Características de las fungemias en un centro de referencia del Perú: retrospectiva de cinco años / Characteristics of fungemia in a peruvian referral center: 5-year retrospective analysis

RESUMEN Con el objetivo de conocer las características de las fungemias en 285 pacientes oncológicos hospitalizados del 2012 al 2016 en el Instituto Nacional de Enfermedades Neoplásicas se realizó un estudio descriptivo retrospectivo. Se evaluó información demográfica, clínica y microbiológica. Las fungemias por C. albicans predominaron en pacientes con tumores sólidos y sin neutropenia, mientras las causadas por C. tropicalis predominaron en pacientes con neoplasias hematológicas y neutropenia. C. tropicalis fue el agente más aislado (47,0%). Las fungemias aumentaron con el tiempo en los pacientes sin neutropenia. Las fungemias causadas por C. albicans aumentan con la edad en pacientes con tumores sólidos sin neutropenia. Se concluye que las fungemias son mayormente causadas por C. tropicalis en pacientes con neoplasias hematológicas con neutropenia y por C. albicans en pacientes con tumores sólidos sin neutropenia. Además, las fungemias en pacientes sin neutropenia aumentan en el tiempo y las causadas por C. albicans, en tumores sólidos sin neutropenia, aumentan con la edad.

ABSTRACT Retrospective descriptive study carried out to determine the characteristics of fungemia in 285 cancer patients hospitalized from 2012 to 2016 at the Instituto Nacional de Enfermedades Neoplásicas (INEN). Demographic, clinical and microbiological information was evaluated. Fungemia by C. albicans predominated in patients with solid tumors and without neutropenia, while those caused by C. tropicalis predominated in patients with hematological neoplasia and neutropenia. C. tropicalis was the agent isolated in most cases (47.0%). Fungemia increased over time in patients without neutropenia. Fungemia caused by C. albicans increases with age in patients with solid tumors without neutropenia. It is concluded that fungemia are mainly caused by C. tropicalis in patients with hematological neoplasia with neutropenia and by C. albicans in patients with solid tumors without neutropenia. In addition, fungemia in patients without neutropenia increases over time; and those caused by C. albicans increase with age in patients with solid tumors without neutropenia.

Candida auris infection in the central catheter of a patient without sepsis symptoms / Infección por Candida auris en el catéter central de un paciente sin síntomas de sepsis

Abstract Background: Candida auris is an emerging yeast frequently reported as resistant to multiple antifungal drugs commonly used to treat Candida infections. This specie can colonize the patient's skin and has great ability for producing outbreaks in hospitals. C. auris is phylogenetically related to other Candida species, can be misidentified using conventional biochemical or commercial methods and requires specific technology for its identification. Case report: We report the first isolate of C. auris in Cali, Colombia, from a central venous catheter in a 37-year-old patient with rheumatoid arthritis and endocarditis who did not have symptoms of sepsis. The yeast was initially misidentified as C. haemulonii using the Phoenix system and subsequently identified as C. auris by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The broth microdilution method was used to determine the minimum inhibitory concentration; the isolate was susceptible to fluconazole, itraconazole, voriconazole and amphotericin B. Conclusions: This report contributes to knowledge of the epidemiology of C. auris infections in individuals with underlying disease and describes an isolate with a behavior different from what is usually reported.

Resumen Antecedentes: Candida auris es una levadura emergente, informada con frecuencia como resistente a diversos antifúngicos usados comúnmente para tratar infecciones por Candida. Esta especie puede colonizar la piel y tiene gran capacidad de producir brotes en ambientes hospitalarios. Está filogenéticamente relacionada con otras especies de Candida, es mal identificada por los métodos bioquímicos o comerciales, y requiere tecnología específica para su identificación. Reporte de caso: Se informa el primer aislamiento de C. auris en Cali, Colombia en un paciente de 37 años con artritis reumatoide y endocarditis, sin síntomas de sepsis, a partir de la punta de catéter venoso central. La levadura inicialmente se identificó como C. haemulonii por el sistema Phoenix® y posteriormente como C. auris por espectrometría de masas desorción/ionización láser asistida por una matriz con detección de masas por tiempo de vuelo (MALDI-TOF MS). Se determinó la concentración inhibitoria mínima por el método de microdilución en caldo que mostró un aislamiento sensible a fluconazol, itraconazol, voriconazol y anfotericina B. Conclusión: Este informe contribuye al conocimiento de la epidemiología de las infecciones por C. auris en individuos con enfermedad subyacente y describe un aislamiento con un comportamiento diferente a lo indicado en otros estudios.

