Evaluation of terrestrial gastropods as possible intermediate hosts of Gurltia paralysans in southern Chile / Avaliação de gastrópodes terrestres como possíveis hospedeiros intermediários de Gurltia paralysans no sul do Chile

Abstract Gurltia paralysans is the causal agent of gurltiosis in domestic cats in South America. Although the life cycle of G. paralysans is unknown, it is thought that gastropods could act as intermediate hosts (IHs), as is the case for several nematodes in the Angiostrongylidae family. The aim of this study was to search for G. paralysans larvae in terrestrial gastropods and determine their role in the life cycle of this nematode species. Terrestrial gastropod samples (n=835) were collected in Punucapa, Valdivia, southern Chile, where cases of gurltiosis had been reported before. The samples included species from the families Arionidae, Limacidae, Helicidae and Milacidae. All gastropods were subjected to enzymatic digestion to isolate G. paralysans larvae. Ten percent of the gastropod samples were analyzed using seminested PCR targeting the 28S rRNA gene, while 2.6% were analyzed by histopathological examination. The results indicated the absence of G. paralysans when using any of the three methods. In conclusion, further studies are needed to evaluate specific species of aquatic or native gastropods acting as possible IHs (in this geographic location).

Resumo Gurltia paralysans é o agente etiológico da gurltiose em gatos domésticos na América do Sul. Embora o ciclo biologico de G. paralysans seja desconhecido, provavelmente é indireto com gastrópodes atuando como hospedeiros intermediários (HIs), como no caso de vários nematoides da família Angiostrongylidae. O objetivo deste estudo foi investigar a presença de larvas de G. paralysans em gastrópodes terrestres para avaliar seu papel no ciclo de vida do parasito. Amostras de gastrópodes terrestres (n = 835) foram coletadas em Punucapa, Valdivia, sul do Chile, onde casos de gurltiose foram relatados anteriormente. As amostras incluíram espécies das famílias Arionidae, Limacidae, Helicidae e Milacidae. Todos os gastrópodes foram submetidos à digestão enzimática para isolar as larvas de G. paralysans. 10% das amostras foram analisadas, utilizando-se seminested PCR para o gen 28S RNAr de G. paralysans, enquanto 2,6% foram analisados ​​por exame histopatológico. Os resultados indicaram ausência de G. paralysans em todos os três métodos. Os dados permitem concluir que são necessários mais estudos para avaliar espécies específicas de gastrópodes aquáticos ou nativos, que atuam como possíveis hospedeiros intermediários nessa localização geográfica.

Abdominal angiostrongyliasis, report of two cases and analysis of published reports from Colombia / Angiostrongiliasis abdominal, reporte de dos casos y análisis de los casos publicados en Colombia

Abdominal angiostrongyliasis is a parasitic zoonosis, endemic in the American continent. Its etiological agent is Angiostrongylus costaricensis, a nematode whose definitive hosts are rats and other rodents and the intermediate hosts, slugs. Mammals acquire the infection by consuming vegetables contaminated with L3 larvae. The disease shows a heterogeneous clinical spectrum and given its low incidence its diagnosis is a great challenge. In Colombia, the first case was reported in 1979 and until 1998, only five additional cases have been reported. However, in the last two decades, no new cases were reported. Here we discuss two cases of children from Huila and Caquetá departments who developed the disease. Both cases required long in-patient care and multiple surgical interventions. The diagnosis was achieved by histopathological observation of parasitic elements inside the mesenteric arteries. One of the children died while the other fully recovered. We discuss the epidemiology, pathogenic cycle, clinical presentation, diagnosis, and prevention strategies of this disease paying particular attention to our patients' features and the Colombian context.

La angiostrongiloidiasis abdominal es una zoonosis parasitaria endémica en el continente americano. Su agente etiológico es el nematodo Angiostrongylus costaricensis, cuyos huéspedes definitivos son los roedores y, los intermediarios, los caracoles y las babosas, por lo que se adquiere al consumir vegetales contaminados con larvas en estadio 3. La presentación clínica es muy variada y, dada su rareza, su diagnóstico es un desafío. En Colombia el primer caso se reportó en 1979 y, desde entonces hasta 1998, se han informado cinco casos más, aunque en las últimas dos décadas no se había reportado ningún caso. Se describen aquí dos casos de angiostrongiloidiasis en niños provenientes de los departamentos de Huila y Caquetá que requirieron una larga hospitalización y múltiples intervenciones quirúrgicas. El diagnóstico se logró al observar en los especímenes quirúrgicos larvas and huevos cuya morfología sugería una infección por nematodos; uno de los pacientes murió y el otro se recuperó satisfactoriamente. Se discuten la epidemiología, la patogenia, la presentación clínica, el diagnóstico y las estrategias de prevención de esta parasitosis, con énfasis en las características particulares de los casos descritos y en el contexto colombiano.

