What happens when the insecticide does not kill? A review of sublethal toxicology and insecticide resistance in triatomines.

Chagas disease is considered one of the most important human parasitosis in the United States. This disease is mainly transmitted by insects of the subfamily Triatominae. The chemical vector control is the main tool for reducing the incidence of the disease. However, the presence of triatomines after pyrethroids spraying has been reported in some regions, as in the case of Triatoma infestans in Argentina and Bolivia. The presence of insects can be explained by the colonization from neighbouring areas, the reduction of insecticide dose to sublethal levels due to environmental factors, and/or by the evolution of insecticide resistance. In the last two scenarios, a proportion of the insects is not killed by insecticide and gives rise to residual populations. This article focuses on the toxicological processes associated with these scenarios in triatomines. Sublethal doses may have different effects on insect biology, that is, sublethal effects, which may contribute to the control. In addition, for insect disease vectors, sublethal doses could have negative effects on disease transmission. The study of sublethal effects in triatomines has focused primarily on the sequence of symptoms associated with nervous intoxication. However, the effects of sublethal doses on excretion, reproduction and morphology have also been studied. Rhodnius prolixus and T. infestans and pyrethroids insecticides were the triatomine species and insecticides, respectively, mainly studied. Insecticide resistance is an evolutionary phenomenon in which the insecticide acts as a selective force, concentrating on the insect population's pre-existing traits that confer resistance. This leads to a reduction in the susceptibility to the insecticide, which was previously effective in controlling this species. The evolution of resistance in triatomines received little attention before the 2000s, but after the detection of the first focus of resistance associated with chemical control failures in T. infestans from Argentina in 2002, the study of resistance increased remarkably. A significant number of works have studied the geographical distribution, the resistance mechanisms, the biological modifications associated with resistance, the environmental influences and the genetic of T. infestans resistant to pyrethroid insecticides. Currently, studies of insecticide resistance are gradually being extended to other areas and other species. The aim of this article was to review the knowledge on both phenomena (sublethal effects and insecticide resistance) in triatomines. For a better understanding of this article, some concepts and processes related to insect-insecticide interactions, individual and population toxicology and evolutionary biology are briefly reviewed. Finally, possible future lines of research in triatomine toxicology are discussed.

Pyrethroid resistance distribution in Triatoma infestans and environmental association along the Argentine endemic zone.

Insecticide resistance is considered a barrier to chemical control of Triatoma infestans, the main vector of Chagas disease in the Southern Cone of South America. Although initiatives to reduce the incidence of the disease in the region have integrated different strategies, they have mainly relied on vector elimination using pyrethroid insecticides such as deltamethrin. Reports of pyrethroid resistance in connection with T. infestans control failures first emerged in northern Argentina and southern Bolivia. Recently, a mosaic pyrethroid-resistant focus has been described in the center of the Argentine Gran Chaco (Department of General Güemes, Chaco Province), characterized by the presence of susceptible and very highly resistant populations in the same area. The involvement of different resistance mechanisms has been proposed, together with the contribution of environmental variables that promote the toxicological heterogeneity described. In the endemic zone of Argentina, however, new questions arise: Are there any other clusters of resistance? Is there a relationship between the distribution of resistance and environmental variables (as has been observed at smaller scale)? We studied toxicological data from insects collected and analyzed at 224 localities between 2010 and 2020 as part of the resistance monitoring conducted by the Chagas National Program. The sites were classified according to the survival rate of insects exposed to a discriminant dose of deltamethrin: 0-0.19 were considered susceptible, 0.2-0.79 low-resistance, and 0.8-1 high-resistance. Localities were georeferenced to describe the spatial distribution of resistance and to identify environmental variables (demographics, land use, urbanization, connectivity, and climate) potentially associated with resistance. We used Generalized Linear Models (GLMs) to examine the association between resistance and environmental predictors, selecting error distributions based on the response variable definition. For the entire period, 197 susceptible localities were distributed across the endemic zone. Localities with different survival rates were found throughout the area; 9 high-resistance localities circled the two previously identified resistant foci, and 18 low-resistance in 6 provinces, highlighting their relevance for control planning. Precipitation variables were linked to resistance in all the GLMs evaluated. Presence/absence models were the most accurate, with precipitation, distance from the capital city, and land use contributing to the distribution of resistance. This information could be valuable for improving T. infestans control strategies in future scenarios characterized by unpredictable changes in land use and precipitation.