Comparison of the identification results of Candida species obtained by BD PhoenixT and Maldi-TOF (Bruker Microflex LT Biotyper 3.1) / Comparación de los resultados de identificación de especies del género Candida obtenidos por BD PhoenixT y Maldi-TOF (Bruker Microflex LT Biotyper 3.1)

In patients with invasive fungal infections, the accurate and rapid identification of the genus Candida is of utmost importance since antimycotic sensitivity is closely related to the species. The aim of the present study was to compare the identification results of species of the genus Candida obtained by BD PhoenixT (Becton Dickinson -#91;BD-#93;) and Maldi-TOF MS (Bruker Microflex LT Biotyper 3.1). A total of 192 isolates from the strain collection belonging to the Mycology Network of the Autonomous City of Buenos Aires, Argentina, were analyzed. The observed concordance was 95%. Only 10 strains (5%) were not correctly identified by the BD PhoenixT system. The average identification time with the Yeast ID panels was 8h 22 min. The BD PhoenixT system proved to be a simple, reliable and effective method for identifying the main species of the genus Candida.

En pacientes con infecciones fúngicas invasoras, la identificación certera y rápida de las especies del género Candida es de suma importancia, ya que la sensibilidad a los antifúngicos está íntimamente relacionada con la especie. El objetivo del presente estudio fue comparar los resultados de identificación de especies del género Candida obtenidos con el equipo comercial BD PhoenixT (Becton Dickinson -#91;BD-#93;) y con la técnica de Maldi-TOF MS (Bruker Microflex LT Biotyper 3.1.) Se analizaron 192 aislamientos provenientes del cepario perteneciente a la Red deMicología de la Ciudad Autónoma de Buenos Aires, Argentina. La concordancia observada fue del 95%. Solo 10 cepas (5%) no fueron identificadas correctamente por el sistema BD PhoenixT. El tiempo promedio de identificación con los paneles Yeast ID fue de 8 h 22 min. El sistema BD PhoenixT demostró ser un método simple, confiable y efectivo para la identificación de las principales especies del género Candida.

Evaluation of the frequency of Candida spp. in hospitalized and non-hospitalized subjects / Avaliação da frequência de candida spp. em indivíduos hospitalizados e não hospitalizados

Abstract The aim of this study was to evaluate the frequency of Candida species between a non-hospitalized and a hospitalized population. For this purpose, samples of saliva were sampled through sterile swabs, moistened in peptone water and rubbed in the oral cavity of 140 individuals, from which, 70 were hospitalized patients from the Medical Clinic of a Teaching Hospital and the other 70 were non-hospitalized subjects. All saliva samples were plated in Sabouraud Dextrose agar added with Chloramphenicol and incubated at 36 °C for 48 hours. The morphology identification was performed through macroscopic and microscopic characterization, the CHROMagar Candida medium and the VITEK® system Yeast Biochemical Card (bio Mérieux SA, France). The results showed a colonization of Candida spp. in 85.7% the hospitalized individuals, where the species found were C. albicans (60%), C. tropicalis (23.4%), C. krusei (3.3%) and Candida spp. (13.3%). In the non-hospitalized individuals the colonization by Candida spp was 47.1%, and the species found were: C. albicans (45.5%), C.krusei (9.1%), C. guilliermondii (9.1% %), C. tropicalis (3.0%), C. famata (3.0%) and Candida spp. (30.3%). In spite of their presence in oral cavity in both groups, Candida spp. was more frequently isolated in hospitalized individuals, who were 6.73 times more likely to have this fungus in the oral cavity and were 3.88 times more likely to have Candida albicans.

