Use of laboratory methodologies to confirm an outbreak of acute diarrheal disease caused by multiple pathogens / Uso de metodologias laboratoriais para confirmação de surto de doença diarreica aguda causada por múltiplos patógenos

The aim of this study was to investigate an outbreak caused by protozoa, which occurred in a municipality in the Brazil southern region. The investigations were carried out analyzing 47 fresh stool samples and 26 water samples by parasitological and molecular methods, as well as, direct immunofluorescence. After the filtrations of water samples and purification of stool samples, the concentrates were evaluated microscopically for presence of parasites. Molecular analyses were performed by polymerase chain reaction (PCR) for DNA detection of Giardia spp., Cryptosporidium parvum, C. hominis and Cyclospora cayetanensis. Out of 26 water samples, 30.8% (8/26) had waterborne protozoa and C. cayetanensis was the most prevalent (15.5%). Out of the 47 stool samples, 23.4% (11/47) were infected with C. cayetanensis and Giardia spp. The results showed that backwash water samples from filters of the Water Treatment Station were contaminated with C. cayetanensis, C. hominis and Giardia spp., suggesting the contamination of water sources with human waste brought by sewage. These results show the importance of protozoa investigation in water and stool samples by laboratory methodologies principally in outbreaks causing acute diarrheal disease (AU).

O objetivo do presente estudo foi investigar um surto causado por protozoários, ocorrido em um município da região sul do Brasil. As investigações foram realizadas analisando 47 amostras de fezes frescas e 26 amostras de água por métodos parasitológicos, moleculares e de imunofluorscência direta. Após as filtrações das amostras de água e purificação das amostras de fezes, os concentrados foram avaliados microscopicamente a procura de parasitas. A seguir, foram analisadas, pela reação em cadeia da polimerase (PCR), a detecção de DNA de Giardia spp., Cryptosporidium parvum, C. hominis e Cyclospora cayetanensis. Das 26 amostras de água, 30,8% (8/26) apresentaram protozoários de veiculação hídrica, sendo que, C. cayetanensis foi o mais prevalente (15,5%). Das 47 amostras de fezes, 23,4% (11/47) estavam infectadas por C. cayetanensis e Giardia spp. Os resultados mostraram que as águas de retrolavagem dos filtros da Estação de Tratamento de Água estavam contaminadas com C. cayetanensis, C. hominis e Giardia spp. sugerindo a contaminação dos mananciais com dejetos humanos trazidos pelo esgoto. Estes resultados mostram a importância da investigação de protozoários em água e fezes por metodologias laboratoriais, principalmente em surtos que causam doença diarreica aguda (AU).

Cyclospora cayetanensis: diagnóstico de un coccidio poco frecuente en la Argentina / Cyclospora cayetanensis: diagnosis of a rare coccidium in Argentina

Resumen Cyclospora cayetanensis es un coccidio intestinal relacionado con brotes epidémicos debido al consumo de alimentos y agua contaminados con ooquistes esporulados. Predomina en regiones tropicales y subtropicales y puede causar síntomas gastrointestinales que son más graves en inmunocomprometidos, en los que puede causar infecciones extraintestinales. El diagnóstico se realiza por la observación microscópica de ooquistes que presentan un tamaño entre 8 y 10 micrones, refringentes y con glóbulos internos. Se confirma con coloración ácido resistente; los ooquistes se tiñen de color fucsia y tienen la capacidad de autofluorescer. En este informe se describe el diagnóstico de Cyclospora cayetanensis en un paciente inmunocomprometido, oriundo de Perú, que reside actualmente en un barrio vulnerable de la Ciudad de Buenos Aires. Este informe representaría el sexto caso diagnosticado en la Argentina.

Abstract Cyclospora cayetanensis is an intestinal coccidium related to epidemic outbreaks due to consumption of food and water contaminated with sporulated oocysts. It predominates in tropical and subtropical regions and may cause gastrointestinal symptoms which are more severe in immunocompromised patients, to whom it may cause extraintestinal infections. Diagnosis is made by the microscopic observation of oocysts between 8 and 10 microns in size, with refractile globules inside of them. This diagnosis is confirmed by acid-fast staining where oocysts are observed red-stained. With ultraviolet epifluorescence, they stand out as bright blue or green staining circles. The aim of this report is to communicate the diagnosis of C. cayetanensis in an immunocompromised patient, born in Peru, but currently living in a vulnerable neighbourhood of Buenos Aires city. To our knowledge, this would be the sixth case of C. cayetanensis diagnosed in Argentina.

