BACKGROUND Chagas disease, caused by Trypanosoma cruzi, affects nearly six million people worldwide. Various serological tests have been developed for its diagnosis. OBJECTIVE Examine the performance of a set of commercial immunological assays in relation to the geographical origin of the patient sample comparing four states of Brazil: Amazonas (AM), Mato Grosso do Sul (MS), Minas Gerais (MG) and Piauí (PI). METHODS Seven immunoassays were employed to detect anti-T. cruzi IgG antibodies in 379 patient samples that had been previously diagnosed using the two-step protocol required by the Brazilian Ministry of Health. FINDINGS A significant variation in the percent reactive was calculated for the samples from AM and MS, while the PI and MG showed a significant variation in the percent non-reactive. The average reactivity index was significantly higher for samples from the states of PI and MG states than AM and MS. MAIN CONCLUSIONS All tests presented a satisfactory performance overall. Yet, variations were observed that were associated to the region of origin of the samples. Our analyses suggest that future evaluations of immunoassays should include a sampling of sera from regions where the test will be applied in addition to the available International Biological Reference Standards.

BACKGROUND Chagas disease, resulting from Trypanosoma cruzi infections, continues to be a health concern mainly in Latin American countries where the parasite is endemic. The laboratory diagnosis of a chronic infection is determined through serological assays for antibodies against T. cruzi and several tests are available that differ in key components, formats and methodologies. To date, no single test meets the criteria of a gold standard. The situation is further complicated by the difficulties associated with performance comparisons between different immunoassays or methodologies executed at different times and geographical areas. OBJECTIVE To improve the diagnosis of Chagas disease, the WHO coordinated the development of two International Biological Reference Standards for antibodies against anti-T. cruzi: NIBSC 09/186 and NIBSC 09/188 that respectively represent geographical regions with the highest prevalence of TcII and TcI lineages of the parasite. METHODS The principle goal of this study was to verify the behavior of these standards when assayed by several commercially available serological tests that employ different methods to capture and detect human anti-T. cruzi antibodies. FINDINGS AND MAIN CONCLUSIONS The results reinforce the recommendation that these standards be considered for performance evaluations of commercialised immunoassays and should be an integral step in the development of new test components or assay paradigms.

Oral transmission of Chagas disease has been increasing in Latin American countries. The present study aimed to investigate changes in hepatic function, coagulation factor levels and parasite load in human acute Chagas disease (ACD) secondary to oral Trypanosoma cruzi transmission. Clinical and epidemiological findings of 102 infected individuals attended in the State of Pará from October 2013 to February 2016 were included. The most common symptoms were fever (98%), asthenia (83.3%), face and limb edema (80.4%), headache (74.5%) and myalgia (72.5%). The hepatic enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of 30 ACD patients were higher compared with controls, and this increase was independent of the treatment with benznidazole. Moreover, ACD individuals had higher plasma levels of activated protein C and lower levels of factor VII of the coagulation cascade. Patients with the highest parasite load had also the most increased transaminase levels. Also, ALT and AST were associated moderately (r = 0.429) and strongly (r = 0.595) with parasite load respectively. In conclusion, the present study raises the possibility that a disturbance in coagulation and hepatic function may be linked to human ACD.

Acute Chagas disease (ACD) has a distinct epidemiological profile in the Amazon Region, with cases and outbreaks of Trypanosoma cruzi infection being possibly related to the ingestion of contaminated food. Data on ACD in the state of Pará retrieved from 2000 to 2016 from the Brazilian Notifiable Diseases Information System (SINAN) were evaluated. During this period, 2,030 of the 16,807 reported cases were confirmed, with a higher incidence between the months of August and December, thus characterising a seasonal pattern of acute infection, and coinciding with the higher production of “açaí”, one fruit likely involved in the oral transmission of the disease. Evaluation of the absolute numbers of confirmed ACD cases secondary to oral infection suggests that infection through this route increased during the 2010-2016 period, differing from what was recorded in terms of vectorial or other infection routes. These findings point to the need of intensifying strategies to prevent or substantially reduce oral transmission.

