Impacto dos fungos entomopatogênicos Metarhizium anisopliae e Beauveria bassiana no controle biológico de Rhodnius neglectus: um estudo realizado em condições de laboratório / Impact of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana on the biological control of Rhodnius neglectus: a study carried out under laboratory conditions

A doença de Chagas é uma doença negligenciada causada pelo protozoário Trypanosoma cruzi que afeta, aproximadamente, seis a sete milhões de pessoas em todo o mundo, resultando, todos os anos, em cerca de 14 mil mortes por complicações cardíacas e digestivas. A principal forma de transmissão dessa enfermidade na América Latina é vetorial, pelos triatomíneos, sendo o controle de vetores a principal medida para mitigar os casos de infecção por T. cruzi. O histórico do controle dos triatomíneos no Brasil se fundamenta, principalmente, no uso de inseticidas, fazendo-se necessário o manejo integrado de pragas como alternativa para a contenção desses vetores. Entre elas, tem sido realizada a utilização de controladores biológicos, visando auxiliar no estabelecimento de estratégias racionais e eficazes de controle de artrópodes vetores, uma vez que causam baixos danos ambientais. Diversos trabalhos consideram os fungos Beauveria bassiana e Metarhizium anisopliae patogênicos para triatomíneos, conferindolhes competência para o controle dos vetores da doença de Chagas. Dessa forma, foi testada a eficácia de produtos comerciais compostos pelos fungos M. anisopliae e B. bassiana no controle biológico de Rhodnius neglectus, como alternativa ao uso de inseticidas. As análises do potencial de B. bassiana resultaram na não eclosão de 28,3% dos ovos, na mortalidade de 8,57% das ninfas de primeiro estágio (N1) e de 24,44% dos adultos. Já M. anisopliae foi responsável pela não eclosão de 25% dos ovos, mortalidade de 21,43% das ninfas N1 e 13,33% dos adultos. Esses resultados, quando comparados ao grupo controle (30% de não eclosão de ovos, nenhuma mortalidade das ninfas e 30% de mortalidade em adultos), demonstraram a ineficácia desses fungos no controle de R. neglectus, uma vez que o grupo controle resultou em eclosão/mortalidade igual ou maior que os tratamentos com os fungos entomopatogênicos.

Chagas disease is a neglected disease caused by the protozoan Trypanosoma cruzi that affects approximately six to seven million people worldwide, resulting, every year, in about 14,000 deaths from heart and digestive complications. The main form of transmission of this disease in Latin America is vector, by triatomines, with vector control being the main measure to mitigate cases of infection by T. cruzi. The history of triatomine control in Brazil is mainly based on the use of insecticides, making integrated pest management necessary as an alternative to contain these vectors. Among them, the use of biological controllers has been carried out, aiming to help in the establishment of rational and effective strategies to control arthropod vectors, since they cause low environmental damage. Several works consider the fungi Beauveria bassiana and Metarhizium anisopliae to be pathogenic for triatomines, giving them competence to control the vectors of Chagas disease. Thus, we tested the effectiveness of commercial products composed of the fungi M. anisopliae and B. bassiana in the biological control of Rhodnius neglectus, as an alternative to the use of insecticides. Analyzes of the potential of B. bassiana resulted in the non-hatching of 28.3% of the eggs, the mortality of 8.57% of the first-stage nymphs (N1) and 24.44% of the adults. M. anisopliae was responsible for the non-hatching of 25% of the eggs, mortality of 21.43% of the N1 nymphs and 13.33% of the adults. These results, when compared to the control group (30% non-hatching of eggs, no nymph mortality and 30% mortality in adults), demonstrate the ineffectiveness of these fungi in the control of R. neglectus, since the control group resulted in hatching/mortality equal to or greater than treatments with entomopathogenic fungi.

Cytokey: desenvolvimento de uma chave de identificação com dados citogenéticos para os vetores da doença de Chagas do estado da Paraíba, Brasil / Cytokey: development of an identification key with cytogenetic data for Chagas disease vectors from the state of Paraíba

Os triatomíneos são insetos hematófagos vetores do protozoário Trypanosoma cruzi, agente etiológico da doença de Chagas (DC). Embora existam diferentes formas de transmissão do parasito, a Organização Mundial de Saúde ressalta que a vetorial seja a principal via de transmissão do T. cruzi na América Latina. No Brasil, existe cerca de 4,6 milhões de pessoas vivendo com DC, sendo os casos mais recentes de contaminação por infecção oral, a partir de barbeiros infectados e/ou fezes contaminadas que foram processados com alimentos consumidos in natura. Diversos casos de infecção oral foram registrados na região Nordeste, associados a surtos causados pelo consumo de caldo de cana ou outros alimentos. Nos estados da Paraíba (PB) e do Rio Grande do Norte (RN), por exemplo, 26 óbitos foram associados com infecção por via oral e 18 casos positivos para DC e duas mortes foram notificadas, respectivamente. Por séculos, as principais chaves de classificação se fundamentaram em chaves dicotômicas baseadas em dados morfológicos. Apenas em 2019, chaves alternativas, com base em dados citogenéticos (CytoKeys), foram propostas. Recentemente, chaves foram desenvolvidas para os estados de PE e RN. Com base no exposto, uma chave dicotômica foi desenvolvida para auxiliar na identificação das espécies de triatomíneos da PB, a partir de dados citogenéticos. Com base na chave dicotômica desenvolvida com dados citogenéticos, todas as espécies de triatomíneos notificadas na PB puderam ser diferenciadas. O desenvolvimento de chaves alternativas mostra-se de grande importância para a comunidade científica e, principalmente, os agentes de saúde que atuam no controle de vetores, auxiliando-os na correta identificação de espécies que apresentam semelhanças morfológicas (como as espécies Triatoma brasilensis e T. petrocchiae notificadas na PB).

