Detection of antigens in paraffin sections of worms and snails infected by Angiostrongylus cantonensis by immunofluorescence reaction / Detecção de antígenos em cortes parafinados de vermes e caramujos infectados por Angiostrongylus cantonensis pela reação de imunofluorescência

The Indirect Immunofluorescence Assay (IFA) was used to identify stage-specific antigenic structures in paraffin sections of female larvae and worms and snails tissues, infected with third stage larvae of Angiostrongylus cantonensis. Sera from eosinophilic meningitis cases were used to assess reactivity. Non-reactive sera from patients with other parasitic diseases and from individuals without other etiologies were used as controls for cross-reactivity. Larvae and worms showed high reactivity to IgG antibodies. IgM antibodies reacted with low intensity only to larvae. Fluorescent reactions were observed in the cuticles and internal structures on worms sections, with a marked reaction in the uterus content. In the snail tissues, the larvae were found exclusively inside the granulomas, with fluorescent markings in the cuticles of the larvae and inside the granulomatous tissues. This fluorescent pattern suggests the presence of excretory/secretory antigens distributed throughout the granulomas. Expressive cross-reactivity occurred in sera from patients with other parasitic diseases, especially strongyloidiasis. The use of IFA applied to paraffin sections to identify structures with antigenic potential and the study of new serological markers, can contribute to the improvement of the laboratory diagnosis of eosinophilic meningitis. (AU)

A Reação de Imunofluorescência Indireta (RIFI) foi utilizada para localizar antígenos em estruturas estágio-específicas em cortes parafinados de vermes fêmeas e em tecidos de caramujos do Gênero Biomphalaria infectados com larvas de terceiro estágio de Angiostrongylus cantonensis. Soros de casos confirmados de meningite eosinfílica foram usados para avaliação da reatividade. Soros não reagentes de casos suspeitos; de pacientes com outras parasitoses e de indivíduos sem outras etiologias foram utilizados como controle de reatividade cruzada. Anticorpos da classe IgG foram reativos para antígenos presentes nos dois estágios e, anticorpos IgM somente para o estágio larvário. Nos cortes de vermes, as marcações fluorescentes foram assinaladas nas cutículas e estruturas internas, com acentuada marcação para os conteúdos uterinos. Nos tecidos dos caramujos as larvas foram encontradas exclusivamente no interior dos granulomas, com marcações fluorescentes nas cutículas das larvas e no interior dos tecidos granulomatosos. O padrão de fluorescência no granuloma sugere a marcação de antígenos excretores/secretores. Reatividade cruzada mais expressiva ocorreu com anticorpos presentes em soros de pacientes com outras parasitoses, com destaque para estrongiloidíase. A RIFI em cortes parafinados abre novas perspectivas para identificação de antígenos e de marcadores sorológicos, que possam ser aplicados no aprimoramento do diagnóstico laboratorial da meningite eosinofílica. (AU)

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.

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.

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.

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.

Comprehensive Review of Ocular Angiostrongyliasis with Special Reference to Optic Neuritis

Angiostrongyliasis, caused by Angiostrongylus cantonensis infection, is a food-borne parasitic disease. Its larvae evoke eosinophilic inflammation in the central nervous system, but can also cause pathological changes in the eyes. Among ocular angiostrongyliasis cases, the incidence of optic neuritis is low and only few sporadic reports exist. Some patients with optic neuritis developed obvious hypopsia or even vision loss, which would seriously influence the quality of life of patients. Prompt treatment of optic neuritis caused by A. cantonensis is the key factor for minimizing the incidence of serious complications of this disease. In this review, we first provide a comprehensive overview of ocular angiostrongyliasis, and then focus on the clinical features of optic neuritis caused by A. cantonensis.

