Ultrastructural and enzymatic alterations in the ovary of Rhodnius prolixus infected with Trypanosoma rangeli

Objective: To investigate the morphological structure of ovarian follicular cells and biochemical parameters of both ovaries and fat bodies (sites of vitellogenesis) from Rhodnius (R.) prolixus infected with Trypanosoma (T.) rangeli. Methods: Adult virgin females of R. prolixus were fed upon a membrane apparatus containing heat-inactivated citrated rabbit blood and a suspension of T. rangeli epimastigotes (Macias strain). Females from the control group and all the males received parasitefree blood. Transmission electron microscopy was used to reveal the morphological aspects of ovarian follicle cells in both control and parasite-infected groups. Protein profile, proteolytic activities and Western blotting analyses were performed in either ovary or fat body samples of control and parasite-infected groups. Results: According to the ultrastructural data, T. rangeli infection elicited a degeneration process in the ovarian follicular cells of R. prolixus. Proteolytic assays indicated a reduction in the activity of aspartic peptidases in the ovary and fat body from parasite-infected group, while a significant increase in the cysteine peptidase activity was measured in both insect organs. Additionally, immunoblotting revealed that vitellogenin was overexpressed in the ovary of parasite-infected insects. Conclusions: T. rangeli infection seems to elicit an early programmed cell death in the ovarian follicle cells as well as induces the modulation on the activities of different peptidase classes in either ovaries or fat bodies and the overexpression of the vitellogenin in the ovary of R. prolixus.

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)

Monitoring Rhodnius neglectus (Lent, 1954) populations' susceptibility to insecticide used in controlling actions in urban areas northwest of São Paulo state

ABSTRACT Background: Chagas disease (CD) is caused by the flagellate protozoan Trypanosoma cruzi and can be carried by different species of triatomines, including Rhodnius neglectus, which is wild, well distributed in Brazil, and has formed colonies in palm trees located in urban areas of municipalities in the state of São Paulo. Chemical control has been routinely used to reduce population density, but each year, there has been an increase in species dispersion and density. This study aimed to evaluate the susceptibility of insects to insecticides used in control. Methods: The reference population was collected from Araçatuba municipality, Nilce Maia. Dilutions of deltamethrin were prepared and applied to the back of the first-stage nymphs, which were biologically synchronized. The control group received pure acetone only. Mortality was assessed after 72 h. Results: The mortality rate with respect to diagnostic dose was 100%. The susceptibility profile observed for this population showed RR50 ranging from 1.76 to 3.632. Conclusions: The populations were susceptible to the insecticides tested. It is possible that the insecticide residual effect on this ecotope has decreased the lifespan, and controlling failures may be the cause of recolonization in this environment.

Effectiveness of fluralaner treatment regimens for the control of canine Chagas disease: A mathematical modeling study

Canine Chagas disease is caused by the protozoan parasite Trypanosoma cruzi and transmitted by insect triatomine vectors known as kissing bugs. The agent can cause cardiac damage and long-term heart disease and death in humans, dogs, and other mammals. In laboratory settings, treatment of dogs with systemic insecticides has been shown to be highly efficacious at killing triatomines that feed on treated dogs. Method We developed compartmental vector-host models of T. cruzi transmission between the triatomine and dog population accounting for the impact of seasonality and triatomine migration on disease transmission dynamics. We considered a single vector-host model without seasonality, and model with seasonality, and a spatially coupled model. We used the models to evaluate the effectiveness of the insecticide fluralaner with different durations of treatment regimens for reducing T. cruzi infection in different transmission settings. Results In low and medium transmission settings, our model showed a marginal difference between the 3-month and 6-month regimens for reducing T. cruzi infection among dogs. The difference increases in the presence of seasonality and triatomine migration from a sylvatic transmission setting. In high transmission settings, the 3-month regimen was substantially more effective in reducing T. cruzi infections in dogs than the other regimens. Our model showed that increased migration rate reduces fluralaner effectiveness in all treatment regimens, but the relative reduction in effectiveness is minimal during the first years of treatment. However, if an additional 10% or more of triatomines killed by dog treatment were eaten by dogs, treatment could increase T. cruzi infections in the dog population at least during the first year of treatment. Conclusion Our analysis shows that treating all peridomestic dogs every three to six months for at least five years could be an effective measure to reduce T. cruzi infections in dogs and triatomines in peridomestic transmission settings. However, further studies at the local scale are needed to better understand the potential impact of routine use of fluralaner treatment on increasing dogs' consumption of dead triatomines.