Morphometric study of the legs of the main Chagas vector, Triatoma infestans (Hemiptera: Reduviidae).

In triatomines, vectors of Chagas disease, active dispersal takes place by walking and flying. Flight has received more attention than walking although the last is the dispersal modality used by nymphs due to their lack of wings and also used by adults, which would facilitate the colonization and reinfestation of houses after vector control actions. The present work studied the morphometrical variation of Triatoma infestans legs, the main vector of Chagas disease the Southern Cone of South America. We described morphometric traits and the natural variation of each leg segment. Different linear, size and shape variables of each component of the three right legs of fifth instar nymphs of T. infestans were analyzed using morphometric tools. We analyzed differentiation, variation and correlation for each segment across the fore-, mid and hind legs using different statistical approaches such as general linear model, canonical variates analysis, test of equality of coefficient of variation and partial least square analysis. We also analyzed variation and correlation between segments within each leg with partial least square and morphometric disparity analyses. Our results showed that the segments differed between legs, as general trends, the dimensions (length, width and/or size) were greater in the hind legs, smaller in the forelegs and intermediate in the mid ones. The femur and tibia (length and/or width) showed differences in morphometric variation between legs and the femur and tibia showed the highest levels of correlation between legs. On the other hand, in the fore- and mid legs, the femur (length or width) showed similar variation with tibia and tarsus lengths, but in the hind legs, the femur showed similar variation with all segments and not with the tibia length, and there were strong correlations between linear measurement within each leg. Our results suggest that the femur and tibia could play a determining role in the coordination between the legs that determines the walking pattern. Considering that these segments would also be linked to the specific function that each leg has, this study suggests a preponderant role of the femur and tibia in the walking locomotion of T. infestans.

Can infection with Trypanosoma cruzi modify the toxicological response of Triatoma infestans susceptible and resistant to deltamethrin?

Chemical control plays a central role in interrupting the vector transmission of Chagas disease. In recent years, high levels of resistance to pyrethroids have been detected in the main vector Triatoma infestans, which were associated with less effectiveness in chemical control campaigns in different regions of Argentina and Bolivia. The presence of the parasite within its vector can modify a wide range of insect physiological processes, including toxicological susceptibility and the expression of resistance to insecticides. This study examined for the first time the possible effects of Trypanosoma cruzi infection on susceptibility and resistance to deltamethrin in T. infestans. Using WHO protocol resistance monitoring assays, we exposed resistant and susceptible strains of T. infestans, uninfected and infected with T. cruzi to different concentrations of deltamethrin in fourth-instar nymphs at days 10-20 post-emergence and monitored survival at 24, 48, and 72 h. Our findings suggest that the infection affected the toxicological susceptibility of the susceptible strain, showing higher mortality than uninfected susceptible insects when exposed to both deltamethrin and acetone. On the other hand, the infection did not affect the toxicological susceptibility of the resistant strain, infected and uninfected showed similar toxic responses and the resistance ratios was not modified. This is the first report of the effect of T. cruzi on the toxicological susceptibility of T. infestans and triatomines in general and, to our knowledge, one of the few on the effect of a parasite on the insecticide susceptibility of its insect vector.

Sublethal effects of a pyrethroid insecticide on cuticle thickness, wing size and shape in the main vector Triatoma infestans.

Exposure to sublethal doses of insecticide may affect biological traits in triatomines. We investigated the effects of toxicological phenotype (pyrethroid resistance status) and exposure to sublethal doses of deltamethrin on two traits of Triatoma infestans Klug (Heteroptera: Reduviidae) using a phenotypic plasticity experimental design. First-instar nymphs from 14 and 10 full-sib families from pyrethroid-susceptible and pyrethroid-resistant populations, respectively, were used. For the susceptible population, we treated first instars topically with acetone (control) or deltamethrin (treatment) once. For the resistant population, instars were treated once, twice and three times as first, third or fifth-instar nymphs, respectively. We measured cuticle thickness, wing size and wing shape of 484 emerging adults, and tested for treatment effects using mixed ANOVA and MANOVA models. Toxicological phenotype, exposure to deltamethrin and full-sib family exerted significant effects on cuticle thickness, wing size and wing shape. Adult triatomines previously treated with deltamethrin developed significantly thicker cuticles than control triatomines only in the resistant population and significantly bigger wings in both populations. Mean cuticle thickness and wing size increased with increasing exposures to deltamethrin. Exposure to sublethal doses of deltamethrin generated morphological modifications that may affect insect survival and flight dispersal, and hence may have evolutionary and epidemiological consequences.

