Prolixus Nymphs Research Articles

Sugar feeding in triatomines: a new perspective for controlling the transmission of Chagas disease.

Introduction: Triatomines are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. Currently, there is no vaccine against this disease. Thus, control of the insect vector population is the main strategy available to reduce the number of cases. Triatomines are considered obligate hematophagous, but different alternative feeding behaviors were described, such as haemolymphagy or plant feeding. Methods: To determine the preference for sugar feeding in nymphs and adults of Rhodnius prolixus, the insects were exposed a piece of cotton containing bromophenol blue plus sucrose. In addition, we offered several sugars for different species of triatomines, and tested sugar meals as a route of delivery of insecticides in first-instar nymphs of R. prolixus. The effect of sugar feeding on the physiology of these different species of triatomines was recorded. Results: First instar nymphs ingested sucrose more strongly than other stages, and showed high mortality rates. In different species of triatomines, sucrose induced an ingestion, but engorgement varied according to the species. R. prolixus nymphs showed an indiscriminate intake of various sugars, with very different physiological effects. Furthermore, ingesting different combinations of insecticides + sugar significantly reduced insect survival. Discussion: In summary, we described for the first-time sugar feeding as a widespread behavior in several species of triatomines, and the possibility of the use of toxic sugar baits for the control of these vectors. The knowledge of feeding behavior in these insects can be fundamental for the development of new strategies to control Chagas disease.

The Rpfor gene modulates the locomotory activity and host-seeking behaviour of Rhodnius prolixus.

The molecular bases of animal behaviour are intricate due to the pleiotropic nature of behaviour-modulating genes, which are often expressed across multiple tissues. The foraging gene (for) encodes a cGMP-dependent protein kinase (PKG), pivotal in regulating downstream target proteins through phosphorylation. In insects, for has been implicated in various behavioural contexts and physiological processes regarding searching for food. Rhodnius prolixus, a hematophagous bug that transmits Trypanosoma cruzi, the causative agent of Chagas disease, exhibits specific activity patterns associated with its hematophagous behaviour. Our previous work demonstrated a correlation between locomotor activity profiles and the expression of Rpfor, suggesting its involvement in modulating triatomine locomotion. In this study, we investigated the impact of Rpfor knockdown on locomotory activity, host-seeking behaviour, feeding performance and lipid metabolism in R. prolixus nymphs. Using RNA interference, we achieved a significant reduction of Rpfor expression in both the brain and fat body of R. prolixus nymphs. Knocked-down nymphs exhibited diminished non-oriented locomotory activity compared with controls, without altering the characteristic bimodal pattern of activity. Additionally, they displayed an increased tendency to approach a host, suggesting a role for Rpfor in modulating host-seeking behaviour. Feeding performance and lipid metabolism remained unaffected by Rpfor knockdown. Our findings underscore the multifaceted role of Rpfor in modulating locomotor activity and host-seeking behaviour in R. prolixus nymphs, shedding light on the molecular mechanisms underlying their hematophagous behaviour and potential implications for disease transmission. Further research is necessary to elucidate the intricate interplay between Rpfor expression, behaviour and physiological processes in triatomine bugs.

Alterations in energy metabolism of Rhodnius prolixus induced by Trypanosoma rangeli infection

Trypanosoma rangeli is a protozoan parasite that infects triatomines and mammals in the Americas, producing mixed infections with Trypanosoma cruzi, the etiological agent of Chagas disease. The former parasite is not pathogenic to humans, but has different levels of pathogenicity, as well as causing physiological and behavioral alterations, to its invertebrate hosts. In this study, we measured locomotory activity, and the glyceride accumulation profile in the hemolymph and fat body, as well as the expression of key genes related to triglyceride metabolism, of Rhodnius prolixus nymphs infected with T. rangeli. We found that the locomotory activity of the insects was correlated with the amount of triglycerides in the fat body. Infected nymphs had increased activity when starved, and also had an accumulation of glycerides in the fat body and hemolymph. These alterations were also associated with a higher expression of the diacylglycerol acyltransferase, lipophorin and lipophorin receptor genes in the fat body. We infer that T. rangeli is able to alter the energetic processes of its invertebrate host, in order to increase the availability of lipids to the parasite, which, in turn modifies the activity levels of the insect. These alterations are discussed with regard to their potential to increase the transmission rate of the parasite.