Cytochrome P450 Genes of the CYP4 Clan and Pyrethroid Resistance in Chagas Disease Vectors

he chemical control of Triatoma infestans, the major Chagas disease vector in southern South America, has been threatened in the last years by the emergence of pyrethroid-resistant bug populations. As an alternative approach, the efficacy of the entomopathogenic fungus Beauveria bassiana to control T. infestans populations (regardless their pyrethroid susceptibility) has been demonstrated. Growing research efforts on the interaction between T. infestans and B. bassiana by molecular, ecological, biochemical and behavioral traits has allowed framing such interaction as an evolutionary arms race. This review will focus on the relationships established in this particular host-pathogen system, compiling available data on the relevance of fungal pathogenesis, insect behavior, population dynamics and human intervention to favor fungal dissemination in bug populations. The current snapshot shows the fungus ahead in the evolutionary arms race and predicts a promissory landscape for the biological control of Chagas disease vectors.

Laboratory Evaluation and Field Feasibility of Micro-Encapsulated Insecticide Effect on Rhodnius prolixus and Triatoma dimidiata Mortality in Rural Households in Boyacá, Colombia

Abstract: Chagas disease is a neglected vector-borne zoonosis caused by the parasite Trypanosoma cruzi that is primarily transmitted by insects of the subfamily Triatominae. Although control efforts targeting domestic infestations of Rhodnius prolixus have been largely successful, with several regions in Boyacá department certified free of T. cruzi transmission by intradomicile R. prolixus, novel native species are emerging, increasing the risk of disease. Triatoma dimidiata is the second most important species in Colombia, and conventional control methods seem to be less effective. In this study we evaluated the efficacy and usefulness of micro-encapsulated insecticide paints in laboratory conditions and its applicability in rural communities to avoid triatomine domiciliation. Laboratory conditions measured mortality at 6 months and 12 months, with an average mortality between 93­100% for T. dimidiata and 100% for R. prolixus. Evaluation of triatomine infestation in rural households was measured after one year, with an overall perception of effectiveness in reducing household domiciliation. Although triatomines were still spotted inside and around the homes, our findings demonstrate the ability of micro-encapsulated insecticide to prevent colonization inside the households when comparing infestation rates from previous years. Current control measures suggest insecticide spraying every six months, which implies great economic cost and logistical effort. Complementary triatomine control measures with insecticide spraying and micro-encapsulated insecticide paint would make public health efforts more efficient and reduce the frequency of treatment.

Monitoramento da suscetibilidade de populações de Rhodnius neglectus (Lent, 1954) a inseticidas utilizados em ações de controle em áreas urbanas do noroeste do estado de São Paulo / Monitoring Rhodnius neglectus (Lent, 1954) populations' susceptibility to insecticide used in controlling actions in urban areas northwest of São Paulo state

Background: Chagas disease (CD) is caused by the flagellate protozoan Trypanosoma cruzi and can be carried by different species of triatomines, including Rhodnius neglectus, which is wild, well distributed in Brazil, and has formed colonies in palm trees located in urban areas of municipalities in the state of São Paulo. Chemical control has been routinely used to reduce population density, but each year, there has been an increase in species dispersion and density. This study aimed to evaluate the susceptibility of insects to insecticides used in control. Methods: The reference population was collected from Araçatuba municipality, Nilce Maia. Dilutions of deltamethrin were prepared and applied to the back of the first-stage nymphs, which were biologically synchronized. The control group received pure acetone only. Mortality was assessed after 72 h. Results: The mortality rate with respect to diagnostic dose was 100%. The susceptibility profile observed for this population showed RR50 ranging from 1.76 to 3.632. Conclusions: The populations were susceptible to the insecticides tested. It is possible that the insecticide residual effect on this ecotope has decreased the lifespan, and controlling failures may be the cause of recolonization in this environment.

