Blood-sucking Bug Research Articles

An antagonist of the neurotransmitter tyramine inhibits the hyperactivating effect of eugenol in the blood-sucking bug, Triatoma infestans

Eugenol is a botanical monoterpene found in the essential oils of several aromatic plants. It has shown to have insecticidal activity, modify insect behavior, and its site of action is most probably in the octopaminergic system. The aim of the present study was to explore whether tyramine receptors are involved in the hyperactivity produced by eugenol in Triatoma infestans, one of the main vectors of Chagas disease. Topical application of tyramine and eugenol increased locomotor activity in third instar nymphs of T. infestans in a dose-dependent way. Yohimbine hydrochloride, a tyramine antagonist, did not modify nymph activity. However, nymphs pretreated with yohimbine and then with tyramine or eugenol showed the same locomotor activity as the controls. Therefore, yohimbine hydrochloride inhibited the hyperactivating effect of both tyramine and eugenol. These results suggest that tyramine receptors are involved in the effect of eugenol on the locomotor activity of T. infestans.

Antimicrobial activity of NK cells to Trypanosoma cruzi infected human primary Keratinocytes.

Infection with the protozoan parasite Trypanosoma cruzi is causative for Chagas disease, which is a highly neglected tropical disease prevalent in Latin America. Humans are primary infected through vectorial transmission by blood-sucking triatomine bugs. The parasite enters the human host through mucous membranes or small skin lesions. Since keratinocytes are the predominant cell type in the epidermis, they play a critical role in detecting disruptions in homeostasis and aiding in pathogen elimination by the immune system in the human skin as alternative antigen-presenting cells. Interestingly, keratinocytes also act as a reservoir for T. cruzi, as the skin has been identified as a major site of persistent infection in mice with chronic Chagas disease. Moreover, there are reports of the emergence of T. cruzi amastigote nests in the skin of immunocompromised individuals who are experiencing reactivation of Chagas disease. This observation implies that the skin may serve as a site for persistent parasite presence during chronic human infection too and underscores the significance of investigating the interactions between T. cruzi and skin cells. Consequently, the primary objective of this study was to establish and characterize the infection kinetics in human primary epidermal keratinocytes (hPEK). Our investigation focused on surface molecules that either facilitated or hindered the activation of natural killer (NK) cells, which play a crucial role in controlling the infection. To simulate the in vivo situation in humans, an autologous co-culture model was developed to examine the interactions between T. cruzi infected keratinocytes and NK cells. We evaluated the degranulation, cytokine production, and cytotoxicity of NK cells in response to the infected keratinocytes. We observed a strong activation of NK cells by infected keratinocytes, despite minimal alterations in the expression of activating or inhibitory ligands on NK cell receptors. However, stimulation with recombinant interferon-gamma (IFN-γ), a cytokine known to be present in significant quantities during chronic T. cruzi infections in the host, resulted in a substantial upregulation of these ligands on primary keratinocytes. Overall, our findings suggest the crucial role of NK cells in controlling acute T. cruzi infection in the upper layer of the skin and shed light on keratinocytes as potential initial targets of infection.

Lessons to be popular: the chemical basis of aggregation in Trypanosoma cruzi-infected and non-infected Chagasic bugs.

Aggregation is one of the most remarkable behaviours in the animal kingdom-a process that is usually governed by pheromones. Triatomines are blood-sucking bugs that act as vectors of Trypanosoma cruzi, the etiological agent of Chagas disease in mammals, including humans. Triatomines usually gather in roosting refuges by using aggregation pheromones of unknown chemical structure. In terms of vector control, one option to reduce triatomine-human contact is via capturing the insects into traps baited with lures based on such aggregation pheromones. As a first step towards this aim, we elucidated the aggregation pheromone in the triatomine Triatoma pallidipennis, using T. cruzi-infected and non-infected bugs. We used different extraction techniques and gas chromatography coupled to mass spectrometry for the identification. Also, two different bioassays were implemented for evaluating the attractant and arrestant activity of the pheromone. We found that T. pallidipennis produced short-chain aldehydes as attractants, and nitrogen-derived compounds as arrestants. We detected differences in the production and perception of these compounds according to whether animals were infected or not. These findings show that T. cruzi may influence triatomine chemical ecology and are promising tools for triatomine control.

