Characterization of Sodium Channel Mutations in the Dengue Vector Mosquitoes Aedes aegypti and Aedes albopictus within the Context of Ongoing Wolbachia Releases in Kuala Lumpur, Malaysia

Abstract

Simple Summary

Mosquitoes, Aedes aegypti and Ae. albopictus are vectors of dengue and must be controlled to prevent and contain outbreaks of this disease. Control by insecticide application is common and pyrethroid insecticides provide rapid knockdown of mosquitoes combined with relatively low mammalian toxicity. However, resistance to pyrethroids and other chemicals is causing problems for mosquito control around the world. In Malaysia, an alternative method of dengue reduction is employed which comprises releases of Ae. aegypti mosquitoes infected with a bacterium, Wolbachia, found naturally in other insects. Wolbachia turns the mosquitoes into incompetent vectors so they do not transmit the disease. Wolbachia mosquitoes are reared in the laboratory before release and must be able to survive in the field where they will encounter insecticides. Our study demonstrates benefits of crossing laboratory mosquitoes to those from the field over generations, so that the mosquito lines acquire field resistance characteristics (mutations in the sodium channel gene). We demonstrate that resistance mutations provide a survival advantage to Wolbachia Ae. aegypti mosquitoes, which must be maintained in laboratory lines by regular backcrossing. We also describe appearance of a sodium channel mutation in Malaysian Ae. albopictus which may indicate that pyrethroid resistance is increasing in this species.

Abstract

Specific sodium channel gene mutations confer target site resistance to pyrethroid insecticides in mosquitoes and other insects. In Aedes mosquito species, multiple mutations that contribute to resistance vary in their importance around the world. Here, we characterize voltage sensitive sodium channel ( Vssc) mutations in populations of Aedes aegypti from Kuala Lumpur, Malaysia, and look at their persistence in populations affected by ongoing Wolbachia releases (a dengue control measure). We also describe a Vssc mutation in Aedes albopictus (F1534L) found for the first time in Malaysia. We show that there are three predominant Vssc haplotypes in Aedes aegypti in this region, which all persist with regular backcrossing, thereby maintaining the original genetic composition of the populations. We identify changes in genotype frequency in closed populations of Ae. aegypti maintained for multiple generations in laboratory culture, suggesting different fitness costs associated with the genotypes, some of which may be associated with the sex of the mosquito. Following population replacement of Ae. aegypti by Wolbachia in the target area, however, we find that the Vssc mutations have persisted at pre-release levels. Mosquitoes in two genotype classes demonstrate a type I pyrethroid resistance advantage over wildtype mosquitoes when exposed to 0.25% permethrin. This resistance advantage is even more pronounced with a type II pyrethroid, deltamethrin (0.03%). The results point to the importance of these mutations in pyrethroid resistance in mosquito populations and the need for regular backcrossing with male mosquitoes from the field to maintain similarity of genetic background and population integrity during Wolbachia releases.