Global characterization of gene expression in the brain of starved immature Rhodnius prolixus.

Rhodnius prolixus is a vector of Chagas disease and has become a model organism to study physiology, behavior, and pathogen interaction. The publication of its genome allowed initiating a process of comparative characterization of the gene expression profiles of diverse organs exposed to varying conditions. Brain processes control the expression of behavior and, as such, mediate immediate adjustment to a changing environment, allowing organisms to maximize their chances to survive and reproduce. The expression of fundamental behavioral processes like feeding requires fine control in triatomines because they obtain their blood meals from potential predators. Therefore, the characterization of gene expression profiles of key components modulating behavior in brain processes, like those of neuropeptide precursors and their receptors, seems fundamental. Here we study global gene expression profiles in the brain of starved R. prolixus fifth instar nymphs by means of RNA sequencing (RNA-Seq). The expression of neuromodulatory genes such as those of precursors of neuropeptides, neurohormones, and their receptors; as well as the enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines were fully characterized. Other important gene targets such as neurotransmitter receptors, nuclear receptors, clock genes, sensory receptors, and takeouts genes were identified and their gene expression analyzed. We propose that the set of neuromodulatory-related genes highly expressed in the brain of starved R. prolixus nymphs deserves functional characterization to allow the subsequent development of tools targeting them for bug control. As the brain is a complex structure that presents functionally specialized areas, future studies should focus on characterizing gene expression profiles in target areas, e.g. mushroom bodies, to complement our current knowledge.

Trypanosoma rangeli infection increases the exposure and predation endured by Rhodnius prolixus.

Trypanosoma rangeli is a protozoan that infects triatomines and mammals in Latin America, sharing hosts with Trypanosoma cruzi, the etiological agent of Chagas disease. Trypanosoma rangeli does not cause disease to humans but is strongly pathogenic to its invertebrate hosts, increasing mortality rates and affecting bug development and reproductive success. We have previously shown that this parasite is also capable of inducing a general increase in the locomotory activity of its vector Rhodnius prolixus in the absence of host cues. In this work, we have evaluated whether infection impacts the insect–vertebrate host interaction. For this, T. rangeli-infected and uninfected R. prolixus nymphs were released in glass arenas offering single shelters. After a 3-day acclimatization, a caged mouse was introduced in each arena and shelter use and predation rates were evaluated. Trypanosoma rangeli infection affected all parameters analysed. A larger number of infected bugs was found outside shelters, both in the absence and presence of a host. Infected bugs also endured greater predation rates, probably because of an increased number of individuals that attempted to feed. Interestingly, mice that predated on infected bugs did not develop T. rangeli infection, suggesting that the oral route is not effective for these parasites, at least in our system. Finally, a smaller number of infected bugs succeeded in feeding in this context. We suggest that, although T. rangeli is not transmitted orally, an increase in the proportion of foraging individuals would promote greater parasite transmission rates through an increased frequency of very effective infected-bug bites.

Effect of salivary CYP4EM1 and CYP4EM2 gene silencing on the life span of Chagas disease vector Rhodnius prolixus (Hemiptera, Reduviidae) exposed to sublethal dose of deltamethrin.

Control of Chagas disease in endemic countries is primarily accomplished through insecticide spraying for triatomine vectors. In this context, pyrethroids are the first-choice insecticide, and the evolution of insect resistance to these insecticides may represent an important barrier to triatomine control. In insects, cytochrome P450s are enzymes involved in the metabolism of xenobiotics and endogenous chemicals that are encoded by genes divided into different families. In this work, we evaluated the role of three Rhodnius prolixus CYP4EM subfamily genes during blood meal and after deltamethrin exposure. CYP4 gene members were expressed in different insect organs (integument, salivary glands (SGs), midgut, fat body and malpighian tubules) at distinct transcriptional levels. CYP4EM1 gene was highly expressed in the SG and was clearly modulated after insect blood meal. Injection of CYP4EM1dsRNA promoted significant reduction in mRNA levels of both CYP4EM1 and CYP4EM2 genes and induced deleterious effects in R. prolixus nymphs subsequently exposed to sublethal doses of deltamethrin (3.4 or 3.8 ng/nymph treated). The higher dose reduced the survival over time and increased susceptibility of R. prolixus nymphs to deltamethrin. A better understanding of this mechanism can help in developing of more efficient strategies to reduce Trypanosoma cruzi vector transmission in Americas.