Resumo Cyclospora cayetanensis é um coccídio intestinal relacionado com surtos epidêmicos devido ao consumo de alimentos e água contaminados com oocistos esporulados. Predomina nas regiões tropicais e subtropicais e pode provocar sintomas gastrointestinais mais graves em pacientes imunodeprimidos, nos quais pode causar infecções extraintestinais. O diagnóstico é feito através da observação microscópica de oocistos com tamanho entre 8 e 10 mícrons, refringentes e com glóbulos internos. O diagnóstico é confirmado com coloração ácido-resistente; os oocistos são tingidos de cor fúcsia e eles têm a capacidade de auto-fluorescer. Este relatório descreve o diagnóstico de Cyclospora cayetanensis em um paciente imunodeprimido, nativo do Peru, atualmente residindo em um bairro vulnerável da cidade de Buenos Aires. Este relatório representaria o sexto caso diagnosticado na Argentina.

Intestinal coccidian: an overview epidemiologic worldwide and Colombia / Coccidias Intestinales: Panorama epidemiológico mundial y en Colombia

Intestinal coccidia have been classified as protozoa of the Apicomplex phylum, with the presence of an intracellular behavior and adaptation to the habit of the intestinal mucosa, related to several parasites that can cause enteric infections in humans, generating especially complications in immunocompetent patients and opportunistic infections in immunosuppressed patients. Alterations such as HIV/AIDS, cancer and immunosuppression. Cryptosporidium spp., Cyclospora cayetanensis and Cystoisospora belli are frequently found in the species. Multiple cases have been reported in which their parasitic organisms are associated with varying degrees of infections in the host, generally characterized by gastrointestinal clinical manifestations that can be observed with diarrhea, vomiting, abdominal cramps, malaise and severe dehydration. Therefore, in this review a specific study of epidemiology has been conducted in relation to its distribution throughout the world and in Colombia, especially, global and national reports about the association of coccidia informed with HIV/AIDS. Proposed revision considering the needs of a consolidated study in parasitology, establishing clarifications from the transmission mechanisms, global and national epidemiological situation, impact at a clinical level related to immunocompetent and immunocompromised individuals, as well as a focus on public health in institutional government policies and scientific information based on the characterization of coccidia in the tropical region and Colombia.

Los coccidios intestinales se han clasificado como protozoos del Apicomplexa phylum, con presencia de un comportamiento intracelular y adaptación al hábito de la mucosa intestinal, relacionado con varios parásitos que pueden causar infecciones entéricas en los humanos, generando especialmente complicaciones en pacientes inmunocompetentes e infecciones oportunistas en pacientes inmunodeprimidos. Alteraciones como el VIH/SIDA, cáncer e inmunosupresión con tratamientos farmacológicos. En las especies que se encuentran frecuentemente se encuentran Cryptosporidium spp., Cyclospora cayetanensis y Cystoisospora belli. Se han reportado múltiples casos en los que sus organismos parásitos se asocian a diversos grados de infecciones en el huésped, generalmente caracterizadas por manifestaciones clínicas gastrointestinales que pueden observarse con diarrea, vómitos, calambres abdominales, malestar general y deshidratación severa. Por lo tanto, en esta revisión se ha realizado un estudio específico de epidemiología con relación a su distribución en todo el mundo y en Colombia, especialmente, reportes a nivel global y nacional a cerca de la asociación de coccidios informados con el VIH/SIDA. Revisión propuesta con el objetivo de considerar las necesidades de un estudio consolidado a nivel del campo de la parasitología, evidenciando literatura actualizada, estableciendo información de los mecanismos de transmisión, situación epidemiológica global y nacional, impacto a nivel clínico relacionadas con individuos inmunocompetentes e inmunocomprometidos, así como un enfoque en salud pública en políticas gubernamentales institucionales y la información científica basada en la caracterización de coccidias en la región tropical y principalmente en Colombia.

Una mirada actual sobre Cyclospora spp. y ciclosporosis / A current look at Cyclospora spp. and cyclosporiasis

RESUMEN Introducción: La ciclosporosis es una enfermedad emergente caracterizada por producir cuadros clínicos de gastroenteritis en humanos luego de la ingesta de agua o de alimentos contaminados, siendo más severa en niños e inmunosuprimidos; es ocasionada por el coccidio Cyclospora cayetanensis cuyo cuerpo de conocimiento está en pleno proceso de construcción. Objetivo: Recopilar y presentar en forma comprimida lo hallado hasta ahora sobre la biología, manifestaciones clínicas, diagnóstico, tratamiento y comportamiento epidemiológico de C. cayetanensis. Metodología: Se trató de un estudio documental con base en la revisión minuciosa de la literatura científica mundial. Resultado y conclusión: Se ofrece a los profesionales de la salud e instituciones sanitarias oficiales, herramientas teóricas necesarias para el control de la parasitosis.