Resumo O objetivo deste estudo foi avaliar a frequência de espécies de Candida entre uma população de indivíduos não-hospitalizados e hospitalizados. Para isto, amostras de saliva foram coletadas através de swabs estéreis, umedecidas em água de peptona e friccionadas na cavidade bucal de 140 indivíduos, dos quais 70 eram pacientes internados em uma Clínica Médica de um Hospital Escola e os outros 70 eram indivíduos não hospitalizados sem contato com ambiente hospitalar. Todas as amostras de saliva foram plaqueadas em ágar Sabouraud dextrose adicionadas de cloranfenicol e incubadas a 36 °C durante 48 horas. A identificação morfológica foi realizada através da caracterização macroscópica e microscópica, com o meio CHROMagar Candida e do sistema VITEK® Biochemical Card (bio Mérieux SA, França). Os resultados mostraram uma colonização de Candida spp. em 85,7% dos indivíduos hospitalizados, onde as espécies encontradas foram: C.albicans (60%), C. tropicalis (23,4%), C. krusei (3,3%) e Candida spp. (13,3%). Nos indivíduos não-hospitalizados a colonização por Candida spp foi de 47,1%, e as espécies encontradas foram: C. albicans (45,5%), C. krusei (9,1%), C. guilliermondii (9,1%), C. tropicalis (3,0%), C. famata (3,0%) e Candida spp. (30,3%). Apesar de sua presença na cavidade oral em ambos os grupos, Candida spp. foi mais freqüentemente isolada em indivíduos hospitalizados, que foram 6,73 vezes mais propensos a ter este fungo na cavidade oral e foram 3,88 vezes mais propensos a ter Candida albicans.

Phenotype-driven strategies for screening Candida parapsilosis complex for molecular identification

Abstract In this study, phenotypic methods presented >80% agreement with the molecular identification of 59 Candida parapsilosis complex. Growth at 15% NaCl or pH 7.0 significantly reduced cfu-counts of Candida orthopsilosis, suggesting these conditions may support the development of phenotypic methods for the differentiation of the cryptic species of C. parapsilosis complex.

Experiencia clínica pediátrica en endocarditis infecciosa por Candida spp / Pediatric clinical experience in infectious endocarditis due to Candida spp

Resumen Introducción: El tratamiento y evolución de endocarditis infecciosa por Candida spp en niños debe basarse en guías de manejo; sin embargo, aún existen controversias al respecto. Objetivo: Describir nuestra experiencia en el tratamiento de EI por Candida spp en pacientes pediátricos. Métodos: Estudio prospectivo, analítico, realizado entre enero de 2006 y diciembre 2017. Análisis paramétrico de variables cuantitativas; razones, proporciones, comparación por medio de χ2 y prueba exacta de Fisher con IC al 95% para variables no paramétricas. Tasa de mortalidad. Resultados: Veinticinco episodios de endocarditis por Candida spp recibieron tratamiento anti fúngico estándar. La mortalidad fue superior en pacientes sometidos a resección de la vegetación endocárdica (66,7%) RR 3,16, χ2 p = 0,029, en niños con síndrome linfo-hemofagocítico SLHF (50%), RR= 1,18 (χ2 NS), en co infección con bacterias multi-resistentes (57,1 %) RR= 2, (χ2 NS) y en EI trombótica (88,9%) RR = 4,74 (χ2 p = 0,004). Conclusiones: Co-infección de EI por Candida sp con bacterias multi-resistentes, SLHF y/o manejo quirúrgico de la vegetación endocárdica, pueden considerarse factores de mal pronóstico.

Background: Treatment and outcome of Candida spp infectious endocarditis in children it most be based on treatment guidelines, however there are some controversies. Aim: To describe our experience on treatment of pediatric candidal infective endocarditis. Methods: Analytic prospective study, from January 2006 to December 2017. Parametric analysis for quantitative variable. Proportions were compared by χ2 and exact Fisher Test CI 95%. Mortality rate. Results: 25 episodes of Candida spp infective endocarditis were treated with standard antifungal drugs. Mortality rate was higher on patients submited to endocardic vegetation resection (66.7%) RR= 3.16, (χ2 p = 0.029), children with lymphohemophagocytic syndrome (LHFS) (50 %) RR= 1.18 (χ2 = N.S.), in multidrug resistant bacterial co infection (57.14%), RR = 2, (χ2 = NS) also thrombotic endocarditis (88.9%) RR= 4.74 (χ2 p = 0.004). Conclusion: Multidrug resistant bacteria co infection with Candida sp IE, LHFS, and/or surgical treatment of endocardic vegetation, might be considered as bad prognostic factors.

