Genetic structure and kdr mutations in Aedes aegypti populations along a road crossing the Amazon Forest in Amapá State, Brazil.

Insecticide resistance in Aedes aegypti poses a significant threat to disease control. One form of resistance, caused by kdr mutations in the NaV gene, hinders vector control efforts in Brazil. Despite genetic differences typically accumulating among isolated populations, this mosquito can actively and passively disperse through human transportation. Our study investigated the genetic structure and spread of kdr mutations in Ae. aegypti populations across six localities in Amapá State, Brazil, within the Amazonian Forest. Using 12 microsatellite loci and qPCR methods, we assessed genetic structure and identified three common kdr mutations (V410L, V1016I, and F1534C). High prevalence of kdr alleles was observed in all localities, indicating widespread distribution in Amapá State. Microsatellite analysis revealed differentiation among mosquito populations, dividing them into two distinct clusters supported by Bayesian and DAPC analyses. Oiapoque, located along the northern border with French Guiana, exhibited the highest kdr frequencies and genetic differentiation compared to other localities. Our findings suggest genetic structure in Ae. aegypti populations in Amapá State, with some passive gene flow between clusters. The study underscores the need for continuous surveillance of Ae. aegypti populations to monitor the spread of insecticide resistance and inform effective vector control strategies.

Efficacy of insecticides used in indoor residual spraying for malaria control: an experimental trial on various surfaces in a "test house".

BACKGROUND: Malaria is a public health problem in the Brazilian Amazon region. In integrated vector management for malaria (anopheline) control, indoor residual spraying (IRS) represents one of the main tools in the basic strategy applied in the Amazonian states. It is essential to understand the residual efficacy of insecticides on different surfaces to determine spray cycles, ensure their rational use, and prevent wastage. This study aimed to evaluate the residual efficacy of six insecticide formulations used in the National Malaria Control Programme on four different types of walls in a field simulation at a "test house". METHODS: The tests were performed as a field-simulating evaluation at a "test house" built in the municipality of Macapá. Six insecticide formulations comprising four pyrethroids, a carbamate, and an organophosphate were used, and evaluated when applied on different wall surfaces: painted wood, unpainted wood, plastered cement, and unplastered cement. The insecticides were applied to the interior walls of the "test house" by a trained technician. RESULTS: In the bioassays performed with pyrethroids, deltamethrin water-dispersible granules (WG) performed particularly well, presenting residual bioefficacy of 8 months on both wood surfaces after the IRS, whereas alpha-cypermethrin suspension concentrate (SC) and etofenprox wettable powder (WP) demonstrated residual bioefficacy of 4 months on at least one of the wood surfaces; however, the pyrethroid lambda-cyhalothrin WP showed a low residual bioefficacy (< 3 months) on all tested surfaces, demonstrating its inefficiency for areas with a long transmission cycle of malaria. For the carbamate-bendiocarb WP, residual bioefficacy for 3 months was achieved only on wood surfaces. In general, the organophosphate pirimifos-methyl capsule suspension (CS) demonstrated the best result, with a mortality rate < 80% over a period of 6 months on all surfaces tested. CONCLUSION: Insecticide efficiency varies among different types of surface; therefore, a "test house" is a valuable evaluation tool. This work highlights the usefulness of associating the residual efficacy of insecticides on the surfaces commonly found in houses in endemic areas, together with knowledge about the transmission cycle duration of the transmission cycle and the insecticide susceptibility of the vector. This association helps in the decision-making for the malaria control intervention regarding.

Spatial and temporal country-wide survey of temephos resistance in Brazilian populations of Aedes aegypti.

The organophosphate temephos has been the main insecticide used against larvae of the dengue and yellow fever mosquito (Aedes aegypti) in Brazil since the mid-1980s. Reports of resistance date back to 1995; however, no systematic reports of widespread temephos resistance have occurred to date. As resistance investigation is paramount for strategic decision-making by health officials, our objective here was to investigate the spatial and temporal spread of temephos resistance in Ae. aegypti in Brazil for the last 12 years using discriminating temephos concentrations and the bioassay protocols of the World Health Organization. The mortality results obtained were subjected to spatial analysis for distance interpolation using semi-variance models to generate maps that depict the spread of temephos resistance in Brazil since 1999. The problem has been expanding. Since 2002-2003, approximately half the country has exhibited mosquito populations resistant to temephos. The frequency of temephos resistance and, likely, control failures, which start when the insecticide mortality level drops below 80%, has increased even further since 2004. Few parts of Brazil are able to achieve the target 80% efficacy threshold by 2010/2011, resulting in a significant risk of control failure by temephos in most of the country. The widespread resistance to temephos in Brazilian Ae. aegypti populations greatly compromise effective mosquito control efforts using this insecticide and indicates the urgent need to identify alternative insecticides aided by the preventive elimination of potential mosquito breeding sites.