ABSTRACT Introduction: Cyclosporiasis is an emergent disease characterized by producing clinical profiles of gastroenteritis in humans after the intake of water or contaminated food, and it is more severe in children and immunosuppressed patients. It is caused by the coccidia Cyclospora cayetanensis, whose body of knowledge is in the process of construction. Objective: To compile and show in compressed form what has been found so far on the biology, clinical manifestations, diagnosis, treatment and epidemiological behavior of C. cayetanensis. Methodology: a documentary study based on a thorough review of the world scientific literature. Results and conclusion: the theoretical tools necessary for the control of parasitosis are offered to health professionals and official health institutions.

Coccidiosis y amibiasis intestinal en niños de edad escolar de un distrito de Lambayeque, Perú / Intestinal coccidiosis and amebiasis in school age children of a district in Lambayeque, Peru

Objetivos: Determinar la prevalencia y los factores asociados a coccidiosis y amibiasis intestinal en niños de edad escolar del distrito de Chongoyape, Chiclayo, Perú. Material y métodos: Estudio transversal de tipo relacional entre noviembre del 2014 y enero del 2015 en 133 escolares. Para recolectar la información sociodemográfica y de saneamiento se usó un cuestionario estructurado. La detección de los parásitos se realizó en muestras seriadas de heces. Para los coccidios intestinales se realizó examen microscópico directo (EMD), técnica ácido resistente modificada (TARM) y ELISA coproantígenos para Cryptosporidium spp., mientras que para la amibiasis se usó EMD y ELISA coproantígenos para Entamoeba hystolitica. Resultados: El 6,8% (9/133) de la muestra presentó coccidiosis intestinal, 3,8% (5/133) con Cryptosporidium spp., y 3,0% (4/133) con Cyclospora cayetanensis. No se detectó Cystoisospora belli. El 4,5% (6/133) de la muestra presentó E. histolytica. El factor asociado en ambas parasitosis fue el consumo de agua insalubre (p=0,001 y p=0,026 respectivamente), mientras que el contacto con animales se asoció a la coccidiosis intestinal (p=0,013). Conclusiones: La coccidiosis y la amibiasis intestinal son frecuentes en niños de la población estudiada, evidenciando su importancia como problema de salud pública y la necesidad de un diagnóstico específico y rutinario en las instituciones de salud de la región. (AU)

Objectives: To determine the prevalence of and risk factors for intestinal coccidiosis and amebiasis in school-age children in the district of Chongoyape, Chiclayo, Peru. Methods: Cross-sectional study carried-out between November 2014 and January 2015 among 133 school-aged children. A structures questionnaire collected socio-demographic and sanitation information. Serial stool samples were required, which were processed under direct microscopy, modified acid-fast staining and ELISA to detect antigens of Cryptosporidium spp, while direct microscopy and ELISA were used to detect Entamoeba hystolitica. Results: 6.8% (9/133) of the samples were positive for coccidian parasites, 3.8% (5/133) were positive for Cryptosporidium and 3% (4/133) for Cyclospora cayetanensis, no Cystoisospora belli was identified. E. hystolitica was found in 4.5% (6/133) samples. Drinking non-potable water was a risk factor for getting both infections, while animal contact was associated with getting a coccidian parasite (p=0.013). Conclusions: Both coccidian and intestinal amebas are frequent protozoan infections in this population. These parasites are public health problem that require routine and specific diagnostic methods in the region. (AU)

Aspectos epidemiológicos de coccidiosis intestinales en comunidad rural de la Península de Paraguaná, estado Falcón, Venezuela / Epidemiological aspects of intestinal coccidiosis in a rural community of Paraguaná peninsula, Falcon state, Venezuela

Resumen Introducción: Las coccidiosis intestinales ocasionadas por Cryptosporidium, Cystoisospora belli y entre abril- junio de 2015, se realizó un estudio prospectivo, descriptivo y transversal para Cyclospora cayetanensis constituyen patologías parasitarias de alta relevancia en la Salud Pública. Objetivo: Determinar la prevalencia y parámetros epidemiológicos de coccidiosis intestinales en 188 habitantes de "El Hato", Estado Falcón, Venezuela. Metodología: Para la identificación de los probables factores de riesgos asociados a las coccidiosis intestinales se usó una ficha encuesta-epidemiológica. El diagnóstico parasitológico se realizó con el método directo y la coloración de Kinyoun. Resultados: La prevalencia global de parasitosis intestinales fue 64,36%, siendo Blastocystis spp. el taxón más frecuentemente observado (39,89%). La prevalencia de coccidios intestinales fue 37,23%, observándose prevalencias de 32,98% para Cyclospora cayetanensis, 26,60% para Cryptosporidium spp. y 3,19% para Cystoisospora belli. Con la aplicación del análisis de regresión logística múltiple se determinó como potenciales factores de riesgo independientes significativamente involucrados en la transmisión de las entero-coccidiosis: lavado inadecuado de manos [Odds Ratio (OR) = 1,89], el consumo de "comidas rápidas" (OR=1,26), empleo del agua más frecuentemente para aseo personal y lavado de vestimentas (OR=2,88), tener un nivel socio-económico bajo (nivel IV/V-Graffar) (OR=1,41), nivel de instrucción de la madre (primaria/secundaria) (OR=0,53), que se realice limpieza del hogar interdiario (OR=2,95), y que se posea animales (OR=2,06). Conclusiones: Los hallazgos de esta investigación muestran que las infecciones ocasionadas por los coccidios intestinales aun representan un problema de salud pública en las áreas rurales de Venezuela.