Nitric oxide effects on Rhodnius prolixus’s immune responses, gut microbiota and Trypanosoma cruzi development

The immune system of Rhodnius prolixus comprehends the synthesis of different effectors that modulate the intestinal microbiota population and the life cycle of the parasite Trypanosoma cruzi inside the vector midgut. One of these immune responses is the production of reactive nitrogen species (RNS) derived by the action of nitric oxide synthase (NOS). Therefore, we investigated the effects of L-arginine, the substrate for nitric oxide (NO) production and Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME), an inhibitor of NOS, added in the insect blood meal. We analyzed the impact of these treatments on the immune responses and development of intestinal bacteria and parasites on R. prolixus nymphs. The L-arginine treatment in R. prolixus nymphs induced a higher NOS gene expression in the fat body and increased NO production, but reduced catalase and antimicrobial activities in the midgut. As expected, L-NAME treatment reduced NOS gene expression in the fat body. In addition, L-NAME treatment diminished catalase activity in the hemolymph and posterior midgut reduced phenoloxidase activity in the anterior midgut and increased the antimicrobial activity in the hemolymph. Both treatments caused a reduction in the cultivatable intestinal microbiota, especially in insects treated with L-NAME. However, T. cruzi development in the insect’s digestive tract was suppressed after L-arginine treatment and the opposite was observed with L-NAME, which resulted in higher parasite counts. Therefore, we conclude that induction and inhibition of NOS and NO production are associated with other R. prolixus humoral immune responses, such as catalase, phenoloxidase, and antibacterial activities in different insect organs. These alterations reflect on intestinal microbiota and T. cruzi development.

Deltamethrin resistance in Chagas disease vectors colonizing oil palm plantations: implications for vector control strategies in a public health-agriculture interface

BackgroundTriatomine bugs are responsible for the vectorial transmission of the parasite Trypanosoma cruzi, etiological agent of Chagas disease, a zoonosis affecting 10 million people and with 25 million at risk of infection. Several triatomine species of the genus Rhodnius have been found inhabiting palm crowns where insects can find shelter in leaves axils and blood from palm-associated vertebrates. Rhodnius prolixus insects have been collected in oil palms in Colombia, and high T. cruzi infection rates were found. Since pest control is carried out in oil palm plantations, continuous exposure to insecticides could be occurring in these triatomines. Some insecticides suggested for pest control in oil palm plantations are also recommended for triatomine control in human dwellings. In this study, our objective was to assess if triatomines inhabiting oil palms exhibit resistance to deltamethrin, an insecticide used for vector control.MethodsRhodnius prolixus nymphs were sampled in oil palms located in Tauramena, Colombia. To determine deltamethrin resistance, biological and biochemical assays were carried out on fifth-instar nymphs from the F1 generation. For biological assays, pure and commercial deltamethrin were used, and in biochemical assays, activities of detoxifying enzymes related to pyrethroid resistance, such as oxidases, esterases and transferases, were quantified.ResultsDeltamethrin lethal dosage 50 and 90 in R. prolixus from oil palms was significantly higher than in those from a susceptible colony suggesting possible deltamethrin resistance. Moreover, mortality with commercial deltamethrin was very low in insects from oil palms. In biochemical assays, the activity of evaluated detoxifying enzymes was significantly higher in R. prolixus from oil palms than in those from the susceptible colony.ConclusionsPossible deltamethrin resistance found in R. prolixus insects from oil palms could threaten traditional vector control strategies in urban settings if insecticide-resistant triatomines can migrate from oil palms plantations. In palm oil producer countries such as Colombia, the oil palm plantations are growing constantly during the last years. We suggest that pest control strategies in oil palm crops should include triatomine surveillance and toxicological monitoring, especially in zones with several Chagas disease cases.