Particularidades de la meningoencefalitis eosinofílica en la era del caracol gigante africano / Particular features of eosinophilic meningoencephalitis in the age of the giant African snail

Introducción: la meningoencefalitis eosinofílica es una enfermedad inflamatoria infecciosa reportada en Cuba desde la década de los ochenta del siglo pasado y actualmente extendida al continente americano. Es producida por el parásito Angiostrongylus cantonensis. Objetivo: determinar si existen diferencias entre los pacientes que sufrieron meningoencefalitis eosinofílica antes y después de la introducción del caracol gigante africano. Métodos: se estudiaron un total de 19 muestras de líquido cefalorraquídeo y suero tomadas simultáneamente a cada paciente diagnosticados con meningoencefalitis eosinofílica, perteneciente a la seroraquioteca del Laboratorio Central del Líquido Cefalorraquídeo (LABCEL). Para la determinación de las proteínas albúmina e IgG se empleó como método de laboratorio la inmunodifusión radial. Resultados: el 14 por ciento de los pacientes fueron adultos antes de la aparición de este molusco, en contraste con el momento actual, donde el 50 por ciento son pacientes con edades superiores a 18 años. El porcentaje de síntesis de IgG intratecal media fue mayor en los enfermos actuales, aunque no de manera significativa. Conclusiones: . existen diferencias antes y después de la aparición del caracol gigante africano dado por la respuesta de síntesis intratecal en los pacientes asociados con el molusco que denota una mayor agresividad del parásito. La edad promedio mayor de los enfermos confirma que estos son los que más manipulan y dispersan el molusco(AU)

Introduction: eosinophilic meningoencephalitis is an infectious inflammatory disease reported in Cuba since the 1980s and currently extended to the American continent. This condition is caused by the parasite Angiostrongylus cantonensis. Objective: determine whether there are differences between the patients suffering from eosinophilic meningoencephalitis before and after the introduction of the giant African snail. Methods: a study was conducted of a total 19 cerebrospinal fluid and serum samples taken simultaneously from each of the patients diagnosed with eosinophilic meningoencephalitis and kept at the sample collection of the Central Cerebrospinal Fluid Laboratory (LABCEL). Radial immunodiffusion was the laboratory method used for determination of the proteins albumin and IgG. Results: of the patients studied, 14 percent were adults before the appearance of this mollusc, in contrast with the present moment, when 50 percent are patients aged over 18 years. The percentage of mean intrathecal synthesis of IgG was higher in the current sufferers, though not significantly. Conclusions: there are differences before and after the appearance of the giant African snail, given the intrathecal synthesis response of patients associated to the mollusc, which denotes greater aggressiveness by the parasite. The higher mean age of sufferers confirms that these are the ones who most often handle and disperse the mollusc(AU)

Frequency of gastrointestinal parasites in Creole horses from the Southern Rio Grande do Sul / Frequência de parasitos gastrintestinais em equinos da raça Crioula do sul do Rio Grande do Sul

A criação de equinos brasileira movimenta anualmente cerca de R$ 16,15 bilhões e impulsiona o setor econômico primário do país. O mercado de animais da raça Crioula vem se expandindo devido ao seu alto potencial zootécnico. No Rio Grande do Sul, a criação é tipicamente extensiva, aliada, muitas vezes, à alta densidade animal, o que favorece as constantes infecções por diversos parasitos. O objetivo deste estudo foi analisar retrospectivamente a frequência de parasitos gastrointestinais em cavalos da raça Crioula naturalmente infectados na região sul do Rio Grande do Sul. Foram utilizadas 585 amostras de fezes de equinos da raça Crioula, adultos, de ambos os sexos, recebidas de criatórios e centros reprodutivos localizados na região. Do total de amostras, 89,74% (525/585) foram positivas para algum helminto. Em 57,60% (337/585) das amostras, observaram-se somente ovos da família Strongylidae. Infecções por Parascaris spp. e Strongyloides sp., apresentaram frequências de 2,22% (13/585) e 1,53% (9/585), respectivamente. Assim, pode-se concluir que os parasitos mais frequentes na população equina da região sul do RS pertencem à família Strongylidae.(AU)

Factores de riesgo asociados a la meningitis eosinofílica causada por Angiostrongylus cantonensis (Nematoda: Metastrongylidae) en Chone, Ecuador / Risk factors associated to eosinophilic meningitis caused by Angiostrongylus cantonensis (Nematoda: Metastrongylidae) in Chone, Ecuador

Introducción: El parásito Angiostrongylus cantonensis es el principal causante de meningitis eosinofílica. En el 2008 se reportó el primer caso en Ecuador y un estudio reciente en la Ciudad de Chone, Manabí, Ecuador determinó una alta prevalencia del parásito en el caracol africano (Achatina fulica). Objetivo: Identificar los factores de riesgo asociados a la meningitis eosinofílica causada por A. cantonensis en la ciudad de Chone. Métodos: Se entrevistaron a 500 personas residentes en la ciudad de Chone y se revisaron las historias clínicas del Hospital General de Chone. Resultados: Se observó que la población de Chone, está expuesta a los siguientes factores de riesgos: consumo de caracoles crudos (7,40 por ciento), el frecuente contacto de las personas con el caracol africano en la casa (67 por ciento), como en el trabajo (51,20 por ciento), el contacto que tienen los niños con caracoles, al jugar en el jardín (76,80 por ciento) y la incorrecta limpieza de los alimentos (54,40 por ciento). En el entorno de la población, se evidenció la presencia del caracol africano cerca de las casas en el 35,20 por ciento y las ratas, en el 50,80 por ciento. Además, el 34 por ciento de la muestra de estudio presentó sintomatología relacionada con meningitis eosinofílica. Se encontró que existía asociación entre la presencia de vectores cerca de las casas y la cantidad de síntomas manifestados por las personas. Conclusiones: Se evidenció que la población de Chone, manifiesta hábitos que exponen a la gente de esta localidad a un alto riesgo de contagio de meningitis eosinofílica(AU)