Abstract Introduction: Intestinal coccidioses caused by Cryptosporidium sp., Cystoisospora belli and Cyclospora cayetanensis are parasitic diseases of major clinical importance in Public Health. Objective: Between April to June 2015, a prospective, descriptive and cross-sectional survey was designed to determine the prevalence and epidemiological profiles of intestinal coccidioses in 188 inhabitants of "El Hato", Falcon State, Venezuela. Methods: Probable risk factors for intestinal coccidioses were identified by using epidemiological questionnaires. The diagnosis of coccidian infection was made by direct wet-mounting and Kinyoun staining. Results: The overall prevalence of intestinal parasitosis was 64.36%, and Blastocystis spp. was the most prevalent taxa (39.89%). Enterococcidioses prevalence was 37.23%, detecting prevalence values of 32.98% for Cyclospora cayetanensis, 26.60% for Cryptosporidium spp. and 3.19% for Cystoisospora belli. Multiple logistic regression analysis allowed us to determine as independent potential risk factors for transmission of these enterococciodioses: inappropriate hand washing [Odds Ratio (OR) = 1.89], fast food consumption (OR=1.26), major use of water for personal and clothes washing (OR=2.88), low socio-economic status (level IV/V-Graffar) (OR=1.41), mother's educational status (primary/secondary school) (OR=0.53), non-daily home cleaning (OR=2.95), and keeping domestic animals (OR=2.06). Conclusions: Findings of this study showed that infections caused by intestinal coccidian infections are still remains as a serious health problem in rural areas of Venezuela.

Development of Molecular Diagnosis Using Multiplex Real-Time PCR and T4 Phage Internal Control to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis from Human Stool Samples

This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler's diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, FAM™, HEX™, Cy5™, and CAL Fluor Red® 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was 2×10 copies for C. parvum and for C. cayetanensis, while it was 2×10³ copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler's diarrhea.

Cyclospora cayetanensis infections among diarrheal outpatients in Shanghai: a retrospective case study / 医学前沿

Cyclospora cayetanensis is a foodborne and waterborne pathogen that causes endemic and epidemic human diarrhea worldwide. A few epidemiological studies regarding C. cayetanensis infections in China have been conducted. During 2013, a total of 291 stool specimens were collected from patients with diarrhea at a hospital in urban Shanghai. C. cayetanensis was not detected in any of the stool specimens by traditional microscopy, whereas five stool specimens (1.72%, 5/291) were positive by PCR. These positive cases confirmed by molecular technology were all in the adult group (mean age 27.8 years; 2.94%, 5/170) with watery diarrhea. Marked infection occurred in the rainy season of May and July. Sequence and phylogenetic analyses of the partial 18S rRNA genes of C. cayetanensis isolated showed intra-species diversity of this parasite. This study showed, for the first time, that C. cayetanensis is a pathogen in outpatients with diarrhea in Shanghai, albeit at a low level. However, the transmission dynamics of this parasite in these patients remain uncertain.

El ciclo biológico de los coccidios intestinales y su aplicación clínica / The biological cycle of the intestinal coccidia and its clinical application

Resumen El estudio de los ciclos biológicos parasitarios que se aprenden en la asignatura de Microbiología y Parasitología en la Facultad de Medicina de la UNAM, brinda a los estudiantes, información importante sobre aspectos fisiopatológicos y clínicos, que les permitirán entender con mayor claridad por qué se realizan ciertas técnicas de diagnóstico específicas y los protocolos terapéuticos a seguir en los pacientes. Se revisa en este artículo el ciclo biológico de los coccidios intestinales y su relación con el cuadro clínico en el huésped.

Abstract The study of the biological cycles of parasites taught in the course of Microbiology and Parasitology in the Faculty of Medicine, UNAM, provides students important information concerning pathophysiological and clinical aspects of the diseases. This will allow them to understand more clearly the diagnostic techniques and the therapeutic protocols to be followed with the patients. This article reviews the biological cycle of the intestinal coccidia and its relationship with the clinical features of the host.