Cellphone picture-based, genus-level automated identification of Chagas disease vectors: Effects of picture orientation on the performance of five machine-learning algorithms

Chagas disease (CD) is a public-health concern across Latin America. It is caused by Trypanosoma cruzi, a parasite transmitted by blood-sucking triatomine bugs. Automated identification of triatomine bugs is a potential means to strengthen CD vector surveillance. To be broadly useful, however, automated systems must draw on algorithms capable of correctly identifying bugs from images taken with ordinary cellphone cameras at varying angles or positions. Here, we assess the performance of five machine-learning algorithms at identifying the main CD vector genera (Triatoma, Panstrongylus, and Rhodnius) based on bugs photographed at different angles/positions with a 72-dpi cellphone camera. Each bug (N = 730; 13 species) was photographed at nine angles representing three positions: dorsal-flat, dorsal-oblique, and front/back-oblique. We randomly split the 6570-picture database into training (80%) and testing sets (20%), and then trained and tested a convolutional neural network (AlexNet, AN); three boosting-based classifiers (AdaBoost, AB; Gradient Boosting, GB; and Histogram-based Gradient Boosting, HB); and a linear discriminant model (LD). We assessed identification accuracy and specificity with logit-binomial generalized linear mixed models fit in a Bayesian framework. Differences in performance across algorithms were mainly driven by AN's essentially perfect accuracy and specificity, irrespective of picture angle or bug position. HB predicted accuracies ranged from ∼0.987 (Panstrongylus, dorsal-oblique) to >0.999 (Triatoma, dorsal-flat). AB accuracy was poor for Rhodnius (∼0.224–0.282) and Panstrongylus (∼0.664–0.729), but high for Triatoma (∼0.988–0.991). For Panstrongylus, LD and GB had predicted accuracies in the ∼0.970–0.984 range. AB misclassified ∼57% of Rhodnius and Panstrongylus as Triatoma, whereas specificity ranged from ∼0.92 to ∼1.0 for the remaining algorithm-genus combinations. Dorsal-flat pictures appeared to improve algorithm performance slightly, but angle/position effects were overall weak-to-negligible. We conclude that, when high-performance algorithms such as AN are used, the angles or positions at which bugs are photographed seem unlikely to hinder cellphone picture-based automated identification of CD vectors, at least at the genus level. Future research should focus on combining mixed-quality pictures and state-of-the-art algorithms to (i) identify triatomine adults to the species level and (ii) distinguish triatomine nymphs (i.e., immature stages) from adults and from other insects.

Evidence that hematophagous triatomine bugs may eat plants in the wild

Blood feeding is a secondary adaptation in hematophagous bugs. Many proteins are secreted in the saliva that are devoted to coping with the host's defense and to process the blood meal. Digestive enzymes that are no longer required for a blood meal would be expected to be eventually lost. Yet, in many strictly hematophagous arthropods, α-amylase genes, which encode the enzymes that digest starch from plants, are still present and transcribed, including in the kissing bug Rhodnius prolixus (Hemiptera, Reduviidae) and its related species, which transmit the Chagas disease. We hypothesized that retaining α-amylase could be advantageous if the bugs occasionally consume plant tissues. We first checked that the α-amylase protein of Rhodnius robustus retains normal amylolytic activity. Then we surveyed hundreds of gut DNA extracts from the sylvatic R. robustus to detect traces of plants. We found plant DNA in 8% of the samples, mainly identified as Attalea palm trees, where R. robustus are usually found. We suggest that although of secondary importance in the blood-sucking bugs, α-amylase may be needed during occasional plant feeding and thus has been retained.

Mechanisms of Systemic Osmoregulation in Insects.

