Encapsulation of benznidazole in nanostructured lipid carriers and increased trypanocidal activity in a resistant Trypanosoma cruzi strain

Abstract Chagas disease is a neglected parasitic disease caused by Trypanosoma cruzi, whose treatment has remained unsatisfactory for over 50 years, given that it is limited to two drugs. Benznidazole (BZN) is an efficient antichagasic drug used as the first choice, although its poor water-solubility, irregular oral absorption, low efficacy in the chronic phase, and various associated adverse effects are limiting factors for treatment. Incorporating drugs with such characteristics into nanostructured lipid carriers (NLC) is a promising alternative to overcome these limiting obstacles, enhancing drug efficacy and bioavailability while reducing toxicity. Therefore, this study proposed NLC-BZN formulations in different compositions prepared by hot-melt homogenization followed by ultrasound, and the optimized formulation was characterized by FTIR, DRX, DSC, and thermogravimetry. Biological activities included in vitro membrane toxicity (red blood cells), fibroblast cell cytotoxicity, and trypanocidal activity against epimastigotes of the Colombian strain of T. cruzi. The optimized NLC-BZN had a small size (110 nm), negative zeta potential (-18.0 mV), and high encapsulation (1.64% of drug loading), as shown by infrared spectroscopy, X-ray diffraction, and thermal analysis. The NLC-BZN also promoted lower in vitro membrane toxicity (<3% hemolysis), and 50% cytotoxic concentration (CC50) for NLC-BZN in L929 fibroblast cells (110.7 µg/mL) was twice the value as the free BZN (51.3 µg/mL). Our findings showed that the NLC-BZN had higher trypanocidal activity than free BZN against the epimastigotes of the resistant Colombian strain, and this novel NLC-BZN formulation proved to be a promising tool in treating Chagas disease and considered suitable for oral and parenteral administration

Systematic review on the biology, ecology, genetic diversity and parasite transmission potential of Panstrongylus geniculatus (Latreille 1811) in Latin America

Panstrongylus geniculatus (Latreille, 1811) is the triatomine with the largest geographic distribution in Latin America. It has been reported in 18 countries from southern Mexico to northern Argentina, including the Caribbean islands. Although most reports indicate that P. geniculatus has wild habitats, this species has intrusive habits regarding human dwellings mainly located in intermediate deforested areas. It is attracted by artificial light from urban and rural buildings, raising the risk of transmission of Trypanosoma cruzi. Despite the wide body of published information on P. geniculatus, many knowledge gaps exist about its biology and epidemiological potential. For this reason, we analysed the literature for P. geniculatus in Scopus, PubMed, Scielo, Google Scholar and the BibTriv3.0 databases to update existing knowledge and provide better information on its geographic distribution, life cycle, genetic diversity, evidence of intrusion and domiciliation, vector-related circulating discrete taxonomic units, possible role in oral T. cruzi transmission, and the effect of climate change on its biology and epidemiology.

Encuesta entomológica y caracterización de las viviendas con el vector de la enfermedad de Chagas, en aldea Chuchuapa, Santa Rosa, Guatemala / Entomological survey and characterization of houses with the vector of Chagas disease, in Chuchuapa village, Santa Rosa, Guatemala

En Guatemala en el 2015, el Ministerio de Salud Pública y Asistencia Social (MSPAS) reportó nuevos casos de la enfermedad de Chagas en la región suroriental del país, siendo Santa Rosa uno de los departamentos endémicos en donde Triatoma dimidiata es el principal transmisor. Se estimó la infestación de T. dimidiata en la aldea Chuchuapa, municipio de Santa María Ixhuatán, de abril a junio de 2019, se calcularon los índices en-tomológicos y se caracterizaron intra y peridomiciliar 149 viviendas (79.67% de las viviendas habitadas). Los datos se obtuvieron mediante un cuestionario mixto, en entrevista cara a cara y evaluación de las viviendas, por un equipo profesional experimentado, mediante el método y protocolo estándar hombre-hora. Se capturaron 20 triatominos en 10 viviendas, con un índice de infestación de 6.71% (10/149), un índice de densidad de 13.42% (20/149), 19 triatominos intradomiciliares y uno peridomiciliar, con un índice de infección natural por Tr y pano-soma cruzi intradomiciliar de 26.3% (5/19), índice de infección natural por T. cr u z i peridomiciliar de 100.0% (1/1) e índice de infección natural por T. c r u z i en los triatominos capturados de 30.0 % (6/20). Las viviendas infestadas presentaron techo de lámina, pared de bajareque con grietas y sin repello, piso de tierra y afiches en la pared, acumulación de leña u otros materiales tanto intra como peridomiciliar, convivencia con animales domésticos y tenencia de animales de corral. Los resultados representan un alto riesgo para la transmisión de la enfermedad de Chagas entre los habitantes de la aldea.