Innate trypanolytic factors in triatomine hemolymph against Trypanosoma rangeli and T. cruzi: a comparative study in eight Chagas disease vectors

There is limited information about the innate immunity of triatomines against Trypanosoma rangeli, an infectious, non-pathogenic human parasite, and T. cruzi, the causative agent of Chagas’ disease. This study aimed at addressing this gap by studying the in vitro trypanolytic hemolymph activity from insects not infected by T. rangeli or T. cruzi. Eight triatomine species were examined including Rhodnius prolixus, R. robustus, R. colombiensis, R. pallescens, R. pictipes, Triatoma dimidiata, T. maculata, and Panstrongylus geniculatus. The hemolymph of R. prolixus and R. robustus laboratory colonies demonstrated strong trypanolytic activity during the first 14 hours of parasite incubation with 81% lysis for the T. rangeli C genotype, 90% for the E genotype, 95% for T. cruzi discrete taxonomic unit (DTU) TcII, 94% for TcV, 96% for TcVI, 94% for Tcbat, and 90% for Tcmarinkellei. No lysis activity was detected 14 or 24 hours after parasite incubation with the hemolymph of insects from R. pictipes R. pallescens, P. geniculatus, and T. maculata colonies. Identical results (absence of lysis) were observed using hemolymph from R. colombiensis and T. dimidiata laboratory colonies and specimens captured in sylvatic environments. The hemolymph lytic activity against recently obtained cultures (95% decrease in live parasite count) and long-term T. cruzi TcII ones (96% decrease) was similar. Lytic activity was similar in hemolymph from R. prolixus nymphs, males and females and insects fed on chicken or mouse blood. This is the first comparative study of the trypanolytic activity of hemolymph from different triatomine species against T. rangeli and T. cruzi genotypes. Although the chemical structures of such lysis factors and the mechanisms determining their expression have not been fully determined, their identification furthers our understanding of triatomines’ innate immunity and their role in Trypanosoma transmission.

Green insecticide against Chagas disease: effects of essential oil from Myrciaria floribunda (Myrtaceae) on the development of Rhodnius prolixus nymphs

ABSTRACTThe main strategy of interrupting Trypanossoma cruzi transmission is to control the vector populations. Studies of new agents with activity against these vectors have a priority interest. This study evaluated the insecticidal activity of the essential oil of Myrciaria floribunda against nymphs of R. prolixus. M. floribunda was collected in Jurubatiba Sandbank (southeastern Brazil). Leaves were extracted by hydrodistillation and the essential oil analyzed by mass spectrometry and flame ionization gas chromatography. The pure essential oil at various dilutions or purified 1,8-cineole was topically applied to R. prolixus. Sesquiterpenes were the main group detected followed by monoterpenes. M. floribunda essential oil, but not 1,8-cineole, significantly increased mortality and disrupted metamorphosis. Overaged nymphs and juvenoid-like insects were also observed. Our data indicate the special metabolites in the essential oil of M. floribunda as promising candidates for studying insecticidal activity and for use as environmentally friendly control compounds to Chagas disease vectors.

Generalist host species drive Trypanosoma cruzi vector infection in oil palm plantations in the Orinoco region, Colombia

BackgroundOil palm plantation establishment in Colombia has the potential to impact Chagas disease transmission by increasing the distribution range of Rhodnius prolixus. In fact, previous studies have reported Trypanosoma cruzi natural infection in R. prolixus captured in oil palms (Elaeis guineensis) in the Orinoco region, Colombia. The aim of this study is to understand T. cruzi infection in vectors in oil palm plantations relative to community composition and host dietary specialization by analyzing vector blood meals and comparing these results to vectors captured in a native palm tree species, Attalea butyracea.MethodsRhodnius prolixus nymphs (n = 316) were collected from A. butyracea and E. guineensis palms in Tauramena, Casanare, Colombia. Vector blood meals from these nymphs were determined by amplifying and sequencing a vertebrate-specific 12S rRNA gene fragment.ResultsEighteen vertebrate species were identified and pigs (Sus scrofa) made up the highest proportion of blood meals in both habitats, followed by house mouse (Mus musculus) and opossum (Didelphis marsupialis). Individual bugs feeding only from generalist mammal species had the highest predicted vector infection rate, suggesting that generalist mammalian species are more competent hosts for T. cruzi infection .ConclusionsOil palm plantations and A. butyracea palms found in altered areas provide a similar quality habitat for R. prolixus populations in terms of blood meal availability. Both habitats showed similarities in vector infection rate and potential host species, representing a single T. cruzi transmission scenario at the introduced oil palm plantation and native Attalea palm interface.