An indirect immunofluorescence antibody test employing whole eggs as the antigen for the diagnosis of abdominal angiostrongyliasis

Abdominal angiostrongyliasis is a potentially fatal zoonotic disease with a broad geographical distribution throughout Central and South America. This study assessed the performance of Angiostrongylus costaricensis eggs as the antigen in an indirect immunofluorescence assay for the determination of parasite-specific IgG and IgG1 antibodies. For prevalence studies, an IgG antibody titre > 16 was identified as the diagnostic threshold with the best performance, providing 93.7 percent sensitivity and 84.6 percent specificity. Cross reactivity was evaluated with 65 additional samples from patients with other known parasitic infections. Cross reactivity was observed only in samples from individuals infected with Strongyloides stercoralis. For clinical diagnosis, we recommend the determination of IgG only as a screening test. IgG1 determination may be used to increase the specificity of the results for patients with a positive screening test.

Impactos socioeconômicos de uma doença emergente / Socioeconomic impacts of an emerging disease

Em 1995, ocorreu uma praga de lesmas identificadas como Sarasinula linguaeformis (Semper, 1885), que prejudicou as lavouras situadas entre os municípios de Nova Itaberaba e Planalto Alegre, no oeste do estado de Santa Catarina, Brasil. Neste artigo analisamos os impactos socioeconômicos provocados pela ocorrência desta praga, que sinalizou a possibilidade, posteriormente verificada, da emergência de casos de angiostrongilíase abdominal. Os dados foram coletados através de observação do estudo soroepidemiológico realizado na região durante o período de agosto de 2000 a agosto de 2001 pela equipe de pesquisa da Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), entrevistas com técnicos da Empresa de Pesquisa Agropecuária e Extensão Rural de Santa Catarina S.A. (Epagri-SC) e com cinquenta famílias locais. Para avaliar esses impactos, elaboramos algumas categorias de análise, como: prejuízo econômico; medidas preventivas e mudança de hábitos; estigma e preconceito, que surgiram à luz das narrativas dos sujeitos entrevistados. Ficou evidente a necessidade de se procederem a estudos sobre os aspectos sociológicos dos problemas epidemiológicos, além dos estritamente médico-sanitários.

In 1955 a population explosion of the slug Sarasinula linguaeformis (Semper, 1885) damaged crop plantations in the municipalities of Nova Itaberaba and Planalto Alegre, western of Santa Catarina State, Brazil. In this article we analyze the socioeconomic impacts caused by the emergence of this plague, which raised the suspicion, eventually confirmed, of the occurrence of human cases of abdominal angyostrongiliasis. The data were collected through the observation of a serological study during the period of August 2000 to August 2001 by a research team from the Catholic University of Rio Grande do Sul (PUCRS) together with field technicians from the Empresa de Pesquisa Agropecuária e Extensão Rural de Santa Catarina S.A. (Epagri-SC), and with the members of 50 local families. In order to analyze these impacts, we elaborated several analytical categories as economic losses; preventive measures, habit change and social prejudice, that emerged from the narratives of the residents interviewed. It became evident the need for sociological analyses of epidemiological problems, in addition to strictly medico-sanitary considerations.

Worms, slugs and humans: the medical and popular construction of an emerging infectious disease / Vermes, caramujos e humanos: as construções médica e popular de uma doença infecciosa emergente

The identification of the worm Angiostrongylus costaricensis parasitizing land snails and humans in Southern Brazil suggests under-diagnosis and under-notification of patients with abdominal angiostrongyliasis. This article analyzes how the concept of abdominal angiostrongyliasis was constructed in different ways in Costa Rica and Brazil and how these changes affected the understanding of its clinical and epidemiological diagnosis. The research shows that abdominal angiostrongyliasis is, de facto, a sociocultural construct, although the parasites and vectors are real. The analisys also shows the importance of an interdisciplinary approach for understanding disease.

A identificação do verme Angiostrongylus costaricensis em humanos e caramujos, no sul do Brasil, sugere a ocorrência de subdiagnóstico e subnotificação de pacientes com angiostrongilíase abdominal. O artigo analisa as diferentes construções do conceito de angiostrongilíase abdominal na Costa Rica e no Brasil e a influência dessas variações em seu diagnóstico clínico e epidemiológico. Demonstra que a angiostrongilíase abdominal é, de facto, um constructo sociocultural, embora parasitas e vetores sejam reais. Também destaca a importância da abordagem interdisciplinar para a compreensão da doença.