In Guatemala in 2015, the Ministry of Public Health and Social Assistance reported new cases of Chagas disease in the southeastern region of the country, with Santa Rosa being one of the endemic departments where Tr ia-toma dimidiata is the main transmitter. The infestation of T. dimidiata was estimated in the Chuchuapa village, municipality of Santa María Ixhuatán, from April to June 2019, the entomological indices were calculated and 149 dwellings were characterized intradomiciliary and peridomiciliary (79.67% of the inhabited dwellings). The data were obtained through a mixed questionnaire, in a face-to-face interview and evaluation of the dwellings, by an experienced professional team, using the standard man-hour method and protocol. 20 triatomines were captured in 10 homes, with an infestation index of 6.71% (10/149), a density index of 13.42% (20/149), 19 intradomiciliary triatomines and one peridomiciliary, with a natural infection index by intradomiciliary Trypanosomacruzi of 26.3% (5/19), natural infection index by peridomiciliary T. c r u z i of 100.0% (1/1) and natural infection index by T. c r u z i in captured triatomines of 30.0% (6/20). The infested dwellings have a sheet roof, awattle and daub wall with cracks and no wall plaster, dirt and posters on the wall, accumulation of firewood and other materials in both intradomiciliary and peridomiciliary, living with domestic animals and keeping poultry animals. The results represent a high risk for the transmission of Chagas disease among the villager.

Application of WHO International Biological Reference Standards to evaluate commercial serological tests for chronic Chagas disease

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.

Síntesis de evidencia: Guía para el diagnóstico y el tratamiento de la enfermedad de Chagas / Synthesis of evidence: Guidance for the diagnosis and treatment of Chagas disease / Síntese de evidências: Guia de diagnóstico e tratamento da doença de Chagas

RESUMEN Introducción. La enfermedad de Chagas o tripanosomiasis americana es causada por el protozoo flagelado Trypanosoma cruzi, transmitido principalmente por insectos vectores (denominados popularmente en las diferentes zonas de la Región de las Américas "vinchucas", "pitos", "chinches", "chirimachas" o "kissing bugs"). El parásito es transmitido por vía transplacentaria, transfusiones y, menos frecuentemente, por vía oral o trasplante de órganos. La implementación de políticas públicas y el manejo de la condición clínica requiere ser fortalecido debido a las dificultades diagnósticas y terapéuticas que presenta esta parasitosis sistémica. Para contribuir con ello, la Organización Panamericana de la Salud (OPS) desarrolló una guía para el manejo de la tripanosomiasis americana. Objetivos. Sintetizar las recomendaciones incluidas en la Guía para el diagnóstico y el tratamiento de la enfermedad de Chagas, publicada por la Organización Panamericana de la Salud en 2018, con el fin de presentar el adecuado diagnóstico y tratamiento de la enfermedad de Chagas y abordar aspectos sobre su implementación. Métodos. Se llevó a cabo una síntesis de la guía y sus recomendaciones. Adicionalmente, se realizó una búsqueda sistemática en PubMed, Lilacs, Health Systems Evidence, Epistemonikos y literatura gris de estudios desarrollados en las Américas con el fin de identificar barreras, facilitadores y estrategias de implementación. Resultados. Se presentan 10 recomendaciones aplicables a pacientes adultos y pediátricos con sospecha de enfermedad de Chagas, exposición a T. cruzi y diagnóstico confirmado de enfermedad de Chagas aguda, crónica y congénita. Se identificaron reportes que abordaron aspectos de implementación. Conclusiones. Las recomendaciones buscan proveer estrategias para el diagnóstico y tratamiento oportunos de la enfermedad de Chagas, así como consideraciones para su implementación.(AU)

ABSTRACT Introduction. Chagas disease or American trypanosomiasis is caused by the flagellate protozoan Trypanosoma cruzi, transmitted mainly by insect vectors (popularly known in the different areas of the Region of the Americas as "kissing bugs", "pitos", "chinches", or "chirimachas" or "vinchuchas"). The parasite is transmitted via the placenta and in transfusions, and less frequently, orally or during organ transplantation. Because it was necessary to strengthen the implementation of public policies and the management of clinical conditions, given the diagnostic and therapeutic difficulties that this systemic parasitosis presents, the Pan American Health Organization (PAHO) developed guidance for the management of American trypanosomiasis. Objectives. Synthesize the recommendations included in the Guide for Diagnosis and Treatment of Chagas Disease, published (in Spanish only) by the Pan American Health Organization in 2018, in order to present the proper diagnostic methods and treatment for Chagas disease and to address aspects of its implementation. Methods. A summary was made of the guide and its recommendations. In addition, a systematic search was conducted in PubMed, Lilacs, Health Systems Evidence, Epistemonikos, and the gray literature for studies carried out in the Americas, in order to identify barriers, facilitators, and implementation strategies. Results. Ten recommendations are made. These are applicable to adult and pediatric patients with suspected Chagas disease, exposure to T. cruzi, or a confirmed diagnosis of acute, chronic, or congenital Chagas disease. Reports that addressed aspects of implementation were identified. Conclusions. The recommendations seek to provide strategies for the timely diagnosis and treatment of Chagas disease, as well as considerations for the implementation of such strategies.(AU)