Introduction: The parasite Angiostrongylus cantonensis is the main causative agent of eosinophilic meningitis. The first case in Ecuador was reported in 2008, and a recent study conducted in the city of Chone, Manabí, Ecuador, determined a high prevalence of the parasite in the African snail (Achatina fulica). Objective: Identify the risk factors associated to eosinophilic meningitis caused by A. cantonensis in the city of Chone. Methods: Interviews were conducted with 500 residents from the city of Chone and a review was carried out of the medical records at Chone General Hospital. Results: It was found that the population of Chone is exposed to the following risk factors: eating raw snails (7.40 percent), frequent contact between people and African snails both at home (67 percent) and at work (51.20 percent), contact between children and snails while playing in gardens (76.80 percent) and washing food items incorrectly (54.40 percent). In the areas surrounding people's households the African snail was found in 35.20 percent and rats in 50.80 percent . Additionally, 34 percent of the study sample had symptoms of eosinophilic meningitis. An association was found between the presence of vectors near the houses and the number of symptoms occurring in people. Conclusions: It was evident that the population of Chone has habits which expose them to a high risk for eosinophilic meningitis(AU)

El riesgo de parasitismo por Angiostrongylus cantonensis: una problemática reemergente en Cuba / The risk of parasitism by Angiostrongylus cantonensis: a re-emerging problem in Cuba / O risco de parasitismo devido a Angiostrongylus cantonensis: um problema reemergente em Cuba

RESUMEN Introducción: El riesgo de parasitismo por Angiostrongylus cantonensis transmitido por el caracol gigante africano es una problemática reemergente en Cuba. Objetivo: Sistematizar contenidos esenciales relacionados con el caracol gigante africano y el parásito Angiostrongylus cantonensis, el riesgo de meningitis eosinofílica y su manejo. Método: En el Hospital General Docente "Dr. Agostinho Neto", entre septiembre y diciembre de 2019 se hizo una revisión narrativa sobre el tema a través de una búsqueda en bases de datos electrónicas (Biblioteca Virtual en Salud): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Resultados: La información se estructuró en caracterización del caracol gigante africano y el riesgo de infección por Angiostrongylus cantonensis en Cuba, patogénesis y epidemiologia de la angiostrongylosis, y diagnóstico y tratamiento de la meningitis eosinofílica por este nemátodo. Se identificaron controversias sobre los criterios diagnósticos, el protocolo de actuación para su prevención y rehabilitación, y la carencia de un instrumento para estratificar el riesgo de esta infección. Conclusiones: Se sistematizaron contenidos esenciales sobre el caracol gigante africano, lo que posibilita la preparación de los profesionales para incrementar la percepción del riesgo de infección por Angiostrongylus cantonensis en la población.

ABSTRACT Introduction: The risk of parasitism by Angiostrongylus cantonensis transmitted by the giant African land snail, Lissachatina fulica is a re-emerging problem in Cuba. Objective: Systematize essential contents related to the giant African land snail and the parasite Angiostrongylus cantonensis, the risk of eosinophilic meningitis and its management. Method: A narrative review was made on the subject in the Teaching General Hospital "Dr. Agostinho Neto", between September and December 2019 through a search in electronic databases (Virtual Health Library): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Results: The information was structured in the characterization of the African giant land snail and the risk of infection by Angiostrongylus cantonensis in Cuba, pathogenesis and epidemiology of angiostrongylosis, and diagnosis and treatment of eosinophilic meningitis by this nematode. Controversies were identified regarding the diagnostic criteria, the protocol of action for its prevention and rehabilitation, and the lack of an instrument to stratify the risk of this infection. Conclusions: Essential content on the giant African land snail was systematized, enabling the preparation of professionals to increase the perception of the risk of infection by Angiostrongylus cantonensis in the population.