Abstract: INTRODUCTION: Few studies have described the risk factors of intestinal parasitic infections in the Amazon. METHODS: A cross-sectional survey was performed in a City of the State of Amazonas (Brazil) to estimate the prevalence of intestinal parasites and determine the risk factors for helminth infections. RESULTS: Ascaris lumbricoides was the most prevalent parasite. The main risk factors determined were: not having a latrine for A. lumbricoides infection; being male and having earth or wood floors for hookworm infection; and being male for multiple helminth infections. CONCLUSIONS: We reported a high prevalence of intestinal parasites and determined some poverty-related risk factors.

INTRODUÇÃO: Foi realizado um estudo soroepidemiológico e clínico em 152 indivíduos residentes no município de Barcelos, Estado do Amazonas. Avaliou-se a soroprevalência da infecção chagásica e a morbidade da doença de Chagas. MÉTODOS: Os testes sorológicos foram a imunofluorescência indireta, ELISA convencional e recombinante e o Tesa-blot. Foram considerados soropositivos 38 pacientes, duvidosos 31 e soronegativos negativos 83. Os 38 casos soropositivos foram pareados com 38 controles soronegativos da mesma idade, sexo e submetidos à avaliação epidemiológica, clínica, eletro e ecocardiográfica, sendo que, 29 pares fizeram exame radiológico do esôfago. RESULTADOS: A soropositividade foi 19,9 vezes mais frequente nos trabalhadores do extrativismo em geral e 10,4 vezes mais frequente no extrativismo da piaçaba. Aplicou-se o teste de reconhecimento com o vetor local do gênero Rhodnius e 86,7% dos pacientes soropositivos o reconheceram, enquanto somente 34,2% dos soronegativos o fizeram. O ECG mostrou-se alterado em 36,8% nos soropositivos e em 21,5% nos soronegativos, enquanto o ecocardiograma mostrou alterações em 31,6% nos soropositivos e 18,4% nos soronegativos. Precordialgia e palpitações foram mais frequentes nos soropositivos. O estudo clínico do aparelho digestivo e radiológico do esôfago não mostrou alterações significativas. CONCLUSÕES. A doença de Chagas na região estudada pode ser considerada uma doença ocupacional.

INTRODUCTION: A seroepidemiological and clinical study was conducted on 152 autochthonous individuals living in the district of Barcelos, State of Amazonas, to evaluate the seroprevalence of Chagas infection and morbidity of Chagas disease. METHODS: The serological tests used were indirect immunofluorescence, conventional and recombinant ELISA and immunoblot (Tesa-blot). Thirty-eight patients were considered seropositive; 31 were considered serodoubtful; and 83 were considered seronegative. The 38 seropositive cases were paired with 38 seronegative controls of the same age and sex, and underwent epidemiological and clinical evaluations, electrocardiograms and echocardiograms. Twenty-nine pairs underwent radiological examinations of the esophagus. RESULTS: Seropositivity was 19.9 times more frequent among workers gathering plant materials from the forests and 10.4 times more frequent among piassaba gatherers. Eighty six point seven percent of the seropositive individuals recognized the genus Rhodnius as the local vector, while only 34.2% of the seronegative individuals recognized this. The EKG was abnormal in 36.8% of the seropositive individuals and in 21.5% of the seronegative individuals, while the echocardiogram showed abnormalities in 31.6% of the seropositive and 18.4% of the seronegative individuals. Precordialgia and palpitation were more frequent among the seropositive individuals. Clinical evaluation on the digestive system and X-ray on the esophagus did not show significant abnormalities. CONCLUSIONS: Chagas disease in the study region can be considered to be an occupational disease.