Variability in the clinical distributions of Candida species and the emergence of azole-resistant non-Candida albicans species in public hospitals in the Midwest region of Brazil

Abstract INTRODUCTION Incidence and antifungal susceptibility of Candida spp. from two teaching public hospitals are described. METHODS The minimum inhibitory concentrations of fluconazole, voriconazole, itraconazole, and amphotericin B were determined using Clinical Laboratory Standard Institute broth microdilution and genomic differentiation using PCR. RESULTS Of 221 Candida isolates, 50.2% were obtained from intensive care unit patients; 71.5% were recovered from urine and 9.1% from bloodstream samples. Candida parapsilosis sensu stricto was the most common candidemia agent. CONCLUSIONS We observed variations in Candida species distribution in hospitals in the same geographic region and documented the emergence of non-C. albicans species resistant to azoles.

Phagocytosis and nitric oxide production by peritoneal adherent cells in response to Candida albicans in aging: a collaboration to elucidate the pathogenesis of denture stomatitis

Abstract Elderly denture wearers are commonly affected by Candida-associated denture stomatitis (DS), an inflammatory process of the oral mucosa strongly associated with Candida spp and other microorganisms, as well as local and systemic factors. The impaired immune response against pathogens is among the inherent host factors that have been also associated with the pathogenesis of DS. Mononuclear phagocytes respond to the pathogens through phagocytosis followed by the production of several substances inside the phagosomes, among them are the reactive nitrogen species (RNS). A failure in these mechanisms may contribute to the DS development. Objective The aim of this study was to investigate the influence of aging on the internalization and the production of nitric oxide (NO) by peritoneal adherent cells (PAC), in response to Candida albicans (C. albicans). Material and methods PAC obtained from young and aged mice were challenged with dead or viable C. albicans by using predetermined proportions (cells:yeast) for 30 and 120 minutes. Phagocytosis was analyzed by acridine orange dye, and NO production by the Griess reaction. Results C. albicans phagocytosis by PAC from aged mice was similar to that of young mice, although the cells from older mice cells present more internalized fungi compared with matched control. In addition, a tendency towards impaired NO production by peritoneal mononuclear phagocytes from aged mice was observed. Conclusions PAC from aged mice may capture and store many fungi, which in turn may mean that these cells are effectively unable to eliminate fungi, probably due to impaired NO production. Therefore, considering the important role of C. albicans overgrowth in the pathogenesis of DS and the aspects observed in this study, aging may favor the onset and severity of local candidosis such as DS and its systemic forms.

Fungoma vesical por Candida tropicalis: un caso clínico pediátrico / Bladder fungus ball by Candida tropicalis: a pediatric case report

Vesical fungus ball is a mobile, oval and echogenic mass as a result of accumulation of long and wide numerous hyphae. Fungal urinary tract infection incidence has increased notoriously and there are isolated yeast in 7 to 8% of urine cultures. Different species of Candida are cause of urinary tract infection. Epidemiologically, the first isolated pathogen is Candida albicans, followed by Candida tropicalis. Bladder poll has been documented as the most important risk factor for candiduria in critical patients into intensive care.

Un fungoma vesical es una masa móvil, oval y ecogénica en la vejiga resultante del acúmulo de hifas largas y anchas. La incidencia de la infección urinaria de etiología fúngica se ha incrementado notablemente. Se aíslan levaduras en 7 a 8% de los urocultivos. Diferentes especies de Candida son causantes de infección urinaria, siendo Candida albicans la más frecuente, seguida de Candida tropicalis. Presentamos el caso de un niño varón de cuatro años, con un síndrome de Guillain Barré, catéter urinario permanente, estadía prolongada en UCI y expuesto a tratamiento antibacteriano de amplio espectro que desarrolló un fungoma vesical, diagnosticado por ecotomografía, con aislamiento de C. tropicalis en orina. Se trató con anfotericina B deoxicolato y extracción del fungoma por cistoscopia, con buena respuesta clínica. El cateterismo vesical se ha documentado como el factor de riesgo más importante para candiduria en pacientes de terapia intensiva.