RESUMO Introdução. A doença de Chagas (ou tripanossomíase americana) é uma infecção causada pelo protozoário flagelado Trypanosoma cruzi, comumente transmitido por insetos vetores (popularmente conhecidos na Região das Américas como "barbeiro", "chupão", "vinchuca", "pito", "chinche", "chirimacha" ou "kissing bug"). O parasita é transmitido por via transplacentária, transfusão de sangue e, menos comumente, por via oral ou transplante de órgãos de doadores infectados. A implementação de políticas públicas e o manejo clínico devem ser reforçados diante das dificuldades de diagnóstico e terapêutica para esta doença parasitária sistêmica. Em apoio a este propósito, a Organização Pan-Americana da Saúde (OPAS) elaborou um guia para o manejo da doença de Chagas. Objetivos. Sintetizar as recomendações contidas no Guía para el diagnóstico y el tratamiento de la enfermedad de Chagas (guia de diagnóstico e tratamento da doença de Chagas, disponível apenas em espanhol), publicado pela Organização Pan-Americana da Saúde em 2018, com o propósito de apresentar o diagnóstico e o tratamento adequados da doença de Chagas e abordar aspectos relativos à implementação. Métodos. Foi realizada uma síntese do guia e das recomendações apresentadas. Foi também conduzida uma busca sistemática nas bases de dados PubMed, LILACS, Health Systems Evidence e Epistemonikos e na literatura cinzenta de estudos realizados na Região das Américas visando identificar barreiras, facilitadores e estratégias de implementação. Resultados. O guia oferece 10 recomendações para pacientes adultos e pediátricos com suspeita de doença de Chagas, exposição ao T. cruzi e diagnóstico confirmado de doença de Chagas aguda, crônica e congênita e discute aspectos relativos à implementação. Conclusões. As recomendações visam fornecer estratégias para o diagnóstico e o tratamento de forma oportuna da doença de Chagas e são feitas considerações para a implementação destas estratégias.(AU)

Estudio histopatológico de tejido cardiaco de roedores infectados con Trypanosoma cruzi capturados en barrios suburbanos de Mérida, México / Histopathological study in cardiac tissue of rodents infected with Trypanosoma cruzi, captured in suburbs of Mérida, México

Resumen Introducción. Trypanosoma cruzi es el agente causal de la tripanosomiasis americana, enfermedad endémica en México. Los roedores Mus musculus y Rattus rattus son reservorios del parásito, el cual invade las fibras cardiacas y desarrolla nidos parasitarios produciendo diversas lesiones. Los estudios histopatológicos en roedores naturalmente infectados son escasos. Objetivo. Describir los tipos y las frecuencias de las lesiones microscópicas en muestras de tejido cardiaco de M. musculus y R. rattus infectados con T. cruzi capturados en Mérida, México. Materiales y métodos. Los roedores se capturaron en los barrios suburbanos de Mérida. Se extrajo el tejido cardiaco y se procesó por la técnica de inclusión en parafina y tinción con hematoxilina y eosina. Su examen se hizo con un microscópico convencional y se determinaron todas las lesiones y su grado de afección. Resultados. Se trabajaron ocho muestras de tejido de M. musculus y siete de R. rattus. Se encontraron nidos parasitarios en siete del total de las muestras: en 3 de las 8 de M. musculus y en 4 de las 7 de R. rattus. Se observaron infiltrados inflamatorios en todas las muestras. Otras lesiones fueron la degeneración de las fibras cardiacas (8/15), la congestión de los vasos sanguíneos (6/15) y la necrosis (5/15). Discusión. Las lesiones observadas están descritas en los modelos animales experimentales y en los humanos con tripanosomiasis americana. Los infiltrados inflamatorios se han descrito como la lesión más significativa en los humanos y en los reservorios en la etapa crónica de la enfermedad. Conclusión. Las lesiones observadas están asociadas con la infección con T. cruzi, lo cual confirma que los roedores estudiados son reservorios de este parásito.

Abstract Introduction: Trypanosoma cruzi is the causal agent of the American trypanosomiasis, an endemic disease in México. The commensal rodents Mus musculus and Rattus rattus are reservoirs of this parasite, which invades cardiac fibers and develops parasite nests causing various lesions. Histopathological studies in naturally infected rodents are scarce. Objective: To describe the types and frequencies of microscopic lesions in cardiac tissue of M. musculus and R. rattus infected with T. cruzi captured in Mérida, México. Materials and methods: The rodents were captured in suburban environments of Mérida. Cardiac tissue was extracted and processed by the paraffin inclusion technique and hematoxylin and eosin stained. The observation was made with a conventional microscope and all the lesions, as well as their degree, were identified. Results: Eight tissue samples of M. musculus and seven of R. rattus were studied. Parasite nests were found in 7/15, specifically 3/8 in M. musculus and 4/7 in R. rattus. The inflammatory infiltrate was the most frequent lesion. Other lesions were: Degeneration of cardiac fibers (8/15), congestion of blood vessels (6/15), and necrosis (5/15). Discussion: The lesions we observed have been described in experimental animal models and in humans with American trypanosomiasis. The inflammatory infiltrate has been identified as the most significant lesion in humans and reservoirs in the chronic stage of the disease. Conclusion: The lesions we described are associated with T. cruzi infection, which confirms that the rodents studied are reservoirs of this parasite.

