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.

Identificación de bacterias y parásitos asociados a la materia fecal de diferentes ejemplares de Equus ferus caballus de Salento, departamento del Quindío / Identification of bacteria and parasites associated with fecal matter from different specimens of Equus ferus caballus from Salento, department of Quindío

Las enfermedades de origen zoonótico han sido gravemente descuidadas en países en vía de desarrollo, sin tener en cuenta que esto puede tener graves implicaciones a nivel de salud pública, particularmente para Colombia, en las últimas décadas se han dado fuertes cambios sociales que han implicado, entre otras cosas, el aumento de la tendencia a la posesión de animales de compañía incluyendo a Equus ferus caballus, para el cual, la microbiota asociada ha sido mal estudiada en comparación con otros animales. Por lo anterior, se planteó como objetivo identificar la microbiota bacteriana y parasitaria asociada a la materia fecal de Equus ferus caballus, para cumplir dicho objetivo, se tomaron muestras independientes para cada uno de los fines, realizando la concentración de parásitos mediante la técnica de Ritchie y cultivo de bacterias para su posterior identificación mediante el Kit BD BBL CRYSTAL E/NF y GP. Para el análisis de los datos se realizó regresión logística mediante Statgraphics. Se lograron aislar 19 bacterias de las cuales seis fueron explicadas por al menos uno de las variables tenidas en cuenta. Nueve de las bacterias aisladas no se habían reportado con anterioridad para equinos. Se reportan tres géneros de parásitos nuevos para la especie (Balantidium sp, Cyclospora sp. y Taenia sp.) y cuatro nuevos para el país (Allantosoma sp, Blepharocorys sp, Cycloposthium sp. y Bundleia sp.) Se concluye que las variables tenidas en cuenta si tienen influencia en la incidencia de algunos microorganismos aislados, de igual manera se reportan nuevas especies de bacterias para Equus ferus caballus así como de parásitos.

Diseases of zoonotic origin have been seriously neglected in developing countries, without taking into account that this may have serious public health implications, particularly for Colombia, in recent decades there have been strong social changes that have involved, among other things, the increase of the tendency to the possession of companion animals including Equus ferus caballus, for which, the associated microbiota has been poorly studied in comparison with other animals. Therefore, we set ourselves the objective of identifying the bacterial and parasitic diversity associated with fecal matter of Equus ferus caballus. For this, independent samples were taken for each of the purposes, making the concentration of parasites using the Ritchie technique; Samples for bacterial isolation were processed and identified with the BD BBL CRYSTAL E / NF and GP Kit. For the analysis of the data, logistic regression was performed using Statgraphics. We were able to isolate 19 bacteria, of which six were explained by at least one of the variables taken into account. Nine of the isolated bacteria had not been previously reported for equines. Three genera of new parasites are reported for the species (Balantidium sp, Cyclospora sp and Taenia sp.) And four new for the country (Allantosoma sp, Blepharocorys sp, Cycloposthium sp. and Bundleia sp.) It is concluded that the variables taken into if they have influence on the incidence of some isolated microorganisms, in the same way new species of bacteria for Equus ferus caballus as well as parasites are reported.

Enteroparasitism and Risk Factors Associated with Clinical Manifestations in Children and Adults of Jalisco State in Western Mexico

OBJECTIVES: To determine the prevalence and risk factors associated with intestinal parasites in the population of San Juan Cosala, Jalisco, Mexico. METHODS: A total of 277 samples from 104 participants were analysed using direct smear, flotation, formaldehyde/ethyl acetate, and modified Kinyoun’s acid-fast stain methods. The Graham method was applied only for samples from children under 12 years of age for the diagnosis of Enterobius vermicularis. RESULTS: The prevalence of parasite infections in the study population was 77.9% including: Entamoeba histolytica/E. dispar/E. moshkovskii/E. bangladeshi (37.5%), Giardia intestinalis (11.5%); commensals: Endolimax nana (44.2%), Entamoeba coli (27.9%), Chilomastix mesnili (6.7%) and Iodamoeba bütschlii, (2.9%); emerging intestinal protozoans: Blastocystis spp. (49%), Cryptosporidium spp. (7.7%) and Cyclospora cayetanensis (2.9%); and helminths: Enterobius vermicularis (18.3%) and Ascaris lumbricoides (5.8%). The results also showed that 58.64% of the studied population presented polyparasitism. A significant association was found between protozoan infections and housewives, and houses that were not built with concrete ceilings, brick walls and cement floors (p < 0.05). CONCLUSION: Polyparasitism was observed in over half the study population. The most prevalent parasite was Blastocystis spp, whilst the prevalence of helminths was less than that of protozoans. The risk factors for infection to intestinal parasites were being a housewife and not having solid brick, cement and concrete materials for house construction.

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.

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.

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.

Identification of Cystoisospora ohioensis in a Diarrheal Dog in Korea

A 3-month-old female Maltese puppy was hospitalized with persistent diarrhea in a local veterinary clinic. Blood chemistry and hematology profile were analyzed and fecal smear was examined. Diarrheal stools were examined in a diagnostic laboratory, using multiplex real-time polymerase chain reaction (PCR) against 23 diarrheal pathogens. Sequence analysis was performed using nested PCR amplicon of 18S ribosomal RNA. Coccidian oocysts were identified in the fecal smear. Although multiplex real-time PCR was positive for Cyclospora cayetanensis, the final diagnosis was Cystoisospora ohioensis infection, confirmed by phylogenetic analysis of 18S rRNA. To our knowledge, this the first case report of C. ohioensis in Korea, using microscopic examination and phylogenetic analysis.