Transcriptomic modulation in response to an intoxication with deltamethrin in a population of Triatoma infestans with low resistance to pyrethroids.

BACKGROUND: Triatoma infestans is the main vector of Chagas disease in the Southern Cone. The resistance to pyrethroid insecticides developed by populations of this species impairs the effectiveness of vector control campaigns in wide regions of Argentina. The study of the global transcriptomic response to pyrethroid insecticides is important to deepen the knowledge about detoxification in triatomines. METHODOLOGY AND FINDINGS: We used RNA-Seq to explore the early transcriptomic response after intoxication with deltamethrin in a population of T. infestans which presents low resistance to pyrethroids. We were able to assemble a complete transcriptome of this vector and found evidence of differentially expressed genes belonging to diverse families such as chemosensory and odorant-binding proteins, ABC transporters and heat-shock proteins. Moreover, genes related to transcription and translation, energetic metabolism and cuticle rearrangements were also modulated. Finally, we characterized the repertoire of previously uncharacterized detoxification-related gene families in T. infestans and Rhodnius prolixus. CONCLUSIONS AND SIGNIFICANCE: Our work contributes to the understanding of the detoxification response in vectors of Chagas disease. Given the absence of an annotated genome from T. infestans, the analysis presented here constitutes a resource for molecular and physiological studies in this species. The results increase the knowledge on detoxification processes in vectors of Chagas disease, and provide relevant information to explore undescribed potential insecticide resistance mechanisms in populations of these insects.

Transcriptomic modulation in response to an intoxication with deltamethrin in a population of Triatoma infestans with low resistance to pyrethroids

Background Triatoma infestans is the main vector of Chagas disease in the Southern Cone. The resistance to pyrethroid insecticides developed by populations of this species impairs the effectiveness of vector control campaigns in wide regions of Argentina. The study of the global transcriptomic response to pyrethroid insecticides is important to deepen the knowledge about detoxification in triatomines. Methodology and findings We used RNA-Seq to explore the early transcriptomic response after intoxication with deltamethrin in a population of T. infestans which presents low resistance to pyrethroids. We were able to assemble a complete transcriptome of this vector and found evidence of differentially expressed genes belonging to diverse families such as chemosensory and odorantbinding proteins, ABC transporters and heat-shock proteins. Moreover, genes related to transcription and translation, energetic metabolism and cuticle rearrangements were also modulated. Finally, we characterized the repertoire of previously uncharacterized detoxification-related gene families in T. infestans and Rhodnius prolixus. Conclusions and significance Our work contributes to the understanding of the detoxification response in vectors of Chagas disease. Given the absence of an annotated genome from T. infestans, the analysis presented here constitutes a resource for molecular and physiological studies in this species. The results increase the knowledge on detoxification processes in vectors of Chagas disease, and provide relevant information to explore undescribed potential insecticide resistance mechanisms in populations of these insects.

Fluctuating asymmetry and exposure to pyrethroid insecticides in Triatoma infestans populations in northeastern Argentina

