Tracking pyrethroid resistance in arbovirus mosquito vectors: mutations I1532T and F1534C in Aedes albopictus across Europe.

BACKGROUND: With the worldwide spread of the Asian tiger mosquito, Aedes albopictus, the number of autochthonous cases of exotic arboviral diseases, such as dengue or chikungunya, is increasing in temperate regions. In Europe, pyrethroids are the only insecticides allowed for the abatement of adult mosquitoes and are thus crucial for limiting ongoing arbovirus transmission. Despite this and the report of resistance rising in vector populations worldwide, information on the pyrethroid resistance status of vector populations and knowledge on resistance mechanisms is widely lacking. Genotyping of knockdown resistance (kdr) mutations situated within the target site of pyrethroids, i.e., the voltage-gated sodium channel (VGSC), and associated with pyrethroid resistance, is a cost-effective approach to investigate the spread of resistance in a population. Herein, we describe the European-wide distribution of two kdr mutations, i.e., I1532T and F1534C, in Ae. albopictus and evaluate their co-occurrence with another well-characterized kdr mutation, V1016G.

METHODS: Genotyping of the kdr mutation F1534C was performed by allele-specific PCR for 1732 Ae. albopictus specimens sampled in 19 European countries; for a subset of 419 specimens mutation I1532T was also genotyped by sequencing. For all samples, information on mutation V1016G was available, allowing evaluation of the co-occurrence of kdr alleles.

RESULTS: Mutation 1534C was detected in nine sites from six countries at an overall frequency close to 5%. Highest frequencies per site were detected in Cyprus (84%) and Greece (45%). Allele 1532 T was identified in 11 sites from 7 countries at frequencies ranging from 4% to 25% per site. Co-occurrence of different kdr alleles (1534C, 1532 T and 1016G) was observed in nine sampling sites from seven countries.

CONCLUSIONS: The present study offers the first map of the occurrence of the major Ae. albopictus kdr alleles across Europe and highlights a differential distribution of the two alleles most strongly associated with pyrethroid resistance, 1016G and 1534C. Our findings also point to the need for enhancing resistance monitoring in the Eastern Mediterranean region, where the two mutations are shown to exist in geographically close areas, with the risk of emergence of highly resistant double mutants.