Anti-microsporidial effect of thymoquinone on Encephalitozoon intestinalis infection in vitro

To evaluate the anti-microsporidial effects of the active component of Nigella sativa seeds, thymoquinone, against Encephalitozoon intestinalis using an in vitro model. Methods: Anti-microsporidial effect of thymoquinone against Encephalitozoon intestinalis was evaluated by using various concentrations of thymoquinone (0, 1, 5, 10, 15, 20, 30, 35, and 40 uM) and sterile dimethyl sulfoxide. Real time PCR was used to evaluate the inhibitory effects of thymoquinone on the life cycle of Encephalitozoon intestinalis. Results: The cytotoxic effect of thymoquinone on HEK293 cell line was observed with 30, 35, and 40 uM concentrations of thymoquinone after 24, 48, and 72 hours of incubation. It was observed that 10, 15, 20, and 30 uM concentrations of thymoquinone decreased the spore density compared with the control; however, it was significant only at 30 uM. Conclusions: Thymoquinone shows potent anti-microsporidial effects against Encephalitozoon intestinalis in the in vitro model; however, the toxic concentrations of thymoquinone are also toxic to the host cells.

Anti-microsporidial effect of thymoquinone on Encephalitozoon intestinalis infection in vitro

Objective: To evaluate the anti-microsporidial effects of the active component of Nigella sativa seeds, thymoquinone, against Encephalitozoon intestinalis using an in vitro model. Methods: Anti-microsporidial effect of thymoquinone against Encephalitozoon intestinalis was evaluated by using various concentrations of thymoquinone (0, 1, 5, 10, 15, 20, 30, 35, and 40 μM) and sterile dimethyl sulfoxide. Real time PCR was used to evaluate the inhibitory effects of thymoquinone on the life cycle of Encephalitozoon intestinalis. Results: The cytotoxic effect of thymoquinone on HEK293 cell line was observed with 30, 35, and 40 μM concentrations of thymoquinone after 24, 48, and 72 hours of incubation. It was observed that 10, 15, 20, and 30 μM concentrations of thymoquinone decreased the spore density compared with the control; however, it was significant only at 30 μM. Conclusions: Thymoquinone shows potent anti-microsporidial effects against Encephalitozoon intestinalis in the in vitro model;however, the toxic concentrations of thymoquinone are also toxic to the host cells.

Agua potable como posible fuente de brote de diarrea por Microsporidium spp. en pacientes inmunocomprometidos en un hospital pediátrico / The water supply of a pediatric hospital as a possible source of an outbreak of diarrhea due to Microsporidium spp. in immunocompromised patients

Introduction: The hospital water supply is a reservoir of a variety of potentially pathogenic microorganisms that can particularly affect children and immunocompromised patients. Potentially pathogenic Microsporidium spp. have been identified in water. Microsporidiosis is an emerging parasitic and opportunistic infection in immunocompromised patients. Objective and Method: to describe an outbreak of nosocomial diarrhea due to Microsporidium, species Encephalitozoon intestinalis. Results: Seven cases of E. intestinalis associated diarrhea were reported between november 2012 and february 2013, in a unit of immunocompromised patients in L. Calvo Mackenna Children's Hospital. Microsporidium spp. was found in the hospital water supply and water reservoir tank. Secondary cases were transmitted by contact. Control measures included contact precautions, not to use faucet water for hand washing, bottled water for drinking and water reservoir tank sanitation. Conclusions: This research is about a nosocomial outbreak associated with water supply. Water quality in Chilean hospitals is an unresolved issue, especially in immunocompromised patient areas. Compliance of cleaning and disinfection of water supply systems in hospitals must be ensured.