Intestinal Parasitic Infections and Environmental Water Contamination in a Rural Village of Northern Lao PDR

A field survey studying intestinal parasites in humans and microbial pathogen contamination at environment was performed in a Laotian rural village to identify potential risks for disease outbreaks. A parasitological investigation was conducted in Ban Lak Sip village, Luang Prabang, Lao PDR involving fecal samples from 305 inhabitants as well as water samples taken from 3 sites of the local stream. Water analysis indicated the presence of several enteric pathogens, i.e., Aeromonas spp., Vibrio spp., E. coli H7, E. coli O157: H7, verocytotoxin-producing E. coli (VTEC), Shigella spp., and enteric adenovirus. The level of microbial pathogens contamination was associated with human activity, with greater levels of contamination found at the downstream site compared to the site at the village and upstream, respectively. Regarding intestinal parasites, the prevalence of helminth and protozoan infections were 68.9% and 27.2%, respectively. Eight helminth taxa were identified in fecal samples, i.e., 2 tapeworm species (Taenia sp. and Hymenolepis diminuta), 1 trematode (Opisthorchis sp.), and 5 nematodes (Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, trichostrongylids, and hookworms). Six species of intestinal protists were identified, i.e., Blastocystis hominis, Cyclospora spp., Endolimax nana, Entamoeba histolytica/E. dispar, Entamoeba coli, and Giardia lamblia. Questionnaires and interviews were also conducted to determine risk factors of infection. These analyses together with a prevailing infection level suggested that most of villagers were exposed to parasites in a similar degree due to limited socio-economic differences and sharing of similar practices. Limited access to effective public health facilities is also a significant contributing factor.

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.

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.

Prevalence of Enteric Protozoan Oocysts with Special Reference to Sarcocystis cruzi among Fecal Samples of Diarrheic Immunodeficient Patients in Iran

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

A New Species of Graphis and New Lichen Records from Vietnam, Including a Second Worldwide Report of Sarcographina cyclospora

Graphis upretii is a new lichen species discovered in Vietnam. The species is characterized by a loosely corticate, rough, whitish grey to greyish green thallus, elongate and irregularly branched lirellae with an apically thin complete thalline margin (negrosina morph), laterally carbonized, entire proper exciple, clear hymenium, hyaline, 16~20 transversely locular ascospores, and about 50~95 x 10~15 microm in size. In addition, members of the taxon produce norstictic and stictic acids. Currently, the lichen flora of Vietnam include Arthonia radiata, Brigantiaea tricolor, Coenogonium implexum, Dirina paradoxa, Herpothallon sipmanii, Pertusaria pertusa, and Sarcographina cyclospora.

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.

Diarrea del viajero / Traveler's diarrhea

La diarrea del viajero es una de las condiciones que con mayor frecuencia afecta a los viajeros de países industrializados que visitan las zonas tropicales y subtropicales del planeta. El 20 a 50% de viajeros se van afectar por esta condición, siendo en ocasiones tan severa como para afectar los planes del viajero en la quinta parte de pacientes. El cuadro se manifiesta por la aparición de diarrea asociada a síntomas entéricos como dolor abdominal, nauseas, vómitos y en caso de diarrea inflamatoria fiebre y deposiciones con sangre. Entre los agentes etiológicos bacterianos más frecuentes están Escherichia coli enterotoxigénica, Salmonella, Shigella, entre otros agentes, aunque en cerca de la mitad de los casos no se aísla un agente etiológico. En caso de diarrea persistente debe descartarse parásitos y en zonas endémicas debe realizarse las pruebas especiales para descartar infección por Cyclospora cayetanensis. En pacientes con diarrea del viajero está indicado el manejo empírico con antibióticos, lo cual disminuye la duración de la enfermedad. En ausencia de fiebre o diarrea con sangre puede usarse loperamida. La prevención es importante en especialmente en pacientes de alto riesgo o en quienes sea importante que no se afecte el viaje.

TravelersÆ diarrhea is one of the most common conditions that affect travelers from industrialized countries who visit tropical and subtropical areas of the world. Twenty to fifty percent of travelers will suffer from this condition, and one fifth of these subjects will have diarrhea severe enough to affect their travel plans. Illness is characterized by the occurrence of diarrhea, associated with other gastrointestinal symptoms such as abdominal pain, nausea, vomiting and sometimes, in association of inflammatory diarrhea, fever and bloody stools. Among the most common etiologic agents Enterotoxigenic Escherichia coli, Salmonella, and Shigella are among the most common bacterial pathogens, although in close to one half of cases culture results are negative. In cases of persistent diarrhea parasites must be ruled out, and in endemic areas special stains must be ordered to rule agents like Cyclospora cayetanensis. In patients with TravelersÆ diarrhea empiric treatment with antibiotics is indicated, reducing the duration of illness. In the absence of fever or bloody diarrhea loperamide can be used. Prevention is important, especially in high risk patients or those in whom the trip must not be affected by the illness.

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.

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.