Evaluation of the insecticide susceptibility profile in Cimex lectularius (Hemiptera: Cimicidae) in Belo Horizonte (Brazil).

INTRODUCTION: Since 2013, major Cimex lectularius infestations have been detected in public shelters in Belo Horizonte (Brazil). Due to this, insecticide resistance has been investigated as one of the possible causes for the failure to control bedbugs. METHODS: Cimicids were subjected to bioassays according to the World Health Organization recommendations using deltamethrin and all commercial insecticides available for control of Cimex in Brazil. RESULTS: Cimicids were deltamethrin resistant and presented indicative of resistance to other insecticides, except for propoxur 1%. CONCLUSIONS: The commercial insecticides have a limited effect on bedbug populations, which may justify the unsatisfactory control observed in the shelters studied.

Evaluation of the insecticide susceptibility profile in Cimex lectularius (Hemiptera: Cimicidae) in Belo Horizonte (Brazil)

Abstract INTRODUCTION: Since 2013, major Cimex lectularius infestations have been detected in public shelters in Belo Horizonte (Brazil). Due to this, insecticide resistance has been investigated as one of the possible causes for the failure to control bedbugs. METHODS: Cimicids were subjected to bioassays according to the World Health Organization recommendations using deltamethrin and all commercial insecticides available for control of Cimex in Brazil. RESULTS: Cimicids were deltamethrin resistant and presented indicative of resistance to other insecticides, except for propoxur 1%. CONCLUSIONS: The commercial insecticides have a limited effect on bedbug populations, which may justify the unsatisfactory control observed in the shelters studied.

Microsatellite variation revealed panmictic pattern for Triatoma brasiliensis (Triatominae: Reduviidae) in rural northeastern Brazil: the control measures implications.

BACKGROUND: Triatoma brasiliensis Neiva, 1911 is the main vector of Trypanosoma cruzi in the caatinga of Northeastern Brazil. Despite of its epidemiological relevance, there are few studies on its genetic variability. Using microsatellite markers, we characterized the variability and dynamics of infestation and reinfestation of T. brasiliensis after residual insecticide spraying in five surveys conducted in a well-defined rural area located in the municipality of Tauá, Ceará, between 2009 and 2015. We evaluated: (1) general variability among local of captures; (2) variability along the time analysis (2009, 2010 and 2015); (3) and reinfestation process. RESULTS: On the analysis (1) global and pairwise FST values suggested absence of clusters among the area. AMOVA indicated that total variation is mainly represented by individual differences. Absence of clustering indicates a panmitic unit, with free gene flow. For (2), Pairwise FST indicated alterations in the genetic profile of the triatomines along the time. (3) Analysis of the reinfestation process showed that the domiciliary units investigated had different sources of infestation despite of its proximity. CONCLUSIONS: Observed homogeneity can be explained by the great dispersal capacity of T. brasiliensis, overlapping the different environments. Persistent house infestation in Tauá may be attributed to the occurrence of postspraying residual foci and the invasion of triatomines from their natural habitats.

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.

Inheritance and heritability of deltamethrin resistance under laboratory conditions of Triatoma infestans from Bolivia.

BACKGROUND: Over the last few decades, pyrethroid-resistant in Triatoma infestans populations have been reported, mainly on the border between Argentina and Bolivia. Understanding the genetic basis of inheritance mode and heritability of resistance to insecticides under laboratory conditions is crucial for vector management and monitoring of insecticide resistance. Currently, few studies have been performed to characterize the inheritance mode of resistance to pyrethroids in T. infestans; for this reason, the present study aims to characterize the inheritance and heritability of deltamethrin resistance in T. infestans populations from Bolivia with different toxicological profiles. METHODS: Experimental crosses were performed between a susceptible (S) colony and resistant (R) and reduced susceptibility (RS) colonies in both directions (♀ x ♂ and ♂ x ♀), and inheritance mode was determined based on degree of dominance (DO) and effective dominance (D(ML)). In addition, realized heritability (h(2)) was estimated based on a resistant colony, and select pressure was performed for two generations based on the diagnostic dose (10 ng. i. a. /nymph). The F1 progeny of the experimental crosses and the selection were tested by a standard insecticide resistance bioassay. RESULTS: The result for DO and D(ML) (< 1) indicates that resistance is an incompletely dominant character, and inheritance is autosomal, not sex-linked. The LD50 for F1 of ♀S x ♂R and ♂S x ♀R was 0.74 and 3.97, respectively, which is indicative of dilution effect. In the resistant colony, after selection pressure, the value of h(2) was 0.37; thus, the LD50 value increased 2.25-fold (F2) and 26.83-fold (F3) compared with the parental colony. CONCLUSION: The inheritance mode of resistance of T. infestans to deltamethrin, is autosomal and an incompletely dominant character; this is a previously known process, confirmed in the present study on T. infestans populations from Bolivia. The lethal doses (LD50) increase from one generation to another rapidly after selection pressure with deltamethrin. This suggests that resistance is an additive and cumulative factor, mainly in highly structured populations with limited dispersal capacity, such as T. infestans. This phenomenon was demonstrated for the first time for T. infestans in the present study. These results are very important for vector control strategies in problematic areas where high resistance ratios of T. infestans have been reported.

Baseline susceptibility to alpha-cypermethrin in Lutzomyia longipalpis (Lutz & Neiva, 1912) from Lapinha Cave (Brazil).