RESUMO Introdução: O risco de parasitismo causado por Angiostrongylus cantonensis transmitido pelo caracol gigante africano é um problema reemergente em Cuba. Objetivo: Sistematizar conteúdos essenciais relacionados ao caracol gigante africano e ao parasita Angiostrongylus cantonensis, ao risco de meningite eosinofílica e seu manejo. Método: No Hospital Geral de Ensino "Dr. Agostinho Neto", entre setembro e dezembro de 2019, foi realizada uma revisão narrativa do assunto através de uma busca em bases de dados eletrônicas (Biblioteca Virtual em Saúde): LILACS, PubMed, SciELO, RedALyC, Scopus, Cochrane. Resultados: As informações foram estruturadas na caracterização do caracol gigante africano e no risco de infecção por Angiostrongylus cantonensis em Cuba, patogênese e epidemiologia da angiostrongilose, diagnóstico e tratamento da meningite eosinofílica por esse nematóide. Foram identificadas controvérsias sobre os critérios diagnósticos, o protocolo de ação para sua prevenção e reabilitação e a falta de um instrumento para estratificar o risco dessa infecção. Conclusões: Foram sistematizados conteúdos essenciais sobre o caracol gigante africano, o que possibilita a preparação de profissionais para aumentar a percepção do risco de infecção por Angiostrongylus cantonensis na população.

Prevalence of Angiostrongylus cantonensis and Angiostrongylus costaricensis in Achatina fulica snails in the municipality of São Bernardo do Campo (SP, Brazil) / Prevalência de Angiostrongylus cantonensis e Angiostrongylus costaricensis em caramujos Achatina fulica na cidade de São Bernardo do Campo (SP, Brasil)

Angiostrongylus cantonensis e Angiostrongylus costaricensis são nematoides parasitas que residem em roedores e usam gastrópodes como hospedeiros intermediários. Achatina fulica, conhecida como caramujo-gigante-africano, é um hospedeiro intermediário que desempenha importante papel na dispersão de A. cantonensis e A. costaricensis, patógenos zoonóticos que causam, respectivamente, meningoencefalite eosinofílica e angiostrongilíase abdominal em humanos. O objetivo deste estudo foi o de coletar caramujos (A. fulica, n=90) de oito regiões diferentes (Rudge Ramos, Baeta Neves, Planalto, Demarchi, Dos Alvarengas, Batistini, Montanhão, Rio Grande) da cidade de São Bernardo do Campo (SP) para detecção molecular de A. cantonensis ou A. costaricensis pela técnica de reação em cadeia da polimerase (PCR). As amostras foram processadas em pools (n=25) segundo a região de coleta. Com exceção da região de Baeta Neves, as demais apresentaram caramujos infectados com larvas de nematoides. Seis regiões apresentaram amostras positivas para A. cantonensis. Quatorze (56%) das 25 amostras em pool apresentaram larvas de nematoides, e 52% delas (13/25) foram positivas para A. cantonensis. Nenhuma amostra positiva para A. costaricensis foi encontrada. A presença de A. fulica naturalmente infectada por A. cantonensis deve servir como um alerta para as autoridades de saúde pública sobre o risco potencial de infecção para humanos.(AU)

Abdominal angiostrongyliasis can be diagnosed with a immunochromatographic rapid test with recombinant galactin from Angiostrongylus cantonensis

Angiostrongylus costaricensis is the causative agent of abdominal angiostrongyliasis, a zoonotic infection that may produce severe eosinophilic enterocolitis or hepatitis in humans. Parasites are usually not released in stools and serology has an important role in diagnosis. Since cross-reactivity is demonstrated between A. costaricensis and another metastrongylid worm, A. cantonensis, we tested heterologous recombinant galectin as a probe in an immunochromatographic rapid diagnostic test (ICT-RDT) for detection of anti-A. costaricensis antibodies. Almost all (11/12) positive control sera from A. costaricensis infected patients were positive at ICT RDT. These are preliminary indications that r-galectin ICT-RDT is useful for diagnosing A. costaricensis infection.

Gastropods as intermediate hosts of Angiostrongylus spp. in the Americas: bioecological characteristics and geographical distribution

BACKGROUND Intermediate hosts are key organisms in maintaining parasite life cycles, because they can act as amplifiers in the transmission from natural reservoirs to humans. One of the most important groups of intermediate hosts for zoonotic nematode infections are gastropods,slugs and snails. These are essential organisms in the larval development of Angiostrongylus species. OBJECTIVES The objective of this paper is to review reports of Angiostrongylus spp. in naturally infected gastropods from the Americas, taking into account the diagnostic methods used in their identification, to be able to provide more accurate list of their intermediate hosts. We also discuss the factors that aid the dispersion of Angiostrongylus spp. in the Americas. METHODS This study reviews scientific publications and book sections on Angiostrongylus spp. in the Americas, including original works assessing larvae of Angiostrongylus in intermediate hosts. The eligible reports were classified accordingly to their geographical location, year of first record, and the larvae identification methodologies used. Digital repositories were used for the search. The bioecological characteristics of the main intermediate hosts are summarised. FINDINGS A total of 29 gastropod species that are naturally infected with Angiostrongylus spp. have been reported as intermediate hosts, 16 of which are land snails, two are freshwater snails, and 11 land slugs. MAIN CONCLUSIONS This study highlights the importance of integrative studies, analysing both the etiological agent and its transmission dynamic in the environment, the biological and ecological characteristics of the hosts, and the impact on host populations. It is necessary to increase interdisciplinary studies to determine the potential epidemiological health risk of angiostrongyliasis in the Americas, and thus be able to establish prevention, monitoring and contingency strategies in the region.