Water is essential to life. Terrestrial insects lose water by evaporation from the body surface and respiratory surfaces, as well as in the excretory products, posing a challenge made more acute by their high surface-to-volume ratio. These losses must be kept to a minimum and be offset by water gained from other sources. By contrast, insects such as the blood-sucking bug Rhodnius prolixus consume up to 10 times their body weight in a single blood meal, necessitating rapid expulsion of excess water and ions. How do insects manage their ion and water budgets? A century of study has revealed a great deal about the organ systems that insects use to maintain their ion and water balance and their regulation. Traditionally, a taxonomically wide range of species were studied, whereas more recent research has focused on model organisms to leverage the power of the molecular genetic approach. Key advances in new technologies have become available for a wider range of species in the past decade. We document how these approaches have already begun to inform our understanding of the diversity and conservation of insect systemic osmoregulation. We advocate that these technologies be combined with traditional approaches to study a broader range of nonmodel species to gain a comprehensive overview of the mechanism underpinning systemic osmoregulation in the most species-rich group of animals on earth, the insects.

Heme protonation affects iron-NO binding in the NO transport protein nitrophorin

The nitrophorins (NPs) comprise an unusual group of heme proteins with stable ferric heme iron nitric oxide (Fe-NO) complexes. They are found in the salivary glands of the blood-sucking kissing bug Rhodnius prolixus, which uses the NPs to transport the highly reactive signaling molecule NO. Nuclear resonance vibrational spectroscopy (NRVS) of both isoform NP2 and a mutant NP2(Leu132Val) show, after addition of NO, a strong structured vibrational band at around 600 cm−1, which is due to modes with significant Fe-NO bending and stretching contribution. Based on a hybrid calculation method, which uses density functional theory and molecular mechanics, it is demonstrated that protonation of the heme carboxyl groups does influence both the vibrational properties of the Fe-NO entity and its electronic ground state. Moreover, heme protonation causes a significant increase of the gap between the highest occupied and lowest unoccupied molecular orbital by almost one order of magnitude leading to a stabilization of the Fe-NO bond.

Microencapsulation of a Native Strain of the Entomopathogenic Fungus Beauveria bassiana and Bioinsecticide Activity against Pyrethroid-Resistant Triatoma infestans to Vector Control of Chagas Disease in the Argentine Gran Chaco Region.

The blood-sucking bug Triatoma infestans is the main Chagas disease vector in the Southern Cone of Latin America. Populations resistant to pyrethroid insecticides have been detected in the early 2000s and then expanded to the endemic area of northern Salta province, Argentina. In this context, the entomopathogenic fungus Beauveria bassiana has been shown to be pathogenic to pyrethroid-resistant T. infestans. In this study, both the bioinsecticidal activity and the residual effect of an alginate-based microencapsulation of a native strain of B. bassiana (Bb-C001) were tested under semi-field conditions against pyrethroid-resistant T. infestans nymphs. Fungal microencapsulated formulation caused higher nymph mortality than the unmicroencapsulated fungus and contributed to maintaining the conidial viability throughout the period evaluated under the tested conditions. These results suggest that alginate microencapsulation is an effective, simple, low-cost method that could be incorporated into the formulation of a bioinsecticide as a strategy to reduce the vector transmission of Chagas disease.

Eugenol and menthol synergize the toxicity of permethrin in the blood-sucking bug, Triatoma infestans

The blood-sucking bug, Triatoma infestans, is one of the main vectors of Chagas disease in America. It is usually controlled with pyrethroids, but the emergence of resistance to these insecticides creates the need to look for alternative products. Eugenol, menthol and menthyl acetate are botanical monoterpenes, which produce lethal and sublethal effects on insects. The purpose of this work was to determine what type of toxicological interactions occur when binary mixtures, formed by the pyrethroid permethrin and sublehtal doses of eugenol, menthol or menthyl acetate, are applied to T. infestans. First instar nymphs were exposed to filter papers impregnated with the insecticides. The number of knocked down insects was registered at different times and Knock Down Time 50% (KT50) values were calculated. The following KT50 values with their corresponding 95% Confidence Intervals were obtained: permethrin, 47.29 (39.92 - 56.32) min; permethrin + eugenol, 34.08 (29.60 - 39.01) min; permethrin + menthol, 27.54 (23.28 - 32.55) min; permethrin + menthyl acetate, 43.62 (39.99 - 47.59) min. Eugenol and menthol increased the speed of action of permethrin (synergism), but menthyl acetate had no effect on it (additivity). These results provide the basis to further explore interactions between conventional insecticides and plant monoterpenes as potential tools for controlling T. infestans.