Triatomines are hematophagous insects vectors of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease (CD). Although there are different forms of transmission of the parasite, the World Health Organization emphasizes that vector transmission is the main route of transmission of T. cruzi in Latin America. In Brazil, there are about 4.6 million people living with CD, with the most recent cases of contamination by oral infection, from infected triatomines and/or contaminated feces that were processed with food consumed in natura. Several cases of oral infection were registered in the Northeast region, associated with outbreaks caused by the consumption of sugarcane juice or other foods. In the states of Paraíba (PB) and Rio Grande do Norte (RN), for example, 26 deaths were associated with oral infection and 18 positive cases for CD and two deaths were reported, respectively. For centuries, the main classification keys were based on dichotomous keys based on morphological data. Only in 2019, alternative keys, based on cytogenetic data (CytoKeys), were proposed. Recently, keys were developed for the states of PE and RN. Based on the above, a dichotomous key was developed to assist in the identification of BP triatomine species, based on cytogenetic data. Based on the dichotomous key developed with cytogenetic data, all triatomine species reported in BP could be differentiated. The development of alternative keys is of great importance for the scientific community and, mainly, for health agents who work in vector control, helping them in the correct identification of species that present morphological similarities (such as the species Triatoma brasilensis and T .petrocchiae notified in BP).

Evaluation of triatomines infestation occurrence in home environments in the city of Taua-Ce, 2012 / Avaliação da ocorrência de infestação por triatomíneos em ambientes domiciliares no município de Tauá-Ce, 2012 / Evaluación de la ocurrencia de infestación por triatominos en ambientes domiciliarios en el municipio de Tauá-Ce, 2012

Background and Objectives: Triatomines are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease, a parasitic disease that affects mammals and humans. The destruction or transformation of natural ecotopes has intensified, resulting in the invasion of triatomines in households, bringing risks to the population. Thus, the objective of the study was to carry out a data survey on triatomine infestation in the countryside of the city of TauaCe, in 2012. Methods: This is a descriptive/exploratory study of a quantitative nature, where the Secondary data were obtained from the Municipal Health Department (Endemic Nucleus). Results: The percentage of infestation in the 15 locations studied was 26.1%, with the most affected locations being: Sítio Central do Incra 50%, Fazenda Brôco 18.2%, Jordão 16.7%, Fazenda Bom Lugar 0%, Fazenda Riacho do Mato 40%, Fazenda Cearauai 37.5%, Sítio Riacho do Mato 30%, Fazenda Cedro 17.4%, Cachoeirinha 27%, Fazenda Várzea do Feijão 24%, Cachoeira do Júlio 41.2%, Fazenda Mutuquinha 9,4%, Sítio Várzea Grande 33.3%, Iparana 9.1% and Cachoeira do Celso 16.7%. Conclusion: The study showed that there is a significant rate of triatomine infestation in the home environment, which represents a great risk to the health of the population of Taua. Therefore, a more frequent entomological control is suggested, going beyond the period of campaigns for a better monitoring of the occurrence of these insects in the area.(AU)

Justificativa e Objetivos: Triatomíneos são vetores de Trypanosoma cruzi, agente etiológico da doença de Chagas, parasitose que atinge mamíferos e humanos. A destruição ou transformação dos ecótopos naturais tem se intensificado, resultando na invasão de triatomíneos em domicílios trazendo riscos a população. Diante do exposto, o objetivo do estudo foi realizar um levantamento de dados sobre a infestação de triatomíneos em localidades do interior do município de Tauá-Ce, em 2012. Métodos: Trata-se de um estudo descritivo/exploratório de natureza quantitativa, onde os dados secundários foram obtidos junto à Secretaria de Saúde (Núcleo de Endemias) do município. Resultados: O percentual de infestação nas 15 localidades estudadas foi de 26,1%, sendo as localidades mais afetadas: Sítio Central do Incra 50%, Fazenda Brôco 18,2%, Jordão 16,7%, Fazenda Bom Lugar 0%, Fazenda Riacho do Mato 40%, Fazenda Cearauai 37,5%, Sítio Riacho do Mato 30%, Fazenda Cedro 17,4%, Cachoeirinha 27%, Fazenda Várzea do Feijão 24%, Cachoeira do Júlio 41,2%, Fazenda Mutuquinha 9,4%, Sítio Várzea Grande 33,3%, Iparana 9,1% e Cachoeira do Celso 16,7%. Conclusão: O estudo mostrou que há um expressivo índice de infestação triatomínica em ambiente domiciliar, o que representa um grande risco a saúde da população tauaense. Sendo assim, sugere-se um controle entomológico mais frequente, ultrapassando o período de campanhas para um melhor acompanhamento da ocorrência desses insetos na região.(AU)

