Screening Aedes aegypti (Diptera: Culicidae) Populations From Pernambuco, Brazil for Resistance to Temephos, Diflubenzuron, and Cypermethrin and Characterization of Potential Resistance Mechanisms.

Resistance to chemical insecticides detected in Aedes aegypti (L.) mosquitoes has been a problem for the National Dengue Control Program (PNCD) over the last years. In order to provide deeper knowledge of resistance to xenobiotics, our study evaluated the susceptibility profile of temephos, diflubenzuron, and cypermethrin insecticides in natural mosquito populations from the Pernambuco State, associating these results with the local historical use of such compounds. Furthermore, mechanisms that may be associated with this particular type of resistance were characterized. Bioassays with multiple temephos and diflubenzuron concentrations were performed to detect and quantify resistance. For cypermethrin, diagnostic dose assays were performed. Biochemical tests were carried out to quantify the activity of detoxification enzymes. In addition, a screening of mutations present in the voltage-gated sodium channel gene (NaV) was performed in samples previously submitted to bioassays with cypermethrin. The populations under study were resistant to temephos and showed a positive correlation between insecticide consumption and the resistance ratio (RR) to the compound. For diflubenzuron, the biological activity ratio (BAR) ranged from 1.3 to 4.7 times, when compared to the susceptible strain. All populations showed resistance to cypermethrin. Altered enzymatic profiles of alpha, p-nitrophenyl acetate (PNPA) esterases and glutathione-S-transferases were recorded in most of these samples. Molecular analysis demonstrated that Arcoverde was the only population that presented the mutated form 1016Ile/Ile. These findings show that the situation is critical vis-à-vis the effectiveness of mosquito control using chemical insecticides, since resistance to temephos and cypermethrin is widespread in Ae. aegypti from Pernambuco.

Insecticide resistance in Aedes aegypti populations from Ceará, Brazil.

BACKGROUND: Organophosphates and pyrethroids are used widely in Brazil to control Aedes aegypti, the main vector of dengue viruses, under the auspices of the National Programme for Dengue Control. Resistance to these insecticides is widespread throughout Brazil. In Ceará the vector is present in 98% of districts and resistance to temephos has been reported previously. Here we measure resistance to temephos and the pyrethroid cypermethrin in three populations from Ceará and use biochemical and molecular assays to characterise resistance mechanisms. RESULTS: Resistance to temephos varied widely across the three studied populations, with resistance ratios (RR(95)) of 7.2, 30 and 192.7 in Juazeiro do Norte, Barbalha and Crato respectively. The high levels of resistance detected in Barbalha and Crato (RR(95) ≥ 30) imply a reduction of temephos efficacy, and indeed in simulated field tests reduced effectiveness was observed for the Barbalha population. Two populations (Crato and Barbalha) were also resistant to cypermethrin, whilst Juazeiro do Norte showed only an altered susceptibility. The Ile1011Met kdr mutation was detected in all three populations and Val1016Ile in Crato and Juazeiro do Norte. 1011Met was significantly associated with resistance to cypermethrin in the Crato population. Biochemical tests showed that only the activity of esterases and GSTs, among the tested detoxification enzymes, was altered in these populations when compared with the Rockefeller strain. CONCLUSIONS: Our results demonstrate that two A. aegypti populations from Ceará are under strong selection pressure by temephos, compromising the field effectiveness of this organophosphate. Our results also provide evidence that the process of reducing resistance to this larvicide in the field is difficult and slow and may require more than seven years for reversal. In addition, we show resistance to cypermethrin in two of the three populations studied, and for the first time the presence of the allele 1016Ile in mosquito populations from northeastern Brazil. A significant association between 1011Met and resistance was observed in one of the populations. Target-site mechanisms seem not to be implicated in temephos resistance, reinforcing the idea that for the studied populations, detoxification enzymes most likely play a major role in the resistance to this insecticide.