Descrição do comportamento sexual de Rhodnius prolixus Stål, 1859 (Hemiptera, Reduviidae) / Description of the sexual behavior of Rhodnius prolixus Stål, 1859 (Hemiptera, Reduviidae)

O estudo de insetos vetores tem fomentado pesquisas sobre sua cópula, reprodução e comportamentos, uma vez que esta informação está diretamente relacionada com a dispersão e colonização potencial desses animais. O presente trabalho teve como objetivo estudar o comportamento sexual de Rhodnius prolixus Stål, 1859, vetor de Trypanosoma cruzi (Chagas, 1909), agente etiológico da doença de Chagas. Foram utilizados 23 casais, alimentados regularmente, obtidos do insetário do Laboratório Interdisciplinar de Vigilância Entomológica em Diptera e Hemiptera. Por meio de observação direta, os principais comportamentos, como rejeição da fêmea à cópula, as tentativas do macho, côrte, cópula e comportamento de guarda foram identificados. Em média, a cópula de R. prolixus dura 40 minutos. O macho foi sempre o primeiro a cortejar sua parceira, que pode aceitar ou não sua investida. A fêmea pode rejeitar o macho ao estridular, se evadir, movimentar o abdômen ou comprimi-lo no substrato. A maioria das fêmeas aceitou ser guardada após a finalização da cópula. Com o presente estudo pudemos concluir que o comportamento sexual de R. prolixus é similar ao de outras espécies do mesmo gênero, e de outras espécies da subfamília Triatominae

The study of insect vectors has fostered research on their copulation and reproduction behaviors, since this information is directly related to the dispersion and colonizing potential of these animals. The present work aimed to study the sexual behavior of Rhodnius prolixus, vector of Trypanosoma cruzi (Chagas, 1909), etiological agent of Chagas disease. Twenty three couples were used, fed regularly, obtained from the insectary of the Interdisciplinary Laboratory of Entomological Surveillance in Diptera and Hemiptera. Through direct observation, the main types of female rejection at copulation, the male's attempts to mate with the same partner, courtship, copulation and guarding behavior were identified. The copulation time of R. prolixus was also registered. The mating time of R. prolixus was also recorded. On average, the copula of R. prolixus lasted 40 minutes. The male has always been the first to court his partner, who may or not accept his advances. The female could reject the male by stridulating, evading, moving her abdomen or pressing him into the substrate. Most deliveries accepted will be kept after the copulation is completed. With the present study we were able to conclude that the sexual behavior of R. prolixus is similar to that of other species of the same genus, and of other species of the Triatominae subfamily

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.

A successful community-base pilot programme to control insect vectors of Chagas disease in rural Guatemala

The adoption of novel integrated vector management (IVM) strategies requires proof-of-concept demonstrations. To implement a community-based intervention, for the control of vectors of Chagas disease in Guatemala, we engaged all relevant stakeholder groups. Based on this and previous experiences of the authors on engaged research and community-based interventions, several key factors can help facilitate effective integration of stakeholders in support of area-wide integrated vector management (AW - IVM) programmes. First and foremost, the diversity of stakeholders needs to be engaged early-on in the participatory action research and implementation processes, to provide ownership and contribute ideas on how to design and implement an intervention. Another important component, situational analysis regarding current pest control policies, practices and relevant stakeholders, is generated through interviews with key informants, at both national and local levels (governmental and non-governmental organizations); it can facilitate the joint identification of strengths, weaknesses, opportunities and threats regarding current pest control strategies and proposing solutions through an AW-IVM approach. In addition, successful AW - IVM can result from identifying locally relevant strategies to implement the proof-of-concept demonstrative project. Further, it is critical to maintain constant communication with the local and national leaders, involving them throughout the implementation and evaluation processes. Flexibility should also be built into the project to allow for community-driven changes in the strategy, through a cyclical joint reflective process. Periodic feedback of project development needs to be scheduled with key stakeholders to maintain rapport. Finally, the results of the evaluation should be shared and discussed with stakeholders to ensure long-term sustainability of the programme, intervention, or project. Here we present the citizen engagement procedures used to integrate community members, health officials, and non-governmental organization staff for Chagas disease control in a region of Guatemala. We demonstrate how these methods can be applied to support AW-IVM programmes, so that communities and authorities are actively involved in the development and implementation of a jointly agreed intervention. In 2012, we developed the IVM intervention in an area of Guatemala with persistentTriatoma dimidiata (Latreille) infestation that is associated with the presence of infected rodents (rats and mice), that act as reservoirs of the Trypanosoma cruzi Chagas parasites inside the households. Nine control communities received only the Ministry of Health insecticide application against the vector and nine intervention communities participated in the AW-IVM intervention. The intervention included a programme for rodent control by the community members, together with education about the risk factors for vector infestation, and insecticide application by the Ministry of Health. Entomological evaluations in 2014 and 2015 showed that vector infestation remained significantly lower in both intervention and control communities. In 2015, we found that there was a higher acceptance of vector surveillance activities in the intervention communities compared to control communities, suggesting that participatory activities increase programme sustainability. Finally, we found that there was a significant increase over time in the number of households with infected vectors in the control group, whereas there was no significant increase in the communities that participated in the programme. Thus, an AW-IVM programme including simultaneous rodent and vector control could reduce the risk of Chagas infection in communities with persistent vector infestation.

