Seasonal dynamics of insecticide resistance, multiple resistance, and morphometric variation in field populations of Culex pipiens

• High frequencies and striking seasonal changes for two kdr mutations were detected.
• Low frequencies of resistance associated mutations in ace-1 and Rdl were detected with no seasonal changes.
• Our study did not reveal the presence of insects that are resistant to all pesticides.
• Morphological analysis displayed a similar pattern for both wings and did not show a clear separation.
• Our results also revealed the presence of genetic exchanges between Cx. pipiens populations.

Resistance to insecticides that impairs nervous transmission has been widely investigated in mosquito populations as insecticides are crucial to effective insect control. The development of insecticide resistance is also of special interest to evolutionary biologists since it represents the opportunity to observe the genetic consequences of a well-characterized alteration in the environment. Although the frequencies of resistance alleles in Culex pipiens populations against different groups of insecticides have been reported, no detailed information is available on the relative change in these allele frequencies over time. In this study, we collected mosquitoes of the Cx. pipiens complex from six locations in three seasons in the Aegean region of Turkey and examined the i) seasonal variations in resistance to four different chemical classes of insecticides, ii) seasonal fluctuations in frequencies of resistance-associated target-site mutations of the three genes (ace-1, kdr, and Rdl), and iii) potential seasonal variations in wing morphometric characters that may be modified in resistant mosquitoes. Our bioassay results indicated the presence of different levels of resistance to all tested insecticides for all three seasons in all locations. The results of the PCR-based molecular analysis revealed low frequencies of mutations in ace-1 and Rdl that are associated with resistance to malathion, bendiocarb, and dieldrin and no obvious seasonal changes. In contrast, we detected high frequencies and striking seasonal changes for two kdr mutations associated with resistance to DDT and pyrethroids. In addition, the evaluation of the field populations from all seasons in terms of the combinations of polymorphisms at four resistance-associated mutations did not reveal the presence of insects that are resistant to all pesticides. Results from the morphological analysis displayed a similar pattern for both wings and did not show a clear separation among the samples from the three different seasons. The results of this study have advanced our knowledge of the potential dynamics of insecticide resistance among populations of the Cx. pipiens complex. The implications of these results to the understanding of the evolution of insecticide resistance and the management of resistance in mosquitoes are discussed.

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