Infection by Angiostrongylus cantonensis in both humans and the snail Achatina (Lissachatina) fulica in the city of Macapá, in the Amazon Region of Brazil

In January and February 2019, a malacological survey was conducted in the area surrounding the residence of a 12-year-old child that had contracted cerebral angiostrongyliasis in the municipality of Macapá, capital of the Amapá State, northern Brazil. The serological examination was positive for Angiostrongylus cantonensis infection, the principal etiological agent of this parasitosis. A sample of 54 molluscs was artificially and individually digested for parasitological analysis, containing 38 specimens of Achatina fulica, nine specimens of Bulimulus tenuissimus and seven specimens of Sarasinula linguaeformis. A. fulica was the most abundant mollusc, and the only species infected with A. cantonensis, as well as presenting co-infections with other nematodes. This is the first report of cerebral angiostrongyliasis in the Amazon Region, and the first record of A. fulica infected with A. cantonensis in Amapá. These findings highlight the potential risks of human angiostrongyliasis, and the need to implement public health measures to control the spread of the disease.

Angiostrongylus (Parastrongylus) cantonensis en huéspedes intermediarios y definitivos en Ecuador, 2014-2017 / Angiostrongylus (Parastrongylus) cantonensis on intermediate and definitive hosts in Ecuador, 2014-2017

Resumen Introducción. Angiostrongylus cantonensis es un serio problema de salud pública y está ampliamente distribuido en el país. Cuando el parásito infecta a los caracoles terrestres Achatina, se enquista en su interior y puede infectar accidentalmente a las personas y otros mamíferos. Objetivo. Establecer la distribución geográfica de A. cantonensis en huéspedes intermediarios (Achatina fulica) y definitivos (Rattus spp.) en Ecuador entre el 2014 y el 2017. Materiales y métodos. Se recolectaron 2.908 ejemplares de A. fulica en 16 provincias utilizando el método de captura por unidad de esfuerzo durante 30 minutos. Se capturaron 211 ejemplares de las especies huéspedes, de los cuales 20 eran Rattus rattus y 191 R. norvegicus. Los ejemplares fueron transportados para su análisis al Instituto Nacional de Salud Pública e Investigación en Guayaquil. En los tejidos del caracol se identificaron y se contabilizaron larvas L3 que luego se inocularon en ratas de laboratorio para reproducir el ciclo de vida. En los roedores se disecaron los cerebros, los corazones, las arterias pulmonares y los pulmones, y se identificaron los parásitos por morfología taxonómica. Resultados. De los caracoles recolectados se encontraron 441 positivos para A. cantonensis (15,2 %) y un total de 6.166 larvas L3. En los ejemplaresRattus spp. capturados (211), 77 (36,5 %) estaban infectados con A. cantonensis, con un total de 220 parásitos (larvas L4-L5 y adultos). Conclusiones. Se constató la presencia de A. cantonensis en varias provincias, lo que confirma el carácter endémico de esta zoonosis en el territorio nacional. La presencia de Rattus spp., huésped definitivo del parásito, y de huéspedes intermediarios, indica el potencial zoonótico de esta infección parasitaria.

Abstract Introduction: Angiostrongylus cantonensis is a serious public health problem and is widely distributed in the country. When the parasite infects the snails, it becomes deeply embedded in their interior and accidentally, it can infect people and other mammals. Objective: To establish the geographical distribution of A. cantonensis intermediate hosts (Achatina fulica) and definitive hosts (Rattus spp.) in Ecuador from 2014 to 2017. Materials and methods: We collected 2,908 A. fulica specimens in 16 provinces using the capture method per unit of effort for 30 minutes. We captured 211 hosts of which 20 were Rattus rattus and 191 R. norvegicus. The specimens were transported to the Instituto Nacional de Salud Pública e Investigación in Guayaquil where the larvae L3 were identified and counted in the tissues of the snail, which were then inoculated in laboratory rats to reproduce the life cycle. In the rodents, the brains, hearts, lung arteries and lungs were dissected, and the parasites were identified by taxonomic morphology. Results: Of the snails harvested, 441 were positive for A. cantonensis (15.2%) and a total of 6,166 L3 larvae were found; 77 (36.5%) specimens of Rattus spp., were infected with A. cantonensis and a total of 220 parasites (L4-L5 larvae and adult worms) were collected. Conclusions: We confirmed the presence of A. cantonensis in several provinces, which ratifies the endemic nature of this zoonosis in the national territory. Rattus spp. specimens constitute the definitive hosts of the parasite, which together with the presence of intermediate hosts, indicates the zoonotic potential of this parasitic infection.