Candida auris: emergence and epidemiology of a highly pathogenic yeast

Candida auris is a multidrug-resistant emerging yeast, which was responsible for healthcare-associated infection outbreaks, and was cataloged as a new species in 2009, after being isolated from a patient's ear canal secretion in Japan. Since the notification of this first occurrence, numerous cases have been reported throughout the world, including Brazil. C. auris affects mainly inpatients, patients in intensive care units, exposed to broad-spectrum antifungal medications and who make use of vascular catheters. Currently, this yeast is one of the main responsible for invasive infections in hospitals and has been cause of concern by authorities and organs due to its rapid dissemination and difficult treatment caused by its low susceptibility to antifungal agents traditionally used in clinical practice. As a contributor to the severity of infections associated with C. auris, the transmission mechanism is still unknown, which implies in a lack of control of the microorganism and high mortality rates. Thus, this literature review presents relevant information in order to alert the importance of C. auris as an etiological agent of systemic infections, as well as its epidemiology and the real challenges of the treatment (AU)

Prevalence of fungemia in a tertiary hospital: Analysis of the last decade / Prevalência de fungemia em um hospital terciário: análise da última década

Summary Introduction: The prevalence of nosocomial fungemia has increased worldwide, and mortality caused by this disease is high. Objective: To assess progress in the last decade, and the prevalence and profile of fungal agents isolated in blood cultures performed in a tertiary university hospital. Method: All the results of blood cultures processed at Hospital das Clínicas, Universidade Federal de Minas Gerais (HC-UFMG), in the time intervals 2001-2003 and 2011-2013 were analyzed retrospectively. For each three-year period, the number of collected blood cultures, the overall positivity rate and the percentage of fungemia were recorded. In addition, all identified fungal species were cataloged. All blood samples were incubated in the BacT/ALERT® (bioMérieux) automation system. Results: In 2001-2003, 34,822 samples were evaluated, with 5,510 (15.8%) positive results. In 2011-2013, the number of blood cultures processed increased to 55,052 samples, with 4,873 (8.9%) positive results. There was an increase in the number of positive cultures for fungi in the analyzed period (2001-2003: 4.16%; 2011-2013: 5.95%; p<0.001). Among the agents, candidemias were predominant, especially those caused by non-albicans Candida species (2001-2003: 57.64%; 2011-2013: 65.17%; p<0.05). There was also an increase in fungemia caused by other genera (2001-2003: 2.62%; 2011-2013: 4.48%; p<0.01). Conclusion: There was an increase in the prevalence of fungemia in the last decade at HC-UFMG. Although candidemias have been responsible for most of the cases, there has been an increase in fungemias caused by other species.

Resumo Introdução: a prevalência de fungemia hospitalar tem aumentado em todo o mundo e a mortalidade por essa afecção é elevada. Objetivo: avaliar a evolução, na última década, da prevalência e do perfil dos agentes fúngicos isolados em hemoculturas realizadas em um hospital universitário terciário. Método: foram analisados retrospectivamente todos os resultados de hemocultura processados no Hospital das Clínicas da Universidade Federal de Minas Gerais (HC-UFMG), entre os períodos de 2001-2003 e de 2011-2013. Para cada triênio foram registrados o número de hemoculturas coletadas, o percentual de positividade geral e o percentual de fungemia. Também foram catalogadas todas as espécies fúngicas identificadas. Todas as amostras sanguíneas foram incubadas no sistema de automação BacT/ALERT® (bioMérieux). Resultados: entre 2001-2003, foram avaliadas 34.822 amostras, sendo 5.510 (15,8%) positivas. Entre 2011-2013, o número de hemoculturas processadas aumentou para 55.052 amostras, sendo 4.873 (8,9%) positivas. Observou-se um aumento do número de culturas positivas para fungos no período analisado (2001-2003: 4,16%; 2011-2013: 5,95%; p<0,001). Dentre os agentes, as candidemias foram predominantes, principalmente por espécies de Candida não albicans (2001-2003: 57,64%; 2011-2013: 65,17%; p<0,05). Houve também aumento da fungemia por outros gêneros (2001-2003: 2,62%; 2011-2013: 4,48%; p<0,01). Conclusão: houve aumento da prevalência de fungemia na última década no HC-UFMG. Embora as candidemias tenham sido responsáveis pela maioria dos casos, houve aumento de fungemias causadas por outras espécies.