Detection of Trypanosoma cruzi strains circulating in Córdoba department (Colombia) isolated from triatomines (Hemiptera: Reduviidae) collected by the community / Detección y caracterización molecular de cepas de Trypanosoma cruzi aisladas de triatominos recolectados por la comunidad en el departamento de Córdoba, Colombia

Abstract Introduction: From 2011 to 2016, 24 cases of Chagas disease were reported in Córdoba according to the national public health surveillance system (Sistema Nacional de Vigilancia en Salud Pública, Sivigila), but the information regarding Trypanosoma cruzi circulating strains and infection rates are unknown. Objectives: To establish the triatomine species with which people come in contact and recognize as Chagas disease vectors, as well as to assess the infection with trypanosomes and make an exploratory approach to host feeding preferences with the participation of the local community. Materials and methods: Triatomines sampling was conducted in 12 municipalities between 2011 and 2016; T. cruzi infection was established by k-PCR, SAT-PCR, while strain genotyping was done by mini-exon and SL-IR (spliced-leader intergenic region) sequence characterization. We also screened for blood sources. Results: Local community members collected the majority of triatomines and we identified three species: Rhodnius pallescens, Panstrongylus geniculatus, and Eratyrus cuspidatus. The overall T. cruzi infection rate in collected triatomines was 66.6% and we detected the TcIDOM and TcI sylvatic strains. Community-based insect collection allowed reporting the presence of P. geniculatus in two new disperse rural settlements, T. cruzi infection of P. geniculatus in Córdoba, and the first report of triatomines infected with T. cruzi in Montería municipality. Conclusions: These results revealed the presence of triatomines infected with T. cruzi inside dwellings in five municipalities of Córdoba. The dominant circulating T. cruzi strain was TcIDOM, a genotype associated with human Chagas disease and cardiomyopathies in Colombia. Our results highlight the importance of local community participation in entomological surveillance tasks.

Resumen Introducción. Entre el 2011 y el 2016, se reportaron 24 casos de enfermedad de Chagas en Córdoba, según el Sistema Nacional de Vigilancia en Salud Pública (Sivigila), pero la información sobre las unidades discretas de tipificación de Trypanosoma cruzi circulantes y las tasas de infección se desconoce. Objetivos. Identificar las especies de triatominos con las cuales las personas entran en contacto y que reconocen como vectores de la enfermedad de Chagas, así como establecer la infección por tripanosomas y explorar posibles fuentes de alimentación de los triatominos con la participación de la comunidad. Materiales y métodos. El muestreo de triatominos se hizo en 12 municipios entre el 2011 y el 2016. T. cruzi se detectó mediante las técnicas de kinetic-polymerase chain reaction (k-PCR) y serial amplification of targets-polymerase chain reaction (SAT-PCR), en tanto que la genotipificación de las cepas se logró mediante la caracterización de secuencias de genes miniexon y de la región intergénica SL-IR (Spliced-Leader Intergenic Region). Se evaluaron, asimismo, las fuentes de alimento. Resultados. La mayoría de los triatominos fue recolectada por miembros de la comunidad y se identificaron tres especies: Rhodnius pallescens, Panstrongylus geniculatus y Eratyrus cuspidatus. La tasa de infección general por T. cruzi fue de 66,6 % y se detectaron las cepas TcIDOM y TcI sylvatic. La participación de la comunidad permitió reportar la presencia de P. geniculatus en dos nuevas localidades, la infección con T. cruzi de P. geniculatus en Córdoba y reportar por primera vez triatominos infectados con T. cruzi en Montería. Conclusiones. Se demostró la presencia de triatominos infectados con T. cruzi dentro de las viviendas en cinco municipalidades. La cepa circulante dominante fue T. cruzi TcIDOM, asociada con la enfermedad de Chagas y con cardiomiopatías en Colombia. Los resultados resaltan la importancia de vincular a miembros de la comunidad en la vigilancia entomológica.

Cardiac Evaluation in the Acute Phase of Chagas' Disease with Post-Treatment Evolution in Patients Attended in the State of Amazonas, Brazil / Avaliação Cardíaca na Fase Aguda da Doença de Chagas com Evolução Pós-Tratamento em Pacientes Atendidos no Estado do Amazonas, Brasil

Abstract Background: In the past two decades, a new epidemiological profile of Chagas' disease (CD) has been registered in the Brazilian Amazon where oral transmission has been indicated as responsible for the increase of acute cases. In the Amazonas state, five outbreaks of acute CD have been registered since 2004. The cardiac manifestations in these cases may be characterized by diffuse myocarditis, with alteration in the electrocardiogram (ECG) and transthoracic echocardiogram (TTE). Objective: To perform a cardiac evaluation in autochthonous patients in the acute phase and at least one year after submitted to treatment for acute CD and evaluate the demographic variables associated with the presence of cardiac alterations. Methods: We evaluated patients diagnosed with acute CD through direct parasitological or serological (IgM) methods from 2007 to 2015. These patients were treated with benznidazole and underwent ECG and TTE before and after treatment. We assumed a confidence interval of 95% (CI 95%, p < 0.05) for all variables analyzed. Results: We observed 63 cases of an acute CD in which oral transmission corresponded to 75%. Cardiac alterations were found in 33% of the cases, with a greater frequency of ventricular repolarization alteration (13%), followed by pericardial effusion (10%) and right bundle branch block and left anterior fascicular block (2%). The follow-up occurred in 48 patients with ECG and 25 with TTE for a mean period of 15.5 ± 4.1 months after treatment. Of these, 8% presented normalization of the cardiac alterations in ECG, 62.5% remained with the normal exams. All of the patients presented normal results in TTE in the post-treatment period. As for the demographic variables, isolated cases presented more cardiac alterations than outbreaks (p = 0.044) as well as cases from Central Amazonas mesoregion (p = 0.020). Conclusions: Although cardiac alterations have not been frequent in most of the studied population, a continuous evaluation of the clinical-epidemiological dynamics of the disease in the region is necessary in order to establish preventive measures.