Los patrones electroforéticos de proteínas salivales permiten diferenciar los grupos transandino y cisandino de las especies de Rhodnius de Colombia / Salivary proteins electrophoretic patterns enabled differentiating Colombian Rhodnius Trans-Andean and Cis-Andean groups

Introducción. Las especies Rhodnius (Hemiptera: Reduviidae: Triatominae) están conformadas por insectos hematófagos vectores de Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, y T. rangeli, parásito infectivo pero no patógeno para el vertebrado. El estudio de la diversidad proteica de la saliva de estos insectos permite la obtención de perfiles electroforéticos unidimensionales característicos de algunas especies de triatominos. Sin embargo, el reporte de los patrones electroforéticos de proteínas salivales de las especies de Rhodnius ha sido escaso. Objetivo. Hacer un análisis comparativo de los perfiles electroforéticos unidimensionales de las proteínas salivales de R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus. Materiales y métodos. Se obtuvieron los perfiles electroforéticos de la saliva de las especies en estudio mediante electroforesis en gel de poliacrilamida con dodecilsulfato sódico (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, SDS-PAGE) y se construyó un fenograma mediante el método UPGMA (Unweighted Pair Group Method Using Arithmetic Averages). Resultados. Los perfiles electroforéticos de las proteínas solubles de saliva presentaron bandas en un rango de masa aproximado de 15 a 45 kDa, los cuales permitieron diferenciar las cinco especies estudiadas. El fenograma reveló la existencia de dos grupos principales: uno conformado por los grupos cisandinos Pictipes y Prolixus y otro constituido por el grupo transandino Pallescens. Conclusiones. Existen diferencias en los perfiles electroforéticos de las proteínas salivales entre R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus, cuya variabilidad permitió construir un fenograma congruente con los grupos del género Rhodnius.

Introduction: Rhodnius (Hemiptera: Reduviidae: Triatominae) species are made up of haematophagous insect vectors for Trypanosoma cruzi (Chagas' disease aetiological agent) and T. rangeli, an infective parasite that is not pathogenic for vertebrate hosts. The study of their salivary protein diversity enables the obtention of characteristic one-dimensional electrophoretic profiles of some triatomine species; however, few reports have dealt with Rhodnius species salivary proteins electrophoretic patterns. Objective: To compare R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus' salivary proteins one-dimensional electrophoretic profiles. Materials and methods: SDS-PAGE was used for obtaining electrophoretic profiles of saliva from the species under study. The unweighted pair group method with arithmetic mean (UPGMA) was used for constructing a phenogram. Results: Electrophoretic profiles of soluble saliva had protein bands ranging from 15 to 45 kDa, thereby enabling the five species studied to be differentiated. The phenogram revealed two main groups, one formed by the Pictipes and Prolixus cis-Andean groups and another consisting of the Pallescens trans-Andean group. Conclusion: Differences were revealed regarding R. colombiensis, R. pallescens, R. pictipes, R. prolixus, and R. robustus electrophoretic profiles of salivary proteins; their variability facilitated constructing a phenogram which was taxonomically congruent with the groups from the genus Rhodnius.