Tuberculosis and fungal co-infection present in a previously healthy patient / Coinfección de tuberculosis y candidiasis pulmonar en paciente previamente sana

Background: The coexistance among fungal pathogens and tuberculosis pulmonary is a clinical condition that generally occurs in immunosuppressive patients, however, immunocompetent patients may have this condition less frequently. Objective: We report the case of an immunocompetent patient diagnosed with coinfection Mycobacterium tuberculosis and Candida albicans. Case Description: A female patient, who is a 22-years old, with fever and a new onset of hemoptysis. Clinical findings and diagnosis: PDiminished vesicular breath sounds in the apical region and basal crackling rales in the left lung base were found in the physical examination. Microbiological tests include: chest radiography and CAT scan pictograms in high resolution, Ziehl-Neelsen stain, growth medium for fungus and mycobacteria through Sabouraudís agar method with D-glucose. Medical examinations showed Candida albicans fungus and Mycobacterium tuberculosis present in the patient. Treatment and Outcome: Patient was treated with anti-tuberculosis and anti-fungal medications, which produced good responses. Clinical relevance: Pulmonary tuberculosis and fungal co-infection are not common in immunocompetent patients. However, we can suspect that there is a presence of these diseases by detecting new onset of hemoptysis in patients.

Antecedentes: La coexistencia entre los hongos patógenos y la tuberculosis pulmonar es una condición clínica que se produce generalmente en pacientes inmunosuprimidos, sin embargo, los pacientes inmunocompetentes puede tener esta condición con menor frecuencia. Objetivo: Presentamos el caso de un paciente inmunocompetente con diagnóstico de una coinfección de tuberculosis Mycobacterium tuberculosis y Candida albicans. Caso clínico: Paciente femenina de 22 años con cuadro abrupto de tos, fiebre y hemoptisis sin antecedentes de enfermedad. Hallazgos clínicos y métodos diagnósticos: Al examen respiratorio se halló disminución del murmullo vesicular en la región apical y estertores crepitantes basales en el pulmón izquierdo. Se realizó estudios microbiológicos de muestras tomadas por expectoración y por fibrobroncoscopia en el que se incluyó la tinción de Ziehl-Neelsen, cultivo para micobacteria y hongos en medio Agar Dextrosa Sabouraud y filamentización en suero obteniéndose positividad para Mycobacterium tuberculosis y Candida albicans. Tratamiento y resultado: Se le realizó manejo con antifímicos de primera categoría y antimicóticos con buena respuesta clínica. Relevancia clínica: La coinfección fúngica y tuberculosis pulmonar no es frecuente en pacientes inmunocompetentes, debe sospecharse en episodios abruptos de hemoptisis.

Candida glabrata among Candida spp. from environmental health practitioners of a Brazilian Hospital

Abstract The incidence of the species Candida albicans and non-albicans Candida was evaluated in a Brazilian Tertiary Hospital from the environment and health practitioners. In a 12-month period we had a total positivity of 19.65% of Candida spp. The most recurring non-albicans Candida species was C. glabrata (37.62%), generally considered a species of low virulence, but with a higher mortality rate than C. albicans. Subsequently, C. parapsilosis (25.74%) and C. tropicalis (16.86%) were the second and third most commonly isolated species. Considering the total samples collected from the emergency room and from the inpatient and the pediatric sector, 19.10% were positive for Candida spp., with the predominance of non-albicans Candida species (89.42%). The high percentage of positivity occurred in the hands (24.32%) and the lab coats (21.88%) of the health care assistants. No sample of C. albicans presented a profile of resistance to the drugs. All the non-albicans Candida species presented a decreased susceptibility to miconazole and itraconazole, but they were susceptible to nystatin. Most of the isolates were susceptible to fluconazole and amphotericin B. As expected, a high resistance rate was observed in C. glabrata and C. krusei, which are intrinsically less susceptible to this antifungal agent. The contamination of environmental surfaces by Candida spp. through hand touching may facilitate the occurrence of Candida infections predominantly in immunocompromised patients. In addition to that, the antifungal agents used should be carefully evaluated considering local epidemiologic trends in Candida spp. infections, so that therapeutic choices may be better guided.