Activity and shelter-related behavior in Rhodnius prolixus: The role of host odours

Triatomine bugs are considered nocturnal insects that feed on the blood of vertebrates and remain hidden inside narrow shelters during daylight hours. Nevertheless, it is not clear whether these insects become active and leave their shelters on a daily basis, less frequently or, even fortnightly. Activity patterns were studied in Rhodnius prolixus Stål, 1859 (Hemiptera: Triatominae) associated with shelters to evaluate whether the decision to leave a shelter depends on bug starvation and the presence of host odours. Experiments were conducted with groups of 5th instar nymphs released in an experimental arena offering an artificial shelter consisting of a piece of corrugated cardboard positioned in its centre. Results indicate that host odours promoted a significant increase in shelter related activity, i.e. shelter-leaving or entering movements, and also in bug locomotion. This increase could only be observed with bugs starved for 30 or 60 days, but not for 21 days. Most R. prolixus nymphs that left shelters and engaged in locomotory activity were starved and in the presence of host odours. Even though R. prolixus is mostly considered a very active and “aggressive” triatomine, our results contradict this perspective and suggest that its main strategy regarding hosts is to wait and carefully evaluate feeding chances before becoming exposed. This behavioural strategy might have arisen through their evolution in palm trees in association with a diverse fauna that may impose predation risks.

Acción insecticida de análogos sintéticos de girgensohnina y de aceites esenciales sobre Rhodnius prolixus (Hemiptera: Reduviidae)

Introduction: The alkaloid girgensohnine has been used as a natural model in the synthesis of new alkaloid-like alpha-aminonitriles with insecticidal effect against disease vectors. Objective: To evaluate the biocide activity of girgensohnine analogues and essential oils of Cymbopogon flexuosus , Citrus sinensis and Eucalyptus citriodora in stage I and stage V Rhodnius prolixus nymphs. Materials and methods: We used a topical application model in tergites and sternites, as well as exposure to treated surfaces with different exploratory doses of each of the molecules and essential oils to determine the lethal doses (LD50 and LD95). Results: Analogue 3 showed the highest insecticidal activity with 83.3±16.7% of mortality when applied on tergites, 38.9±4.8% on sternites and 16.7±0% on treated surfaces in stage I nymphs at 72 hours (h) and 500 mg.L-1. In stage V nymphs, the compounds induced mortality only in sternums (11.1±9.6% for analogue 6 and 5.5±4.7% for analogues 3 and 7 at 72 h and 1500 mg.L-1). The lethal doses for molecule 3 on tergites in stage I nymphs were LD50 225.60 mg.L-1 and LD95 955.90 mg.L-1. The insecticidal effect of essential oils was observed only in stage I nymphs, with 11.1±4.8% for C. flexuosus when applied in sternites, while using exposure to surfaces treated it was 5.6±4.8% for C. sinensis applied on tergites and 8.3±0% on sternites at 72 h and 1000 mg.L-1. Conclusion: Synthetic girgensohnine analogues, and C. flexuosus and C. sinensis essential oils showed insecticidal activity in R. prolixus . Analogue 3 showed the greatest insecticidal activity among all molecules and oils evaluated under our laboratory conditions.

Insecticidal action of synthetic girgensohnine analogues and essential oils on Rhodnius prolixus (Hemiptera: Reduviidae)

The alkaloid girgensohnine has been used as a natural model in the synthesis of new alkaloid-like alpha-aminonitriles with insecticidal effect against disease vectors. To evaluate the biocide activity of girgensohnine analogues and essential oils of Cymbopogon flexuosus, Citrus sinensis and Eucalyptus citriodora in stage I and stage V Rhodnius prolixus nymphs. We used a topical application model in tergites and sternites, as well as exposure to treated surfaces with different exploratory doses of each of the molecules and essential oils to determine the lethal doses (LD50 and LD95). Analogue 3 showed the highest insecticidal activity with 83.3±16.7% of mortality when applied on tergites, 38.9±4.8% on sternites and 16.7±0% on treated surfaces in stage I nymphs at 72 hours (h) and 500 mg.L-1. In stage V nymphs, the compounds induced mortality only in sternums (11.1±9.6% for analogue 6 and 5.5±4.7% for analogues 3 and 7 at 72 h and 1500 mg.L-1). The lethal doses for molecule 3 on tergites in stage I nymphs were LD50 225.60 mg.L-1 and LD95 955.90 mg.L-1. The insecticidal effect of essential oils was observed only in stage I nymphs, with 11.1±4.8% for C. flexuosus when applied in sternites, while using exposure to surfaces treated it was 5.6±4.8% for C. sinensis applied on tergites and 8.3±0% on sternites at 72 h and 1000 mg.L-1. Synthetic girgensohnine analogues, and C. flexuosus and C. sinensis essential oils showed insecticidal activity in R. prolixus. Analogue 3 showed the greatest insecticidal activity among all molecules and oils evaluated under our laboratory conditions.