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.

Efficacy of a topical combination of fipronil-permethrin against Rhodnius prolixus on dogs

A controlled clinical trial was carried out to assess the mortality and repellency of a new topical combination of f ipronil-permethrin (Effitix® Virbac, Mexico) against Rhodnius prolixus in dogs. Ten medium-size dogs (10−15kg) with short hair were used. The dogs were exposed to 8 adult triatomines once weekly for 7 weeks. On the control day (D0), the dogs were exposed to the insects without treatment. On D7, the dogs were immediately treated with a spot-on 2.2ml pipette containing 134mg of fipronil and 1200mg permethrin after exposure to the insects. The dose was repeated after 4 weeks following the manufacturer's instructions. Repellency at D0 was, 0 % and the insects had a high blood content. After 12h post-contact, repellency was 86.3 % and slowly decrease though D21 and D28. On D7, none of the insects survived after 3h of feeding on the treated dogs. On D14, D35 and D42, all insects died within 12h post-feeding, whereas no mortality was observed in the control D0 (P < 0.05). The results of this study indicated that administration of the product following the manufacturer's instructions was efficacious at inducing rapid mortality of R. prolixus and therefore could be useful to prevent the transmission of American trypanosomiasis in dogs.

Studies on the biology and control of the Chagas disease vectors in Guatemala and their control in the Central America

The main vectors of the Chagas disease in the Central America were Triatoma dimidiata, T. nitida and Rhodnius prolixus. The former two species inhabited cracks and crevice on the mud walls, especially closed to beds, while R. prolixus lived only inside thatched roofs, especially above beds. The parasitic rate of the protozoa, Tripanosoma cruzi, was 20% or higher in the vector and human during investigation periods (1994‒1999) in Guatemala. The vectors distributed over 21 departments (=prefectures) out of 22 in Guatemala; however, the vectors could not be collected over 1600 m above sea level. In the nine departments located in the south-west region, the infestation indexes were high. T. cruzi was found serologically positive in the blood of the domestic or wild animals such as chickens, dogs, cats, opossums and cows; whereas, most in human blood. The number of persons at risk were estimated approximately 330,000 according to the vector infestation index, natural infection of T. cruzi, risk houses in each department in 1994 National CENSUS in Guatemala. The vectors were highly susceptible to pyrethroid insecticides. The nine departments were selected for vector control project from 2000 in Guatemala. After spraying pyrethroid insecticides, R. prolixus was dramatically reduced and could not be recovered again. T. dimidiata required more than two time treatments to the walls. The vector control program established in Guatemala has spread to El Salvador, Honduras and Nicaragua in the Central America from 2003. R. prolixus has been eliminated from the Central America by 2009, while T. dimidiata remained in some area after insecticide spraying.

First report of Rhodnius neglectus (Hemiptera, Reduviidae, Triatominae) from the State of Acre, Brazil, and the Brazilian Western Amazon Region

Abstract INTRODUCTION This communication reports the occurrence of Rhodnius neglectus in the State of Acre, Brazil. METHODS An adult male R. neglectus specimen was collected from the Catuaba Experimental Reserve, which is located in the municipality of Senador Guiomard. RESULTS This increases the number of triatomine species reported from the State of Acre to eight and is also the first report of R. neglectus from the Brazilian Western Amazon Region. CONCLUSIONS The occurrence of R. neglectus is alarming because even though the species is wild, it can invade and colonize human dwellings and peridomiciles, thereby increasing Chagas disease transmission.

