Co-occurrence of Point Mutations in the Voltage-Gated Sodium Channel of Pyrethroid-Resistant Aedes aegypti Populations in Myanmar

Abstract

Background

Single amino acid substitutions in the voltage-gated sodium channel associated with pyrethroid resistance constitute one of the main causative factors of knockdown resistance in insects. The kdr gene has been observed in several mosquito species; however, point mutations in the para gene of Aedes aegypti populations in Myanmar have not been fully characterized. The aim of the present study was to determine the types and frequencies of mutations in the para gene of Aedes aegypti collected from used tires in Yangon City, Myanmar.

Methodology/Principal Findings

We determined high pyrethroid resistance in Aedes aegypti larvae at all collection sites in Yangon City, by using a simplified knockdown bioassay. We showed that V1016G and S989P mutations were widely distributed, with high frequencies (84.4% and 78.8%, respectively). By contrast, we were unable to detect I1011M (or I1011V) or L1014F mutations. F1534C mutations were also widely distributed, but with a lower frequency than the V1016G mutation (21.2%). High percentage of co-occurrence of the homozygous V1016G/S989P mutations was detected (65.7%). Additionally, co-occurrence of homozygous V1016G/F1534C mutations (2.9%) and homozygous V1016G/F1534C/S989P mutations (0.98%) were detected in the present study.

Conclusions/Significance

Pyrethroid insecticides were first used for malaria control in 1992, and have since been constantly used in Myanmar. This intensive use may explain the strong selection pressure toward Aedes aegypti, because this mosquito is generally a domestic and endophagic species with a preference for indoor breeding. Extensive use of DDT for malaria control before the use of this chemical was banned may also explain the development of pyrethroid resistance in Aedes aegypti.

Author Summary

The use of pyrethroids with high killing activity has accelerated the development of pyrethroid resistance in vector mosquitoes. The knockdown resistance ( kdr) allele contains a single amino acid substitution in the voltage-gated sodium channel and is one of the main causative factors of pyrethroid resistance in insects. We investigated the distribution of the kdr gene in Aedes aegypti larvae collected from used tires in Yangon City, Myanmar. We detected three patterns of co-occurrence of point mutations, namely, V1016G/S989P with wide distribution, and small number of V1016G/F1534C and V1016G/F1534C/S989P, both of which were first found in the field collected Ae. aegypti population. The use of pyrethroids and/or DDT for malaria control is thought to be one of the main causes of pyrethroid resistance in Aedes aegypti. Insecticide treatment for malaria vector control seems to have been intensively conducted in the interior and along the periphery of human habitation areas, where the breeding and resting sites of Aedes aegypti are located.