Simultaneous Molecular Detection of Cryptosporidium and Cyclospora from Raw Vegetables in Korea

Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, Cryptosporidium and Cyclospora were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, Cryptosporidium was isolated from all 6 kinds of vegetables, whereas Cyclospora was detected in 4 kinds of vegetables (except perilla leaves and chives). Cryptosporidium (17.8%) and Cyclospora (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. Cryptosporidium was detected all year long; however, Cyclospora was detected only from October to January. In 2 samples (sprout and blueberry), both Cryptosporidium and Cyclospora were detected. Further investigations using TaqI restriction enzyme fragmentation and nested PCR confirmed Cryptosporidium parvum and Cyclospora cayetanensis, respectively. In conclusion, we detected C. cayetanensis in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.

Coccidiosis intestinal en niños admitidos en un hospital de Perú y comparación de dos métodos para la detección del Cryptosporidium spp / Intestinal coccidiosis in children admitted to a hospital in Peru and comparison of two methods for detecting Cryptosporidium spp

Con el objetivo de estimar la frecuencia de infección por coccidios intestinales en niños admitidos en un hospital de Perú, y comparar la tinción ácido-resistente modificada (TARM) y el ELISA para la detección de Cryptosporidium spp.; se realizó un estudio transversal entre octubre de 2014 y junio de 2015. Los coccidios se detectaron mediante la TARM y ELISA Cryptosporidium (kit r-Biopharm) en muestras seriadas de heces. De un total de 325 niños, el 5,5% tuvieron algún coccidio intestinal, 3,7% Cryptosporidium spp. (usando ambas técnicas) y 1,8% Cyclospora cayetanensis (TARM). La TARM y ELISA mostraron una concordancia de 0,955 en la detección de Cryptosporidium spp. Se concluye que los coccidios intestinales son frecuentes en niños de la población estudiada; asimismo, ambas técnicas pueden usarse para la detección de Cryptosporidium spp., sin embargo, el menor costo y la capacidad de detectar otros coccidios ofrecen una ventaja a la TARM en la práctica diaria.

A cross-sectional study was done between October 2014 and June 2015 to estimate the frequency of infection due to intestinal coccidiosis in children admitted to a hospital in Peru, and compare the modified acid-fast staining procedure with the enzyme-linked immunoassay (ELISA) method for the detection of Cryptosporidium spp. Coccidia were detected using the modified acid-fast staining procedure and the Cryptosporidium ELISA method (kit r-Biopharm) on seriated stool samples. Out of a total of 325 children, 5.5% had some type of intestinal coccidiosis: 3.7% involved Cryptosporidium spp. (using both techniques) and 1.8% involved Cyclospora cayetanensis (using the modified acid-fast staining procedure). The modified acid-fast staining procedure and ELISA method revealed a 0.955 consistency in the detection of Cryptosporidium spp. In conclusion, intestinal coccidiosis is frequent in children based on the population studied; similarly, both techniques can be used to detect Cryptosporidium spp., but the modified acid-fast staining procedure has an advantage over the other because it is cost-effective and can detect other coccidia in everyday practice.

Primer reporte de Cyclospora cayetanensis en una comunidad indígena Kariña del municipio Sucre, estado Sucre, Venezuela / First report of Cyclospora cayetanensis in an indigenous Kariña A community in Sucre state, Venezuela

Cyclospora cayetanensis es un protozoario emergente, considerado un patógeno importante causante de diarrea endémica y epidémica en niños y adultos en todo el mundo. En el estado Sucre, son escasas las investigaciones dedicadas al diagnóstico de los coccidios intestinales y se desconocen los aspectos epidemiológicos asociados con estos parásitos. En este sentido, se realizó un estudio en la comunidad indígena Kariña de Piñantal, estado Sucre, Venezuela, con la finalidad de determinar la prevalencia de las enteroparasitosis haciendo énfasis en los coccidios intestinales. Para ello, se recolectaron 141 muestras fecales de individuos de ambos géneros con edades comprendidas entre 0 y 60 años. El diagnóstico parasitológico se realizó mediante el examen directo, técnica de filtración de formol al 5%, dicromato de potasio al 2,5%, coloración de Kinyoun y micrometría ocular. La prevalencia de enteroparasitosis fue de 82,98%, destacándose el cromista Blastocystis spp. (60,80%), Endolimax nana (24,70%) y Ascaris lumbricoides (25,53%). De los coccidios intestinales evaluados, sólo se encontró Cyclospora cayetanensis (10%), resultando los individuos menores de 20 años los más afectados (85,71%); esta alta prevalencia de C. cayetanensis refleja precarias condiciones de vida asociadas con insalubridad y falta de educación en la población indígena Kariña. De los individuos con ciclosporiosis, sólo el 36% presentó diarrea y el 64% restante resultaron portadores asintomáticos, lo que representa un riesgo epidemiológico que debe ser tomado en cuenta por los organismos sanitarios competentes para la implementación del diagnóstico de coccidios de forma rutinaria, contribuyendo así a la prevención de brotes.