Introducción: Los sistemas de suministro de agua potable de los hospitales constituyen un reservorio de una variedad de microorganismos potencialmente patógenos que pueden afectar especialmente a niños y pacientes inmunocomprometidos. Especies de Microsporidium spp. potencialmente patógenos para el hombre han sido identificadas en el agua potable. La microsporidiosis es una infección parasitaria oportunista en pacientes inmunocomprometidos. Objetivos y Método: Describir un brote de diarrea nosocomial por Microsporidium de la especie Encephalitozoon intestinalis. Resultados: Se registraron siete casos de diarrea por E. intestinalis, entre noviembre de 2012 y febrero de 2013, en una unidad de pacientes inmunocomprometidos del Hospital de Niños Luis Calvo Mackenna, comprobándose la presencia de Microsporidium spp. abundante en el agua potable y estanques del hospital. Los casos secundarios pudieron transmitirse por contacto. Las medidas de control fueron precauciones de contacto, no usar agua de grifos para lavado de manos, ingesta de agua envasada y desinfección de estanques. Conclusiones: Esta investigación corresponde a un brote nosocomial transmitido por agua potable. La importancia de la calidad del agua en los hospitales de nuestro país es un tema no resuelto, especialmente en áreas que atienden pacientes inmunocomprometidos. Debe asegurarse el cumplimiento de limpieza y desinfección de los sistemas de suministro de agua en los hospitales.

Prevalencia de microsporidiosis intestinal en niños con desnutrición severa de un hospital de la ciudad de Maracaibo / Intestinal Microsporidiosis Prevalence in Children With Severe Malnutrition at a Hospital in the City of Maracaibo

Con la finalidad de comparar la prevalencia de Microsporidiosis intestinal en niños con desnutrición severa y niños eutróficos de la Unidad de Recuperación Nutricional del Hospital Chiquinquirá de Maracaibo, Estado Zulia, se realizó la técnica de reacción en cadena de la polimerasa (PCR) para la identificación de Encephalitozoon intestinalis y Enterocytozoon bieneusi en las muestras de heces de 50 niños desnutridos graves y 50 niños eutróficos, cuyas edades fueron estratificadas de la siguiente manera: lactantes menores (0-11 meses), lactantes mayores (12-23 meses), preescolares (2-6 años), y escolares (7 a 12 años). De las especies de microsporidios investigadas, se evidenció Enterocytozoon bieneusi en el 14% de los niños desnutridos graves y 8% en los niños eutróficos, no se obtuvieron amplificaciones para Encephalitozoon intestinalis en los grupos estudiados; al aplicar la prueba estadística X², resultó no significativa para las variables microsporidiosis intestinal y desnutrición. Se concluye que la medida en la que se relacionan la desnutrición, y la microsporidiosis es difícil de esclarecer, ya que la desnutrición es una condición multifactorial y aún no está bien establecido cuál de estos dos factores es la causa y cual la consecuencia.

In order to compare the prevalence of intestinal microsporidiosis in children with severe malnutrition and eutrophic children at the Nutritional Recovery Unit, Chiquinquirá Hospital in Maracaibo, State of Zulia, a chain reaction technique was performed on the polymerase (PCR) to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi in stool samples from 50 healthy and 50 severely malnourished children, whose ages were stratified as follows: younger infants (0-11 months), older infants (12-23 months), preschool (2-6 years) and school children (7-12 years). Of the microsporidia species investigated, Enterocytozoon bieneusi was evident in 14% of severely malnourished children and 8% of the eutrophic children. No amplifications for Encephalitozoon intestinalis were obtained in the groups studied. On applying the chi-square statistical test, the result was not significant for the variables intestinal microsporidiosis and malnutrition. Conclusions are that the extent to which malnutrition and microsporidiosis relate is difficult to clarify, because malnutrition is a multifactorial condition, and it is not yet well established which of these two factors is the cause and which is the consequence.

Detection of Encephalitozoon spp. from Human Diarrheal Stool and Farm Soil Samples in Korea

Microsporidia are eukaryotic organisms that cause zoonosis and are major opportunistic pathogens in HIV-positive patients. However, there is increasing evidence that these organisms can also cause gastrointestinal and ocular infections in immunocompetent individuals. In Korea, there have been no reports on human infections with microsporidia to date. In the present study, we used real-time PCR and nucleotide sequencing to detect Encephalitozoon intestinalis infection in seven of 139 human diarrheal stool specimens (5%) and Encephalitozoon hellem in three of 34 farm soil samples (8.8%). Genotype analysis of the E. hellem isolates based on the internal transcribed spacer 1 and polar tube protein genes showed that all isolates were genotype 1B. To our knowledge, this is the first report on human E. intestinalis infection in Korea and the first report revealing farm soil samples as a source of E. hellem infection. Because microsporidia are an important public health issue, further large-scale epidemiological studies are warranted.

