Grasielle Caldas DÁvila Pessoa

History of insecticide resistance of Triatominae vectors.

In the last 15 years, different types of Triatominae resistance to different insecticides have been reported; thus, resistance may be more widespread than known, requiring better characterization and delimitation, which was the aim of this review. This review was structured on a literature search of all articles from 1970 to 2015 in the PubMed database that contained the keywords Insecticide resistance and Triatominae . Out of 295 articles screened by title, 33 texts were selected for detailed analysis. Insecticide resistance of Triatomines is a complex phenomenon that has been primarily reported in Argentina and Bolivia, and is caused by different factors (associated or isolated). Insecticide resistance of Triatominae is a characteristic inherited in an autosomal and semi-dominant manner, and is polygenic, being present in both domestic and sylvatic populations. The toxicological profile observed in eggs cannot be transposed to different stages of evolution. Different toxicological profiles exist at macro- and microgeographical levels. The insecticide phenotype has both reproductive and developmental costs. Different physiological mechanisms are involved in resistance. Studies of Triatomine resistance to insecticides highlight three deficiencies in interpreting the obtained results: I) the vast diversity of methodologies, despite the existence of a single guiding protocol; II) the lack of information on the actual impact of resistance ratios in the field; and III) the concept of the susceptibility reference lineage. Research on the biological and behavioral characteristics of each Triatominae species that has evolved resistance is required in relation to the environmental conditions of each region.

Deltamethrin pyrethroid susceptibility characterization of Triatoma sordida Stål, 1859 (Hemiptera: Reduviidae) populations in the Northern Region of Minas Gerais, Brazil

INTRODUCTION Triatoma sordida is the most captured Triatomine species in the Brazilian artificial environment. In 2008, the discovery of three Triatomine populations with altered susceptibilities to deltamethrin highlighted the importance of investigating the genetic potential for resistance in triatomines. The purpose of this study was to characterize the susceptibility to deltamethrin of peridomestic T. sordida populations in Minas Gerais, Brazil. METHODS A susceptibility reference lineage derived from Uberaba, Minas Gerais, Brazil was used. Serial dilutions of deltamethrin were prepared and applied to the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed mortality in response to a diagnostic dose of 1xLD99 of the susceptibility reference lineage. RESULTS Susceptibility profile characterization of T. sordida populations revealed resistance ratios (RR50s) ranging from 0.42 to 3.94. The percentage mortality in response to the diagnostic dose varied from 70% to 100%. A comparison of the results obtained in the quantitative and qualitative assays demonstrated a lack of correspondence for some populations. CONCLUSIONS We demonstrated that only T. sordida populations that present a RR50>1.0 have altered susceptibility, and the execution of simultaneous field and laboratory tests is required to understand the actual effect of vector control. A possible cause of the observed resistance ratios might be the continuous use of pyrethroids in Brazil since the 1980s.

Standardization of laboratory bioassays for the study of Triatoma sordida susceptibility to pyrethroid insecticides

BackgroundIncreasing reports of high-resistant Triatominae populations concerns scientists and sanitarians as little is known about the factors behind the occurrence of such phenotype and its real impact on vector control strategies. Moreover, the utilization of a large variety of methodologies hinder the comparison of the reported studies.MethodsThis work aims to review laboratory bioassays, redefining the assessed biological features (age, generation and insecticide application area) and technical procedures (mortality recording time and the ideal diagnostic dose).ResultsResults were not influenced by the insecticide application area in nymphs or by their generation. Three days-old specimen’s revealed lower susceptibility to the tested insecticide. We determined that it is more appropriate to record mortality 72 h after treatment with insecticide, as well as using a diagnostic dose of 1xDL99.ConclusionThis work suggests more adequate methodological parameters for assessing insecticide resistance in triatomines, which also allows the comparison of results obtained by different research groups. For laboratory bioassays, we recommend: 1) the use of first instar nymphs from first or second generation; 2) 3 day-old specimens; 2) application of insecticide in the dorsal or ventral abdomen area; 3) mortality recording 72 h after treatment with pyrethroids and 4) a diagnostic dose of 1x LD99.

