientes geriátricos / Morphological and physiological characterization of the candida species isolated in the oral cavity of geriatric patients

Caracetrizar las especies de Cándida aisladas de la cavidad bucal de pacientes geriátricos del Hogar San José de la Montaña Maracaibo, Venezuela. Se tomaron 35 muestras orales, a las cuales se les realizaron pruebas morfológicas y fisiológicas que permitieron identificar las especies involucradas. Las especies más frecuentes fueron: C. albicans (42.1 por ciento) y C. parasilopsis (30.2 por ciento). La candidiasis pseudomembranosa fue la forma clínica más frecuente (72 por ciento), los factores locales y sistémicos predisponentes fueron: la mala higiene oral (28.3 por ciento), la ausencia total de dientes (16 por ciento) y la diabetes mellitus (46.1). El grupo etareo más afectado fue el de 65.1 a 70 (37 por ciento). En la candidiasis pseudomembranosa las especies mayormente involucrada fueron C. parasilopsis (27.9 por ciento) y C. albicans (20.9 por ciento). En la condición local "mala higiene oral", Las especies más comunes fueron c. parasilopsis (10.3 por ciento) y C. albicans (9.4) y en condición sistémica "Diabetes mellitus", C. albicans (25 por ciento). C. albicans constituye la especie más frecuentemente aislada, sin embargo, existe una importante participación de otras especies en la etiología de la candidiasis principalmente de C. parasilopsis. Así mismo, las condiciones locales y sistémicas constituyen factores predisponentes en la aparición de esta enfermedad.

Isosporiasis como primera infección oportunista en un paciente con infección por virus de inmunodeficiencia humana: reporte de un caso / Isosporiasis first oportunist infection in a patient with human inmunodeficiency virus: report of a case

Se presenta el caso de un paciente masculino, bisexual, con diarrea crónica intermitente, a quien por esta sintomatología, se investiga para infección por el Virus de Inmunodeficiencia Humana, resultando positivo por las técnicas de ELISA y WESTERN BLOTT; y diagnosticandosee Isosporiasis, después de varios exámenes copro-parasitologicos. El paciente fue tratado con trimetoprin-sulfametoxazole, observándose recurrencia de su sintomatología

Uso del medio chromagar cándida para identificar levaduras en muestras clínicas / The use of chromagar medium for the identification of clinical samples of yeast

Con la finalidad de evaluar el uso del medio chromagar candida en la rutina de identificación de levaduras, se estudiaron 237 cepas obtenidas de sangre, orina, secreción vaginal y otras muestras clínicas en el centro de referencia bacteriológica del Hospital Universitario de Maracaibo, Estado de Zulia (Venezuela), provenientes de 116 pacientes ambulatorios u hospitalizados, durante el período Marzo a Diciembre de 1999. Para la preparación del medio de cultivo y la lectura del color de las colonias, se siguió la recomendación del fabricante (chromagar candida, París, France). De manera simultánea se realizaron pruebas de filamentización, fermentación de carbohidratos y asimilación de fuentes carbonadas y nitrogenadas; en algunos casos se utilizó el sistema api 20c. Los resultados mostraron una especifidad del 100 porciento del medio chromagar candida para la identificación de candida albicans (colonia verde), candida tropicalis (colonia azul) y candida krusei (colonia rosada). En tres muestras se pudo separar mas de una especie, esta diferenciación se hizo fácilmente visible por los diferentes colores de las colonias. Este estudio corrobora la utilidad de este medio para la identificación rápida de las principales especies del género aisladas de material clínico