Estandarización de una prueba múltiple de reacción en cadena de la polimerasa en tiempo real para la identificación de Angiostrongylus cantonensis, A. costaricensis y A. vasorum / Standardization of a multiplex real-time PCR test for the identification of Angiostrongylus cantonensis, A. costaricensis and A. vasorum

Resumen Introducción. En el mundo, las angiostrongilosis de mayor impacto en salud humana y animal son ocasionadas por Angiostrongylus cantonensis, A. costaricensis y A. vasorum. En las personas, las formas clínicas son la meningitis eosinofílica y la angiostrongilosis abdominal, y, en los mamíferos cánidos, el daño cardiopulmonar. Se las consideran enfermedades emergentes debido a la propagación mundial del caracol africano Lissachatina fulica, un huésped intermediario de los parásitos. Los escasos métodos de identificación de Angiostrongylus spp. no son muy específicos ni sensibles y son costosos. Se necesita urgentemente una herramienta diagnóstica asequible, sensible y específica para el manejo de las angiostrongilosis humana y la animal. Objetivo. Desarrollar una prueba de PCR múltiple en tiempo real (qPCR) para identificar las tres especies patógenas de Angiostrongylus. Materiales y métodos. Mediante un análisis bioinformático se seleccionó una secuencia del genoma ITS-2 de Angiostrongylus para garantizar la especificidad del cebador y las sondas. El ADN de los parásitos adultos (control positivo) y de las larvas se extrajo con el estuche DNeasyBlood & Tissue®. Las reacciones de la PCR cuantitativa se ejecutaron en un termociclador Smartcycler Cepheid®, usando el estuche de mezcla maestra QuantiTect®. Como control negativo, se utilizó ADN humano, de otros parásitos y del caracol africano. Resultados. Los valores del ciclo umbral para los controles positivos de ADN fueron: 21 para Angiostrongylus cantonensis, 22 para A. costaricensis y 31 para A. vasorum. En los controles negativos, el ciclo umbral fue cero. La qPCR mostró una eficiencia de amplificación de 2 (100 %). Conclusiones. En el laboratorio se estandarizó una qPCR múltiple para tres especies clínicamente significativas de Angiostrongylus.

Abstract Introduction: Angiostrongyliasis is a disease caused by Angiostrongylus nematodes that is present worldwide. The infections with the highest impact on human and animal health are caused by A. cantonensis, A. costaricensis, and A. vasorum. Clinical forms of the disease in humans are eosinophilic meningitis and abdominal angiostrongyliasis, while the most common effect on dogs are cardiopulmonary damages. It is deemed as an emerging disease as the result of the global dissemination of the African snail Lissachatina fulica, an intermediary host of these parasites. The few diagnostic methods for Angiostrongylus spp. are unspecific, costly, and not very sensitive. It is urgent to develop a sensitive, specific and accessible diagnostic tool for the control of human and animal angiostrongyliasis. Objective: To develop a qPCR multiple test to identify the three pathogenic species of Angiostrongylus. Materials and methods: Through a bio-informatic analysis, we selected a sequence of the ITS-2 region of the Angiostrongylus genome to guarantee the specificity of primers and probes. We extracted DNA from adult parasites as positive control, and from larvae using the DNeasy Blood&Tissue® kit. Quantitative PCR reactions were conducted on a Smartcycler Cepheid® thermocycler using a master mix QuantiTect® kit. DNA from human beings, other parasites and the African snail was used as negative control. Results: The threshold cycle values for positive DNA controls were: 21 for Angiostrongylus cantonensis, 22 for A. costaricensis, and 31 for A. vasorum. In negative controls, the threshold cycle was zero. qPCR showed an amplification efficiency of 2 (100%). Conclusions: A multiple qPCR was standardized at the laboratory for three clinically significant species of Angiostrongylus.

Angiostrongylus spp. in the Americas: geographical and chronological distribution of definitive hosts versus disease reports

BACKGROUND Angiostrongyliasis is an infection caused by nematode worms of the genus Angiostrongylus. The adult worms inhabit the pulmonary arteries, heart, bronchioles of the lung, or mesenteric arteries of the caecum of definitive host. Of a total of 23 species of Angiostrongylus cited worldwide, only nine were registered in the American Continent. Two species, A. cantonensis and A. costaricensis, are considered zoonoses when the larvae accidentally parasitise man. OBJECTIVES In the present study, geographical and chronological distribution of definitive hosts of Angiostrongylus in the Americas is analysed in order to observe their relationship with disease reports. Moreover, the role of different definitive hosts as sentinels and dispersers of infective stages is discussed. METHODS The study area includes the Americas. First records of Angiostrongylus spp. in definitive or accidental hosts were compiled from the literature. Data were included in tables and figures and were matched to geographic information systems (GIS). FINDINGS Most geographical records of Angiostrongylus spp. both for definitive and accidental hosts belong to tropical areas, mainly equatorial zone. In relation to those species of human health importance, as A. cantonensis and A. costaricensis, most disease cases indicate a coincidence between the finding of definitive host and disease record. However, in some geographic site there are gaps between report of definitive host and disease record. In many areas, human populations have invaded natural environments and their socioeconomic conditions do not allow adequate medical care. MAIN CONCLUSIONS Consequently, many cases for angiostrongyliasis could have gone unreported or unrecognised throughout history and in the nowadays. Moreover, the population expansion and the climatic changes invite to make broader and more complete range of observation on the species that involve possible epidemiological risks. This paper integrates and shows the current distribution of Angiostrongylus species in America, being this information very relevant for establishing prevention, monitoring and contingency strategies in the region.