Susceptibility characterization of residual Brazilian populations of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae) to deltamethrin pyrethroid

INTRODUCTION Despite years of efforts towards the elimination of Triatoma infestans in Brazil, residual foci still persist in some areas of the States of Bahia and Rio Grande do Sul. The persistence of these T. infestans populations in the country has two different origins of equal concern: operational failures or insecticide resistance. Thus, the objective of this study was to characterize the susceptibility profile of the residual Brazilian populations of T. infestans to deltamethrin. METHODS The susceptibility reference lineage was derived from Cipein/Argentina. The populations studied were manually collected using a dislodging agent in peridomiciles in the States of Bahia (Novo Horizonte) and of Rio Grande do Sul (Santa Rosa and Doutor Maurício Cardoso). Serial dilutions of deltamethrin were prepared and applied at the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed the mortality of a diagnostic dose of 1xLD99 (2.76ng a.i./nymph) determined for the susceptibility reference lineage. RESULTS The susceptibility profile characterization of the T. infestans populations revealed an RR50 ranging from 1.73 to 3.26. The mortality percentage in response to a diagnostic dose was 100%. The results obtained in the quantitative and qualitative assays corresponded for all populations. CONCLUSIONS The results of this study indicate that the persistence of residual foci of T. infestans in Bahia and Rio Grande do Sul is not related to insecticide resistance but may be associated with operational failures. In Rio Grande do Sul, we must consider the possibility of continuous reinfestation by Argentinian individuals, which justifies active and efficient epidemiological surveillance.

Markers for the population genetics studies of Triatoma sordida (Hemiptera: Reduviidae).

BackgroundTriatoma sordida, a vector of Trypanosoma cruzi, is native of Brazil, Bolivia, Paraguay, Argentina, and Uruguay, and occurs primarily in peridomiciles. Currently, it is the species most frequently captured by the Chagas Disease Control Program in Brazil. For this reason, population genetic studies attract great interest, as they can provide further information about the dispersal and household invasion processes of this species. In the absence of suitable markers, the objective of this study was to test the cross amplification of microsatellite primers.Findings23 primers were tested for microsatellite loci already described for other species of the genus Triatoma sp. Forty four specimens of T. sordida captured in the north of Minas Gerais were used to validate the use of standardized loci for population genetic analyses. It was possible to amplify 10 of the 23 loci tested for T. sordida.ConclusionsThis is the first study that provides 10 microsatellite markers for population analysis of this triatomine species. Cross-amplification of primers can be used among other phylogenetically related species whose loci are already available for study.

Toxicological profile of deltamethrin in Triatoma brasiliensis (Hemiptera: Reduviidae) in State of Ceará, Northeastern Brazil

INTRODUCTION Triatoma brasiliensis is the species of greatest epidemiological relevance in the semi-arid region of Brazil. This species is predominantly found in domestic environments, and it has the ability to build large colonies with high levels of natural infection via Trypanosoma cruzi. Thus, T. brasiliensis is one of the most efficient transmitters of Chagas disease (CD) to humans. Despite household spraying with residual insecticides, many areas report persistent reinfestations for reasons that remain poorly understood. Therefore, this study sought to characterize the toxicological profile of deltamethrin in T. brasiliensis from areas with persistent reinfestation in State of Ceará, Brazil. METHODS The susceptibility reference lineage (SRL) was derived from Umari. Serial dilutions of deltamethrin were prepared and applied to the dorsal abdomen of first instar nymphs. The control group received only pure acetone. Mortality was evaluated after 72h. Qualitative tests assessed mortality in response to a diagnostic dose of 1xLD99 (0.851 nanograms of active ingredient per treated nymph) of the SRL. RESULTS The susceptibility profile characterization of the T. brasiliensis populations revealed 50% resistance ratios (RR50) that ranged from 0.32 to 1.21. The percentage of mortality in response to the diagnostic dose was 100%. CONCLUSIONS We demonstrated that T. brasiliensis was highly susceptible to deltamethrin. The control difficulties found might be related to the recolonization of the triatomines originating from neighboring environments and the possible operational failures related to the process of spraying that enabled specimens less susceptible to deltamethrin to survive.

Insights from tissue-specific transcriptome sequencing analysis of Triatoma infestans.

ABSTRACT Triatoma infestans is an insect of subfamily Triatominae (Hemiptera: Reduviidae) and an important vector of Trypanosoma cruzi, the etiologic agent of human Chagas disease. In this work we reported a transcriptome assembly and annotation of T. infestans heads obtained by Next Generation Sequencing (NGS) technologies.

Variability of susceptibility to deltamethrin in peridomestic Triatoma sordida from Triângulo Mineiro, State of Minas Gerais, Brazil.