Spatial and temporal country-wide survey of temephos resistance in Brazilian populations of Aedes aegypti

The organophosphate temephos has been the main insecticide used against larvae of the dengue and yellow fever mosquito (Aedes aegypti) in Brazil since the mid-1980s. Reports of resistance date back to 1995; however, no systematic reports of widespread temephos resistance have occurred to date. As resistance investigation is paramount for strategic decision-making by health officials, our objective here was to investigate the spatial and temporal spread of temephos resistance in Ae. aegypti in Brazil for the last 12 years using discriminating temephos concentrations and the bioassay protocols of the World Health Organization. The mortality results obtained were subjected to spatial analysis for distance interpolation using semi-variance models to generate maps that depict the spread of temephos resistance in Brazil since 1999. The problem has been expanding. Since 2002-2003, approximately half the country has exhibited mosquito populations resistant to temephos. The frequency of temephos resistance and, likely, control failures, which start when the insecticide mortality level drops below 80%, has increased even further since 2004. Few parts of Brazil are able to achieve the target 80% efficacy threshold by 2010/2011, resulting in a significant risk of control failure by temephos in most of the country. The widespread resistance to temephos in Brazilian Ae. aegypti populations greatly compromise effective mosquito control efforts using this insecticide and indicates the urgent need to identify alternative insecticides aided by the preventive elimination of potential mosquito breeding sites.

Surveillance of Aedes aegypti: comparison of house index with four alternative traps.

INTRODUCTION: The mosquito Aedes aegypti, vector of dengue, chikungunya and yellow fever viruses, is an important target of vector control programs in tropical countries. Most mosquito surveillance programs are still based on the traditional household larval surveys, despite the availability of new trapping devices. We report the results of a multicentric entomological survey using four types of traps, besides the larval survey, to compare the entomological indices generated by these different surveillance tools in terms of their sensitivity to detect mosquito density variation. METHODS: The study was conducted in five mid-sized cities, representing variations of tropical climate regimens. Surveillance schemes using traps for adults (BG-Sentinel, Adultrap and MosquiTRAP) or eggs (ovitraps) were applied monthly to three 1 km(2) areas per city. Simultaneously, larval surveys were performed. Trap positivity and density indices in each area were calculated and regressed against meteorological variables to characterize the seasonal pattern of mosquito infestation in all cities, as measured by each of the four traps. RESULTS: The House Index was consistently low in most cities, with median always 0. Traps rarely produced null indices, pointing to their greater sensitivity in detecting the presence of Ae. aegypti in comparison to the larval survey. Trap positivity indices tend to plateau at high mosquito densities. Despite this, both indices, positivity and density, agreed on the seasonality of mosquito abundance in all cities. Mosquito seasonality associated preferentially with temperature than with precipitation even in areas where temperature variation is small. CONCLUSIONS: All investigated traps performed better than the House Index in measuring the seasonal variation in mosquito abundance and should be considered as complements or alternatives to larval surveys. Choice between traps should further consider differences of cost and ease-of-use.

Assessing the effects of Aedes aegypti kdr mutations on pyrethroid resistance and its fitness cost.

Pyrethroids are the most used insecticide class worldwide. They target the voltage gated sodium channel (NaV), inducing the knockdown effect. In Aedes aegypti, the main dengue vector, the AaNaV substitutions Val1016Ile and Phe1534Cys are the most important knockdown resistance (kdr) mutations. We evaluated the fitness cost of these kdr mutations related to distinct aspects of development and reproduction, in the absence of any other major resistance mechanism. To accomplish this, we initially set up 68 crosses with mosquitoes from a natural population. Allele-specific PCR revealed that one couple, the one originating the CIT-32 strain, had both parents homozygous for both kdr mutations. However, this pyrethroid resistant strain also presented high levels of detoxifying enzymes, which synergistically account for resistance, as revealed by biological and biochemical assays. Therefore, we carried out backcrosses between CIT-32 and Rockefeller (an insecticide susceptible strain) for eight generations in order to bring the kdr mutation into a susceptible genetic background. This new strain, named Rock-kdr, was highly resistant to pyrethroid and presented reduced alteration of detoxifying activity. Fitness of the Rock-kdr was then evaluated in comparison with Rockefeller. In this strain, larval development took longer, adults had an increased locomotor activity, fewer females laid eggs, and produced a lower number of eggs. Under an inter-strain competition scenario, the Rock-kdr larvae developed even slower. Moreover, when Rockefeller and Rock-kdr were reared together in population cage experiments during 15 generations in absence of insecticide, the mutant allele decreased in frequency. These results strongly suggest that the Ae. aegypti kdr mutations have a high fitness cost. Therefore, enhanced surveillance for resistance should be priority in localities where the kdr mutation is found before new adaptive alleles can be selected for diminishing the kdr deleterious effects.

Comparison of different uses of adult traps and ovitraps for assessing dengue vector infestation in endemic areas.

This report presents a set of field experiments designed to assess different protocols for the use of ovitrap and MosquiTRAP, a promising new trap for dengue vector monitoring. Percentage of positive houses, calculated either by the use of 2 traps (outside + inside) or 1 trap (outside) per house, tended to increase with time of exposure, at similar rates. When the aim was either to obtain a qualitative index (Aedes aegypti-positive site) or to determine percentage of positive houses in a selected neighborhood, the use of 1 ovitrap per house with only 5 days of exposure at the peridomestic area was quite sensitive. Seven days of exposure was too long, as saturation is expected in some places. The number of eggs collected per site increased with the time of exposure in all sites. At the 3-day trap exposure, we were not able to discriminate neighborhoods in terms of egg productivity per house. At the 5-day trap exposure, a rank of 4 sites was achieved. There was no correlation between the number of adults caught in MosquiTRAPs and number of eggs collected in ovitraps, neither per neighborhood nor per house where both traps were simultaneously exposed for 7 days in the peridomestic area.