Functional expression of a thrombin exosite I inhibitor triabin in Escherichia coli and elucidation of the role of key residues in its inhibitory activity

Triabin, a lipocalin-like thrombin inhibitor from the saliva of the blood-sucking triatomine bug Triatoma pallidipennis, exhibits effective inhibition comparable to hirudin despite binding exclusively at exosite I. Interestingly, it was reported that higher triabin doses would not inhibit thrombin completely, which makes it a promising antithrombotic candidate agent with a larger therapeutic window. However, few structural and functional studies about triabin have been reported in the past three decades, mostly due to the lack of a reliable and practicable recombinant expression technology for this seemingly small protein. In this work, we have adopted the SUMO fusion technology for the expression of triabin in E. coli cells—with facile refolding and purification procedures—and the bioactive triabin was produced in ∼12 mg/L culture medium. Subsequently, the structure-function studies through extensive site-directed mutagenesis reveal that triabin's Phe-106 involved in the hydrophobic contacts plays a surprisingly important role in the thrombin inhibition, in contrast to the negatively charged residues Asp-135 or Glu-128 involved in the salt-bridge interaction. As such, this study complements our understanding of the interaction mechanism of natural thrombin inhibitors, which should facilitate the development of anticoagulant drugs with a novel mode of action against thrombin.

Empty-spiracles is maternally expressed and essential for neurodevelopment and early embryo determination in Rhodnius prolixus

Since empty-spiracles (ems) was identified and characterized in Drosophila melanogaster as a head-gap gene, several studies have been carried out in other insect orders to confirm its evolutionary conserved function. Using the blood-sucking bug Rhodnius prolixus as biological model, we found an ems transcript with three highly conserved regions: Box-A, Box-B, and the homeodomain. R. prolixus embryos silenced by parental RNAi for two of these ems conserved regions showed both maternal and zygotic defects. Rp-emsB fragment results in early lethal embryogenesis, with eggs without any embryonic structure inside. Rp-emsB expression pattern is only maternally expressed and localized in the ovary tropharium, follicular cells, and in the unfertilized female pronucleus. Rp-emsA fragment is zygotically expressed during early blastoderm formation until late developmental stages in two main patterns: anterior in the antennal segment, and in a segmentary in the neuroblast and tracheal pits. R. prolixus knockdown embryos for Rp-emsA showed an incomplete larval hatching, reduced heads, and severe neuromotor defects. Furthermore, in situ hybridization revealed a spatial and temporal expression pattern that highly correlates with Rp-ems observed function. Here,Rp-ems function in R. prolixus development was validated, showing that empty-spiracles does not act as a true head-gap gene, but it is necessary for proper head development and crucial for early embryo determination and neurodevelopment.

Molecular and functional basis of high-salt avoidance in a blood-sucking insect

SummarySalts are essential nutrients required for many physiological processes, and accordingly, their composition and concentration are tightly regulated. Taste is the ultimate sensory modality involved in resource quality assessment, resulting in acceptance or rejection. Here we found that high salt concentrations elicit feeding avoidance in the blood-sucking bug Rhodnius prolixus and elucidate the molecular and neurophysiological mechanisms involved. We found that high-salt avoidance is mediated by a salt-sensitive antennal gustatory receptor neuron (GRN). Using RNAi, we demonstrate that this process requires two amiloride-sensitive pickpocket channels (PPKs; RproPPK014276 and RproPPK28) expressed within these cells. We found that antennal GRNs project to the insect primary olfactory center, the antennal lobes, revealing these centers as potential sites for the integration of taste and olfactory host-derived cues. Moreover, the identification of the gustatory basis of high-salt detection in a hematophagous insect suggests novel targets for the prevention of biting and feeding.