Resumo Fundamento: Nas últimas duas décadas, um novo perfil epidemiológico da Doença de Chagas (DC) foi registrado na Amazônia brasileira, onde a transmissão oral foi indicada como responsável pelo aumento dos casos agudos. No estado do Amazonas, foram registrados cinco surtos da doença desde 2004. As manifestações cardíacas nesses casos podem ser caracterizadas por miocardite difusa, com alteração nos resultados eletrocardiograma (ECG) e ecocardiografia transtorácica (ETT). Objetivo: avaliar parâmetros cardíacos em pacientes autóctones com DC na fase aguda e em um ano ou mais após tratamento, e avaliar as variáveis demográficas associadas com a presença de alterações cardíacas. Métodos: Avaliamos os pacientes diagnosticados com DC aguda por método direto parasitológico e exame sorológico (IgM) entre 2007 e 2015. Os pacientes foram tratados com benzonidazol e submetidos à ECG e ETT antes e após tratamento. Assumimos um intervalo de confiança de 95% (p < 0,05) para todas as variáveis analisadas. Resultados: Observamos 63 casos de DC aguda em que a transmissão oral ocorreu em 75% dos casos. Alterações cardíacas foram encontradas em 33% dos casos, com maior frequência de repolarização ventricular (13%), seguida de derrame pericárdico (10%), e bloqueio do ramo direito e bloqueio fascicular anterior esquerdo (2%). O acompanhamento foi realizado com 48 pacientes com ECG e 25 com ETT por um período médio de 15,5±4,1 meses após o tratamento. Desses pacientes, observou-se normalização das alterações eletrocardiográficas em 8% dos pacientes, e 62,5% continuaram com os parâmetros normais. Todos os pacientes apresentaram resultados da ETT normais no período pós-tratamento. Quanto às variáveis demográficas, os casos isolados apresentaram mais alterações cardíacas em comparação aos casos de surtos (p=0,044) e os casos identificados na mesorregião do Amazonas Central (p = 0,020). Conclusões: Apesar de as alterações cardíacas não terem sido frequentes na maioria da população do estudo, é necessária uma avaliação contínua da dinâmica clínica-epidemiológica da doença na região para se estabelecer medidas preventivas.

Seroprevalence of trypanosoma cruzi infection among blood donors in the state of Bahia, Brazil

Abstract INTRODUCTION: Chagas disease (CD) affects 5.7-7.0 million individuals worldwide, and its prevalence reached 25.1% in the state of Bahia, Brazil. There is an association between the prevalence of CD, the socioeconomic status of the population, and the risk of re-emergence due to non-vectorial transmission, such as blood transfusion. This study determined the seroprevalence of T. cruzi infection among blood donors in the state of Bahia, located in northeastern Brazil, and their epidemiological profile during a 10-year period. METHODS: We performed a descriptive cross-sectional study involving a database review. Data were collected from patients with non-negative results for T. cruzi infection during a 10-year period. RESULTS: A total of 3,084 (0.62%) samples were non-negative for T. cruzi infection in an initial serological screening, and 810 (0.16%) samples were non-negative in the second screening. The correlation between infection and age (30 years or older) and between infection and lower educational level (12 years or less) in the first and second screening was statistically significant. The seroprevalence of T. cruzi infection was higher in men in the first screening. In addition, 99.52% of the municipalities of Bahia had at least one case of CD. Livramento de Nossa Senhora and Salvador presented the highest disease prevalence and recurrence, respectively. CONCLUSIONS: The seroprevalence of T. cruzi infection in these populations was lower than that found in other studies in Brazil but was comparatively higher in densely-populated areas. The demographic characteristics of our population agreed with previous studies.

Synanthropic triatomines (hemiptera: reduviidae): infestation, colonization, and natural infection by trypanosomatids in the state of Rio Grande do Norte, Brazil

Abstract INTRODUCTION The ecoepidemiological situation in the State of Rio Grande do Norte, Brazil is characterized by frequent invasion and colonization of domiciliary units (DUs) by several triatomine species, with high rates of natural infection by Trypanosoma cruzi. METHODS: We evaluated the possibility of vector transmission of T. cruzi based on records of the occurrence of domiciled triatomines collected by the Secretariat of State for Public Health from 2005 to 2015. During this period, 67.7% (113/167) of municipalities conducted at least one active search and 110 recorded the presence of insects in DUs. These activities were more frequent in municipalities considered to have a high and medium-level risk of T. cruzi transmission. RESULTS Of 51,569 captured triatomines, the most common species were Triatoma brasiliensis (47.2%) and T. pseudomaculata (40.2%). Colonies of T. brasiliensis, T. pseudomaculata, T. petrocchiae, Panstrongylus lutzi, and Rhodnius nasutus were also recorded in the intradomicile and peridomicile. Natural infection by trypanosomatids was detected in 1,153 specimens; the highest rate was found in R. nasutus (3.5%), followed by T. brasiliensis (2.5%) and T. pseudomaculata (2.4%). There have been high levels of colonization over the years; however, not all infested DUs have been sprayed. CONCLUSIONS: This is the first report of intradomicile and peridomicile colonization by P. lutzi. These results demonstrate the risk of new cases of infection by T. cruzi and reinforce the need for continuous entomological surveillance in the State of Rio Grande do Norte.