The risk that Chagas disease becomes established as a major endemic threat in Amazonia (the world's largest tropical biome, today inhabited by over 30 million people) relates to a complex set of interacting biological and social determinants. These include intense immigration from endemic areas (possibly introducing parasites and vectors), extensive landscape transformation with uncontrolled deforestation, and the great diversity of wild Trypanosoma cruzi reservoir hosts and vectors (25 species in nine genera), which maintain intense sylvatic transmission cycles. Invasion of houses by adventitious vectors (with infection rates > 60%) is common, and focal adaptation of native triatomines to artificial structures has been reported. Both acute (~ 500) and chronic cases of autochthonous human Chagas disease have been documented beyond doubt in the region. Continuous, low-intensity transmission seems to occur throughout the Amazon, and generates a hypoendemic pattern with seropositivity rates of ~ 1-3%. Discrete foci also exist in which transmission is more intense (e.g., in localized outbreaks probably linked to oral transmission) and prevalence rates higher. Early detection-treatment of acute cases is crucial for avoiding further dispersion of endemic transmission of Chagas disease in Amazonia, and will require the involvement of malaria control and primary health care systems. Comprehensive eco-epidemiological research, including prevalence surveys or the characterization of transmission dynamics in different ecological settings, is still needed. The International Initiative for Chagas Disesae Surveillance and Prevention in the Amazon provides the framework for building up the political and scientific cooperation networks required to confront the challenge of preventing Chagas disease in Amazonia.

The occurrence of autochthonous cases of Chagas disease in the Amazon region of Brazil over recent decades has motivated an intensification of studies in this area. Different species of triatomines have been identified, and ten of these have be proven to be carriers of the parasite Trypanosoma cruzi or " cruzi-like " parasites. Studies conducted in the municipalities of Santa Isabel do Rio Negro and Barcelos, located on the Upper and Middle of the Negro River, microregion of Negro River, state of Amazonas have confirmed not only that Rhodnius brethesi is present in the palm tree Leopoldinia piassaba, but also that this insect was recognized by palm fiber collectors. A morphological study of eyes, inter-ocular and inter-ocellar regions, antennae, buccula, labrum, rostrum, stridulatory sulcus and feet, including the apex of the tibia, spongy fossette and ctenidium was conducted by scanning electron microscopy. The buccula and the stridulatory sulcus presented notable differences in specimens of different genera and also of different species. These data make it possible to suggest that the details presented in these structures can be included as diagnostic characteristics to be used in new dichotomous keys, thereby contributing towards studies of taxonomy and systematics and furnishing backing for comparative analysis of specimens collected from different localities.

A new conglomerate family sample of 194 dwellings with 996 resident persons were studied in the town of Barcelos, State of Amazonas, in order to re-evaluate the risk of Chagas disease. During the survey the persons were interviewed and in this occasion we showed to them a collection of Panstrongylus, Rhodnius and Triatoma, asking if they recognized and eventually have been bitten by this kind of bugs. At this time we collected 500 ul of blood in microtainer® tubes from 886 interviewed persons who gave permission after informed consent. A screening test for T. cruzi antibodies based on agglutination of colored polymer particles, sensitized with three different synthetic peptides of T. cruzi (ID-PaGIA Chagas Test)®, showed 13.2% of sera positivity, but only 6.8% were confirmed by indirect immunofluorescence, and ELISA with purified T. cruzi antigens. Two hundred and six interviewed persons (20.7%) recognized the triatomines, as "piaçavas' lice" and 62 (30%) confirmed that have been bitten by the bugs, 25.8% of them had a positive serology for T. cruzi infection. Electrocardiographic alterations were shown in 9.3% of the seropositives and in 11.9% of the seronegative cases. This was considered not statistically significant.

