The Pattern of Knockdown Resistance Mutations Correlated to the Annual Average Temperature in Field Populations of Aedes Albopictus (Diptera: Culicidae) in China

Abstract Background: Aedes albopictus is the main vector of dengue fever in China, distributed from north to south in China. Insecticides are an important method to control the mosquitoes, especially in the outbreak of dengue fever, but insecticide resistance raises the risk of failure to control vector-borne diseases. Knockdown resistance (kdr) caused by point mutations in the VGSC gene is a key mechanism that confers resistance to pyrethroids. To explore the characteristics and possible evolution trend of kdr mutation in Ae. albopictus, we analyzed the kdr mutations of field populations in China in this study.Methods: A total of 1 549 Ae. albopictus were collected from 18 sites in China from 2017 to 2019, as well as 50 individuals from three sites in the 1990s. A fragment of approximately 350 bp from part of S6 segment in the VGSC gene domain III was amplified and sequenced. The haplotypes of VGSC gene were recorded and the parsimony network was constructed using TCS 1.21. The data of annual average temperatures (AAT) of collection sites was acquired from national database. The correlation between AAT of the collection site and the kdr mutation rate was analyzed by Pearson Correlation using SPSS 21.0. Results: The overall frequency of mutant allele F1534 is 45.62%. Nine mutant alleles were detected at codon 1534 in fifteen field populations, namely TCC/TCG (S) (38.86%), TTG/CTG/CTC/TTA (L) (3.71%), TGC (C) (2.68%), CGC (R) (0.27%) and TGG (W) (0.10%). Only one mutant allele ACC (T) was found at codon 1532 with frequency of 6.39% in ten field populations. Moreover, multiple mutations at I1532 and F1534 in a sample appeared in five populations. The 1534 mutation rate was significantly positive related to AAT (Coefficient=0.624, p=0.0056), while the 1532 mutation rate was significantly negative related to AAT (Coefficient=-0.645, p=0.0038). Thirteen haplotypes were inferred, in which six mutant haplotypes were formed by one step, and the other six haplotypes were formed by one more mutations. In the samples from 1990s, no mutant allele was detected at codon 1532 of VGSC gene. However, F1534S/TCC was found in HNHK94 with an unexpected frequency of 100%.ConclusionsKdr mutations are widespread in the field populations of Ae. albopictus in China. Two novel mutant alleles F1534W/TGG and F1534R/CGC were the detected. The 1534 kdr mutation appeared in the population of Ae. albopictus no later than 1990s. F1534 mutation rate is positive correlated to AAT, while I1532 mutation rate is negative correlated to AAT. Insecticide using should be carefully managed to slow down the spread of high-resistance Ae. albopictus populations.