Use of heterologous antigens for the immunodiagnosis of abdominal angiostrongyliasis by an enzyme-linked immunosorbent assay

Angiostrongylus costaricensis has a broad geographic distribution spanning from North to South America and the infections of vertebrates with this nematode can result in abdominal complications. Human infections are diagnosed by histological or serological methods because the isolation of larvae from feces is not feasible, as most parasites become trapped in intestinal tissues due to intense eosinophilic inflammation. Because A. costaricensis is difficult to maintain in the laboratory, an immunodiagnostic IgG enzyme-linked immunosorbent assay (ELISA) using antigens from the congeneric Angiostrongylus cantonensis species was evaluated against a panel of serum samples from patients who were histologically diagnosed with A. costaricensis infections. Sera from uninfected individuals and individuals infected with other parasites were used as controls. The sensitivity and specificity of the assay were estimated at 88.4 percent and 78.7 percent, respectively. Because the use of purified or cloned antigens has not been established as a reliable diagnostic tool, the use of heterologous antigens may provide a viable alternative for the development of an ELISA-based immunodetection system for the diagnosis of abdominal angiostrongyliasis.

Peripheral eosinophilia as an indicator of meningitic angiostrongyliasis in exposed individuals

The diagnosis of meningitic angiostrongyliasis (MA) is based on clinical criteria. A lumbar puncture is used as a diagnostic tool, but it is an invasive procedure. The blood eosinophil levels are also assessed and used in the diagnosis of this disease. We enrolled 47 patients with serologically proven MA and 131 controls with intestinal parasite infections. An absolute eosinophil count model was found to be the best marker for MA. An eosinophil count of more than 798 cells led to sensitivity, specificity, positive predictive and negative predictive values of 76.6 percent, 80.2 percent, 58.1 percent and 90.5 percent, respectively. These data support the use of testing for high blood eosinophil levels as a diagnostic tool for MA in individuals that are at risk for this disease.

Betametasona e extrato aquoso de Arctium lappa no tratamento da angiostrongilíase / Betamethasone and aqueous extract of Arctium lappa for treating angiostrongyliasis

Angiostrongylus costaricensis é um parasita que causa angiostrongilíase abdominal em humanos, seu tratamento inclui o uso de antiinflamatórios apesar da falta de estudos que justifiquem esta conduta. O objetivo deste artigo é avaliar o efeito da betametasona e da Arctium lappa na evolução de lesões intestinais induzidas pelo parasita. Utilizou-se camundongos Swiss, machos, adultos, distribuídos em 4 grupos: infectados tratados com betametasona; com Arctium lappa; não tratados e grupo controle. Os tratamentos iniciaram no 15º dia de infecção e permaneceram por 15 dias. Infiltrado eosinofílico e granuloma foram avaliados (1-leve; 2-moderado; 3-severo). A betametasona permitiu a evolução das lesões para formas mais graves, enquanto o extrato não interferiu na progressão da patologia. As substâncias empregadas não mostraram eficácia na proteção das lesões induzidas pelo Angiostrongylus costaricensis em camundongos. Estes achados desmotivam o uso de betametasona e Arctium lappa em humanos acometidos por angiostrongilíase abdominal.

Angiostrongylus costaricensis is a parasite that causes abdominal angiostrongyliasis in humans. The treatment for it includes the use of anti-inflammatory drugs, despite the lack of studies to justify this approach. The objective of this paper was to evaluate the effect of betamethasone and Arctium lappa on the evolution of intestinal lesions induced by this parasite. Adult male Swiss mice were used, distributed into four groups: infected and treated with betamethasone; infected and treated with Arctium lappa; infected and not treated; and control group. The treatments were started on the 15th day after infection and continued for 15 days. The presence of eosinophilic infiltration and granuloma was evaluated (1-mild; 2-moderate; 3-severe). Betamethasone allowed the lesions to evolve into more severe forms, while the extract did not interfere with disease progression. The substances applied were ineffective for protection against the lesions induced by Angiostrongylus costaricensis in mice. These findings discourage the use of betamethasone and Arctium lappa for humans affected by abdominal angiostrongyliasis.