Chagas: The Silent Disease

Chagas disease, also known as American trypanosomiasis, is a potentially life-threatening illness caused by the protozoan parasite Trypanosoma cruzi. An estimated 6 to 7 million people worldwide are infected with T. cruzi. Chagas disease is found mainly in endemic areas of 21 continental Latin American countries, where it is mostly transmitted when humans come into contact with faeces and/or urine of infected blood-sucking triatomine bugs (vector-borne transmission)1. Chagas disease is named after the Brazilian physician Carlos Chagas, who discovered the disease in 19092. Trypanosoma cruzi infection is curable if treatment is initiated soon after infection. In chronic patients, antiparasitic treatment can potentially prevent or curb disease progression and prevent transmission. Chagas disease patients are at risk of severe COVID-19 manifestations and should be a priority group to be vaccinated because the SARS-CoV-2 can cause myocarditis and the chronic Chagas disease typically originates a pro-thrombotic status, cardiac alterations and secondary thrombotic strokes3.

Bacterial symbionts influence host susceptibility to fenitrothion and imidacloprid in the obligate hematophagous bed bug, Cimex hemipterus

The use of insecticides remains important in managing pest insects. Over the years, many insects manifested physiological and behavioral modifications resulting in reduced efficacy of insecticides targeted against them. Emerging evidence suggests that bacterial symbionts could modulate susceptibility of host insects against insecticides. Here, we explore the influence of host microbiota in affecting the susceptibility of insect host against different insecticides in the blood-sucking bed bug, Cimex hemipterus. Rifampicin antibiotic treatment resulted in increased susceptibility to fenitrothion and imidacloprid, but not against deltamethrin. Meanwhile, the host fitness parameters measured in the present study were not significantly affected by rifampicin treatment, suggesting the role of bacterial symbionts influencing susceptibility against the insecticides. 16S metagenomics sequencing revealed a drastic shift in the composition of several bacterial taxa following rifampicin treatment. The highly abundant Alphaproteobacteria (Wolbachia > 90%) and Gammaproteobacteria (Yersinia > 6%) in control bed bugs were significantly suppressed and replaced by Actinobacteria, Bacilli, and Betaproteobacteria in the rifampicin treated F1 bed bugs, suggesting possibilities of Wolbachia mediating insecticide susceptibility in C. hemipterus. However, no significant changes in the total esterase, GST, and P450 activities were observed following rifampicin treatment, indicating yet unknown bacterial mechanisms explaining the observed phenomena. Re-inoculation of microbial content from control individuals regained the tolerance of rifampicin treated bed bugs to imidacloprid and fenitrothion. This study provides a foundation for a symbiont-mediated mechanism in influencing insecticide susceptibility that was previously unknown to bed bugs.

The Effects of a Male Audience on Male and Female Mating Behaviour in the Blood-Sucking Bug Rhodnius prolixus

Social context is a key factor affecting sexual behaviour and cannot be neglected in gregarious species, such as triatomine blood-sucking bugs. Here we study the influence of the social context on the sexual reproductive behaviour of males and females of Rhodnius prolixus Stål. Specifically, we identify and compare the frequencies and sequence of sexual behaviours exhibited by a focal pair in presence or absence of a male audience. We expect that in presence of a male audience females increase their selectiveness level since the risk of losing infertile eggs decreases in presence of more candidates and females can benefit from copulating with a better quality male. Besides, in presence of potential rivals, we expect changes in the sexual behaviour of focal males, associated to a reduction in the risk of sperm competition. As expected, in presence of a male audience, females significantly increased the exhibition of rejection behaviour. Moreover, focal males exhibited shorter latencies to mount the female, longer duration of copula, and differences in their stereotyped behaviour exhibited during copula. We discuss the influence of the social context on the reproductive behaviour of females and males of R. prolixus.