Amines from vertebrates guide triatomine bugs to resources

Most triatomine bugs (Heteroptera: Reduviidae) are nest-living insects that require vertebrate blood or invertebrate haemolymph to complete their life cycle. Vertebrates accumulate excretory products in or near their nesting sites and we hypothesize that triatomines use emanations from such host wastes when searching for resources. Here we recount how triatomine bugs increase upwind locomotion on a servosphere in response to volatile amine constituents of vertebrate excretions. Fresh chicken faeces is strongly attractive to Rhodnius prolixus nymphs. Ammonia induces attraction and an increase in both speed and total path length by R. prolixus on the servosphere. Whereas ethylamine and dimethylamine attract R. prolixus, Triatoma infestans and Panstrongylus geniculatus, other amine constituents of vertebrate excretions such as isobutylamine and hexylamine induce R. prolixus nymphs to walk faster and for a longer period. These amines are derived from generally occurring metabolites of vertebrates and from gut flora metabolism. We conclude that amines and other products associated with nesting hosts serve as signals for foraging triatomines.

Evaluation of a CO2-free commercial mosquito attractant to capture triatomines in the laboratory

Efforts have been made to develop vertebrate odor-based attractants to lure hematophagous triatomines into traps. However, more work is needed to reach a practical, cheap, and efficient odor lure. We carried out attraction and capture tests in a dual-choice olfactometer and a pitfall trap. Here we report that a three-component, CO2 -free, synthetic blend of vertebrate odor (consisting of ammonia, L(+) lactic acid and hexanoic acid, and known as Sweetscent®) significantly induces 3(rd) -instar Rhodnius prolixus and Triatoma infestans nymphs to fall into the test capture-tube of the olfactometer. Blend constituents presented singly or in two-component blends did not evoke a response and, therefore, we propose that the insects respond specifically to the three-component blend in a synergistic way. When tested in a pitfall trap in an experimental arena, this blend induced capture in 37.5% of the lured traps, whereas 9% of the nymphs tested were captured in a single night. No insects were captured in control traps. Our work represents a proof-of-concept regarding capture of triatomines using host odor-based, CO2 -free synthetic mixtures as lures for pitfall traps. CO2 -free lures are more practical for field work than natural or CO2 -containing synthetic blends.

The interaction between Trypanosoma rangeli and the nitrophorins in the salivary glands of the triatomine Rhodnius prolixus (Hemiptera; Reduviidae)

The parasite Trypanosoma rangeli develops in the intestinal tract of triatomines and, particularly in species of the genus Rhodnius, invades the hemolymph and salivary glands, where subsequent metacyclogenesis takes place. Many aspects of the interaction between T. rangeli and triatomines are still unclear, especially concerning the development of the parasite in the salivary glands and how the parasite interacts with the saliva. In this work, we describe new findings on the process of T. rangeli infection of the salivary glands and the impact of infection on the saliva composition. To ensure a complete infection (intestinal tract, hemolymph and salivary glands), 3rd instar Rhodnius prolixus nymphs were fed on blood containing T. rangeli epimastigotes using an artificial feeder. After molt to the 4th instar, the nymphs were inoculated with epimastigotes in the hemolymph. The results showed that the flagellates started to invade the salivary glands by the 7th day after the injection. The percentage of trypomastigotes inside the salivary glands continuously increased until the 25th day, at which time the trypomastigotes were more than 95% of the T. rangeli forms present. The salivary contents from T. rangeli-infected insects showed a pH that was significantly more acidic (<6.0) and had a lower total protein and hemeprotein contents compared with non-infected insects. However, the ratio of hemeprotein to total protein was similar in both control and infected insects. qPCR demonstrated that the expression levels of three housekeeping genes (18S rRNA, β-actin and α-tubulin) and nitrophorins 1–4 were not altered in the salivary glands after an infection with T. rangeli. In addition, the four major nitrophorins (NPs 1–4) were knocked down using RNAi and their suppression impacted T. rangeli survival in the salivary glands to the point that the parasite burden inside the R. prolixus salivary glands was reduced by more than 3-fold. These results indicated that these parasites most likely non-specifically incorporated the proteins that were present in R. prolixus saliva as nutrients, without impairing the biosynthesis of the antihemostatic molecules.