Epidemiological features, echocardiographic findings, and parasite load in patients with Chagas disease

Abstract INTRODUCTION Chagas disease is a major public health problem that is endemic in Brazil and Latin America. This study aimed to determine the socioeconomic, demographic, and clinical characteristics of 171 patients (mean age, 45 years; female, 65%) with Chagas disease at Hospital Universitário de Brasília, Federal District, Brazil. METHODS We implemented this cross-sectional study using a clinical epidemiological questionnaire, electrocardiography, echocardiography, and quantitative detection of Trypanosoma cruzi DNA in blood using qRT-PCR. RESULTS Among the patients, 26.3% had a full elementary education, and 13.2% were illiterate. Most (63.6%) were economically classified as class C, and 51.5% were born in Bahia state. A total of 62.0% participants reported previous contact with the triatomine bug. The clinical forms of the disease were indeterminate (69.51%), cardiac (15.24%), digestive (10.37%), and mixed (4.88%). The most common electrocardiographic abnormality was complete right bundle branch block in association with a divisional anterosuperior block. Only 14.6% of the patients complied with benznidazole medication for at least 60 days, and 164 of them were assessed by echocardiography. The parasite load was positive in 56% of the patients. CONCLUSIONS: Chagas disease affected mostly women, with the indeterminate chronic form of the disease.

Comparison of methodologies for detecting Trypanosoma cruzi parasites by microscopic observation of microhematocrit capillary tubes

Abstract INTRODUCTION: The microscopic examination of microhematocrit tubes (mHCT) has been proposed as the gold standard for acute and congenital Chagas disease diagnosis. We compared different mHCT methodologies detecting T. cruzi parasites in the blood. METHODS: The rotating method, water mount, and immersion oil methods were compared for their suitability, sensitivity, and specificity. RESULTS: The rotating method was easier, faster, and more sensitive than the others with 100% specificity. CONCLUSIONS: The rotating method is feasible for laboratory technicians with standard training in microscopic techniques and is recommended for the diagnosis of acute Chagas disease in primary health care facilities.

Occurrence of triatomines (Hemiptera: Reduviidae) in domestic and natural environments in Novo Remanso, Itacoatiara, Amazonas, Brazil

Abstract INTRODUCTION The present study reports the presence of triatomines in natural, peridomestic, and intradomicile environments in Itacoatiara municipality, state of Amazonas, a non-endemic region for Chagas disease. METHODS Active search was performed inside tree trunks, and palm trees, residences, and peridomiciles localized near the forest area. RESULTS: Twenty adults and ten triatomines nymphs were collected, fifteen of which were from natural forests, thirteen from intradomiciles, and two from peridomicile areas. CONCLUSIONS: The new records of adults and nymphs of triatomines in the intra- and peridomiciles suggest the adoption of prophylactic measures for vector surveillance in the study area.

Infección natural por Trypanosoma cruzi en triatominos que habitan en la palma corozo ( Acrocomia aculeta ) en regiones del oriente de Venezuela / Triatominae in Palm Trees and Their Natural Infection by Trypanosoma cruzi in Regions of Eastern Venezuela

RESUMEN Objetivos Conocer la infestación natural por triatominos y su infección por Trypanosoma cruzi (T. cruzi) en Acrocomia aculeata (A. aculeata) o palma corozo en el estado Anzoátegui, Venezuela. Materiales y métodos Se estudió la infestación triatomínica y su infección por T. cruzi en A. aculeata desafectadas en campañas fitosanitarias. La presencia del parásito se determinó por microscopia y PCR-kDNA, y se realizó su caracterización mediante marcadores moleculares. Resultados Se encontraron 14 palmeras con infestación triatomínica, el 48,8 % de los ejemplares correspondieron a Rhodnius prolixus y el 48,2 % a Triatoma maculata, con desarrollo ontogénico hacia el adulto. Las pruebas parasitológicas y moleculares, su morfología típica y la infección en el modelo murino revelaron la presencia de T. cruzi en 54,8 % en promedio, para ambas especies de triatominos, con circulación del genotipo TcI de T. cruzi. Conclusiónes Se reportó para el estado Anzoátegui en Venezuela, la infestación de palma corozo con Rhodnius prolixus y Triatoma maculata y la presencia de subpoblaciones TcI de T. cruzi, siendo esta palma el hábitat peridomiciliar del binomio triatominos-T. cruzi y posible bioindicador de riesgo de infección para poblaciones humanas circunvecinas.