Sympatry influence in the interaction of Trypanosoma cruzi with triatomine

Abstract INTRODUCTION: Trypanosoma cruzi, the etiologic agent of Chagas disease, is widely distributed in nature, circulating between triatomine bugs and sylvatic mammals, and has large genetic diversity. Both the vector species and the genetic lineages of T. cruzi present a varied geographical distribution. This study aimed to verify the influence of sympatry in the interaction of T. cruzi with triatomines. Methods: The behavior of the strains PR2256 (T. cruzi II) and AM14 (T. cruzi IV) was studied in Triatoma sordida (TS) and Rhodnius robustus (RR). Eleven fifth-stage nymphs were fed by artificial xenodiagnosis with 5.6 × 103 blood trypomastigotes/0.1mL of each T. cruzi strain. Every 20 days, their excreta were examined for up to 100 days, and every 30 days, the intestinal content was examined for up to 120 days, by parasitological (fresh examination and differential count with Giemsa-stained smears) and molecular (PCR) methods. Rates of infectivity, metacyclogenesis and mortality, and mean number of parasites per insect and of excreted parasites were determined. RESULTS: Sympatric groups RR+AM14 and TS+PR2256 showed higher values of the four parameters, except for mortality rate, which was higher (27.3%) in the TS+AM14 group. General infectivity was 72.7%, which was mainly proven by PCR, showing the following decreasing order: RR+AM14 (100%), TS+PR2256 (81.8%), RR+PR2256 (72.7%) and TS+AM14 (36.4%). CONCLUSIONS: Our working hypothesis was confirmed once higher infectivity and vector capacity (flagellate production and elimination of infective metacyclic forms) were recorded in the groups that contained sympatric T. cruzi lineages and triatomine species.

Blood-feeding of Rhodnius prolixus

Triatomines (Hemiptera: Reduviidae) are blood-sucking insect vectors of the protozoan Trypanosoma cruzi which is the causative agent of Chagas' disease. Rhodnius prolixus is the most epidemiologically important vector of T. cruzi in Colombia. Triatomines are regarded to be vessel-feeders as they obtain their blood meals from vertebrate hosts by directly inserting their mouthparts into vessels. Microscopic techniques are useful for visualizing and describing the morphology of biological structures. Here, we show images of the blood-feeding of R. prolixus, including some histological features by light microscopy and scanning electron microscopy of the mouthparts of R. prolixus when feeding on a laboratory mouse.

Los triatominos (Hemiptera: Reduviidae) son insectos hematófagos vectores del protozooTrypanosoma cruzi, el cual causa la enfermedad de Chagas. Rhodnius prolixus es el vector de T. cruzi de mayor importancia epidemiológica en Colombia. Para alimentarse, los triatominos introducen su probóscide directamente en los vasos sanguíneos de los huéspedes vertebrados. La microscopía es una técnica útil para visualizar y describir la morfología de estructuras biológicas. Se presentan imágenes de la hematofagia de R. prolixus, incluidas algunas características histológicas visibles por microscopía de luz y microscopía electrónica de barrido de las partes bucales de R. prolixus al alimentarse de un ratón de laboratorio.

Lethal and sublethal effects of essential oil of Lippia sidoides (Verbenaceae) and monoterpenes on Chagas´ disease vector Rhodnius prolixus

The aim of this study was to identify the composition of the essential oil from leaves of Lippia sidoides (EOLS), a typical shrub commonly found in the dry northeast of Brazil, popularly known as “alecrim-pimenta”. Additionally, we investigated the nymphicidal, ovicidal, phagoinhibitory and excretion effects of EOLS, its major constituent thymol and its isomer carvacrol, on fourth instar nymphs and eggs of Rhodnius prolixus, the Chagas’ disease vector. The nymphicidal and ovicidal activity of thymol, carvacrol, and EOLS was assessed by tests using impregnated Petri dishes. The lethal concentration values (LC50) for EOLS, carvacrol, and thymol were 54.48, 32.98, and 9.38 mg/cm2, respectively. The ovicidal test showed that both carvacrol and thymol (50 mg/cm2) inhibited hatching (50% and 23.3%, respectively), while treatments with 10 mg/cm2 or 50 mg/cm2 EOLS did not affect the hatching rate at all (80% and 90%, respectively). We observed an anti-feeding effect in insects fed with blood containing natural products at the higher concentrations (100 µg/mL). Finally, excretion rate was affected by EOLS and carvacrol, but not by thymol. These findings offer novel insights into basic physiological processes that make the tested natural compounds interesting candidates for new types of insecticides.