Levels of testosterone, progesterone and oestradiol in pregnant-lactating does in relation to aggression during group housing

The neuroendocrine regulation of rabbit maternal behaviour has been explored in detail. However, little is yet known about the hormonal regulation of aggression in concurrently pregnant-lactating does, a reproductive condition that prevails during group housing of rabbits on farms. Therefore, in this study we determined the relation between a) the levels of progesterone, testosterone, and oestradiol during lactation; b) the anogenital distance at artificial insemination; and c) the timing of grouping with the intensity of agonistic behaviour, published previously. We performed four consecutive trials, where three groups of eight does each were artificially inseminated on day 10 postpartum (pp) and grouped on either day 12, 18 or 22 pp. Using Dipetalogaster maxima, a reduviid blood-sucking bug, we collected blood samples during the pregnant-lactating phase (days 13, 15, 17, 19, 21, 23 pp) on one or two randomly chosen does per treatment group. Testosterone levels varied little across the pregnant-lactating phase, agreeing with results from pregnant-only rabbits, while progesterone levels increased from day 3 (=13 dpp) to day 7 (=17 dpp) and remained unchanged until day 13 (=23 dpp) of pregnancy. All oestradiol concentrations fell below the limit of detection. Overall, all concentrations were slightly lower in comparison to rabbit studies with pregnantonly does. The agonistic behaviour was not related to the respective hormonal concentrations at grouping. In conclusion, the time point of grouping does after artificial insemination (AI) in the semi-group housing system only had a weak influence on aggression and the hormonal profile did not indicate an optimum time for grouping.

Deforestation effects on Attalea palms and their resident Rhodnius, vectors of Chagas disease, in eastern Amazonia.

Attalea palms provide primary habitat to Rhodnius spp., vectors of Trypanosoma cruzi. Flying from palms, these blood-sucking bugs often invade houses and can infect people directly or via food contamination. Chagas disease (CD) risk may therefore increase when Attalea palms thrive near houses. For example, Attalea dominate many deforested landscapes of eastern Amazonia, where acute-CD outbreaks are disturbingly frequent. Despite this possible link between deforestation and CD risk, the population-level responses of Amazonian Attalea and their resident Rhodnius to anthropogenic landscape disturbance remain largely uncharted. We studied adult Attalea palms in old-growth forest (OGF), young secondary forest (YSF), and cattle pasture (CP) in two localities of eastern Amazonia. We recorded 1856 Attalea along 10 transects (153.6 ha), and detected infestation by Rhodnius spp. in 18 of 280 systematically-sampled palms (33 bugs caught). Distance-sampling models suggest that, relative to OGF, adult Attalea density declined by 70–80% in CP and then recovered in YSF. Site-occupancy models estimate a strong positive effect of deforestation on palm-infestation odds (βCP-infestation = 4.82±1.14 SE), with a moderate decline in recovering YSF (βYSF-infestation = 2.66±1.10 SE). Similarly, N-mixture models suggest that, relative to OGF, mean vector density sharply increased in CP palms (βCP-density = 3.20±0.62 SE) and then tapered in YSF (βYSF-density = 1.61±0.76 SE). Together, these results indicate that disturbed landscapes may support between ~2.5 (YSF) and ~5.1 (CP) times more Attalea-dwelling Rhodnius spp. per unit area than OGF. We provide evidence that deforestation may favor palm-dwelling CD vectors in eastern Amazonia. Importantly, our landscape-disturbance effect estimates explicitly take account of (i) imperfect palm and bug detection and (ii) the uncertainties about infestation and vector density arising from sparse bug data. These results suggest that incorporating landscape-disturbance metrics into the spatial stratification of transmission risk could help enhance CD surveillance and prevention in Amazonia.

Under pressure: phenotypic divergence and convergence associated with microhabitat adaptations in Triatominae