Two of the major problems facing the Amazon - human migration from the other areas and uncontrolled deforestation - constitute the greatest risk for the establishment of endemic Chagas disease in this part of Brazil. At least 18 species of triatomines had been found in the Brazilian Amazon, 10 of them infected with Trypanosoma cruzi, associated with numerous wild reservoirs. With wide-range deforestation, wild animals will perforce be driven into other areas, with tendency for triatomines to become adapted to alternative food sources in peri and intradomicilies. Serological surveys and cross-sectional studies for Chagas disease, carried out in rural areas of the Rio Negro, in the Brazilian Amazon, showed a high level of seropositivity for T. cruzi antibodies. A strong correlation of seroreactivity with the contact of gatherers of piaçava fibers with wild triatomines could be evidenced.

Uma paciente com síndrome de imunodeficiência adquirida (SIDA) e doença de Chagas, com xenodiagnóstico positivo, estava em uso prolongado de cetoconazol com o objetivo de suprimir a parasitemia e prevenir a reativação da doença de Chagas. O cetoconazol foi suspenso inadvertidamente após 6 meses de uso. Um mês após, a paciente foi internada com febre, cefaléia, vômitos, taquicardia e hepatoesplenomegalia. Tanto o xenodiagnóstico como o exame de sangue a fresco demonstraram a presença de Trypanosoma cruzi. O tratamento com benzonidazol foi instituído, com supressão da parasitemia. A paciente desenvolveu concomitantemente uma provável neurotoxoplasmose, evoluindo para o óbito em septicemia. À necropsia, não foram encontrados parasitas.

A patient with AIDS and asymptomatic Chagas's disease and positive xenodiagnosis was taking ketoconazole in order to supress parasitemia and prevent reactivation of Chagas's disease. Ketoconazole was unplanned suspended after 6 months, and the patient was admitted with fever, headache, vomiting, tachycardia, postural hypotension, hepatosplenomegaly, and positive xenodiagnosis one month later. Treatment with benzonidazole was begun leading to supression of parasitemia. The patient had probabily a neurotoxoplasmosis associated and progressed to coma and death with sepsis. No parasite was found in autopsy.

Foram estudados 186 pares de indivíduos sorologicamente positivos e negativos para infecção chagásica, da mesma idade e sexo, do Sertão da Paraíba e 200 indivíduos também sorologicamente positivos nos municípios de Oeiras e Colônia do Piauí. Depois de confirmados por pelo menos dois outros testes sorológicos: imunofluorescência indireta quantitativa, ELISA, hemaglutinação ou fixação do complemento,foi feito o exame clínico, eletrocardiográfico e radiológico nos indivíduos selecionados para o estudo exenodiagnóstico, hemocultura e PCR em amostras representativas dos casos soropositivos. As manifestações clínicas predominantes entre os soropositivos em ambas as áreas foram palpitações, dispnéia aos esforços, disfagia, odinofagia, pirose e obstipação. As freqüências das alterações eletrocardiográficas sugestivas da doença de Chagas foram, respectivamente, na Paraíba e no Piauí: BAV = 3,8 % e 2 %, BRD III = 6,4 % e 7 %, BRD III + HBAE = 10,7 % e 10,5 % e extra-sístoles ventriculares complexas = 2,7 % e 3,% . O xenodiagnóstico foi positivo em 13 % dos casos soropositivos da Paraíba e em 34 % dos casos do Piauí, enquanto que o PCR foi positivo, respectivamente, em 44,6 e 59,5 %. A hemocultura realizada apenas no Piauí foi positiva em 25,7 % dos casos estudados. Foram realizados inquéritos triatomínicos em 132 domicílios e peridomicílios no Sertão da Paraíba e em 159 na Caatinga do Piauí, sendo capturados 16 exemplares de T. brasiliensis não infectados no peridomicílio na Paraíba e 750 exemplares no Piauí, dos quais 625 foram examinados: 49 de T. pseudomaculata não infectados com T. cruzi (19 no intradomicílio e 30 no peridomicílio) e 576 de T. brasiliensis (371 no intradomicílio e 205 no peridomicílio) entre os quais 32 (5,5% ) estavam infectados com T. cruzi (31 no intradomicílio e um no peridomicílio).