Exposure to sublethal doses of insecticide may affect biological traits in triatomines. We investigated the effects of toxicological phenotype (pyrethroid resistance status) and exposure to sublethal doses of deltamethrin on two traits of Triatoma infestans Klug (Heteroptera: Reduviidae) using a phenotypic plasticity experimental design. First-instar nymphs from 14 and 10 full-sib families from pyrethroid-susceptible and pyrethroid-resistant populations, respectively, were used. For the susceptible population, we treated first instars topically with acetone (control) or deltamethrin (treatment) once. For the resistant population, instars were treated once, twice and three times as first, third or fifth-instar nymphs, respectively. We measured cuticle thickness, wing size and wing shape of 484 emerging adults, and tested for treatment effects using mixed ANOVA and MANOVA models. Toxicological phenotype, exposure to deltamethrin and full-sib family exerted significant effects on cuticle thickness, wing size and wing shape. Adult triatomines previously treated with deltamethrin developed significantly thicker cuticles than control triatomines only in the resistant population and significantly bigger wings in both populations. Mean cuticle thickness and wing size increased with increasing exposures to deltamethrin. Exposure to sublethal doses of deltamethrin generated morphological modifications that may affect insect survival and flight dispersal, and hence may have evolutionary and epidemiological consequences.

Phenotypic plasticity, canalisation and developmental stability of Triatoma infestans wings: effects of a sublethal application of a pyrethroid insecticide.

BACKGROUND: Triatomine control campaigns have traditionally consisted of spraying the inside of houses with pyrethroid insecticides. However, exposure to sublethal insecticide doses after the initial application is a common occurrence and may have phenotypic consequences for survivors. Here, using Triatoma infestans (the main vector of Chagas disease in the Southern Cone of South America) as a model species, we quantified the effects of exposure to a sublethal dose of pyrethroid insecticide on wing morphology. We tested if the treatment (i) induced a plastic effect (change in the character mean); (ii) altered environmental canalisation (higher individual variation within genotypes); (iii) altered genetic canalisation (higher variation among genotypes); and (iv) altered developmental stability (higher fluctuating asymmetry [FA]). METHODS: Each of 25 full-sib families known to be susceptible to pyrethroid insecticides were split in two groups: one to be treated with a sublethal dose of deltamethrin (insecticide-treated group) and the other to be treated with pure acetone (control group). Wings of the emerging adults were used in a landmark-based geometric morphometry analysis to extract size and shape measurements. Average differences among treatments were measured. Levels of variation among families, among individuals within families and among sides within individuals were computed and compared among treatments. RESULTS: Wing size and shape were affected by a sublethal dose of deltamethrin. The treated insects had larger wings and a more variable wing size and shape than control insects. For both wing size and shape, genetic variation was higher in treated individuals. Individual variations and variations in FA were also greater in deltamethrin-treated insects than in control ones for all full-sib families; however, the patterns of shape variation associated with genetic variation, individual variation and FA were different. CONCLUSIONS: Insects exposed to a sublethal dose of deltamethrin presented larger, less symmetrical and less canalised wings. The insecticide treatment jointly impaired developmental stability and genetic and environmental canalisation. The divergent patterns of shape variation suggest that the related developmental buffering processes differed at least partially. The morphological modifications induced by a single sublethal exposure to pyrethroids early in life may impinge on subsequent flight performance and consequently affect the dynamics of house invasion and reinfestation, and the effectiveness of triatomine control operations.

Estado de la resistencia a insecticidas en Triatoma infestans de Argentina

Los insectos triatominos son los vectores de la enfermedad de Chagas. El control químico de estos insectos es la principal herramienta utilizada para reducir la incidencia de la enfermedad. Los piretroides son los principales insecticidas utilizados para el control de los triatominos en todos los países endémicos. La evolución de resistencia a insecticidas convierte en inefciente una exitosa estrategia de control químico. En Argentina se han detectado diferentes focos de resistencia en Triatoma infestans, el vector más importante en el Cono Sur de Sudamérica, asociados a defciencias en el control. Los estudios realizados sobre esta especie muestran que la resistencia es un problema complejo, ya que evolucionó en varias zonas de la distribución geográfca de la especie, se identifcaron diferentes mecanismos de resistencia, está asociada a modifcaciones en diversos procesos biológicos, está vinculada a variaciones ambientales y hay escasez de alternativas de control. En este manuscrito presentamos una revisión del conocimiento sobre la resistencia a piretroides en T. infestans en Argentina.

Genetic structure of deltamethrin-resistant populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco.