Cyclospora cayetanensis is an emerging protozoan pathogen considered as a major cause of endemic and epidemic diarrhea in children and adults worldwide. In Sucre state, Venezuela, few investigations have focused on the diagnosis of intestinal coccidia, and the epidemiological aspects associated with these intestinal parasites are unknown. The prevalence of enteroparasitosis, in particular intestinal coccidian, was studied in an indigenous Kariña community in Piñantal, Sucre state, Venezuela. A total of 141 fecal specimens were collected from individuals of both sexes, aged between 0 and 60 years. Parasitological diagnosis was done by direct examination, filtration technique with 5% formalin, 2.5% potassium dichromate, Kinyoun stain and ocular micrometry. The total prevalence of intestinal parasites was 82.98% with the chromista Blastocystis spp. (60.8%), Endolimax nana (24.7%) and Ascaris lumbricoides (25.53%), being the most abundant species. Of the evaluated intestinal coccidia, only Cyclospora cayetanensis (10%) was found, with individuals under 20 years old being the most affected (85.71%). The high prevalence of C. cayetanensis reflects inadequate living conditions associated with poor health and a lack of education among the indigenous Kariña population. Of the individuals infected with Cyclosporiosis, only 36% had diarrhea and the remaining 64% were asymptomatic carriers, these latter representing an epidemiological risk. This should be taken into account by the competent health authorities and the diagnosis of coccidia conducted routinely in order to reduce outbreaks.

Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler's Diarrhea

This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler’s diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10³ oocysts for C. parvum, >1×10⁴ cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

<i>Cyclospora</i> Infection among School Children in Kathmandu, Nepal: Prevalence and Associated Risk Factors

Background: The intestinal coccidian protozoa <i>Cyclospora cayetanensis</i> has emerged as an important cause of parasitic diarrhea among children living in developing countries. This study aimed to determine the prevalence of <i>Cyclospora</i> among the school children of Kathmandu with reference to various associated risk factors.Methodology: A total of five hundred and seven stool samples from students between the age of 3–14 years, studying in 13 different schools in Kathmandu were collected during the study period (May–November, 2014) and processed at the Public Health Research Laboratory, Institute of Medicine, Kathmandu, Nepal. A modified acid fast staining technique (Kinyoun’s method) was used to detect oocyst of <i>Cyclospora</i> from the formal-ether concentrated stool samples.Results: <i>Cyclospora</i> was detected in 3.94% (20/507) of the stool samples examined. The prevalence was found to be highest among the students in the 3–5 year age group i.e. 10.15% (13/128), peaking during the rainy season (June–August). The detection rate was found to be significantly higher (p < 0.05) among children presenting with diarrheal symptoms, household keeping livestock and consumers of raw vegetables/fruits, showing a prevalence of 10.57% (11/104), 10.11% (9/89) and 7.25% (14/193) respectively.Conclusion: Consumption of untreated drinking water, fresh produce (raw fruits/vegetables) without proper washing and the presence of livestock at home were found to be predisposing factors for higher susceptibility of infection due to <i>Cyclospora</i>. This finding confirms the existence of a public-health issue with potentially serious consequences whereby children can be infected through exposure to oocysts in contaminated food and water and get ill as a result.

Parasitos oportunistas intestinais em pacientes submetidos à hemodiálise - revisão sistemática da literatura / Opportunistic intestinal parasites in hemodialysis patients - a systematic literature review / Parásitos intestinales oportunistas en pacientes sometidos a hemodiálisis - revisión sistemática de la literatura

Realizou-se uma revisão sistemática da literatura com o objetivo de identificar a ocorrência de parasitos entéricos oportunistas em pacientes nefropatas crônicos submetidos à hemodiálise. A revisão consistiu de busca de artigos publicados nas bases de dados MEDLINE, LILACS, SciELO, PubMed entre 1991 e 2013. Foram identificados 178 artigos, dos quais dez foram considerados relevantes para o presente trabalho. Nos trabalhos os pesquisadores demonstraram que os pacientes imunossuprimidos submetidos à hemodiálise são potencialmente infectados por agentes entéricos oportunistas. Há necessidade de maior quantidade de estudos sobre o tema uma vez que existe uma ocorrência mundial crescente das doenças renais crônicas e o potencial acometimento de doenças oportunistas nesta população podendo, inclusive, ocasionar a contaminação de ambientes hospitalares por protozoários entéricos oportunistas.