A clinical and electrocardiographic case control study was carried out with 186 pairs of persons with positive and negative serology for T. cruzi infection from the Sertão Paraíba and in 200 seropositive cases from the region of Caatinga in the State of Piauí, North-eastern Brazil. The predominant clinical manifestations in seropositive cases in both areas were: palpitations, dyspnea on effort, precordial pain, dysphagia, odynophagia, pyrosis and intestinal constipation. The EKG abnormalities rates suggestive of chronic chagasic cardiopathy were respectively in Paraíba and Piauí: AV block 3.8% and 2%, RBBB 111 6.4% and 7%, RBBB III + LAB 10.7% and 10.5%, and multifocal extrasystoles 2.7% and 3%. Xenodiagnosis in a sample of 54 seropositive individuais in the Sertão of Paraíba and in 120 in the Caatinga of Piauí was revealed 13% and 34 % positive; PCR tests in a sample of 47 seropositives in Paraíba and 101 in Piauí revealed positives in 44.6 % and 59.5% respectively. Blood culture in LIT media of 101 seropositive cases from the Caatinga of Piauí was positive for T. cruzi in 25.7%. A triatomine survey carried out in a sample of 132 domiciles and peridomiciles in the Sertão of Paraíba and in a sample of 159 in the Caatinga of Piauí showed the following results: In Paraíba, 16 specimens of T. brasiliensis, not infected with T. cruzi, were captured. In Piauí, 750 triatomines were captured, of these 625 were examined: 49 were T. pseudomaculata, not infected with T. cruzi (19 in peridomiciles and 30 in the domiciles), and 576 were T. brasiliensis (371 in the domiciles and 205 in the peridomiciles) and of this latter specie 32 (5.5%) were infected with T. cruzi (31 in the domiciles and one in the peridomicile).

Pelo menos dezoito espécies de triatomíneos foram encontradas na Amazônia brasileira, nove das quais infectadas com Trypanosoma cruzi ou semelhante ("cruzi-like"), associadas com numerosos reservatórios silvestres. A despeito do pequeno número de casos humanos da doença de Chagas descritos até agora na Amazônia brasileira, o risco que essa doença se torne endêmica é cada vez maior, pelas seguintes razões: a) desmatamentos e colonização descontrolados, alterando o balanço entre reservatórios e vetores; b) adaptação de reservatórios e vetores silvestres com T.cruzi ao peridomicílio, como única alternativa alimentar; c) migração de populações humanas infectadas com T.cruzi acompanhadas de reservatórios domésticos (cães e gatos) ou de vetores de suas regiões de origem na bagagem, já adaptados ao domicílio. Em resumo, o risco de que a doença de Chagas se torne endêmica na Amazônia brasileira está ligado à transposição do ciclo silvestre para o doméstico na própria área ou do ciclo doméstico já estabelecido em áreas endêmicas para a Amazônia.

At least eighteen species of triatominae have been found in the Brazilian Amazon, nine of them naturally infected with Trypanosoma cruzi or "cruzi-like" trypanosomes and associated with numerous wild reservoirs. Despite the small number of human cases of Chagas' disease described to date in the Brazilian Amazon the risk that the disease will become endemic in this area is increasing for the following reasons: a) uncontrolled deforestation and colonization altering the ecological balance between reservoir hosts and wild vectors; b) the adaptation of reservoir hosts of T.cruzi and wild vectors to peripheral and intradomiciliary areas, as the sole feeding alternative; c) migration of infected human population from endemic areas, accompanied by domestic reservoir hosts (dogs and cats) or accidentally carrying in their baggage vectors already adapted to the domestic habitat. In short, risks that Chagas' disease will become endemic to the Amazon appear to be linked to the transposition of the wild cycle to the domestic cycle in that area or to transfer of the domestic cycle from endemic areas to the Amazon.