Adaptation of the repellency response to DEET in Rhodnius prolixus

For many years it has been accepted that DEET interferes with the detection of odours from the host instead of having a repellent effect. However, recent work showed that DEET acts as an odorant molecule and elicits a behavioural response in the absence of other stimuli. Therefore, DEET must promote some phenomenon connected with the stimuli–sensory system interaction, such as a sensory adaptation, where the sensory system regulates its sensitivity to different stimuli intensities during continuous or repetitive exposure. In this work, we studied different aspects of the insect–DEET interaction through behavioural observations. Previous exposure of fifth instar Rhodnius prolixus nymphs to DEET decreased the behavioural response to this repellent. We observed a decrease in repellence after different times of continuous stimulation with DEET in a time-dependent manner. The response to DEET was recovered 10 min after exposure, when insects were continuously stimulated during 5 or 10 min; maximum repellence was recovered 20 min after exposure when insects were stimulated for 20 min. DEET produced a repellent effect when nymphs were exposed only to its vapours. These results suggest that exposure to DEET produces adaptation in R. prolixus nymphs, and that the behavioural response elicited by DEET occurs via olfaction when no other stimuli are present.

Laboratory evaluation of the effects of triflumuron on the development of Rhodnius prolixus nymph

Studies were carried out to evaluate the efficacy of the growth regulator, triflumuron (TFM) (Starycide sc 480 Bayer), for disrupting the development of Rhodnius prolixus fifth-instar nymph by oral, topical or continuous treatment. All treatments were able to induce high levels of mortality, delay development and molt inhibition. Oral treatment induced molt inhibition in all insects that survived at doses of 0.25, 0.50 and 5.0 mg/mL of a blood meal. The highest levels of both mortality in 24 h and molt inhibition were always observed after topical treatment. The lowest doses needed to obtain considerable biological effects were always observed after continuous treatment. In this way, the highest levels of mortality within 30 days were detected after continuous treatment, which also induced an extended inter-molting period, a lower number of over-aged nymphs and the highest level of molting in nymphs that survived. Moreover, the effects of TFM on insects were often displayed in a dose response manner. These results indicate that TFM acts as a potent growth inhibitor of R. prolixus nymphs and has the potential to be used in integrated vector control programs against hematophagous triatomine species.

Microclimatic factors influencing refugium suitability for Rhodnius prolixus

Abstract Rehydration in the triatomine bug Rhodnius prolixus (Stål) is dependent on the blood meal, and water balance regulation is crucial for survival of starving bugs. In an experimental arena with zones at different climatic conditions, starved R. prolixus nymphs prefer a cooler and more humid zone, stopping there more often and for longer periods. This is probably a behavioural adaptation to limit water loss and reduce metabolic rate. In the Venezuelan State of Portuguesa, temperature and humidity were monitored in three kinds of potential refugia during the dry season: (i) in a palm roof; (ii) in a crack of the wall of a house; and (iii) in a palm tree crown. Fluctuations in temperature and saturation deficit are not very different inside and outside the palm roof except during a few hours of the day when the sun is at its zenith. In the crack of the wall, the diurnal temperature range is reduced to 6.5 °C compared with 12.4 °C outside, and the saturation deficit varies by only 7.6 hPa compared with 28.6 hPa outside. In the palm tree crown, the daily temperature range is only 4.2 °C compared with 13.8 °C outside, and the saturation deficit is permanently &lt; 5 hPa. The microclimatic conditions in the palm tree crown would appear to be ideal for starving R. prolixus, but this kind of refugium generally harbours low densities of bugs, probably related to a combination of predation, pathogens and lower mean temperature within the crown. Such biotic and abiotic constraints play a lesser role in the less ideal palm roof and wall crack refugia where bugs can proliferate as long as hosts are readily available nearby.