Aedes aegypti is an important vector for dengue virus and thus has been targeted with pyrethroid insecticides in many areas of the world. As such, resistance has been detected to several of these insecticides, including in China, but the mechanisms of the resistance are not well understood in this country. Using the World Health Organization larval mosquito bioassay, five field populations of Aedes aegypti from Southern China were characterized for their resistance to cypermethrin and cyhalothrin. RNA extraction with PCR amplification, cloning and sequencing of the sodium channel gene was followed by comparisons of susceptible and wild mosquito strains Additionally, genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect S989P, V1016G and F1534C mutations and allow for correlation analysis of resistance expression for the different mutations. All wild strains expressed resistance to cypermethrin and cyhalothrin and the resistance expression between the two insecticides was highly correlated suggesting cross-resistance between these two pyrethroids. The AS-PCR technique effectively distinguished individual genotypes for all three mutations. Among the five wild strains tested, two strains carried all three mutations. Although the S989P and V1016G mutations were positively correlated to resistance expression of both pyrethroids, the F1534C mutation was negatively correlated. Our methodology proved highly reliable and will aid future detection of kdr mutations. The three sodium channel mutations were common in the Ae. aegypti strains sampled from Southern China. The V1016G mutation appears to be the most important kdr mutation in Ae. aegypti strains in Southern China.
Culex pipiens pallens and Cx. p. quinquefasciatus are important vectors of many diseases, such as West Nile fever and lymphatic filariasis. The widespread use of insecticides to control these disease vectors and other insect pests has led to insecticide resistance becoming common in these species. In this study, high throughout Illumina sequencing was used to identify hundreds of Cx. p. pallens and Cx. p. quinquefasciatus genes that were differentially expressed in response to insecticide exposure. The identification of these genes is a vital first step for more detailed investigation of the molecular mechanisms involved in insecticide resistance in Culex mosquitoes.
There was no record of Aedes aegypti in Yunnan Province, China, until 2002, but this species is now continuously found in nine cities (or counties). Until now, little was known about the genetic diversity and population structure of this invasive species. Thus, a detailed understanding of the invasion strategies, colonisation and dispersal of this mosquito from a population genetics perspective is urgently needed for controlling and eliminating this disease vector. The genetic diversity and population structure of Ae. aegypti communities were analysed by screening nine microsatellite loci from 833 Ae. aegypti mosquitoes sampled from 28 locations in Yunnan Province. In total, 114 alleles were obtained, and the average polymorphic information content (PIC) value was 0.672. The value of the alleles per locus ranged from 2.90 to 5.18, with an average of 4.04. The value of He ranged from 0.353 to 0.681, and the value of Ho within populations ranged from 0.401 to 0.689. Of the 28 locations, two showed significant departures from the Hardy-Weinberg equilibrium (HWE) with P-values less than 0.05, and a bottleneck effect was detected among locations from Ruili and the border areas with the degree of 60% and 50%, respectively. Combined with the F-statistics (FIT = 0.222; FCT = 0.145), the analysis of molecular variance (AMOVA) revealed that there was substantial molecular variation among individuals, accounting for 77.76% of the sample, with a significant P-value (<0.0001). The results suggest that genetic differences in Ae. aegypti originated primarily among individuals rather than among populations. Furthermore, the STRUCTURE and UPGMA cluster analyses showed that Ae. aegypti from the border areas were genetically isolated compared to those from the cities Ruili and Jinghong, consistent with the results of the Mantel test (R 2 = 0.245, P < 0.0001). Continuous invasion contributes to the maintenance of Ae. aegypti populations’ genetic diversity and different invasion accidents result in the genetic difference among Ae. aegypti populations of Yunnan Province.
Aedes albopictus is an indigenous primary vector of dengue and Zika viruses in China. Understanding the population spatial genetic structure, migration, and gene flow of vector species is critical to effectively preventing and controlling vector-borne diseases. The genetic variation and population structure of Ae. albopictus populations collected from 22 cities along the Yangtze River Basin were investigated with nine microsatellite loci and the mitochondrial CoxI gene. The polymorphic information content (PIC) values ranged from 0.534 to 0.871. The observed number of alleles (Na) values ranged from 5.455 to 11.455, and the effective number of alleles (Ne) values ranged from 3.106 to 4.041. The Shannon Index (I) ranged from 1.209 to 1.639. The observed heterozygosity (Ho) values ranged from 0.487 to 0.545. The FIS value ranged from 0.047 to 0.212. All Ae. albopictus populations were adequately allocated to three clades with significant genetic differences. Haplotype 2 is the most primitive molecular type and forms 26 other haplotypes after one or more site mutations. The rapid expansion of high-speed rail, aircraft routes and highways along the Yangtze River Basin have accelerated the dispersal and communication of mosquitoes, which appears to have contributed to inhibited population differentiation and promoted genetic diversity among Ae. albopictus populations.
Aedes aegypti is the primary vector of mosquito-borne viruses and threatens human health globally. To monitor temporal changes in the population genetics of Ae. aegypti in Yunnan Province, we collected specimens from five sites between 2016 and 2018 and analyzed them at nine microsatellite loci and the mitochondrial cox I gene. Genetic diversity in these populations rose initially and then declined over the study period. The inbreeding coefficient shifted from a heterozygous state toward moderate inbreeding. Ten cox I haplotypes were identified, with haplotypes H06 and H09 prevalent across regions, while other haplotypes were limited to specific regions or years. Clustering analyses demonstrated significant population shifts of Ae. aegypti across the five sampling sites from 2016 to 2018. AMOVA analysis of all Ae. aegypti populations indicated that from 2016 to 2018, the main source of variation was predominantly among individuals, accounting for 76.43%, 65.15%, and 80.36%, respectively. Population dispersion patterns across locations initially increased and then decreased over time. Therefore, continuous monitoring is essential, and particular attention should be paid to intra-provincial spread, especially between port and urban areas.
Aedes aegypti is a major invasive vector in China, where prolonged pyrethroid use has induced resistance, complicating dengue control. This study evaluated the resistance levels of Ae. aegypti to the pyrethroid beta-cypermethrin in Yunnan and explored the underlying mechanisms to inform control strategies. Mosquitoes were collected from five regions of Yunnan in 2015–2016. Larval bioassays, adult diagnostic dose determination, and adult bioassays were conducted to assess resistance to beta-cypermethrin. Mixed-function oxidase (MFO) amount was measured in larvae, and PCR amplification was used to detect mutations in the voltage-gated sodium channel (VGSC) gene. Correlations between enzyme activity, mutations, and phenotypic resistance were analyzed. Larval bioassays indicated that all five populations exhibited resistance to beta-cypermethrin, with resistance ratios ranging from 11.31 to 41.56. The adult diagnostic dose was determined as 0.74668 g/L, with mortality rates ranging from 8.89% to 58.89%, confirming resistance. A significant negative correlation was found between adult mortality and larval LC50 values. MFO amount was closely correlated with resistance levels. The F1534C mutation was the only VGSC mutation detected, and its frequency showed a significant positive correlation with beta-cypermethrin resistance. Our data were collected in 2015–2016; resistance levels may have changed since then, and no confirmatory bioassays were performed by us after 2020. The findings demonstrate that the F1534C mutation and increased MFO amount contribute to enhanced beta-cypermethrin resistance of Ae. aegypti in Yunnan. By linking phenotype resistance to key molecular and biochemical mechanisms, these findings support the continued monitoring of resistance and provide a basis for the evidence-based optimization of dengue vector control strategies.