ABSTRACT Introduction To know the natural infestation by triatominae and their infection by Trypanosoma cruzi (T. cruzi) in Acrocomia Aculeata (A. aculeata) or coyol palm in the state of Anzoátegui, Venezuela. Materials and Methods Triatominic infestation and its infection by T. cruzi was studied in non-affected A. aculeata in phytosanitary campaigns. The presence of the parasite was determined by microscopy and PCR-kDNA, and its characterization was made by means of molecular markers. Results Fourteen palm trees with triatominic infestation were found; 48.8% of the individuals corresponded to Rhodnius prolixus and 48.2% to Maculata Triatoma, with ontogenetic development towards adult. The parasitology and molecular tests, their typical morphology and the infection in the murine model revealed the presence of T. cruzi in an average of 54,8%, for both species of triatominae, with circulation of the TcI genotype of T. cruzi. Conclusions The infestation of coyol palm trees with Rhodnius prolixus and Maculata Triatoma was reported for the state of Anzoátegui in Venezuela, as well as the presence of TcI sub-populations of T. cruzi, being this palm tree the peridomicilar habitat of the triatominae-T. cruzi binomial and possible bioindicador of risk of infection for surrounding human populations.

Trypanosoma cruzi in Triatomines and wild mammals in the National Park of Serra das Confusões, Northeastern Brazil

Abstract INTRODUCTION The National Park of Serra das Confusões (NPSC) is a protected area of natural landscape located in Southern Piauí, Brazil, and it is considered as one of the largest and most important protected areas in the Caatinga biome. METHODS The natural occurrences of trypanosomatids from hemocultures on small mammals and cultures from intestinal contents triatomines were detected through molecular diagnoses of blood samples, and phylogenetic relationship analysis of the isolates parasites using the trypanosome barcode (V7V8 SSUrDNA) were realized. RESULTS Only two Galea spixii (8.1%) and six Triatoma brasiliensis (17.6%) were positive by hemoculture, and the isolates parasites were cryopreserved. All the isolates obtained were positioned on the Trypanosoma cruzi DTU TcI branch. CONCLUSIONS Research focused on studying the wild animal fauna in preserved and underexplored environments has made it possible to elucidate indispensable components of different epidemiological chains of diseases with zoonotic potential.

First report of Triatoma sordida Stål, 1859 (Hemiptera, Reduviidae, Triatominae) in the State of Acre and Brazilian Western Amazon

Abstract INTRODUCTION The occurrence of Triatoma sordida in the Brazilian Western Amazon is reported for the first time. METHODS Triatoma sordida specimens were collected from a Gallus gallus nest in a peridomiciliary area of Senador Guiomard municipality in the state of Acre. RESULTS The number of triatomine species in Acre increased from six to seven with this first report of T. sordida in the Brazilian Western Amazon. CONCLUSIONS The occurrence of T. sordida in Acre is of concern since it is among the most captured triatomines in peridomiciliary environments in Brazil, and carries a high potential for vector transmission.

Investigação de surto de doença de Chagas aguda na região extra-amazônica, Rio Grande do Norte, Brasil, 2016 / Investigation of an outbreak of acute Chagas disease outside the Amazon Region, in Rio Grande do Norte State, Brazil, 2016 / Investigación del brote de la enfermedad de Chagas aguda en la región extra-amazónica, Rio Grande do Norte, Brasil, 2016

Resumo: O objetivo do artigo foi confirmar e descrever um surto da doença de Chagas aguda por transmissão oral na mesorregião Oeste Potiguar, Rio Grande do Norte, Brasil. Trata-se de um estudo descritivo do tipo série de casos, tendo como fonte de dados os registros de atendimentos médicos e entrevistas com os casos suspeitos entre 16 de setembro e 19 de novembro de 2015. Realizou-se pesquisa entomológica nas prováveis localidades de infecção dos casos de doença de Chagas aguda. Foram confirmados 18 casos de doença de Chagas aguda em residentes de quatro municípios do Rio Grande do Norte. Os sinais e sintomas mais relatados foram febre e fraqueza (n = 18), seguidos de mialgia (n = 17), prostração, inapetência e edema de membros inferiores (n = 15). A mediana de duração da febre foi de 20 dias (intervalo: 6 a 45 dias). Quinze casos foram confirmados por critério laboratorial e três por vínculo epidemiológico com clínica compatível. Todos os casos confirmados relataram ter consumido caldo de cana da mesma procedência. Foram capturados 110 triatomíneos na fazenda onde ocorreu a moagem da cana consumida. Os insetos estavam no peridomicílio, em amontoados de lenha e próximos ao engenho. A maioria dos exemplares capturados era da espécie Triatoma brasiliensis, e apresentou um índice de infecção natural de 63%. Foi confirmado surto da doença de Chagas de transmissão oral por ingestão de caldo de cana contaminado por triatomíneo infectado, evidenciado pelo vínculo epidemiológico entre os casos investigados e a pesquisa entomológica no local provável de infecção.

Abstract: The aim of this article was to confirm and describe an outbreak of acute Chagas disease involving oral transmission in the western region of Rio Grande do Norte State, Brazil. This was a descriptive case series study in which the data sources were medical records and interviews with suspected cases from September 16 to November 19, 2015. An entomological investigation was conducted in the probable sites of infection for acute Chagas disease cases. Eighteen cases of acute Chagas disease were confirmed in residents of four municipalities (counties) in Rio Grande do Norte State. The most frequently reported signs and symptoms were fever and weakness (n = 18), followed by myalgia (n = 17), prostration, loss of appetite, and edema of the lower limbs (n = 15). Median duration of fever was 20 days (range: 6 to 45 days). Fifteen cases were confirmed by the laboratory criterion and three by epidemiological link with consistent clinical characteristics. All confirmed cases reported having consumed sugar cane juice from the same mill. A total of 110 triatomines were captured on the plantation where the sugar cane had been crushed for juice. The insects were found in the peridomicile, in stacks of firewood and close to the sugar cane mill. The majority of the captured specimens were Triatoma brasiliensis and showed a natural infection rate of 63%. The Chagas disease outbreak was confirmed with oral transmission via ingestion of sugar cane juice contaminated with infected triatomines, as evidenced by the epidemiological link between the investigated cases and the entomological survey in the probable site where the infection occurred.