Justificación y Objetivos: Los triatomíneos son vectores de Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, parasitosis que afecta a mamíferos y humanos. La destrucción o transformación de los ecótopos naturales se ha intensificado, resultando en la invasión de triatomíneos en domicilios trayendo riesgos a la población. Ante lo expuesto, el objetivo del estudio fue realizar un levantamiento de datos sobre la infestación de triatomíneos en localidades del interior del municipio de Tauá-Ce, en 2012. Métodos: Se trata de un estudio descriptivo/exploratorio de naturaleza cuantitativa, donde los datos secundarios fueron obtenidos junto a la Secretaría de Salud (Núcleo de Endemias) del municipio. Resultados: El porcentaje de infestación en las 15 localidades estudiadas fue de 26,1%, siendo las localidades más afectadas: Sítio Central do Incra 50%, Fazenda Brôco 18,2%, Jordão 16,7%, Fazenda Bom Lugar 0%, Fazenda Riacho do Mato 40%, Fazenda Cearauai 37,5%, Sítio Riacho do Mato 30%, Fazenda Cedro 17,4%, Cachoeirinha 27%, Fazenda Várzea do Feijão 24%, Cachoeira do Júlio 41,2%, Fazenda Mutuquinha 9,4%, Sítio Várzea Grande 33,3%, Iparana 9,1% e Cachoeira do Celso 16,7%. Conclusión: El estudio mostró que hay un expresivo índice de infestación triatomínica en ambiente domiciliar, lo que representa un gran riesgo para la salud de la población tauaense. Siendo así, se sugiere un control entomológico más frecuente, superando el período de campañas para un mejor seguimiento de la ocurrencia de esos insectos en la región.(AU)

Hemolymph of triatomines presents fungistatic activity against Cryptococcus neoformans and improves macrophage function through MCP-I/TNF-α increase

Triatomines are blood-feeding arthropods belonging to the subfamily Triatominae (Hemiptera; Reduviidae), capable of producing immunomodulatory and water-soluble molecules in their hemolymph, such as antimicrobial peptides (AMPs). In this work, we evaluated the antifungal and immunomodulatory activity of the hemolymph of Meccus pallidipennis (MPH) and Rhodnius prolixus (RPH) against Cryptococcus neoformans. Methods: We assessed the activity of the hemolymph of both insects on fungal growth by a minimum inhibitory concentration (MIC) assay. Further, RAW 264.7 macrophages were cultivated with hemolymph and challenged with C. neoformans. Then, their phagocytic and killing activities were assessed. The cytokines MCP-1, IFN-γ, TNF-α, IL-10, IL-12, and IL-6 were measured in culture supernatants 4- and 48-hours post-infection. Results: Both hemolymph samples directly affected the growth rate of the fungus in a dose-dependent manner. Either MPH or RPH was capable of inhibiting fungal growth by at least 70%, using the lowest dilution (1:20). Treatment of RAW 264.7 macrophages with hemolymph of both insects was capable of increasing the production of MCP-I and TNF-α. In addition, when these cells were stimulated with hemolymph in the presence of C. neoformans, a 2- and a 4-fold increase in phagocytic rate was observed with MPH and RPH, respectively, when compared to untreated cells. For the macrophage killing activity, MPH decreased in approximately 30% the number of viable yeasts inside the cells compared to untreated control; however, treatment with RPH could not reduce the total number of viable yeasts. MPH was also capable of increasing MHC-II expression on macrophages. Regarding the cytokine production, MCP-I and TNF-α, were increased in the supernatant of macrophages treated with both hemolymphs, 4 and 48 hours after stimulation. Conclusion: These results suggested that hemolymph of triatomines may represent a source of molecules capable of presenting antifungal and immunomodulatory activity in macrophages during fungal infection.(AU)

Avaliação do conhecimento sobre a doença de Chagas em escolares das zonas rural e urbana de municípios endêmicos em Minas Gerais / Assessment of knowledge about Chagas disease in schoolchildren in rural and urban areas of endemic municipalities in Minas Gerais state, Brazil

Resumo O objetivo deste estudo foi avaliar o conhecimento de escolares do ensino médio de três escolas localizadas em municípios mineiros endêmicos para a doença de Chagas e extrair informações relativas à sua percepção em relação aos aspectos gerais, biológicos, clínicos e epidemiológicos da doença, do vetor e controle entomológico da doença de Chagas, além de comparar o conhecimento dos participantes em função da localização das escolas (zona rural ou urbana). Para isso, aplicou-se um questionário semiestruturado a 68 alunos. Todos os participantes já ouviram falar a respeito da doença de Chagas, afirmando ter adquirido esse conhecimento, sobretudo, com familiares e escola. Além disso, muitos participantes demonstraram conhecer os hábitos dos vetores, tais como hematofagia e fontes alimentares. Não houve associação clara entre o perfil de urbanização e o conhecimento dos participantes. No entanto, alunos da zona urbana apresentaram menos conhecimento prático a respeito dos triatomíneos e aspectos relativos à transmissão da doença, em relação aos alunos da zona mais "ruralizada". Sugere-se o aprofundamento do tema nos currículos escolares do ensino médio e a necessidade de elaboração de ações de educação em saúde voltadas à vigilância entomológica da doença de Chagas nesses municípios e, provavelmente, em outras regiões do Brasil.

Abstract This study aimed to evaluate the knowledge of high school students from three schools located in endemic municipalities of Minas Gerais to Chagas disease and to extract information related to their perception regarding the general, biological, clinical and epidemiological aspects of the disease, vector and entomological control of Chagas disease, in addition to comparing the knowledge of participants according to the location of schools (rural or urban area). For this, a semi-structured questionnaire was applied to 68 students. All participants have heard about Chagas disease, stating that they have acquired this knowledge, especially with family members and school. In addition, many participants demonstrated to know the habits of vectors, such as hematophagy and dietary sources. There was no clear association between the urbanization profile and the participants' knowledge. However, students from the urban area presented less practical knowledge about triatoins and aspects related to the transmission of the disease, in relation to students in the most "ruralized" area. It is suggested the deepening of the theme in high school curricula and the need to develop health education actions aimed at entomological surveillance of Chagas disease in these municipalities and, probably, in other Brazilian regions.

Actualización de la distribución geográfica de triatominos en el departamento del Putumayo / Update to the geographical distribution of triatomines in the Department of Putumayo / Atualização da distribuição geográfica de triatomíneos no Departamento de Putumayo

La enfermedad de Chagas es una zoonosis producida por la infección del Trypanosoma cruzi (T. cruzi) (1), cuya principal vía de transmisión es vectorial (2). Esta enfermedad se caracteriza por ser una infección crónica que puede ocasionar daños cardiacos, digestivos y neurológicos irreversibles (3). En el departamento del Putumayo, de acuerdo con los datos del Sistema de Vigilancia Epidemiológica (SIVIGILA), entre el año 2015 y el 2020, se han notificado 19 casos de Chagas crónico y 4 casos de Chagas agudos (4). Por este motivo resulta de gran interés compartir con los lectores de la revista MedUNAB la actualización de la distribución geográfica de los triatominos, vectores de la enfermedad de Chagas, y establecer el riesgo epidemiológico que representan para la población Putumayense, donde hay hallazgos de gran importancia porque se identifican especies en municipios y localidades donde antes no se conocían. Palabras claves: Triatominae; Enfermedad de Chagas; Infección por Trypanosoma cruzi; Infecciones; Insectos Vectores; Colombia.

Chagas Disease is a zoonotic disease produced by infection with Trypanosoma cruzi (T. cruzi) (1), whose main route of transmission is vectorial (2). This disease is characterized by being a chronic infection that can cause irreversible cardiac, digestive, and neurological damage (3). In the department of Putumayo, according to data from the epidemiological surveillance system (SIVIGILA, for the Spanish original), between 2015 and 2020, 19 chronic cases and 4 acute cases of Chagas were reported (4). For this reason, it is important to share with the readers of the MedUNAB journal the update to the geographical distribution of triatomines, vectors of Chagas Disease, and to establish the epidemiological risk that they represent for the population of Putumayo, where there are findings of great importance as species are identified in municipalities and localities where they were not previously found. Keywords: Triatominae; Chagas Disease; Trypanosoma cruzi; Infections; Insect Vectors; Colombia.

A doença de Chagas é uma zoonose causada por infecção com Trypanosoma cruzi (T. cruzi) (1), cujo principal modo de transmissão é vetorial (2). Esta doença é uma infecção crônica que pode causar danos cardíacos, digestivos e neurológicos irreversíveis (3). No Departamento de Putumayo, de acordo com dados do Sistema de Vigilância Epidemiológica (SIVIGILA), entre 2015 e 2020, foram notificados 19 casos de Chagas crônica e quatro casos de Chagas aguda (4). Por esta razão, é de grande interesse compartilhar com os leitores da revista MedUNAB a atualização da distribuição geográfica dos triatomíneos, vetores da doença de Chagas, e estabelecer o risco epidemiológico que representam para a população de Putumayo, onde há descobertas de grande importância, pois são identificadas espécies em municípios e localidades onde antes eram desconhecidas. Palavras-chave: Triatominae; Doença de Chagas; Infecção por Trypanosoma cruzi; Infecções; Insetos Vetores; Colômbia.

Validation of a documentary on Chagas disease by a population living in an endemic area / Validação de um documentário sobre a doença de Chagas por uma população de área endêmica

Abstract Educational interventions may trigger actions that contribute to prevent parasitic diseases, such as Chagas disease (CD). This study aimed at investigating the impact of an instructional video named "Documentary on Chagas Disease" on knowledge about CD and its vectors displayed by a population that lives in an endemic area in Brazil, so as to validate it as an educational tool. The video was shown to 226 subjects, divided into two groups. Group 1 was composed of users of Basic Health Units (BHU) in Pelotas and Pinheiro Machado, cities located in Rio Grande do Sul (RS) state, Brazil, where CD is endemic. Group 2 consisted of students who attend three public schools located in the rural area in Pinheiro Machado, RS. Two questionnaires with questions about their knowledge about triatomines and CD were applied, before and after the documentary was shown. After the video was shown, there was significant increase in individuals' knowledge (in both groups) about "kissing bugs", their notification, the disease and its prevention. Besides, watchers considered that the quality of the material was satisfactory. Since the "Documentary on CD" can be easily accessed on the internet and was effective in teaching the population that lives in endemic areas, its use should be encouraged in places and meetings connected to health that aim at fighting against triatominae and at exposing an updated view of CD.

Resumo Intervenções educacionais podem desencadear ações que contribuam para a prevenção de doenças parasitárias, como a doença de Chagas (DC). Este estudo teve como objetivo investigar o impacto de um vídeo instrucional denominado "Documentário Doença de Chagas" no conhecimento sobre DC e seus vetores exibidos a uma população que vive em área endêmica no Brasil, para validá-lo como uma ferramenta educacional. O vídeo foi exibido para 226 indivíduos, divididos em dois grupos. O grupo 1 foi composto por usuários das Unidades Básicas de Saúde (UBS) de Pelotas e Pinheiro Machado, cidades do estado do Rio Grande do Sul (RS), Brasil, onde a DC é endêmica. O grupo 2 foi formado por estudantes que frequentam três escolas públicas localizadas na zona rural de Pinheiro Machado, RS. Dois questionários com perguntas sobre conhecimentos sobre triatomíneos e DC foram aplicados, antes e depois da exibição do documentário. Após a exibição do vídeo, houve um aumento significativo no conhecimento dos indivíduos (em ambos os grupos) sobre "triatomíneos", sua notificação, a doença e sua prevenção. Além disso, os observadores consideraram que a qualidade do material era satisfatória. Como o "Documentário DC" pode ser facilmente acessado na Internet e ser eficaz no ensino à população que vive em áreas endêmicas, seu uso deve ser incentivado em locais e reuniões relacionadas à saúde, que visam combater os triatomíneos e exibir uma informação atualizada sobre a DC.

Metaciclogénesis de Trypanosoma cruzi en Belminus ferroae (Reduviidae: Triatominae) y capacidad infectiva de las heces en condiciones de laboratorio / Metacyclogenesis of Trypanosoma cruzi in B. ferroae (Reduviidae: Triatominae) and feces infectivity under laboratory conditions

Resumen | Introducción. Belminus ferroae es un triatomino de comportamiento entomófago, sin embargo, puede alimentarse de vertebrados ocasionalmente. No se ha demostrado infección natural por Trypanosoma cruzi en esta especie, como tampoco la metaciclogénesis del parásito. Objetivo. Examinar la metaciclogénesis de T. cruzi en B. ferroae y la capacidad infectiva de las heces o sus contenidos intestinales en roedores. Materiales y métodos. Se analizaron las heces y la orina expulsadas espontáneamente por los insectos o mediante compresión abdominal o extracción del contenido intestinal a los 10, 20, 30, 40, 50 y 60 días. Se cuantificó la carga parasitaria de T. cruzi y sus formas evolutivas se identificaron con tinción de Giemsa. Asimismo, se evaluó en ratones albinos la capacidad infectiva de los tripomastigotes metacíclicos de T. cruzi obtenidos de las heces o contenidos intestinales de los especímenes infectados. Resultados. El análisis parasitológico reveló tres (15 %) insectos infectados con T.cruzi a los 30 (n=1), 40 (n=1) y 50 (n=1) días después de la infección con cargas parasitarias de hasta 1,62 x 105 tripanosomas/mm3 y porcentajes de metaciclogénesis entre el 3,5 y el 6,78 %. Conclusiones. Se demuestra por primera vez, en una especie del género Belminus, la metaciclogenésis de T. cruzi en condiciones de laboratorio y la capacidad infectiva de las heces para un huésped vertebrado.

Abstract | Introduction: Belminus ferroae is a triatominae with entomophagous behavior. However, it may occasionally feed on vertebrates. Currently, there is no evidence of natural infection with Trypanosoma cruzi or the occurrence of metacyclogenesis in this species. Objective: To test T. cruzi metacyclogenesis in B. ferroae and the infectivity of their feces or intestinal contents in rodents under laboratory conditions. Materials and methods: Twenty nymphs of B. ferroae were infected with an autochthonous strain of T. cruzi (M/HOM/VE/09/P6). Fecal and urine samples were collected from spontaneous droppings or by compressing the bugs' abdomens and, eventually, by removing their gut contents, and then examined at 10, 20, 30, 40, 50, and 60 days. We quantified T. cruzi parasitic load, as well as the evolutionary forms in feces, urine, and intestinal contents by Giemsa staining. Similarly, we evaluated the infectivity of T. cruzi metacyclic trypomastigotes in albino mice. Results: The parasitological analysis showed three insects (15%) infected with T. cruzi at 30 (n=1), 40 (n=1), and 50 (n=1) days post-infection. We observed parasitic loads of up to 1.62 x 105 trypanosomes/mm3 and metacyclogenesis percentages between 3.5% and 6.78%. Conclusions: This is the first time that T. cruzi metacyclogenesis is reported in a species of the genus Belminus under laboratory conditions and the infectivity of Belminus' feces is demonstrated on a vertebrate host.

First report of colonization by Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil / Primeiro relato de colonização por Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) na Região Metropolitana de São Paulo, Brasil

Abstract We report the first known occurrence of Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) in the Metropolitan Region of São Paulo, Brazil. In 2018, adult specimens were sent by residents to the competent authorities and, in the inspection of the property, a large focus associated with a marsupial nest was found. This finding reinforces the importance of the species in the state of São Paulo, serves as an alert for epidemiological surveillance and extends the species colonization area in the state of São Paulo.

Resumo Relatamos a primeira ocorrência de Panstrongylus megistus (Burmeister, 1835) (Hemiptera, Reduviidae, Triatominae) na Região Metropolitana de São Paulo, Brasil. Em 2018, espécimes adultos foram enviados por residentes e a pesquisa na propriedade constatou a presença de um grande foco associado a um ninho marsupial. Este achado reforça a importância da espécie no estado de São Paulo, serve de alerta para a vigilância epidemiológica e amplia a área de colonização da espécie no estado de São Paulo.

Panstrongylus rufotuberculatus (Champion, 1899) (Hemiptera, Reduviidae, Triatominae) in Rondônia, Brazil: A novel report

Abstract INTRODUCTION: This short communication presents a novel report on the occurrence of Panstrongylus rufotuberculatus in the Brazilian state of Rondônia. METHODS: Two specimens were collected inside dwellings and identified using dichotomous keys. RESULTS: The present study showed the extensive geographic distribution of P. rufotuberculatus and the increased number of species in the state of Rondônia. CONCLUSIONS: This new record of P. rufotuberculatus is important for understanding the epidemiology of Chagas disease because this species is found naturally infected with Trypanosoma cruzi. Studies on the ecology, biology, and vector-host-parasite interactions of this species are essential for surveillance programs.

First report of Microtriatoma borbai Lent & Wygodzinsky, 1979 (Hemiptera, Reduviidae, Triatominae) in the state of Espírito Santo, Brazil: would M. borbai be living in eucalyptus crops?

Abstract INTRODUCTION: The occurrence of Microtriatoma borbai in the state of Espírito Santo, Brazil is reported by the first time. METHODS: A triatomine specimen collected in a hybrid eucalyptus crop in the municipality of Aracruz, Espírito Santo state was found to be a male M. borbai. RESULTS: This finding expands the geographical distribution of M. borbai from four to five Brazilian states. It is the first report of M. borbai occurrence inside a eucalyptus crop. CONCLUSIONS: The occurrence of M. borbai in the state of Espírito Santo broadens the geographical distribution of this species in southeastern Brazil.

Do the new triatomine species pose new challenges or strategies for monitoring Chagas disease? An overview from 1979-2021

Chagas disease persists as one of the most important, and yet most neglected, diseases in the world, and several changes in its epidemiological aspects have been recorded since its discovery. Currently, some of the most relevant changes are related to: (i) the reduction in the incidence of the endemic due to the control of the most important vectors, Triatoma infestans and Rhodnius prolixus, in many countries; (ii) the migration of human populations spreading cases of the disease throughout the world, from endemic to non-endemic areas, transforming Chagas disease into a global threat; and (iii) new acute cases and deaths caused by oral transmission, especially in the north of Brazil. Despite the reduction in the number of cases, new challenges need to be responded to, including monitoring and control activities aiming to prevent house infestation by the secondary vectors from occurring. In 1979, Lent & Wygodzinsky(1) published the most complete review of the subfamily Triatominae, encompassing 111 recognised species in the taxon. Forty-two years later, 46 new species and one subspecies have been described or revalidated. Here we summarise the new species and contextualise them regarding their ecology, epidemiologic importance, and the obstacles they pose to the control of Chagas disease around the world.

Triatominae (Hemiptera, Reduviidae) in homes: Report of their occurrence in the municipality of Cruzeiro do Sul, Acre, South Western Amazon

Abstract INTRODUCTION: Triatomines are hematophagous insects that are important to public health since they are the vectors of American Trypanosomiasis. The objective of this study was to describe the occurrence of triatomines in homes in Cruzeiro do Sul, Acre, Brazil. METHODS The specimens were collected by an active search inside homes and also by a passive search by the residents. RESULTS: A total of 55 triatomines were captured comprising of 5 species each of the genera Rhodnius, Eratyrus, and Panstrongylus. No colonies were detected, ruling out the possibility of domiciliation. CONCLUSIONS: Information on regional epidemiological dynamics contributes to the prevention and control of disease.

The rediscovery of Rhodnius domesticus Neiva & Pinto, 1923 (Hemiptera: Reduviidae: Triatominae) in the state of Espírito Santo, Brazil

Abstract INTRODUCTION: Rhodnius domesticus Neiva & Pinto, 1923 is a rare sylvatic triatomine endemic to the Atlantic Forest, with one known record for Espírito Santo (ES), Brazil from 1969. We present here its rediscovery in ES, 42 years after its first record. METHODS: In January 2011, a triatomine specimen was collected from a rural area of the municipality of Santa Teresa, ES. RESULTS: We confirmed this as a new record of R. domesticus in the Baixo Caldeirão locality. CONCLUSIONS: This finding supports the possibility of a wild population of R. domesticus in the mountainous region of the Atlantic forest of ES.

Colonization by Panstrongylus megistus (Hemiptera, Reduviidae, Triatominae) in an urban park in the city of São Paulo

Abstract INTRODUCTION: This communication reports on the occurrence of colonization by Panstrongylus megistus in an urban park in the municipality of São Paulo, Brazil. METHODS: Entomological research includes active search for vectors based on notifications by the population and identification and examination of insects. RESULTS: A colony of triatomines was found to be associated with enclosed birds. CONCLUSIONS: The occurrence of P. megistus has already been reported in the city of São Paulo; however, reports of colonization by this species provide evidence of its potential for the occupation of artificial ecotopes, which may pose a risk to the human population.

Análise da percepção de acadêmicos de medicina de São José do Rio Preto, São Paulo, Brasil sobre a doença de Chagas e seus vetores / Perception analysis of medical students from São José do Rio Preto, São Paulo, Brazil about Chagas disease and its vectors

As doenças negligenciadas (DN) são um grupo diversificado de enfermidades que prevalecem em condições tropicais e subtropicais de 149 países e afetam mais de um bilhão de pessoas que vivem em situação de pobreza em todo o mundo. Atualmente, estima-se que oito milhões de pessoas estejam infectadas com a doença de Chagas (DC). Embora São José do Rio Preto, São Paulo, Brasil seja uma região do estado de São Paulo conhecida pelo alto grau de incidência de dengue, existem, pelo menos, 700 pacientes chagásicos registrados no Hospital de Base de São José do Rio Preto e alta incidência do vetor, sendo que mais de 14 mil triatomíneos foram coletados no noroeste paulista entre 2004 e 2011. Assim, levando em consideração que a DC é clinicamente curável se o tratamento for realizado no estágio inicial de contaminação com o Trypanosoma cruzi e que a ação dos profissionais da saúde é fundamental para o diagnóstico laboratorial dessa doença na fase aguda da infecção (período em quem os fármacos anti-tripanosomatídeos são efetivos), avaliou-se a percepção de acadêmicos de Medicina de São José do Rio Preto, São Paulo, Brasil sobre questões gerais relacionadas com essa enfermidade, por meio de uma pesquisa transversal, descritiva, prospectiva e quantitativa. A maioria dos alunos demonstrou conhecer o agente etiológico da DC. Além disso, os alunos de medicina demonstraram conhecimento da principal forma de transmissão do parasita, embora uma parcela deles não associou a transmissão da DC às fezes de triatomíneos. Outra dificuldade observada por esses discentes foi em relação ao tratamento da DC, pois mais da metade dos alunos respondeu erroneamente que a DC não tem cura ou tem cura na fase crônica. Levando-se em consideração que a DC foi descrita por um médico que caracterizou todos os aspectos da doença, há necessidade dos cursos de graduação de medicina abordarem as DNs de forma holística, pois as Diretrizes Curriculares Nacionais exigem a formação de profissionais de saúde competentes e capazes de integrarem dimensões biológicas, psicológicas, sociais e ambientais.

Neglected Diseases (ND) are a diverse group of diseases that are prevalent in tropical and subtropical conditions in 149 countries and affect more than one billion people living in poverty worldwide. Currently, it is estimated that eight million people are infected with Chagas disease (CD). Although São José do Rio Preto, São Paulo, Brazil is a region of the state of São Paulo known for its high incidence of dengue, there are at least 700 patients with CD registered at the Base Hospital of São José do Rio Preto and high incidence of the vector, as more than 14 thousand triatomines were collected in the northwest of São Paulo between 2004 and 2011. Thus, taking into account that CD is clinically curable if the treatment is carried out at the initial stage of contamination with Trypanosoma cruzi and that the action of health professionals is essential for the clinical diagnosis of this disease in the acute phase of the infection (period in which anti-trypanosomatids are effective), was evaluated the perception of medical students from São José do Rio Preto, São Paulo, Brazil about general questions related to this disease, through a cross-sectional, descriptive, prospective and quantitative research. Most students demonstrated knowledge of the etiological agent of CD. In addition, medical students demonstrated knowledge of the main form of transmission of the parasite, although a portion of them did not associate CD transmission with triatomine feces. Another difficulty observed by these students was related to the treatment of CD, as more than half of the students answered erroneously that CD has no cure or can be cured in the chronic phase. Taking into account that CD was described by a doctor who characterized all aspects of the disease, there is a need for undergraduate medical courses to address NDs in a holistic way, as the National Curriculum Guidelines require the training of competent health professionals capable of integrating biological, psychological, social and environmental dimensions.

Avaliação externa da qualidade da identificação entomológica de triatomíneos realizada na Rede de Laboratórios Públicos em Pernambuco, 2017 / Evaluación externa de la calidad en la identificación entomológica de triatominos realizada en la red de laboratorios públicos en Pernambuco, Brasil, 2017 / External quality assessment of entomological identification of triatomines carried out in the Pernambuco Public Health Laboratory network, Brazil, 2017

Objetivo: Avaliar a qualidade da identificação de triatomíneos na rede de laboratórios do estado de Pernambuco, Brasil. Métodos: Os nove laboratórios participantes receberam material de apoio com as chaves dicotômicas e um painel composto por sete insetos triatomíneos conhecidos no estado, para identificação da situação do espécime no recebimento (estrutura completa ou danificada), espécie e sexo. Resultados: Nove laboratórios de 12 aderiram ao estudo. A proporção de acerto para identificação do sexo foi de 56/63, e para espécie, 45/63, não apresentando relação direta com a ocorrência de danos nas estruturas morfológicas durante o transporte dos insetos. Para Panstrongylus megistus, houve acerto em todos os espécimes (9/9), enquanto para espécies do gênero Rhodnius a proporção foi menor (3/9). Conclusão: Apesar do bom desempenho na identificação entomológica, as fragilidades observadas poderão orientar ações para melhoria na rede de laboratórios e serão essenciais para os programas de controle vetorial da doença de Chagas.

Objetivo: Evaluar la calidad de la identificación de triatominos en la red de laboratorios del estado de Pernambuco, Brasil. Métodos: Los nueve laboratorios participantes recibieron material de apoyo con claves dicotómicas y un panel conformado por siete insectos triatominos conocidas en el estado, para identificar la situación del espécimen al recibirlo (estructura completa o dañada), especie y sexo. Resultados: La adherencia al laboratorio fue del 9/12. La proporción de aciertos para la identificación del sexo fue del 56/63, y para las especies 45/63, no mostraron relación directa con la ocurrencia de daños en las estructuras morfológicas durante el transporte de insectos. Solo Panstrongylus megistus logró el 9/9 de precisión, mientras que para las especies del género Rhodnius fue el más pequeño (3/9). Conclusión: A pesar del buen desempeño en identificación entomológica, la evaluación indicó debilidades que permitirán implementar acciones correctivas para mejorar la red de laboratorios, esenciales para los programas de control vectorial de la enfermedad de Chagas.

Objective: To assess the quality of triatomine identification in the laboratory network of the state of Pernambuco, Brazil. Methods: The participating laboratories received support material with dichotomous keys and a panel made up of seven triatomine insects known in the Pernambuco, in order to identify specimen status on receipt (intact or damaged structure), as well as species and sex. Results: Nine out of 12 laboratories took part of the study. The proportion of correct answers was 56/63 for sex identification, and 45/63, for species. The answers did not present a direct relationship with occurrence of damage to morphological structures during insect transportation. Panstrongylus megistus identification was correct for all specimens (9/9 laboratories), while correct identification of species of the Rhodnius genus was the lowest (3/9 laboratories). Conclusion: Despite the good performance in entomological identification, the weaknesses observed may guide improvements in the laboratory network and will be essential for Chagas disease vector control programs.

First report of Microtriatoma borbai Lent & Wygodzinsky, 1979 (Hemiptera, Reduviidae, Triatominae) in the state of Espírito Santo, Brazil: would M. borbai be living in eucalyptus crops?

Abstract INTRODUCTION: The occurrence of Microtriatoma borbai in the state of Espírito Santo, Brazil is reported by the first time. METHODS: A triatomine specimen collected in a hybrid eucalyptus crop in the municipality of Aracruz, Espírito Santo state was found to be a male M. borbai. RESULTS: This finding expands the geographical distribution of M. borbai from four to five Brazilian states. It is the first report of M. borbai occurrence inside a eucalyptus crop. CONCLUSIONS: The occurrence of M. borbai in the state of Espírito Santo broadens the geographical distribution of this species in southeastern Brazil.

Zuccagnia punctata Cav. Essential Oil into Poly(ε-caprolactone) Matrices as a Sustainable and Environmentally Friendly Strategy Biorepellent against Triatoma infestans (Klug) (Hemiptera, Reduviidae)

The main strategies against Triatoma infestans (primary vector responsible for the Chagas disease transmission) are the elimination or reduction of its abundance in homes through the application of insecticides or repellents with residual power, and environmental management through the improvement of housing. The use of plant-derived compounds as a source of therapeutic agents (i.e., essential oils from aromatic plants and their components) is a valuable alternative to conventional insecticides and repellents. Essential oil-based insect repellents are environmentally friendly and provide reliable personal protection against the bites of mosquitoes and other blood-sucking insects. This study investigates, for the first time to our knowledge, the potential repellent activity of Zuccagnia punctata essential oil (ZEO) and poly(ε-caprolactone) matrices loaded with ZEO (ZEOP) prepared by solvent casting. The analysis of its essential oil from aerial parts by GC­FID and GC-MS, MS allowed the identification of 25 constituents representing 99.5% of the composition. The main components of the oil were identified as (−)-5,6-dehydrocamphor (62.4%), alpha-pinene (9.1%), thuja-2, 4 (10)-diene (4.6%) and dihydroeugenol (4.5%). ZEOP matrices were homogeneous and opaque, with thickness of 800 ± 140 µm and encapsulation efficiency values above 98%. ZEO and ZEOP at the lowest dose (0.5% wt./wt., 96 h) showed a repellency of 33 and 73% respectively, while at the highest dose (1% wt./wt., 96 h) exhibited a repellent activity of 40 and 66 %, respectively. On the other hand, until 72 h, ZEO showed a strong repellent activity against T. infestans (88% repellency average; Class V) to both concentrations, compared with positive control N-N diethyl-3-methylbenzamide (DEET). The essential oils from the Andean flora have shown an excellent repellent activity, highlighting the repellent activity of Zuccagnia punctata. The effectiveness of ZEO was extended by its incorporation in polymeric systems and could have a potential home or peridomiciliary use, which might help prevent, or at least reduce, Chagas' disease transmission.