First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilm-growing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

Essential oils and major compounds of Hedychium coronarium Koenig (Zingiberaceae) against pathogenic yeast of Candida and Cryptococcus genus / Óleos essenciais e compostos majoritários de Hedychium coronarium Koenig (Zingiberaceae) contra leveduras patogênicas dos gêneros Candida e Cryptococcus

Among the major causative agents of invasive fungal infections stands out the opportunistic yeasts of Candida and Cryptococcus. Regarding the problem of the high incidence of infections by these agents and the difculty of treating the low stockpile of antifungal drugs and the high toxicity of most therapies, the search for new antifungal compounds has been highlighted in recent decades. Hedychium coronarium, popularly known as "lírio-do-brejo" or "gengibre-branco" features several previously reported biological activities, including antimicrobial activity. Compound 1.8-cineole is the major compound in essential oil extracted from roots of H. coronarium, while caryophyllene oxide presents itself as the major in essential oil extracted from leaves of this plant. Our data show strong antifungal activity of compounds, against species of Candida albicans, Candida parapsilosis, Candida krusei, Cryptococcus neoformans and Cryptococcus gattii, with minimal inhibitory concentration and minimal fungicidal concentration equal to 0.2 % (v/v) for essential oil extracted from roots, while the essential oil extracted from leaves showed no activity against yeasts. The caryophyllene oxide showed higher antifungal activity for Cryptococcus spp. Thus, our results showed that the essential oil of rhizome is a promising antifungal agent against pathogenic yeasts.(AU)

Candida spp e Cryptococcus spp estão classifcadas entre os maiores causadores de infecções fúngicas invasivas em pacientes imunocomprometidos. Diante a alta incidência destas infecções por estes agentes e a difculdade do sucesso no tratamento, decorrente do baixo arsenal de fármacos antifúngicos e da alta toxicidade presente na maioria dos esquemas terapêuticos, a busca por novos compostos antifúngicos tem sido alvo de diversos estudos nas últimas décadas. Hedychium coronarium, popularmente conhecido como "lírio-do-brejo" ou "gengibre-branco", apresenta diversas atividades biológicas já descritas, entre elas a atividade antimicrobiana. O composto 1.8-Cineol é o composto majoritário presente no óleo essencial extraído de raízes de H. coronarium e o composto óxido de cariofleno é o composto majoritário extraído das folhas desta planta. Nossos resultados mostram que os compostos extraídos de H. coronarium apresentam forte atividade contra Candida albicans, Candida parapsilosis, Candida krusei, Cryptococcus neoformans e Cryptococcus gattii, com valores de concentração inibitória minima e concentração fungicida minima igual a 0,2 % (v/v) para o óleo essencial extraído das raízes, enquanto que o óleo essencial extraído das folhas, não mostrou atividade contras as leveduras. O composto óxido de cariofleno mostrou maior atividade antifúngica para Crytopcoccus spp. Assim, nossos dados mostraram que o óleo essencial extraído das raízes de H. coronarium, é um agente antifúngico promissor contra leveduras patogênicas.(AU)

Distribución de especies y susceptibilidad antifúngica de Candida spp. causantes de micosis superficiales. Coro, estado Falcón, Venezuela / Species distribution and antifungal susceptibility of Candida spp. causing superficial mycosis. Coro, Falcon state, Venezuela.

Las especies de Candida distintas a C. albicans se describen con frecuencia como agentes causales de micosis superficial y presentan una mayor resistencia al tratamiento con los azoles. Con la finalidad de determinar la distribución de especies y la susceptibilidad antifúngica in vitro de Candida spp., se realizó un estudio ambispectivo donde se analizaron 18 aislados de levaduras obtenidas de muestras de pacientes con diagnóstico de micosis superficial. La identificación taxonómica se realizó mediante la visualización de las características macroscópicas de crecimiento en agar cromogénico y por métodos convencionales. La susceptibilidad a fluconazol y voriconazol se evaluó por el método de difusión en disco. El 88,8% de los aislados provenía de muestras de uñas. C. parapsilosis fue la especie más frecuente, seguida de C. tropicalis, C. albicans y C. krusei, lo cual confirmó el predominio de especies no albicans como causa de micosis superficial. El patrón de susceptibilidad a fluconazol y voriconazol fue similar: todos los aislados de C. parapsilosis y C. albicans resultaron sensibles, mientras que el 83,3% de C. tropicalis mostró sensibilidad a ambos antifúngicos. C. krusei, especie resistente a fluconazol, presentó sensibilidad intermedia al voriconazol. El uso de agar cromogénico permitió detectar infecciones mixtas en muestras de uñas, involucrando en uno de los casos a Candid spp. y C.tropicalis, esta última con resistencia tanto a fluconazol como a voriconazol. Los resultados obtenidos demuestran la importancia de la identificación de especies y la realización de pruebas de susceptibilidad con el fin de evitar fracasos terapéuticos en micosis superficiales. Autor de correspondencia: Leyla Humbría-García. Laboratorio de Micología