BackgroundTriatomine bugs, the vectors of Chagas disease, associate with vertebrate hosts in highly diverse ecotopes. It has been proposed that occupation of new microhabitats may trigger selection for distinct phenotypic variants in these blood-sucking bugs. Although understanding phenotypic variation is key to the study of adaptive evolution and central to phenotype-based taxonomy, the drivers of phenotypic change and diversity in triatomines remain poorly understood.Methods/resultsWe combined a detailed phenotypic appraisal (including morphology and morphometrics) with mitochondrial cytb and nuclear ITS2 DNA sequence analyses to study Rhodnius ecuadoriensis populations from across the species’ range. We found three major, naked-eye phenotypic variants. Southern-Andean bugs primarily from vertebrate-nest microhabitats (Ecuador/Peru) are typical, light-colored, small bugs with short heads/wings. Northern-Andean bugs from wet-forest palms (Ecuador) are dark, large bugs with long heads/wings. Finally, northern-lowland bugs primarily from dry-forest palms (Ecuador) are light-colored and medium-sized. Wing and (size-free) head shapes are similar across Ecuadorian populations, regardless of habitat or phenotype, but distinct in Peruvian bugs. Bayesian phylogenetic and multispecies-coalescent DNA sequence analyses strongly suggest that Ecuadorian and Peruvian populations are two independently evolving lineages, with little within-lineage phylogeographic structuring or differentiation.ConclusionsWe report sharp naked-eye phenotypic divergence of genetically similar Ecuadorian R. ecuadoriensis (nest-dwelling southern-Andean vs palm-dwelling northern bugs; and palm-dwelling Andean vs lowland), and sharp naked-eye phenotypic similarity of typical, yet genetically distinct, southern-Andean bugs primarily from vertebrate-nest (but not palm) microhabitats. This remarkable phenotypic diversity within a single nominal species likely stems from microhabitat adaptations possibly involving predator-driven selection (yielding substrate-matching camouflage coloration) and a shift from palm-crown to vertebrate-nest microhabitats (yielding smaller bodies and shorter and stouter heads). These findings shed new light on the origins of phenotypic diversity in triatomines, warn against excess reliance on phenotype-based triatomine-bug taxonomy, and confirm the Triatominae as an informative model system for the study of phenotypic change under ecological pressure.Graphical

Regulation of a Trehalose-Specific Facilitated Transporter (TRET) by Insulin and Adipokinetic Hormone in Rhodnius prolixus, a Vector of Chagas Disease.

Using the blood-sucking kissing bug, Rhodnius prolixus as an experimental model, we have studied the involvement of insulin-like peptides (ILPs) and adipokinetic hormone (AKH) signaling in carbohydrate metabolism, focusing on the regulation of the trehalose-specific facilitated transporter (Rhopr-TRET), particularly in the ovaries. We find that trehalose stores in ovaries increase after feeding, synchronously with the beginning of vitellogenesis, but that the transcript expression of enzymes involved in trehalose synthesis show no changes between unfed and blood-fed animals. However, an eightfold increase in Rhopr-TRET transcript expression is observed in the ovaries post-blood meal. In vivo and ex vivo assays using exogenous insulins and Rhopr-AKH, reveal that Rhopr-TRET is up-regulated in ovaries by both peptide families. In accordance with these results, when ILP and AKH signaling cascades are impaired using RNA interference, Rhopr-TRET transcript is down-regulated. In addition, trehalose injection induces an up-regulation of Rhopr-TRET transcript expression and suggests an activation of insulin signaling. Overall, the results support the hypothesis of a direct trehalose uptake by ovaries from the hemolymph through Rhopr-TRET, regulated by ILP and/or AKH. We also show that Rhopr-TRET may work cooperatively with AKH signaling to support the release of trehalose from the ovaries into the hemolymph during the unfed (starved) condition. In conclusion, the results indicate that in females of R. prolixus, trehalose metabolism and its hormonal regulation by ILP and AKH play critical roles in adapting to different nutritional conditions and physiological states.

Mimicry in Cretaceous Bugs.

SummaryMimicry is ubiquitous in nature, yet understanding its origin and evolution is complicated by the scarcity of exceptional fossils that enable behavioral inferences about extinct animals. Here we report bizarre true bugs (Hemiptera) that closely resemble beetles (Coleoptera) from mid-Cretaceous amber. The unusual fossil bugs are described as Bersta vampirica gen. et sp. nov. and Bersta coleopteromorpha gen. et sp. nov. and are placed into a new family, Berstidae fam. nov. The specialized mouthparts of berstids indicate that they were predaceous on small arthropods. Their striking beetle-like appearance implies that they were either involved in defensive mimicry or mimicked beetles to attack unsuspecting prey. The latter would represent the first case of aggressive mimicry in the invertebrate fossil record. These findings enrich our understanding of the paleoecological associations and extinct behavioral strategies of Mesozoic insects.