The objective of this systematic literature review was to identify the occurrence of opportunistic enteric parasites in chronic kidney patient undergoing hemodialysis. The review consisted on searching articles published on MEDLINE, LILACS, SciELO, and PubMed databases between 1991 and 2013. A total 178 articles were identified, ten of which were considered relevant for the present study. In the referred studies, the researchers demonstrated that immunosuppressed patients undergoing hemodialysis are potentially infected by opportunistic enteric agents. Further studies are needed on this topic, as there is a growing global concern with chronic kidney diseases and the potential for these patients contracting opportunistic diseases, which, inclusively, could contaminate hospital environments with opportunistic enteric protozoa.

Se realizó revisión sistemática de la literatura con el objetivo de identificar la ocurrencia de parásitos entéricos oportunistas en pacientes de nefropatía crónica sometidos a hemodiálisis. La revisión consistió en la búsqueda de artículos publicados en las bases de datos MEDLINE, LILACS, SciELO y PubMed, entre 1991 y 2013. Fueron identificados 178 artículos, de los cuales diez fueron considerados relevantes para el presente estudio. En los trabajos, los investigadores demostraron que los pacientes inmunosuprimidos sometidos a hemodiálisis son potencialmente infectados por agentes entéricos oportunistas. Hay necesidad de mayor cantidad de estudios sobre el tema, toda vez que existe presencia creciente a nivel mundial de enfermedades renales crónicas y el potencial agravamiento por enfermedades oportunistas en ésta población, pudiendo, inclusive, dar lugar a la contaminación de ambientes hospitalarios por protozoarios entéricos oportunistas.

Cyclospora cayetanensis: Un coccidio emergente / Cyclospora cayetanensis: An emerging coccidian / Cyclospora cayetanensis: Um coccídio emergente

Cyclospora cayetanensis es un protozoo coccidio con diversa prevalencia a nivel mundial que causa cuadros con diarrea acuosa y voluminosa tanto en pacientes inmunocompetentes como inmunocomprometidos. En estos últimos, además de producir cuadros entéricos más severos, puede cursar con compromisos biliares. Su distribución es cosmopolita, jugando un rol importante en su transmisión, el suelo, el agua y los alimentos contaminados, especialmente las verduras ingeridas crudas. El hombre, único reservorio constatado hasta el presente, elimina con las heces ooquistes inmaduros, que evolucionan en el ambiente transformándose en ooquistes maduros infectivos. Como otros coccidios, presenta un ciclo evolutivo complejo con formas sexuada y asexuada de reproducción en un único hospedador. El diagnóstico se basa fundamentalmente en el hallazgo de ooquistes ácido alcohol resistentes variables en materia fecal. Se han desarrollado métodos moleculares para su detección tanto en muestras fecales como ambientales y de alimentos.

Cyclospora cayetanensis is a coccidian protozoon with several prevalence worldwide that causes watery and voluminous diarrhea conditions both in immunocompetent and immunocompromised patients. In the latter, apart from originating more severe enteric diseases, it can occur with billiary involvement. Its distribution is cosmopolitan, with soil, water and contaminated food, especially consumption of raw vegetables, playing an important role in its transmission. Man, the only reservoir recorded so far, eliminates immature oocytes in feces, which evolve in the environment becoming infective- mature oocytes. Like other coccidians, it presents a complex evolutionary cycle with sexed and unsexed forms of reproduction in a single host. Diagnosis is mainly based on findings of variable acid-alcohol resistant variables in feces. Molecular methods have been developed for its detection in fecal samples and in environmental and food samples as well.

Cyclospora cayetanensis é um protozoo coccídio com diversa prevalência em nível mundial que causa quadros com diarreia aquosa e volumosa tanto em pacientes imunocompetentes como imunocomprometidos. Nestes últimos, além de produzir quadros entéricos mais severos, pode cursar com compromissos biliares. Sua distribuição é cosmopolita, tendo um papel importante na sua transmissão o solo, a água e os alimentos contaminados, especialmente as verduras ingeridas cruas. O homem, único reservatório constatado até o presente, elimina com as fezes oocistos imaturos, que evoluem no ambiente transformando-se em oocistos maduros infectivos. Como outros coccídios apresenta um ciclo evolutivo complexo com formas sexuada e assexuada de reprodução num único hospedeiro. O diagnóstico se baseia fundamentalmente no achado de oocistos ácido-álcool resistentes variáveis em matéria fecal. Foram desenvolvidos métodos moleculares para sua detecção tanto em amostras fecais quanto ambientais e de alimentos.

Principales aspectos de los coccidos asociados a diarrea en pacientes VIH positivos / Key aspects of coccidia associated with diarrhea in HIV patients

Las parasitosis intestinal afectan sobre todo a los países en vías de desarrollo y constituyen un problema de salud pública, relacionado muchas veces con la falta de sistemas de salud eficientes, o fuentes de agua potable, que también se acentúan con enfermedades de fondo como Sida, que se presentan también en los países desarrollados. La bibliografía describe que Cryptosporidium spp, Isospora belli y Cyclospora cayetanensis son los parásitos con más frecuencia asociados con diarrea persistente en casos avanzados de pacientes con VIH/Sida. Este grupo de protozoarios requiere exámenes específicos para su diagnóstico, siendo la coloración de Ziehl-Neelsen, uno de los exámenes no rutinarios que permiten su identificación y que, en la mayoría de las ocaciones, no se realiza en el laboratorio si no es solicitado por el médico de manera explícita...

