Caracterización molecular de los aislamientos de Cryptococcus neoformans de pacientes con HIV, Guayaquil, Ecuador

Introduction: Neurocryptococcosis is an opportunistic fungal infection that represents a high cost in human lives and for the economy of countries. Its causative agent, the Cryptococcus neoformans/Cryptococcus gattii species complex, has a sexual and an asexual phase, four major serotypes and seven molecular varieties with phenotypic, clinical-epidemiological and antifungal susceptibility differences.Objective: To characterize by molecular methods clinical isolates of C. neoformans from Guayaquil, Ecuador.Materials and methods: We determined mating types, serotypes and molecular varieties by PCR and RFLP in 27 yeast isolates previously identified as C. neoformans by conventional methods. The isolates were recovered from cerebrospinal fluid of HIV seropositive patients with neurological syndrome admitted at “Dr. José Daniel Rodríguez Maridueña” Hospital from December, 2013, to January, 2015.Results: We established a wide prevalence of C. neoformans serotype A, MATα and genotype VNI among the studied isolates.Conclusions: These data are similar to those obtained in other countries and the first identified by molecular characterization in Guayaquil, Ecuador. Therefore, they constitute an important contribution to the knowledge on cryptococcosis in this country.Introducción. La neurocriptococosis es una infección fúngica oportunista que representa un alto costo en vidas humanas y para la economía de los países. Sus agentes causales, las especies del complejo Cryptococcus neoformans/Cryptococcus gattii, tienen una fase sexuada y otra asexuada, cuatro serotipos principales y siete variedades moleculares con diferencias clínico-epidemiológicas, fenotípicas y de sensibilidad a los antifúngicos.Objetivo. Caracterizar molecularmente los aislamientos clínicos de C. neoformans de Guayaquil, Ecuador.Materiales y métodos. Se determinó el tipo de apareamiento, el serotipo y la variedad molecular mediante reacción en cadena de la polimerasa y análisis del polimorfismo de los fragmentos de restricción de 27 aislamientos levaduriformes previamente identificados como C. neoformans mediante métodos convencionales. Los aislamientos fueron recuperados del líquido cefalorraquídeo de pacientes con síndrome neurológico seropositivos para HIV, internados en el Hospital de Infectología “Dr. José Daniel Rodríguez Maridueña”, entre diciembre de 2013 y enero de 2015.Resultados. Se demostró el amplio predominio de C. neoformans del serotipo A, MATα y el genotipo VNI entre los aislamientos estudiados.Conclusiones. Estos datos son similares a los obtenidos en otros países y son los primeros de su tipo en Guayaquil, Ecuador, por lo cual constituyen un aporte importante al conocimiento de la criptococosis en esta ciudad

The status of cryptococcosis in Latin America

Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.Fil: Firacative, Carolina. University of Sydney; AustraliaFil: Lizarazo, Jairo. Universidad de Pamplona; EspañaFil: Illnait Zaragozí, María Teresa. Tropical Medicine Institute Pedro Kourí; CubaFil: Castañeda, Maria Elizabeth. Instituto Nacional de Salud; Colombia. Latin American Cryptococcal Study Group; BrasilFil: Arechavala, Alicia. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Córdoba, Susana Beatríz. Latin American Cryptococcal Study Group; Brasil. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; ArgentinaFil: Mazza, Mariana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Taverna, Constanza Giselle. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Isla, Guillermina. Latin American Cryptococcal Study Group; Brasil. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; ArgentinaFil: Chiapello, Laura Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Vergara, Mario León Silva. Universidade Federal do Triangulo Mineiro; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Melhem, Marcia S. C.. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Szeszs, Maria Walderez. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Martins, Marilena dos Anjos. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Bonfietti, Lucas Xavier. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Oliveira, Rogério Antonio de. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Oliveira, Lidiane de. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Santos, Dayane Christine Silva. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Lazera, Marcia S.. Latin American Cryptococcal Study Group; Brasil. Fundación Oswaldo Cruz; BrasilFil: Wanke, Bodo. Fundación Oswaldo Cruz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Díaz, María Cristina. Latin American Cryptococcal Study Group; Brasil. Universidad de Chile; ChileFil: Escandón, Patricia. Instituto Nacional de Salud; Colombia. Latin American Cryptococcal Study Group; BrasilFil: Noguera, María Clara. Latin American Cryptococcal Study Group; Brasil. Universidad Metropolitana; ColombiaFil: Andreu, Carlos Manuel Fernández. Latin American Cryptococcal Study Group; BrasilFil: Castril­Lón, Laura. Universidad Nacional Autónoma de México; México. Latin American Cryptococcal Study Group; BrasilFil: Bustamante, Beatriz. Hospital Cayetano Heredia. Instituto de Medicina Tropical Alexander von Humboldt; Perú. Hospital Cayetano Heredia; Perú. Latin American Cryptococcal Study Group; BrasilFil: Dolande, Maribel. Universidad Central de Venezuela; Venezuela. Latin American Cryptococcal Study Group; BrasilFil: Ferrara, Giussepe. Universidad Central de Venezuela; Venezuela. Latin American Cryptococcal Study Group; Brasi

The status of cryptococcosis in Latin America

<div><p>Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.</p></div

The status of cryptococcosis in latin America

Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America. © 2018, Fundacao Oswaldo Cruz. All rights reserved