The genetic structure of natural populations offers insight into the complexities of their dynamics, information that can be relevant to vector control strategies. Microsatellites are useful neutral markers to investigate the genetic structure and gene flow in Triatoma infestans, one of the main vectors of Chagas disease in South America. Recently, a heterogeneous pyrethroid-resistant hotspot was found in the Argentine Gran Chaco, characterized by the highest levels of deltamethrin resistance found at the present time. We applied population genetics analyses to microsatellite and village data and search for associations between the genetic variability and the heterogeneous toxicological pattern previously found. We genotyped 10 microsatellite loci in 67 T. infestans from 6 villages with no, low, and high pyrethroid resistance. The most genetically diverse populations were those susceptible or with low values of resistance. In contrast, high-resistance populations had lower herozygosity and some monomorphic loci. A negative association was found between variability and resistant ratios. Global and pairwise FSTs indicated significant differentiation between populations. The only susceptible population was discriminated in all the performed studies. Low-resistance populations were also differentiated by a discriminant analysis of principal components (DAPC) and were composed mostly by the same two genetic clusters according to STRUCTURE Bayesian algorithm. Individuals from the high-resistance populations were overlapped in the DAPC and shared significant proportions of a genetic cluster. These observations suggest that the resistant populations might have a common origin, although more genetic markers and samples are required to test this hypothesis more rigorously.

Resistance to deltamethrin in Triatoma infestans: microgeographical distribution, validation of a rapid detection bioassay and evaluation of a fumigant canister as control alternative strategy.

Triatoma infestans (Klug) (Hemiptera: Reduviidae) is the main vector of Chagas disease in the Southern Cone of America and resistance to pyrethroid insecticides has been detected in several areas from its geographical distribution. Pyrethroid resistance presents a complex geographical pattern at different spatial scales. However, it is still unknown if the toxicological variability is a common feature within villages of the Gran Chaco were high resistance was descripted. The objectives of this study were to determine: (a) the microgeographical distribution of the deltamethrin-resistance in insects from Pampa Argentina village, (b) the performance of the insecticide impregnated paper bioassay to evaluate deltamethrin-resistance in field collected insects and (c) the lethal activity of the fumigant canister containing DDVP against insects resistant to deltamethrin. High survival of T. infestans exposed to discriminant dose was observed in the samples of all the evaluated dwellings, suggesting that the resistance to deltamethrin is homogeneous at the microgeographical level. Resistance determination by impregnated paper bioassay was similar to traditional topical determination, highlighting the use of this rapid methodology in field large-scale monitoring. The fumigant canister was not effective against resistant insects, remarking the need to develop suitable formulations that ensure minimal toxicological risk and high effectivity.

Behavioral response mediated by feces in Triatoma infestans (Hemiptera: Reduviidae: triatominae) susceptible and resistant to deltamethrin.

Chemical cues from feces promote aggregation behavior in Triatoma infestans nymphs and adults. Given the importance of T. infestans resistant to pyrethroids in several areas of Argentina and Bolivia, it would be important to know if there is an association with specific attraction and aggregation behaviors. These behaviors, to and surrounding refuges, play an important role in triatomine population dynamics, an important factor to consider and model for vector control strategies. The aim of the present study was to analyze the behavior of orientation to chemical signals emitted by feces from deltamethrin resistant (R) and susceptible (S) T. infestans. The behavioral assays were performed in a circular glass arena divided in two equal sectors. Fecal signals emitted by both S and R feces are attractants to fifth-instar nymphs of both S and R populations. Both toxicological phenotypes remained significantly longer on R feces, as compared to S feces. This is the first evidence in a triatomine, for the association of an aggregation behavior and insecticide resistance and may be the result of pleiotropic effects surrounding resistance genes.

Insecticide Resistance Mechanisms in Triatoma infestans (Reduviidae: Triatominae): The Putative Role of Enhanced Detoxification and Knockdown Resistance (kdr) Allele in a Resistant Hotspot From the Argentine Chaco.

Chagas disease affects around 6 million people in the world, and in Latin America, it is mainly transmitted by the kissing bug. Chemical control of the vector with pyrethroid insecticides has been the most frequently used tool to reduce the disease incidence. Failures of field control have been detected in areas of the Argentinian Gran Chaco that correlate with high levels of insecticide resistance. Here, we provide evidence of the mechanisms involved in the resistance to insecticides of field populations of T. infestans from General Güemes Department (Chaco Province, Argentina). The biochemical analysis suggests the increase in the activity of the degradative enzymes P450 oxidases and esterases as a minor contributive mechanism in low-resistance populations. The molecular study revealed high frequencies of the kdr L925I mutation at the voltage-gated sodium channel as responsible for the high resistance ratios detected. This knowledge contributes to the generation of comprehensive vector control strategies that reduce the incidence of the disease.