Opportunistic Parasites among Immunosuppressed Children in Minia District, Egypt

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children.

Reporte de cinco casos de cyclosporiosis en un centro penitenciario en Venezuela / Cyclosporiosis five cases report in Venezuelan penitentiery center

Cyclospora cayetanensis, es un protozoario intestinal humano, productor de diarrea en individuos inmunocompetentes e inmunocomprometidos. La transmisión ocurre por ingestión de ooquistes que han esporulado en el ambiente. La transmisión del parásito ocurre a través de un vehículo ambiental. En los países en vías de desarrollo, la ciclosporiosis se ha relacionado con el agua y los alimentos contaminados, contacto con la tierra o animales, y variables asociadas con bajas condiciones socioeconómicas. En junio de 2004 se realizó un estudio parasitológico en un centro pe nitenciario en Venezuela, donde se presentaron varios casos con diarrea. A 7 personas del sexo masculino, con edades entre 19-39 años y con diarrea de 2 a 10 días de du ración, se les realizó examen de heces con S.S. 0,85%, lugol, Quesel, Sudán III, ziehl Neelsen, Micrometría y Esporulación en K2Cr2O4 2,5%. Se encontró ooquistes de C. cayetanensis en 5 (71, 42%) y huevos de Ancylos tomídeos en 1 (14,3%). El número de casos, en los cuales se observó C. cayetanensis, en individuos con diarrea, en este centro penitenciario en Venezuela, resulta muy llamativo en virtud de ser este un microorganismo de baja prevalencia a escala mundial y nacional. No obstante, las condiciones de hacinamiento y probablemente medidas higiénicas inadecuadas, tanto en las áreas físicas como en la conservación y manipulación del agua y alimentos, favorecen la transmisión de parásitos que ocasionan diarrea. Lo cual demuestra, la necesidad de establecer medidas de control y educación sanitaria a los manipuladores de alimentos, para evitar la transmisión y aparición de diarrea por diversos agentes infecciosos entre estas poblaciones.

Cyclospora cayetanensis is a human intestinal protozoan, causing diarrhea in immunocompetent and immunocompromised individuals. Transmission occurs by ingestion of oocysts that have sporulated in the environment. In developing countries, ciclosporosis has been associated with contaminated food and water, contact with soil or animals and low socioeconomic conditions. In June 2004, a parasitological survey in a venezuelan prison in, where several diarrhea cases appeared. All seven (7) people were male, age ranging 19-39 year-old with diarrhea during between 2 and 10 days; they were evaluated by stool examination with ss 0,85%, iodine, quensel, sudan III, ziehl Neelsen stain/mi - crometry and sporulation in K2Cr2O4 2,5%. C. cayetanensis oocysts were found in 5 (71,42%) and hookworm eggs in one (14,3%). The number of diarrhea cases at this prison in Venezuela in which C. cayetanensis was observed, is very striking, knowing the premise for this microorganism to be global and regional low prevalent protozoan. However, overcrowding and poor hygienic measures, probably both, in the physical areas and in the storage and food/water handling, contribute to diarrhea-causing parasites transmission. This shows the need to implement control measures and health education for food handlers to prevent transmission and diarrhea outbreaks caused by infectious agents among this population.

Infection of Cyclospora cayetanensis in Diarrhoeal Children of Nepal.

Introduction: Cyclospora cayetanensis is a coccidian parasite that causes recurrent gastroenteritis among children living under poor sanitary condition and adults from industrialized countries who lived or traveled in endemic developing countries. Methods: A total of 1842 stool specimens from gastroenteritis patients were examined between March 2006 to February 2007 and collected various types of faecal specimens from human, animals and samples of water and green leafy vegetables. Results: Of 1842 stool specimens collected from different areas of Nepal, 146 (7.9 %) were found to be positive for Cyclospora cayetanensis. with the majority were from children 2-9 years of age. The highest rate of infections was found in the month of June; (31.6%). Cyclospora cayetanensis were found to be contaminated in green vegetables including leaves of basil, mint leaves, water sources and feces of domestic animals and these findings were presented in the paper. Conclusion: Cyclospora-like oocysts were detected in vegetables leaves, feces of domestic animals and water sources. The results suggest that water, vegetables and domestic animals are possible sources of infection in Nepal. The study also obtained more information on Cyclospora cayetanensis a coccidian parasite that infect humans especially children and causes prolonged diarrhea and life threatening infestation.