INTRODUCTION Despite chemical and physical vector control strategies, persistent infestations of Triatoma sordida have been reported in a large part of Minas Gerais, Brazil, and the cause for this is little investigated. We aimed to characterize the deltamethrin toxicological profile in peridomestic T. sordidapopulations from Triângulo Mineiro area of Minas Gerais. METHODS Susceptibility to deltamethrin was assessed in seventeen peridomestic T. sordida populations. Serial dilutions of deltamethrin in acetone (0.2µL) were topically applied on the first instar nymphs (F1; five days old; fasting weight, 1.2 ± 0.2mg). Dose response results were analyzed using Probit software, and the lethal doses, slope and resistance ratios were determined. Qualitative tests were also performed. RESULTS The deltamethrin susceptibility profile of T. sordida populations revealed resistance ratios ranging from 0.84 to 2.8. The percentage mortality in response to a diagnostic dose was 100.0% in all populations. CONCLUSIONS From our results, the lack of resistance to insecticides but persistent T. sordida infestations in the Triângulo Mineiro area may be because of: 1) environmental degradation facilitating dispersion of T. sordida, allowing colonization in artificial ecotopes; 2) operational failures; and 3) complexity of the peridomicile in the study area.These variables are being investigated.

Resistance to deltamethrin by domestic and wild Triatoma infestans populations in the municipality of Toro Toro, Potosi, Bolivia

BackgroundChemical control with pyrethroid insecticides has been effective in reducing endemic areas of distribution of Triatoma infestans in the Southern Cone, as well as Bolivia; this had considerably reduced the infestation of households in a large part of the territory. Nowadays, areas such as the Chaco and the Inter-Andean Valleys are regions where the reach of vector control strategies is becoming limited, and infestations of insecticide-treated households are reported more often. The objective of this study was to determine if the persistence of T. infestans stems from changes in the susceptibility of its toxicological profile in four communities in the municipality of Toro Toro, Potosi, Bolivia.MethodsSusceptibility to deltamethrin of wild and domestic populations of T. infestans was evaluated in two stages (16 populations before and 13 populations after spraying) among DUs (structures in the intra- and peridomicile) and wild ecotopes, in four communities. Serial dilutions of deltamethrin in acetone (0.2 μl) were applied topically on standardized first-stage nymphs. Dose-response results were analyzed with the software PoloPlus and the relationships between lethal doses (LD) and resistance ratios (RR50) were determined.ResultsDifferent degrees of RR50 were detected among the populations before and after spraying (25.66–54.70 and 21.91–40.67, respectively), as well as in different ecotopes within a DU (DU JC 3, 28.06–36.13, in mixed structures of corrals and chicken coops; and DU JG 3, 46.27–25.70, in kitchen roofs), or in the wild environment of the community JG Sil (29.21–40.67). The mortality of insects undergoing diagnostic dose (DD) was never higher than 34%.ConclusionThe results obtained in this study showed resistance of T. infestans to deltamethrin in four communities, hence the complexity of this phenomenon is not only limited to the level of communities, but also applies to the microgeographical level, as in different ecotopes present within the DUs. This phenomenon should be considered while planning the activities of control programs.

Genetic characterization of residual Triatoma infestans populations from Brazil by microsatellite

In spite of long-term efforts to eliminate Triatoma infestans (Klug 1834) from Brazil, residual foci still persist in the states of Bahia and Rio Grande do Sul. Data on the genetic variability and structuring of these populations are however lacking. Using nine microsatellite loci, we characterized one residual T. infestans population from Bahia and four from Rio Grande do Sul, and compared them with bugs originally from an older focus in São Paulo; 224 bugs were genotyped. The number of alleles per locus ranged from 5 to 11. Observed and expected heterozygosities per locus ranged, respectively, from 0 to 0.786 and from 0 to 0.764. Significant departures from Hardy–Weinberg equilibrium, mainly due to heterozygote deficits, were detected in all loci and in most populations. Global indices estimated by AMOVA were: Fis was 0.37; Fst was 0.28; and Fit was 0.55; overall indices with p = 0.00 indicated substantial differentiation. Inter-population Fst ranged from 0.118 to 0.562, suggesting strong genetic structuring and little to no gene flow among populations. Intra-population Fis ranged from 0.301 to 0.307. Inbreeding was apparent in all populations except that from Bahia—which might be either linked by gene flow to nearby unsampled populations or part of a relatively large local population. The overall pattern of strong genetic structuring among pyrethroid-susceptible residual T. infestans populations suggests that their persistence is probably due to operational control failures. Detection and elimination of such residual foci is technically feasible and must become a public health priority in Brazil.