Resumen: El objetivo de este artículo fue confirmar y describir un brote de la enfermedad de Chagas aguda por transmisión oral en la mesorregión oeste de Rio Grande do Norte, Brasil. Se trata de un estudio descriptivo del tipo serie de casos, contando como fuente de datos los registros de atención médica y entrevistas en los casos sospechosos entre el 16 de septiembre y el 19 de noviembre de 2015. Se realizó una investigación entomológica en las probables localidades de infección de los casos de enfermedad de Chagas aguda. Fueron confirmados 18 casos de enfermedad de Chagas aguda en residentes de cuatro municipios de Rio Grande do Norte. Los signos y síntomas más informados fueron fiebre y debilidad (n = 18), seguidos de mialgia (n = 17), postración, inapetencia y edema de miembros inferiores (n = 15). La media de duración de la fiebre fue de 20 días (intervalo: 6 a 45 días). Quince casos fueron confirmados por criterio de laboratorio y tres por vínculo epidemiológico con clínica compatible. Todos los casos confirmados informaron haber consumido caldo de caña de la misma procedencia. Fueron capturados 110 triatominos en la hacienda donde se produjo la molienda de la caña consumida. Los insectos estaban en el peridomicilio, en montones de leña y cerca del ingenio azucarero. La mayoría de los ejemplares capturados era de la especie Triatoma brasiliensis, y presentó un índice de infección natural de un 63%. Se confirmó un brote de la enfermedad de Chagas de transmisión oral por ingestión de caldo de caña contaminado por triatomino infectado, evidenciado por el vínculo epidemiológico entre los casos investigados y la investigación entomológica en el lugar probable de infección.

Prevalência da doença de Chagas entre doadores de sangue do Estado do Piauí, Brasil, no período de 2004 a 2013 / Prevalence of Chagas disease among blood donors in Piauí State, Brazil, from 2004 to 2013 / Prevalencia de la enfermedad de Chagas entre donadores de sangre del estado de Piauí, Brasil, durante el período de 2004 a 2013

Apesar do declínio na prevalência da doença de Chagas no Brasil, a Região Nordeste apresenta condições propícias ao seu recrudescimento. O objetivo deste estudo foi analisar a positividade por doença de Chagas entre doadores de sangue do Piauí entre 2004 e 2013. A prevalência de sorologia reagente para doenças de Chagas na triagem de doadores foi de 1%, variando de 0,4% na Regional de Saúde de Uruçuí a 2,4% na Regional de Saúde de São Raimundo Nonato. Dos 220 municípios, 58,6% apresentaram casos. Apenas 34,5% das amostras positivas na triagem foram encaminhadas para testes complementares e entre estes, 84,4% apresentaram resultados negativos. Nossos resultados indicam a possibilidade da manutenção da transmissão vetorial em áreas do Estado do Piauí e a necessidade da implantação de ações que melhorem o índice de realização dos testes complementares referentes aos casos positivos na triagem.

Despite the decline in prevalence of Chagas disease in Brazil, the Northeast region of the country has favorable conditions for its resurgence. The study aimed to analyze positive Chagas disease serology rates among blood donors in Piauí State from 2004 to 2013. Prevalence of positive Chagas disease serology in blood donor screening was 1%, ranging from 0.4% in the Uruçuí Regional Health District to 2.4% in the São Raimundo Nonato Regional Health District. Of the state's 220 municipalities, 58.6% reported cases. Only 34.5% of the positive samples in screening were referred for complementary tests, and 84.4% of these showed negative results. Our findings suggest the possibility of persistent vector-borne transmission in areas of Piauí State and the need for measures to improve complementary testing in positive cases detected by screening.

A pesar del declive en la prevalencia de la enfermedad de Chagas en Brasil, la Región Nordeste presenta condiciones propicias para su recrudecimiento. El objetivo de este estudio fue analizar la positividad por enfermedad de Chagas entre donadores de sangre de Piauí entre 2004 y 2013. La prevalencia de la serología reactiva para la enfermedad de Chagas en la clasificación de donadores fue de un 1%, variando de 0,4% en la Regional de Salud de Uruçuí, a 2,4% en la Regional de Salud de São Raimundo Nonato. De los 220 municipios, un 58,6% presentaron casos. Sólo un 34,5% de las muestras positivas en la clasificación fueron dirigidas a test complementarios y entre estos, un 84,4% presentaron resultados negativos. Nuestros resultados indican la posibilidad del mantenimiento de la transmisión vectorial en áreas del Estado de Piauí y la necesidad de la implantación de acciones que mejoren el índice de realización de los test complementarios referentes a los casos positivos en la clasificación.