Abstract

BackgroundAedes aegypti is an important vector for dengue virus and thus has been targeted with pyrethroid insecticides in many areas of the world. As such, resistance has been detected to several of these insecticides, including in China, but the mechanisms of the resistance are not well understood in this country.MethodsUsing the World Health Organization larval mosquito bioassay, five field populations of Aedes aegypti from Southern China were characterized for their resistance to cypermethrin and cyhalothrin. RNA extraction with PCR amplification, cloning and sequencing of the sodium channel gene was followed by comparisons of susceptible and wild mosquito strains Additionally, genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect S989P, V1016G and F1534C mutations and allow for correlation analysis of resistance expression for the different mutations.ResultsAll wild strains expressed resistance to cypermethrin and cyhalothrin and the resistance expression between the two insecticides was highly correlated suggesting cross-resistance between these two pyrethroids. The AS-PCR technique effectively distinguished individual genotypes for all three mutations. Among the five wild strains tested, two strains carried all three mutations. Although the S989P and V1016G mutations were positively correlated to resistance expression of both pyrethroids, the F1534C mutation was negatively correlated.ConclusionsOur methodology proved highly reliable and will aid future detection of kdr mutations. The three sodium channel mutations were common in the Ae. aegypti strains sampled from Southern China. The V1016G mutation appears to be the most important kdr mutation in Ae. aegypti strains in Southern China.

Highlights

  • Aedes aegypti is an important vector for dengue virus and has been targeted with pyrethroid insecticides in many areas of the world

  • Identification of sodium channel gene mutations Three mutations were identified from the cDNA of a pooled sample of F1 specimens of wild Ae. aegypti: S989P (TCC-CCC), V1016G (GTA-GGA), and F1534C (TTC-TGC), compared to the susceptible strain

  • We found that the F1534C mutation was negatively correlated with resistance to cypermethrin and cyhalothrin, which suggests that the higher the frequency of this mutation in mosquito strains, the lower their resistance to these two insecticides; the presence of this mutation does not enhance resistance to cypermethrin and cyhalothrin but reduces it

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Summary

Introduction

Aedes aegypti is an important vector for dengue virus and has been targeted with pyrethroid insecticides in many areas of the world. Because there is currently no dengue vaccine, mosquito control has become the most important method of preventing transmission of these diseases [1] and insecticides are expected to remain a key component of dengue control for the foreseeable future [2] Both pyrethroids and DDT target the voltage-gated sodium channel of insect neurons and single amino acid substitutions in the sodium channel have been associated with resistance to both these insecticides [5] This form of resistance, known as knockdown resistance (kdr), has been observed in several insect species including the mosquitoes Anopheles gambiae Giles [6], An. stephensi Liston [7], Culex quinquefasciatus Say [8] and Ae. aegypti [9]. A number of kdr mutations have been identified in Ae. aegypti (S989P, I1011M/V, V1016G/I, F1269C, F1534C) [9, 14, 15] and at least two of these are known to be related to pyrethroid resistance; 1016 (Val to Ile or Gly) and 1534 (Phe to Cys) in the IIS6 and IIIS6 segments of the Vssc gene, respectively [9, 14, 16, 17]

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