Behavioral fever response in Rhodnius prolixus (Reduviidae: Triatominae) to intracoelomic inoculation of Trypanosoma cruzi

Abstract: INTRODUCTION: Behavioral fever is a response to infections with microorganisms observed in some poikilothermic animals. Rhodnius prolixus is involved in the transmission of two parasites: Trypanosoma cruzi (pathogenic for humans and transmitted in feces) and Trypanosoma rangeli (non-pathogenic for humans, pathogenic for Rhodnius and transmitted by the bite of an infected individual). Only T. rangeli is found in the hemolymph of Rhodnius as it travels to the salivary glands. METHODS: To study vector-parasite interactions, we evaluated possible behavioral fever responses of R. prolixus to intracoelomic inoculation with T. cruzi or T. rangeli. Temperature preferences of fifth-instar nymphs of R. prolixus were evaluated after inoculation with T. rangeli KP1(+), KP1(-), T. cruzi I, or the Trypanosome culture medium. Four different fixed temperatures (25, 30, 35, and 40°C) in two simultaneous experiments (enclosed and free-moving insects) were evaluated. Free-moving insects were marked daily according to their temperature preferences on each of the 15 days after inoculation. Numbers of insects in each temperature shelter and daily mortality were compared with those enclosed shelters of different temperatures. RESULTS: Rhodnius prolixus inoculated with both strains of T. rangeli and with the trypanosome culture medium showed preferences for the lowest temperatures (25°C). However, R. prolixus inoculated with T. cruzi I showed significant preferences for temperatures around 35°C. CONCLUSIONS: This is the first known investigation to demonstrate a behavioral fever response in R. prolixus injected intracoelomically with T. cruzi I.

Estrategia para determinar la línea base en áreas de interrupción vectorial de la enfermedad de Chagas / Strategy for determining a baseline in areas of vector interruption for Chagas disease

RESUMEN Objetivos Presentar una estrategia para determinar la línea de base en áreas endémicas en el proceso de interrupción vectorial de la enfermedad de Chagas (ECh). Métodos Con un cuestionario socioambiental y una encuesta entomológica, se evaluaron las condiciones físicas de los domicilios, el conocimiento de los habitantes sobre la ECh, los indicadores entomológicos de triatominos y la relación estadística entre estas variables. Resultados Existe colonización e infección natural con Trypanosoma cruzi en Rhodnius prolixus, el principal vector de la ECh en Colombia. La colonización estuvo relacionada con casas de techos de hojas de palma y paredes de adobe o bahareque. Se encontró el vector Panstrongylus geniculatus con hábitos de colonización. Casi 50% de la población encuestada asociaba el término ECh con una enfermedad humana y 37%, con los triatominos. Conclusiones R. prolixus puede considerarse el principal vector de T. cruzi en ambientes domésticos y el proceso de interrupción vectorial es factible dentro del municipio priorizado. Se necesitan nuevos estudios que comprueben la existencia de poblaciones silvestres de R. prolixus que puedan afectar las etapas futuras del proceso y demostrar si hay una participación de P. geniculatus en la dinámica de transmisión de T. cruzi. Los anteriores escenarios pueden ser viabilizados con la inclusión de los habitantes en todo el proceso, ya que han sido altamente sensibles en la detección de vectores dentro de sus casas. El diseño del estudio aquí presentado puede ser adaptado en otras áreas endémicas de la Región de las Américas.

ABSTRACT Objectives Present a strategy to determine the baseline in endemic areas in the process of vector interruption for Chagas disease (CHD). Methods A social and environmental questionnaire and an entomological survey evaluated the physical conditions of dwellings, the inhabitants’ knowledge of CHD, the entomological triatomine indicators and the statistical relationship among these variables. Results Colonization and natural infection with Trypanosoma cruzi exist in Rhodnius prolixus, the principal vector of CHD in Colombia. Colonization was related to palm-thatched houses constructed with adobe or wattle and daub. The Panstrongylus geniculatus vector was found to be colonizing. Almost 50% of the surveyed population associated the term CHD with human disease and 37%, with triatomines. Conclusions R. prolixus can be considered to be the principal vector of T. cruzi in domestic environments and the process of interruption is feasible within the prioritized municipality. New studies are needed to verify the existence of wild populations of R. prolixus that could affect future stages of the process and demonstrate whether P. geniculatus is a factor in the transmission of T. cruzi. These scenarios can be made viable by including the inhabitants throughout the process, since they have been highly sensitive in vector detection inside their houses. The study design presented here can be adapted to other endemic areas of the Region of the Americas.