Candida species other than C. albicans are often described as causative agents of superficial mycosis and are more resistant to treatment with azoles. In order to determine the distribution of species and in vitro antifungal susceptibility of Candida spp., one ambispective study, which analyzed 18 yeast isolates obtained from samples from patients diagnosed with superficial mycosis, was performed. Taxonomic identification was performed by macroscopic visualization of the growth characteristics in chromogenic agar and by conventional methods. The susceptibility to fluconazole and voriconazole was evaluated by the disc diffusion method. Most of the isolates (88.8%), came from nail samples. C. parapsilosis was the most common species, followed by C. tropicalis, C. albicans and C. krusei, which confirmed the prevalence of non-albicans species as a cause of superficial mycoses. The pattern of susceptibility to fluconazole and voriconazole was similar: all isolates of C. parapsilosis and C. albicans were susceptible, while 83.3% of C. tropicalis showed sensitivity to both antifungals. C. krusei, fluconazole-resistant species showed intermediate susceptibility to voriconazole. The use of chromogenic agar allowed to detect mixed infections in nail samples, involving Candida spp. and C. tropicalis in one case, the latter with resistance to both fluconazole and voriconazole. The results demonstrate the importance of species identification and susceptibility testing to avoid therapeutic failures in superficial mycoses.

Nosocomial candidiasis in Rio de Janeiro State: Distribution and fluconazole susceptibility profile

One hundred and forty-one Candida species isolated from clinical specimens of hospitalized patients in Rio de Janeiro, Brazil, during 2002 to 2007, were analized in order to evaluate the distribution and susceptibility of these species to fluconazole. Candida albicans was the most frequent species (45.4%), followed by C. parapsilosis sensu lato (28.4%), C. tropicalis (14.2%), C. guilliermondii (6.4%), C. famata (2.8%), C. glabrata (1.4%), C. krusei (0.7%) and C. lambica (0.7%). The sources of fungal isolates were blood (47.5%), respiratory tract (17.7%), urinary tract (16.3%), skin and mucous membrane (7.1%), catheter (5.6%), feces (2.1%) and mitral valve tissue (0.7%). The susceptibility test was performed using the methodology of disk-diffusion in agar as recommended in the M44-A2 Document of the Clinical and Laboratory Standards Institute (CLSI). The majority of the clinical isolates (97.2%) was susceptible (S) to fluconazole, although three isolates (2.1%) were susceptible-dose dependent (S-DD) and one of them (0.7%) was resistant (R). The S-DD isolates were C. albicans, C. parapsilosis sensu lato and C. tropicalis. One isolate of C. krusei was resistant to fluconazole. This work documents the high susceptibility to fluconazole by Candida species isolated in Rio de Janeiro, Brazil.

Synergistic effects of tacrolimus and azole antifungal compounds in fluconazole-susceptible and fluconazole-resistant Candida glabrata isolates

In vitro interaction between tacrolimus (FK506) and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole) against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%), followed by that of the combination with ketoconazole (37%), against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%), itraconazole (73%), voriconazole (63%) and fluconazole (60%). The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.

Candida species biofilm and Candida albicans ALS3 polymorphisms in clinical isolates

Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Candida isolates obtained from patients attended at a Brazilian tertiary public hospital (Botucatu, Sao Paulo). C. albicans ALS3 gene polymorphism was also evaluated by determining the number of repeated motifs in the central domain. Of the 198 total biofilm-positive isolates, 72 and 126 were considered as low and high biofilm producers, respectively. Biofilm production by C. albicans was significantly lower than that by non-albicans isolates and was most frequently observed in C. tropicalis. Biofilm production was more frequent among bloodstream isolates than other clinical sources,in urine, the isolates displayed a peculiar distribution by presenting two distinct peaks, one containing biofilm-negative isolates and the other containing isolates with intense biofilm production. The numbers of tandem-repeat copies per allele were not associated with biofilm production, suggesting the evolvement of other genetic determinants.