Intravitreal Angiostrongylus cantonensis: first case report in South America / Angiostrongylus cantonensis intravítreo: primeiro relato de caso na América do Sul

ABSTRACT This study reports the first case of intravitreal angiostrongyliasis in South America treated with posterior worm removal via pars plana vitrectomy. This was a retrospective, observational case study. Data from medical charts, wide-field digital imaging, ocular ultrasound, and visual evoked potential studies were reviewed. A 20-month-old boy presented with eosinophilic meningitis and right eye exotropia. Polymerase chain reaction analysis of the cerebrospinal fluid showed a positive result for Angiostrongylus cantonensis. Fundus examination revealed a pale optic disc, subretinal tracks, vitreous opacities, peripheral tractional retinal detachment, and a dead worm in the vitreous cavity. The patient underwent pars plana vitrectomy with worm removal. This case report illustrates the first case of intravitreal angiostrongyliasis in South America, possibly related to the uncontrolled spread of an exotic invasive species of snail.

RESUMO O objetivo deste estudo foi relatar o primeiro caso na América do Sul de angiostrongilíase intravítrea tratada com vitrectomia posterior via pars plana e remoção do verme. Este foi um relato de caso observacional. O prontuário médico, sistema de imagem digital de campo amplo, ultrassonografia ocular, e potenciais evocados visuais foram revistos. Um menino de 1 ano e 8 meses de idade manifestou meningite eosinofílica e exotropia olho direito. A análise de PCR do liquor foi positiva para Angiostrongylus cantonensis. O exame de fundo de olho revelou disco óptico pálido, faixas sub-retinianas, opacidades vítreas, descolamento de retina tracional periférico e um verme morto no vítreo. O paciente foi submetido a vitrectomia posterior via pars plana com a remoção do verme. Concluindo, este é o primeiro relato de caso de angiostrongilíase intravítrea na América do Sul, possivelmente relacionado com a disseminação de uma espécie de lesma exótica neste continente.

Características de angiostrongilosis addominal en Honduras / Characteristics of abdominal angiostrongylosis in Honduras

La angiostrongilosis abdominal es una patología descrita en Honduras desde 1972, se caracteriza por abdomen agudo con eosinofilia, existen pocos datos publicados en el país. Objetivo: caracterizar aspectos generales de la angiostrongilosis abdominal y la distribución de los casos estudiados en Honduras Material y métodos: se revisaron publicaciones nacionales e internacionales en las bases de datos: Biblioteca Virtual Salud Honduras, PubMed, SciELO, Google Académico; en inglés y español, con publicaciones desde 1972 hasta 2016, utilizando como descriptores: angiostrongilosis abdominal, angiostrongylus costaricensis en Honduras. Resultados: se encontraron casos publicados en Cortés, Francisco Morazán, Comayagua y El Paraíso, y babosas infectadas en los departamentos mencionados y Olancho. La mayoría de casos son niños, con abdomen agudo, fiebre, hiporexia e irritación peritoneal con leucocitosis y eosinofilia. Hay estudios de serología pero no disponibles en Honduras, el estándar de oro es la biopsia y el tratamiento es quirúrgico. Conclusión: en Honduras existen publicaciones de la existencia de esta zoonosis tanto en humanos como en roedores y los parásitos se han encontrado en babosa. Se considera una enfermedad parasitaria de difícil diagnóstico y prevención, por tanto, existen aspectos que no se conocen, por lo que, se deben realizar estudios serológicos en humanos, búsqueda de parásitos en babosas y roedores, para conocer las áreas afectadas y su distribución geográfica. Palabras clave: An

Susceptibility of Biomphalaria glabrata submitted to concomitant infection with Angiostrongylus costaricensis and Schistosoma mansoni / Suscetibilidade de Biomphalaria glabrata submetida à infecção concomitante com Angiostrongylus costaricensis e Schistosoma mansoni

Abstract The easy adaptation of Angiostrongylus costaricensis, nematode responsible for abdominal angiostrongyliasis to several species of terrestrial and freshwater molluscs and the differences observed in the interactions of trematodes with their intermediate hosts have induced us to study the concomitant infection of Biomphalaria glabrata with Schistosoma mansoni and A. costaricensis. Prior exposure of B. glabrata to A. costaricensis (with an interval of 48 hours), favored the development of S. mansoni, observing higher infection rate, increased release of cercariae and increased survival of molluscs, when compared to molluscs exposed only to S. mansoni. Prior exposure of B. glabrata to A. costaricensis and then to S. mansoni also enabled the development of A. costaricensis since in the ninth week of infection, higher amount of A. costaricensis L3 larvae was recovered (12 larvae / mollusc) while for molluscs exposed only to A. costaricensis, the number of larvae recovered was lower (8 larvae / mollusc). However, pre-exposure of B. glabrata to S. mansoni (with an interval of 24 hours), and subsequently exposure to A. costaricensis proved to be very harmful to B. glabrata, causing extensive mortality of molluscs, reduced pre-patent period to release cercariae and greater recovery of L3 A. costaricensis larvae.