Cryptosporidium spp., Giardia lamblia y encephalitozoon intestinalis, oportunistas emergentes / Cryptosporidium spp., Giardia lamblia y encephalitozoon intestinalis emergent opportunists

Es un hecho conocido que las infecciones oportunistas por protozoos y hongos han aumentado en los últimos años, debido especialmente al aumento de las infecciones por VIH. Cryptosporidium spp., Giardia lamblia y Encephalitozoon intestinalis son protozoos y hongo, respectivamente, mundialmente reconocidos como agentes oportunistas emergentes, responsables de brotes epidémicos provocados por la ingestión de agua potable contaminada, incluso después de una correcta desinfección. La ingestión de estos protozoos puede provocar diferentes grados de enfermedad, entre aguda o leve (población sana) hasta situaciones más graves y agresivas, hasta a veces mortales (pacientes inmunocomprometidos y/o inmunodeprimidos). A pesar de ser responsables de muchos brotes epidémicos, su diagnóstico de laboratorio permanece arduo y trabajoso, incluso utilizando las nuevas técnicas desarrolladas en los últimos años. En esta revisión se resumen las consideraciones generales de estos oportunistas emergentes, así como los métodos de diagnóstico más usuales, incluso los más recientes y específicos.

Epidemiological data, regarding parasitic and fungi opportunist infections, have changed in the last years, especially due to HIV infection. Cryptosporidium spp., Giardia lamblia and Encephalitozoon intestinalis are protozoan and fungi, respectively, worldwide known as opportunistic emergent agents, being responsible by epidemic outbreaks after ingestion of contaminated water, even following a correct disinfection treatment. Its ingestion can cause different effects on individuals’ health, from light or acute among the healthy population, to serious, aggressive or even deadly among the immunodepressed or immunocompromised patients. Contaminated water ingestion can result in outbreaks but protozoa laboratory diagnosis still remains very laborious, even after the development of more sensitive and specific techniques in the last years. In this paper, a revision of these emergent opportunists, their main characteristics and diagnostic tools are described, including the most recent and specific techniques.

Prevalencia de Encephalitozoon intestinalis y Enterocytozoon bieneusi en pacientes VIH positivos de Maracaibo, Venezuela / Prevalence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in HIV positive patients to Maracaibo, Venezuela

Los microsporidios pueden provocar infecciones emergentes y oportunistas en individuos inmunocomprometidos de todo el mundo. Se realizó éste estudio para identificar las especies de microsporidios intestinales presentes en pacientes con VIH-SIDA del Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM). Se recolectaron 50 muestras fecales de individuos con diagnóstico confirmado de VIH durante los años 2007-2008; se obtuvieron las cifras de CD4 de solo 42 pacientes. Las muestras se analizaron mediante PCR separadas para la identificación de Encephalitozoon intestinalis y Enterocytozoon bieneusi. Las especies de microsporidios presentaron un 36% de prevalencia, 10 pacientes presentaron Encephalitozoon intestinalis, 4 Enterocytozoon bieneusi y 4 ambas especies. Se determinó una relación inversamente proporcional y estadísticamente significativa entre el contaje de CD4 y la presencia de microsporidios en la muestra fecal. Es destacable la elevada prevalencia de especies de microsporidios observada en los pacientes VIH estudiados, donde predominó E. intestinalis.

Microsporidioses are considered emerging and opportunistic infections in immunocompromised individuals worldwide. The purpose of this study was to identify the species of intestinal microsporidia in patients with HIV-AIDS from the Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela (SAHUM). Fecal samples were collected from 50 patients with confirmed diagnosis of HIV, during the years 2007 and 2008; the CD4 values were obtained from 42 patients. The samples were analyzed by separate PCRs to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi. Microsporidia species showed a 36% prevalence: ten patients had Encephalitozoon intestinalis, four Enterocytozoon bieneusi and four both species. An inverse and statistically significant relationship between the CD4 count and the presence of microsporidia in the fecal sample was also found. It is remarkable the high prevalence of microsporidia species observed in the HIV patients studied, with a predominance of E. intestinalis.