Insecticide Resistance Mechanisms in Triatoma infestans (Reduviidae: Triatominae): The Putative Role of Enhanced Detoxification and Knockdown Resistance (kdr) Allele in a Resistant Hotspot From the Argentine Chaco / Mecanismos de resistência a inseticidas em Triatoma infestans (Reduviidae: Triatominae): o papel putativo do alelo de desintoxicação aprimorada e resistência à queda ( kdr ) em um hotspot resistente do Chaco argentino

Chagas disease affects around 6 million people in the world, and in Latin America, it is mainly transmitted by the kissing bug. Chemical control of the vector with pyrethroid insecticides has been the most frequently used tool to reduce the disease incidence. Failures of field control have been detected in areas of the Argentinian Gran Chaco that correlate with high levels of insecticide resistance. Here, we provide evidence of the mechanisms involved in the resistance to insecticides of field populations of T. infestans from General Güemes Department (Chaco Province, Argentina). The biochemical analysis suggests the increase in the activity of the degradative enzymes P450 oxidases and esterases as a minor contributive mechanism in low-resistance populations. The molecular study revealed high frequencies of the kdr L925I mutation at the voltage-gated sodium channel as responsible for the high resistance ratios detected. This knowledge contributes to the generation of comprehensive vector control strategies that reduce the incidence of the disease.

Resistance to deltamethrin in Triatoma infestans: microgeographical distribution, validation of a rapid detection bioassay and evaluation of a fumigant canister as control alternative strategy / Resistência à deltametrina em Triatoma infestans: distribuição microgeográfica, validação de um bioensaio de detecção rápida e avaliação de um recipiente fumigante como estratégia alternativa de controle

Triatoma infestans (Klug) (Hemiptera: Reduviidae) is the main vector of Chagas disease in the Southern Cone of America and resistance to pyrethroid insecticides has been detected in several areas from its geographical distribution. Pyrethroid resistance presents a complex geographical pattern at different spatial scales. However, it is still unknown if the toxicological variability is a common feature within villages of the Gran Chaco were high resistance was descripted. The objectives of this study were to determine: (a) the microgeographical distribution of the deltamethrin-resistance in insects from Pampa Argentina village, (b) the performance of the insecticide impregnated paper bioassay to evaluate deltamethrin-resistance in field collected insects and (c) the lethal activity of the fumigant canister containing DDVP against insects resistant to deltamethrin. High survival of T. infestans exposed to discriminant dose was observed in the samples of all the evaluated dwellings, suggesting that the resistance to deltamethrin is homogeneous at the microgeographical level. Resistance determination by impregnated paper bioassay was similar to traditional topical determination, highlighting the use of this rapid methodology in field large-scale monitoring. The fumigant canister was not effective against resistant insects, remarking the need to develop suitable formulations that ensure minimal toxicological risk and high effectivity.

Genetic structure of deltamethrin-resistant populations of Triatoma infestans (Hemiptera: Reduviidae) in the Gran Chaco / Estrutura genética de populações de Triatoma infestans (Hemiptera: Reduviidae) resistentes à deltametrina no Gran Chaco

The genetic structure of natural populations offers insight into the complexities of their dynamics, information that can be relevant to vector control strategies. Microsatellites are useful neutral markers to investigate the genetic structure and gene flow in Triatoma infestans, one of the main vectors of Chagas disease in South America. Recently, a heterogeneous pyrethroidresistant hotspot was found in the Argentine Gran Chaco, characterized by the highest levels of deltamethrin resistance found at the present time. We applied population genetics analyses to microsatellite and village data and search for associations between the genetic variability and the heterogeneous toxicological pattern previously found. We genotyped 10 microsatellite loci in 67 T. infestans from 6 villages with no, low, and high pyrethroid resistance. The most genetically diverse populations were those susceptible or with low values of resistance. In contrast, high-resistance populations had lower herozygosity and some monomorphic loci. A negative association was found between variability and resistant ratios. Global and pairwise FSTs indicated significant differentiation between populations. The only susceptible population was discriminated in all the performed studies. Low-resistance populations were also differentiated by a discriminant analysis of principal components (DAPC) and were composed mostly by the same two genetic clusters according to STRUCTURE Bayesian algorithm. Individuals from the high-resistance populations were overlapped in the DAPC and shared significant proportions of a genetic cluster. These observations suggest that the resistant populations might have a common origin, although more genetic markers and samples are required to test this hypothesis more rigorously.