Resumo A facilidade de adaptação do Angiostrongylus costaricensis, nematódeo responsável pela angiostrongiliase abdominal, a diversas espécies de moluscos terrestres e dulciaquícolas e as divergências observadas nas interações dos trematódeos com seus hospedeiros intermediários nos induziu a estudar a infecção concomitante de Biomphalaria glabrata com Schistosoma mansoni e A. costaricensis. A exposição prévia de B. glabrata ao A. costaricensis (com intervalo de 48 horas), favoreceu o desenvolvimento do S. mansoni observando-se elevação da taxa de infecção, maior liberação de cercárias e maior sobrevivência dos moluscos, quando comparado com os moluscos expostos somente ao S. mansoni. A exposição de B. glabrata previamente ao A. costaricensis e posteriormente ao S. mansoni também facilitou o desenvolvimento do A. costaricensis uma vez que na nona semana de infecção foi recuperada maior quantidade de larvas L3 de A. costaricensis, enquanto nos moluscos expostos somente ao A. costaricensis, o número de larvas recuperadas foi menor . Entretanto a pré-exposição de B. glabrata ao S. mansoni (com intervalo de 24 horas), e posteriormente a exposição ao A. costaricensis mostrou-se muito prejudicial à B. glabrata provocando grande mortalidade dos moluscos, redução do período pré-patente para liberação de cercárias e maior recuperação de larvas L3 de A. costaricensis.

Characterisation of the vascular pathology in Sigmodon hispidus (Rodentia: Cricetidae) following experimental infection with Angiostrongylus costaricensis (Nematoda: Metastrongylidae)

BACKGROUND Angiostrongylus costaricensis is a nematode that causes human abdominal angiostrongyliasis, a disease found mainly in Latin American countries and particularly in Brazil and Costa Rica. Its life cycle involves exploitation of both invertebrate and vertebrate hosts. Its natural reservoir is a vertebrate host, the cotton rat Sigmodon hispidus. The adult worms live in the ileo-colic branches of the upper mesenteric artery of S. hispidus, causing periarteritis. However, there is a lack of data on the development of vasculitis in the course of infection. OBJECTIVE To describe the histopathology of vascular lesions in S. hispidus following infection with A. costaricensis. METHODS Twenty-one S. hispidus were euthanised at 30, 50, 90 and 114 days post-infection (dpi), and guts and mesentery (including the cecal artery) were collected. Tissues were fixed in Carson’s Millonig formalin, histologically processed for paraffin embedding, sectioned with a rotary microtome, and stained with hematoxylin-eosin, resorcin-fuchsin, Perls, Sirius Red (pH = 10.2), Congo Red, and Azan trichrome for brightfield microscopy analysis. FINDINGS At 30 and 50 dpi, live eggs and larvae were present inside the vasa vasorum of the cecal artery, leading to eosinophil infiltrates throughout the vessel adventitia and promoting centripetal vasculitis with disruption of the elastic layers. Disease severity increased at 90 and 114 dpi, when many worms had died and the intensity of the vascular lesions was greatest, with intimal alterations, thrombus formation, iron accumulation, and atherosclerosis. CONCLUSION In addition to abdominal angiostrongyliasis, our data suggest that this model could be very useful for autoimune vasculitis and atherosclerosis studies.

Angiostrongylus cantonensis infection in molluscs in the municipality of São Gonçalo, a metropolitan area of Rio de Janeiro, Brazil: role of the invasive species Achatina fulica in parasite transmission dynamics

The aim of this study was to analyse the infection dynamics ofAngiostrongylus cantonensisin its possible intermediate hosts over two years in an urban area in the state of Rio de Janeiro where the presence ofA. cantonensis had been previously recorded in molluscs. Four of the seven mollusc species found in the study were exotic.Bradybaena similariswas the most abundant, followed byAchatina fulica, Streptaxissp., Subulina octona, Bulimulus tenuissimus, Sarasinula linguaeformisand Leptinaria unilamellata. Only A. fulicaand B. similariswere parasitised by A. cantonensis and both presented co-infection with other helminths. The prevalence of A. cantonensisin A. fulicawas more than 50% throughout the study. There was an inverse correlation between the population size ofA. fulicaand the prevalence of A. cantonensisand abundance of the latter was negatively related to rainfall. The overall prevalence of A. cantonensisin B. similariswas 24.6%. A. fulicawas the most important intermediary host of A. cantonensisin the studied area andB. similariswas secondary in importance for A. cantonensistransmission dynamics.

A Case of Ocular Angiostrongyliasis with Molecular Identification of the Species in Vietnam

A 23-year-old female residing in a village of Cao Bang Province, North Vietnam, visited the Hospital of Hanoi Medical University in July 2013. She felt dim eyes and a bulge-sticking pain in her left eye for some days before visiting the hospital. In the hospital, a clinical examination, an eye endoscopy, and an operation were carried out. A nematode specimen was collected from the eye of this patient. The body of this worm was thin and long and measured 22.0x0.3 mm. It was morphologically suggested as an immature female worm of Angiostrongylus cantonensis. By a molecular method using 18S rRNA gene, this nematode was confirmed as A. cantonensis. This is the first molecular study for identification of A. cantonensis in Vietnam.