Abdominal angiostrongyliasis: report of two cases with different clinical presentations / Angiostrongilíase abdominal: relato de dois casos com diferentes apresentações clínicas

Abdominal angiostrongyliasis is a sporadic infectious disease caused by the nematode Angiostrongylus costaricensis. It usually presents as acute abdomen, secondary to mesenteric ischemia, and pronounced eosinophilia. In some cases its course is insidious and transient, and the diagnosis is suspicious. The disease is confirmed by the detection of A. costaricensis elements in surgical specimen. The treatment is supportive, with avoidance of antihelminthic administration due to a possible erratic migration followed by worsening of the disease. We report two cases, both with intense eosinophilia and serum IgG-ELISA positive to A. costaricensis. The first case presented ileal perforation and was surgically treated. The second one showed hepatic nodules at ultrasound and was only symptomatically treated, evolving to an apparent protracted resolution. These two cases exemplify different clinical forms of the disease, one of them with liver involvement.

A angiostrongilíase abdominal é doença esporádica decorrente da infecção pelo nematódeo Angiostrongylus costaricensis. Costuma manifestar-se como abdome agudo secundário a isquemia mesentérica, além de marcada eosinofilia. Pode também apresentar-se de forma insidiosa e transitória, exigindo alta suspeita clínica para o diagnóstico. A doença é confirmada pela identificação de elementos do A. costaricensis em peças cirúrgicas. O tratamento é apenas de suporte, devendo-se evitar o uso de anti-helmínticos pela possibilidade de migração errática do verme com piora do quadro. Aqui foram apresentados dois casos, ambos com acentuada eosinofilia e ELISA-IgG sérico positivo para A. costaricencis. O primeiro caso cursou com perfuração ileal e foi tratado cirurgicamente. O segundo caso apresentou nódulos hepáticos ao ultrassom e foi tratado sintomaticamente, evoluindo para lenta resolução. Estes dois casos exemplificam diferentes formas de apresentação clínica da doença, uma delas com envolvimento hepático.

Individual serological follow-up of patients with suspected or confirmed abdominal angiostrongyliasis

Abdominal angiostrongyliasis (AA) is a zoonotic nematode infection caused by Angiostrongylus costaricensis, with widespread occurrence in the Americas. Although the human infection may be highly prevalent, morbidity is low in Southern Brazil. Confirmed diagnosis is based on finding parasitic structures in pathological examination of biopsies or surgical resections. Serology stands as an important diagnostic tool in the less severe courses of the infection. Our objective is to describe the follow up of humoral reactivity every 2-4 weeks up to one year, in six individuals with confirmed (C) and ten suspected (S) AA. Antibody (IgG) detection was performed by ELISA and resulted in gradually declining curves of reactivity in nine subjects (56 percent) (4C + 5S), that were consistently negative in only three of them (2C + 1S) after 221, 121 and 298 days. Three individuals (2C + 1S) presented with low persistent reacitivity, other two (1C + 1S) were serologically negative from the beginning, but also presenting a declining tendency. The study shows indications that abdominal angiostrongyliasis is usually not a persistent infection: although serological negativation may take many months, IgG reactivity is usually declining along time and serum samples pairing may add valuable information to the diagnostic workout.

Angiostrongiliasis: Reporte de un caso / Angiostrongiliasis: Report of a case

Se presenta el caso de un niño de 9 años, costarricense, con angioestrongiliasis, el cual consultó por dolor abdominal de un año de evolución. La anamnesis demostró deficiencias en hábitos de higiene. El examen físico revela dolor en fosa iliaca derecha sin masas palpables. El examen serológico demostró la existencia de una importante eosinofilia, los análisis coproparasitoscópicos fueron negativos, presentaba anemia leve. La prueba de laboratorio test de Morera fue positiva.

The following is the case of a 9 yearoldCosta Rican male child withangiostrongyliasis, who consulteddue to an abdominal pain with yearof evolution. The medical history ofthe child demonstrated deficienciesin his hygiene habits. The physicalexamination revealed pain in theright iliac fossa without any palpablemass. The serological test showed theexistence of a relevant eosinophilia; the fecal stool analysis was negative.Yet, the boy suffered from anemia.The result of Morera Blood forangiostrongilos test was positive.