Effect of reproductive state on active dispersal in Triatoma infestans (Klug, 1834) (Hemiptera: Reduviidae: Triatominae) susceptible and resistant to deltamethrin

Triatoma infestans (Klug, 1834) is the main vector of the Chagas´ disease in Argentina. The presence of insects in the domestic environment after application of pyrethroid insecticides was reported. The colonization and the evolution of insecticide resistance are processes that can explain these control failures. In both processes, the active dispersal of insects is a determining factor. The aim of the present study was to determine whether the reproductive state modulate the dispersal capacity of females of T. infestans susceptible and resistant to deltamethrin. For this, different variables associated with active dispersal were measured in virgin and copulated females both susceptible and resistant to deltamethrin. The experimental arena was 10 m long and contained two shelters. Groups of 12 females of each experimental group were released in one of the shelters and were able to move during 3 days/nights. The results showed that the females mainly dispersed by walking regardless of the reproductive state and the toxicological phenotype. On the other hand, the copula affected the posterior dispersal capacity in females susceptible and resistant to deltamethrin. The copulated females dispersed more times, leaved more times from the shelters and showed a higher proportion of dispersed individuals than the virgin females. Finally, the insecticide resistance reduced the dispersal capacity of females. The resistant insects showed lower number of dispersal events, a lower proportion of dispersed individuals, and lower exit and entry events from/to shelter than susceptible insects. This is the first report of the effect of copula on the capacity of active dispersion in vectors of Chagas disease susceptible and resistant to insecticides.

Interaction between two aggregation chemical signals in Triatoma infestans (Hemiptera: Reduviidae).

The nymphs and adults of Triatoma infestans spend much of their time aggregated among themselves within narrow and dark shelters. The search for a suitable shelter depends in part on the recognition of chemical signals coming from the feces and the cuticle of the other individuals who use the refuge. The aim of this study was determine the possible interaction between the chemical signals associated to the feces and to the cuticle of T. infestans. The results showed that the insects remained significantly more time on the feces that had contact with legs and the feces plus footprints than feces or footprints alone, demonstrating the interaction between evaluated signals. These results demonstrates also that feces extracted a chemical stimulus from the legs. Understanding the interaction feces-legs as an interaction feces-cuticle of legs, the results suggest that the feces could extract some cuticular compound with activity on the behavior of the insects. This is the first report of the interaction between the two aggregation signals recognized in T. infestans and of the increase in the behavioral response of insects exposed to feces that had contact with a cuticular structure.

Excretion/defecation patterns in Triatoma infestans populations that are, respectively, susceptible and resistant to deltamethrin

Pyrethroid resistance has been detected in Triatoma infestans (Klug) (Hemiptera: Reduviidae) specimens from different areas of Argentina and Bolivia. Genes conferring resistance can have a pleiotropic effect with epidemiological and evolutionary consequences. This research studied excretion/defecation patterns in deltamethrin-resistant T. infestans in order to elucidate its biological performance, adaptive consequences and role in the transmission of Chagas' disease. One deltamethrin-susceptible strain and two deltamethrin-resistant strains were used. Fifth-instar nymphs were fed ad libitum and their defecations recorded during and after the first or second feeding in the stadium. Resistant insects began to defecate later, defecated less, showed a lower proportion of defecating individuals and lower defecation indices compared with susceptible insects during the first hour after feeding. The number of bloodmeals in the stadium did not affect the main variables determining the pattern of defecation. The present study suggests that alterations in the excretion/defecation pattern in resistant insects entail an adaptive cost and, considering only this pattern, determine a lower capacity for transmission of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) compared with susceptible insects.