BACKGROUND: Given the increase in cases of visceral leishmaniasis in recent years, associated with the socio-economic impact of this disease, as well as the wide distribution of Lutzomyia longipalpis in Brazil and the likelihood that this vector may develop resistance to insecticides used for control, the Ministry of Health considers as crucial the creation of a network in order to study and monitor the resistance of this vector to insecticides used for control. In this sense, this study aimed: 1) to characterize the susceptibility of L. longipalpis from Lapinha Cave (Lagoa Santa, MG - Brazil) to Alfateck SC200 in field bioassays, and 2) to define the susceptibility baseline to alpha-cypermethrin in laboratory bioassays, checking the possibility of using it as susceptibility reference lineage (SRL). FINDINGS: The field bioassays revealed that the tested population was highly susceptible to alpha-cypermethrin in all time periods with high mortality (~100 %) in all treated surfaces before six months after spraying. In the laboratory bioassays, the studied population presented LD50, LD95 and LD99 to 0.78013, 10.5580 and 31.067 mg/m(2), respectively. The slope was 1.454121. CONCLUSIONS: The studied population of L. longipalpis was considered as adequate for SRL according criterion recommended by Pan-American Health Organization and has proven susceptibility to tested insecticide in the field. One cannot rule out the possibility of finding populations of L. longipalpis more susceptible to alpha-cypermethrin; therefore, further research is necessary on other populations with potential use as a SRL.

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.

Susceptibility of Triatoma sordida Stal, 1859 (Hemiptera: Reduviidae) to alpha-cypermethrin under natural climatic conditions.

INTRODUCTION: Despite the recommendations by interpretation of resistance ratios obtained in laboratory bioassays, little is known about the actual impact of these results in the effectiveness of vector control activities in the field. In this context, our objective was to determine the mean value of different resistance ratios obtained by laboratory bioassays performed as part of the chemical control strategies of Triatoma sordida in the field. METHODS: Field bioassays were developed in Monte Azul and Coração de Jesus (Southeast, Brazil). In each location, samples were formed with three domestic units treated with alpha-cypermethrin 20.0% (Alfatek (r) 200 SC). One day after spraying, 10 fifth-instar nymphs remained in contact with the surfaces treated (adobe with plaster, adobe without plaster, or wood) with insecticide in plastic cones for 72h. Three cones were exposed inside the intradomicile and the peridomicile. The insects in the control group were exposed to an insecticide-free piece of cardboard. Mortality was measured 72h after removal of the insects from the treated surfaces. The tests were realized in triplicate. RESULTS: Mortality was 100.0% in all locations, except for Monte Azul; Landinho (96.6%) and Coração de Jesus; Barriguda (96.6%). CONCLUSIONS: Although the resistant populations in laboratory tests proved to be susceptible in the field, this observation is not sufficient to suggest that the cut-off points used to justify the resistance ratio should be changed. In this sense, we recommend that laboratory and field bioassays are carried out with a greater number of Triatominae populations to allow more in-depth consideration of the subject.

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.

Susceptibility of Triatoma sordida Stal, 1859 (Hemiptera: Reduviidae) to alpha-cypermethrin under natural climatic conditions

Despite the recommendations by interpretation of resistance ratios obtained in laboratory bioassays, little is known about the actual impact of these results in the effectiveness of vector control activities in the field. In this context, our objective was to determine the mean value of different resistance ratios obtained by laboratory bioassays performed as part of the chemical control strategies of Triatoma sordida in the field.

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

BACKGROUND: Triatoma 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. FINDINGS: 23 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. CONCLUSIONS: This 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.

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.

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. .

Susceptibility to deltamethrin of wild and domestic populations of Triatoma infestans of the Gran Chaco and the Inter-Andean Valleys of Bolivia.

BACKGROUND: The persistence of Triatoma infestans and the continuous transmission of Trypanosoma cruzi in the Inter-Andean Valleys and in the Gran Chaco of Bolivia are of great significance. Coincidentally, it is in these regions the reach of the vector control strategies is limited, and reports of T. infestans resistance to insecticides, including in wild populations, have been issued. This study aims to characterize the susceptibility to deltamethrin of wild and domestic populations of T. infestans from Bolivia, in order to better understand the extent of this relevant problem. METHODS: Susceptibility to deltamethrin was assessed in nine, wild and domestic, populations of T. infestans from the Gran Chaco and the Inter-Andean Valleys of Bolivia. Serial dilutions of deltamethrin in acetone (0.2 µL) were topically applied in first instar nymphs (F1, five days old, fasting, weight 1.2 ± 0.2 mg). Dose response results were analyzed with PROBIT version 2, determining the lethal doses, slope and resistance ratios (RR). Qualitative tests were also performed. RESULTS: Three wild T. infestans dark morph samples of Chaco from the Santa Cruz Department were susceptible to deltamethrin with RR50 of <2, and 100% mortality to the diagnostic dose (DD); however, two domestic populations from the same region were less susceptible than the susceptibility reference lineage (RR50 of 4.21 and 5.04 respectively and 93% DD). The domestic population of Villa Montes from the Chaco of the Tarija Department presented high levels of resistance (RR50 of 129.12 and 0% DD). Moreover, the domestic populations from the Valleys of the Cochabamba Department presented resistance (RR50 of 8.49 and 62% DD), the wild populations were less susceptible than SRL and T. infestans dark morph populations (RR50 < 5). CONCLUSION: The elimination of T. infestans with pyrethroid insecticides in Brazil, Uruguay, Chile, and its drastic reduction in large parts of Paraguay and Argentina, clearly indicates that pyrethroid resistance was very uncommon in non-Andean regions. The pyrethroid susceptibility of non-Andean T. infestans dark morph population, and the resistance towards it, of Andean T. infestans wild and domestic populations, indicates that the Andean populations from Bolivia are less susceptible.