Evaluación de cuatro métodos de extracción del ADN de Histoplasmacapsulatum y su uso en reacciones de PCR

Histoplasmacapsulatum es un hongo endémico de Cuba y el agente causal de la histoplasmosis. Esta micosis se presenta en forma de brotes epidémicos, afecta a las personas, independiente de su estado inmunológico. Su diagnóstico se realiza mediante técnicas microbiológicas, serológicas e histológicas que tienen una sensibilidad moderada y son demoradas. Por lo que se hace necesario la estandarización de métodos moleculares rápidos y eficientes para el diagnóstico de esta micosis. Se evaluaron cuatro métodos de extracción del ADN (enzimático, mecánico y dos químicos: tiocianato de guanidinio y Tritón X-100) a partir de una cepa de H. capsulatum en fase filamentosa. Se determinó la concentración, la pureza, el rendimiento y la integridad del ADN mediante espectrofotometría y electroforesis en gel de agarosa. Se evaluaron dos técnicas de PCR con el ADN obtenido, con cebadores específicos de especie y otro con un cebador arbitrario. Los mejores resultados de concentración, pureza, rendimiento e integridad se obtuvieron con el método de extracción enzimático. Se logró la estandarización de las PCR con cebadores específicos, confirmándose así las 16 cepas estudiadas como H. capsulatum. Con el cebador arbitrario se amplificaron secuencias diferentes en las cepas ensayadas. Se demostró que el método de extracción enzimática brindó un ADN de buena calidad para su utilización en PCR. Este trabajo facilita el diagnóstico rápido y confiable de H. capsulatum en Cuba y brinda la posibilidad de identificar cepas con estructuras relacionadas genéticamente con los marcadores de patogenicidad y la virulencia, útiles para conformar un inmunógenovacunal

Etiological Diagnosis and Correlation of Chromoblastomycosis with Climatic Variables of Weather Stations in Villa Clara, Cuba

In Villa Clara province, Cuba, microorganisms and climatic conditions coexist for the development of the infectious entity chromoblastomycosis. Besides, there is a high percentage of agricultural and handicraft workers exposed to acquiring this infection. The objective of the research consisted in describing the etiological diagnosis and correlating chromoblastomycosis with average climatic variables of the meteorological stations located in Villa Clara. Between 2018-2022, a descriptive, cross-sectional study was conducted in a population of 25 patients diagnosed with chromoblastomycosis by anatomopathological and microbiological studies. Absolute frequencies were used as a summary measure, and for correlation with climatic data from the five meteorological stations, inferential statistics. Of the total number of patients, 92% were diagnosed by anatomopathological studies and only 2/8% had mycological studies, in which the species Fonsecaea pedrosoi was isolated. There was a significant correlation between mean temperature (p=0.045) and mean predicted wind (p=0.028) and the number of cases. This was not the case with relative air humidity (p&gt;0.05). It is concluded that in Villa Clara province, only the etiological agent F. pedrosoi was the cause of the disease during the period studied, with a significant relationship between the disease and the average climatic variables mean temperature and mean predicted wind

Screening for M220 mutation in azole-resistant Aspergillus fumigatus isolates from clinical and environmental specimens in Cuba

Aspergillosis is a fungal disease caused by different Aspergillus species and have high morbidity/mortality rates. Aspergillus fumigatus is the main aetiological agent related to most of Aspergillus infections. The use of triazoles is recommended as first line of treatment, however, the reports of azole-resistant A. fumigatus due to mutations in CYP51 have increased around the world. The goals were to determine the antifungal susceptibility patterns for triazoles in thirty A. fumigatus isolates from clinical and environmental samples and to detect mutation M220 in the resistant ones. Determination of minimum inhibitory concentrations of every isolate was performed by using the Etest commercial method for itraconazole, voriconazole and posaconazole. For the resistant isolates, a PCR was performed for amplifying a 173 bp fragment of cyp51A and the resulting amplicons were sequenced. Nine out of 30 isolates were resistant to itraconazole and none were resistant to voriconazole or posaconazole. The alignment of sequences with a CYP51 A. fumigatus wild type strain did not show any modifications at codon 220. Other molecular mechanisms of resistance are probably related to the resistant phenotypes isolated

Chronic Pulmonary Aspergillosis in Patients with Underlying Respiratory Disorders in Cuba—A Pilot Study

Chronic pulmonary aspergillosis (CPA) is a fungal infection with high mortality and morbidity rates. This disease is caused by several Aspergillus species and affects patients with an underlying respiratory condition. This pilot study aims to recognize CPA among patients with different respiratory diseases. Twenty-one out of 47 patients were classified as CPA based on the examination of clinical signs and symptoms, radiological findings, mycological culture of respiratory samples and analysis of Aspergillus IgG antibodies. There was a close association between high levels of Aspergillus IgG antibodies and the presence of cavities. Although Aspergillus flavus was the predominant species among clinical isolates, the number of isolates was small to reach conclusions on the prevalence of this species as main cause of CPA in Cuba. From the eleven evaluable patients for the treatment with itraconazole (Lozartil&#174;), nine improved their health status while two did not show any recovery. This drug is included in the therapy schemes for aspergillosis in Cuba

The status of cryptococcosis in latin America

© 2018, Fundacao Oswaldo Cruz. All rights reserved. Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification o