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BACKGROUND: The study of the ecology of Trypanosoma cruzi is challenging due to its extreme adaptive plasticity, resulting in the parasitism of hundreds of mammal species and dozens of triatomine species. The genetic analysis of blood meal sources (BMS) from the triatomine vector is an accurate and practical approach for gathering information on which wild mammal species participate in a local transmission network. South American coatis, Nasua nasua, act as important reservoir host species of T. cruzi in the Pantanal biome because of their high rate of infection and elevated parasitemia, with the main discrete typing unit (DTU) lineages (TcI and TcII). Moreover, the carnivore coati is the only mammal species to build high arboreal nests for breeding and resting that can be shared by various vertebrate and invertebrate species. Herein, we applied the sensitive and specific methodology of DNA barcoding and molecular cloning to study triatomines found in a coati nest to access the diversity of mammal species that explore this structure, and therefore, may be involved in the parasite transmission network. METHODS: Twenty-three Triatoma sordida were collected in one coati's nest in the subregion of Nhecolândia, Pantanal. The DNA isolated from the gut of insects was subjected to BMS detection by PCR using universal primers that flank variable regions of the cytochrome b (cytb) and 12S rDNA mitochondrial genes from vertebrates. The Trypanosoma spp. diagnosis and DTU genotyping were based on an 18S rDNA molecular marker and also using new cytb gene primers designed in this study. Phylogenetic analyses and chord diagrams were constructed to visualize BMS haplotypes, DTU lineages detected on vectors, and their interconnections. RESULTS: Twenty of 23 triatomines analyzed were PCR-positive (86.95%) showing lineages T. cruzi DTU TcI (n = 2), TcII (n = 6), and a predominance of TcI/TcII (n = 12) mixed infection. Intra-DTU diversity was observed mainly from different TcI haplotypes. Genetic analyses revealed that the southern anteater, Tamandua tetradactyla, was the unique species detected as the BMS of triatomines collected from the coati's nest. At least three different individuals of T. tetradactyla served as BMS of 21/23 bugs studied, as indicated by the cytb and 12S rDNA haplotypes identified. CONCLUSIONS: The identification of multiple BMS, and importantly, different individuals of the same species, was achieved by the methodology applied. The study demonstrated that the southern anteaters can occupy the South American coati's nest, serving as the BMS of T. sordida specimens. Since anteaters have an individualist nonsocial behavior, the three individuals detected as BMS stayed at the coati's nest at different times, which added a temporal character to BMS detection. The TcI and TcII infection, and significantly, a predominance of TcI/TcII mixed infection profile with different TcI and TcII haplotypes was observed, due to the discriminatory capacity of the methodology applied. Tamandua tetradactyla, a host which has been little studied, may have an important role in the T. cruzi transmission in that Pantanal subregion. The data from the present study indicate the sharing of coatis' nests by other mammal species, expanding the possibilities for T. cruzi transmission in the canopy strata. We propose that coatis' nests can act as the true hubs of the T. cruzi transmission web in Pantanal, instead of the coatis themselves, as previously suggested.

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BACKGROUND: Chagas disease (American trypanosomiasis) is an important neglected tropical illness, which has the flagellate protozoan Trypanosoma cruzi as etiological agent and blood-feeding insects of the Triatominae subfamily as vectors. Despite its importance for disease epidemiology, field studies targeting microgeographic dispersal of triatomines in endemic areas are rare. The ability wingless nymphs have to move (crawl) within peridomestic settings is a key component regarding the design and development of rational control strategies. METHODS: We double-marked Triatoma sordida fourth-instar nymphs (N4) with a reliable fluorescent dye and a trace element. This new methodology allowed us to simultaneously evaluate (i) nymph dispersal and (ii) the effectiveness of copper (Cu), chromium (Cr), and cadmium (Cd) trace elements as potential new markers. In the mark-release-recapture (MRR) experiment, 390 T. sordida N4 were released in the peridomicile of a single rural household, 130 individuals at each of three release points, at distances of 2, 5, and 10 m from the chicken coop (CC) and 27, 32, and 35 m away from the horse corral (HC). All specimens were double marked (Cu/blue, Cr/orange, Cd/green). Recaptures occurred in two intervals: 1-3 days and 15-17 days after release. RESULTS: Specimens were successfully recaptured at all distances up to 10 m. A total of 19, 23, and 10 specimens were able to disperse 2, 5, and 10 m, respectively, to reach the CC. No insects were recaptured at the HC. Of the three analyte/paint combinations tested, Cr/orange gave the most promising results; Cu/blue marker and Cd/green marker performed very poorly with only 4/19 and 0/10 analyte/paint ratios, respectively. CONCLUSIONS: Triatoma sordida N4 could cover a distance of 10 m in 17 days. This indicates that nymphs seem to have a reduced dispersal capability compared to adults. Ninety-one percent of the 22 recaptured orange-marked nymphs were still Cr positive after the 17-day period evaluated. This makes this analyte a good candidate for future investigations that will apply this marking method in MRR studies.

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Chagas disease is caused by Trypanosoma cruzi, which has several forms of transmission. Among them, the vector route requires the constant action of the Health Surveillance in the control of triatomines. Our objective is to describe the diversity of triatomines and the rate of natural infection by T. cruzi in these insects, as well as to analyse entomological indicators, through secondary data generated by the Health Surveillance of the municipality of Barra, in the period from 2009 to 2019. The secondary data were subjected to descriptive analysis and the entomological indicators calculated. Williams' G test and Fisher's exact test were used to analyse the categorical data. A total of 10,913 triatomines of the genera Rhodnius sp., Panstrongylus sp., Eratyrus sp. and Triatoma sp. were captured, represented by 12 species. Triatoma sordida was the most prevalent 98.66% (10,767/10,913), reflected in the overall infestation index 7.61% (2,555/33,544) and triatomine density 0.24 (8,247/33,544). While the overall infection rate was 0.81% (41/5,048) and remained low during all years. There was significant association for T. sordida in relation to "T. cruzi infection index" and "Developmental stage" and "Capture site". It is concluded that the municipality of Barra has a high occurrence of triatomines of various species, with T. sordida being the most prevalent species and responsible for the rates of infestation and infection by T. cruzi. The presence of these insects in households is a risk for the domestic cycle of the parasite, and thus can transmit T. cruzi to various domestic animals, including humans.(AU)

doença de Chagas é causada pelo Trypanosoma cruzi, que possui diversas formas de transmissão. Dentre elas, a via vetorial, requer a atuação constante da Vigilância em Saúde no controle dos triatomíneos. Nosso objetivo é descrever a diversidade de triatomíneos e o índice de infecção natural por T. cruzi nestes insetos, bem como analisar os indicadores ento-mológicos, por meio de dados secundários gerados pela Vigilância em Saúde do município de Barra, no período de 2009 a 2019. Os dados secundários foram submetidos a análise descritiva e os indicadores entomológicos calculados. Os testes G de Williams e exato de Fisher foram utilizados para a análise dos dados categóricos. Foram capturados 10.913 triatomíneos, dos gêneros Rhodnius sp., Panstrongylus sp., Eratyrus sp. e Triatoma sp., representados por 12 espécies. Triatoma sordida foi a mais prevalente 98,66% (10.767/10.913), refletindo no índice de infestação geral 7,61% (2.555/33.544) e densidade triatomínea 0,24 (8.247/33.544). Enquanto que o índice de infecção geral foi de 0,81% (41/5.048) e manteve-se baixo durante todos os anos. Houve associação significativa para T. sordida em relação ao "Índice de infecção por T. cruzi" e "Estágio de desenvolvi-mento" e "Local de captura". Conclui-se que o município de Barra apresenta uma alta ocorrência de triatomíneos de diversas espécies, sendo T. sordida a espécie mais prevalente e responsável pelos índices de infestação e infecção por T. cruzi. A presença desses insetos nos domicílios é um risco para o ciclo doméstico do parasito, podendo assim transmitir o T. cruzi para diversos animais domésticos, inclusive humanos.(AU)

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Chagas disease (CD) is a parasitic zoonosis endemic in Brazil. Despite virtual control of Triatoma infestans, the main domesticated vector of Trypanosoma cruzi, vectorial transmission by other triatomine species persists in some rural communities. This study aims to characterize triatomines role in transmitting T. cruzi to dogs and humans in the district of Santo Inácio, located in the northwest region of the state of Bahia, Brazil. It also describes environmental factors in housings associated with insect occurrence and assesses the perception, knowledge, and preventive practices adopted by the population regarding CD. Blood samples of humans and dogs, and biological samples of triatomines, were collected between November 2018 and February 2019 and subjected to the detection of T. cruzi by serological and molecular biology tests. Also, we applied a questionnaire to research the perception, knowledge, and local practices of people related to CD. The capture of triatomines in households was associated with exploratory variables of the questionnaires using multivariate logistic regression (p < 0.05). The 155 triatomines captured in the wild and domestic environment were of the species Triatoma sherlocki (n = 151), Panstrongylus sherlocki (n = 1) and Triatoma sordida (n = 3), and had a natural infection rate for T. cruzi by PCR of 18.5%, 100% and 0%, respectively. District residents (n = 126) were seronegative for T. cruzi, while 17.5% (7/40) of the dogs were seropositive. The fact that residents are aware that triatomines can "cause" CD was configured as a protection factor for residents according to the fitted logistic regression model (p = 0.04). However, respondents have limited perception and knowledge about the CD, prevention and control practices for triatomines in a household. The results suggest the existence of a domestic cycle of transmission of T. cruzi between triatomines and dogs, configuring a latent risk of infection to the human population of Santo Inácio. Studies that clarify the potential for the establishing of intrusive triatomines in households, surveillance actions for triatomines, and health education in rural communities are indispensable to prevent the reemergence of CD in vulnerable regions of Brazil and other American countries with similar epidemiological characteristics.

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BACKGROUND: Chagas disease is endemic throughout most of Bolivia, with prevalence rates of 25% observed in some geographic areas located mainly in the sub-Andean region. METHODS: Community-based entomological surveillance was carried out in the sub-Andean departments of Cochabamba (municipalities of Cochabamba, Punata and Sacaba), Tarija (municipality of Tarija) and Chuquisaca (municipality of Sucre). The surveillance parameters evaluated were: (i) the proportion of cards with the presence of triatomines; (ii) the distribution of positive cards by area; and (iii) the proportion of cards with the presence of infected triatomines. RESULTS: Of the cards returned, in 852 (3.1%) there was a mention of the presence of triatomines. The species Triatoma infestans, Triatoma sordida and Triatoma guasayana were identified in 812 (95.3%), 39 (4.6%) and 1 (0.1%), respectively. The median monthly positivity rate of the cards during 2011-2018 was higher in Punata (9.1%; IQR=3.2-15.4%). The median monthly rate was highest in 2012 (2.7%; IQR=0-5.6%). Fifty positive cards (5.8%) presented insects that were positive for trypanosomatids, mainly in Cochabamba and Punata. CONCLUSIONS: The report of triatomines foci by inhabitants represents an effective surveillance system coordinated by a network of specialized and multidisciplinary health centers. These strategies, which should be included in the health policies of endemic countries, enable extending and deepening the dialogue among technicians, communities and their local authorities.

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A juvenile specimen of a mermithid (Nematoda) was found parasitizing a third instar nymph of Triatoma sordida from Mato Grosso, Brazil. This is the first record of mermithid parasitism in a triatomine species. The Mermithidae represents a family of nematodes that are specialized insect parasites. Entomonematodes are one of the highly influential agents regulating the population dynamics of insects. This report introduces the opportunity to think about mermithids as a possible candidate for use as triatomine biological control.

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Chagas disease is one of the most important insect-vectored diseases in Brazil. The entomopathogenic fungus Metarhizium anisopliae was evaluated against nymphs and adults of Panstrongylus megistus, Triatoma infestans, and T. sordida. Pathogenicity tests at saturated humidity demonstrated high susceptibility to fungal infection. The shortest estimates of 50% lethal time (LT50 ) for P. megistus varied from 4.6 (isolate E9) to 4.8 days (genetically modified strain 157p). For T. infestans, the shortest LT50 was 6.3 (E9) and 7.3 days (157p). For T. sordida, the shortest LT50 was 8.0 days (157p). The lethal concentration sufficient to kill 50% of T. infestans (LC50 ) was 1.9 × 107 conidia/ml for strain 157p. In three chicken coops that were sprayed with M. anisopliae, nymphs especially were well controlled, with a great population reduction of 38.5% after 17 days. Therefore M. anisopliae performed well, controlling Triatominae in both laboratory and field studies.

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Triatoma sordida is an endemic species to South America, currently considered the species most frequently found in the peridomestic environment in Brazil. This triatomine has a wide ecological tolerability that allows it to inhabit several ecotopes and use different food sources. Although the species is considered predominantly peridomestic, it is also capable of colonizing households and forming numerous intra-household colonies, leading to its inclusion among the priorities for triatomine control campaigns in Brazil. All Brazilian populations of T. sordida are considered as T. sordida sensu stricto by chromosomal analyses (which highlights their epidemiological importance), although molecular studies that characterize the genetic diversity of these populations are scarce, being restricted only to the region of Minas Gerais. Thus, several populations of this vector distributed in the states of Bahia, Goiás, Mato Grosso do Sul, and Minas Gerais were analyzed using mitochondrial markers (cyt b and nd1). Low nucleotide diversity, high haplotypic diversity, low genetic distance, and high FST value were observed, as well as the formation of a monophyletic clade of the Brazilian populations of T. sordida, which confirms that this species has low genetic variability, with all specimens in Brazil grouped in T. sordida sensu stricto. In addition to the genetic and evolutionary importance for the knowledge of the biology of these vectors, these results are important from an epidemiological point of view, thus being able to direct vector control programs.

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Background: The detection of residual foci inside the houses, or the reinfestation of triatomines is one of the main entomological surveillance challenges. Actions aimed at increasing the probability of vector detection and to detect re-infestation when the density of vector populations is low, is a priority objective for Chagas control programs. Methods: Families belonging to local communities were responsible of triatomine specimens capture, following a strategic methodology based on entomological surveillance with community participation developed by the National Chagas Programme of the Ministry of Health of Bolivia. The main objective of the study is to evaluate the entomological surveillance strategy with community engagement implemented in Chagas Platform Centers for comprehensive care (CPs). The degree of intradomicillary residual vector infestation, the main seasonal period of triatomines capture, and natural infection by trypanosomatids rates were evaluated. Results: In rural and peri-urban Punata, in the Department of Cochabamba, the houses infestation rate by triatomines exceeds the national average and is above the recommendations of PAHO / WHO. The observations during the seasons of the year showed that Spring season (September to December) was the period where there was a higher average of T. infestans positive houses detected by the families participating in the study in the three departments of Bolivia. The presence of infected triatomines with Trypanosomatideos in positive houses was 6% in the study area, still finding active domestic cycles in rural and peri-urban areas and not in urban areas where triatomines with parasites were not reported during the seven years of monitoring. Conclusions: Reporting infestation foci by the inhabitants is the simplest and most direct way of participation of the community in entomological surveillance. These strategies should be included in the health policies of the countries, as well as extending and deepening the dialogue between technicians, communities and their local authorities.

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BACKGROUND: The Sordida subcomplex (Triatominae) comprises four species, Triatoma garciabesi, T. guasayana, T. patagonica and T. sordida, which differ in epidemiological importance and adaptations to human environments. Some morphological similarities among species make taxonomic identification, population differentiation and species delimitation controversial. Triatoma garciabesi and T. sordida are the most similar species, having been considered alternatively two and a single species until T. garciabesi was re-validated, mostly based on the morphology of male genitalia. More recently, T. sordida from Argentina has been proposed as a new cryptic species distinguishable from T. sordida from Brazil, Bolivia and Paraguay by cytogenetics. We studied linear and geometric morphometry of the head, wings and pronotum in populations of these species aiming to find phenotypic markers for their discrimination, especially between T. sordida and T. garciabesi, and if any set of variables that validates T. sordida from Argentina as a new species. RESULTS: Head width and pronotum length were the linear variables that best differentiated species. Geometric morphometry revealed significant Mahalanobis distances in wing shape between all pairwise comparisons. Triatoma patagonica exhibited the best discrimination and T. garciabesi overlapped the distribution of the other species in the morphometric space of the first two DFA axes. Head shape showed differentiation between all pairs of species except for T. garciabesi and T. sordida. Pronotum shape did not differentiate T. garciabesi from T. guasayana. The comparison between T. garciabesi and T. sordida from Argentina and T. sordida from Brazil and Bolivia revealed low differentiation based on head and pronotum linear measurements. Pronotum and wing shape were different between T. garciabesi and T. sordida from Brazil and Bolivia and T. sordida from Argentina. Head shape did not differentiate T. garciabesi from T. sordida from Argentina. CONCLUSIONS: Wing shape best delimited the four species phenotypically. The proposed cryptic species, T. sordida from Argentina, differed from T. sordida from Brazil and Bolivia in all measured shape traits, suggesting that the putative new species may not be cryptic. Additional studies integrating cytogenetic, phenotypic and molecular markers, as well as cross-breeding experiments are needed to confirm if these three entities represent true biological species.

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