目的 了解辽宁省白纹伊蚊对3种拟除虫菊酯类杀虫剂的抗药性水平及其击倒抗性基因型分布特点,为该地区防控白纹伊蚊及规范使用杀虫剂提供科学依据。方法 2023年7－8月,在辽宁省大连和营口市白纹伊蚊孳生地采集蚊虫,经形态学方法鉴定为白纹伊蚊后,采用成蚊接触筒法测定其对氯菊酯、溴氰菊酯和高效氯氰菊酯的抗药性。提取单只蚊虫的基因组DNA,PCR扩增电压门控钠离子通道（VGSC）基因部分片段,测序后对白纹伊蚊击倒抗性基因突变情况进行描述性统计分析。结果 大连地区白纹伊蚊成蚊对氯菊酯、溴氰菊酯、高效氯氰菊酯为可能抗性种群。营口地区白纹伊蚊成蚊对氯菊酯、溴氰菊酯、高效氯氰菊酯均已产生抗性。共检测2个野外种群110只白纹伊蚊,获得220条基因序列,长度约为400 bp,白纹伊蚊在1016、1532和1534位点均检测到突变。1016位点有2种等位基因,分别为野生型GTA/V（82.73%）和突变型GGA/G（17.27%）；有3种基因型,分别为野生型纯合子V/V（68.18%）、野生/突变型杂合子V/G（29.09%）和突变型纯合子G/G（2.73%）。1532位点有2种等位基因,分别为野生型ATC/I（86.82%）和突变型ACC/T（13.18%）；有3种基因型,分别为野生型纯合子I/I（76.36%）、野生/突变型杂合子I/T（20.91%）和突变型纯合子T/T（2.73%）。1534位点有4种等位基因,分别为野生型TTC/F（50.46%）、突变型TCC/S（23.64%）、突变型TGC/C（15.45%）和突变型CTC/L（10.45%）；有8种基因型,分别为野生型纯合子F/F（24.55%）、野生/突变型杂合子F/S（24.55%）、F/C（11.82%）和F/L（15.45%）,突变型纯合子S/S（9.09%）、C/C（7.27%）和L/L（2.73%）,突变型杂合子S/C（4.54%）。结论 辽宁省大连地区白纹伊蚊成蚊对3种拟除虫菊酯类杀虫剂为可能抗性种群,营口地区白纹伊蚊成蚊为抗性种群,大连和营口市白纹伊蚊均发生击倒抗性基因突变,其中1534位点突变率高且突变情况复杂多样,应持续关注白纹伊蚊抗药性水平,减少拟除虫菊酯类杀虫剂的使用,以延缓杀虫剂抗药性产生和发展。
Objective To investigate the resistance level of Aedes albopictus to three pyrethroid insecticides and the distribution of their knockdown resistance genotypes in Liaoning Province, China, so as to provide a scientific basis for the prevention and control of Ae. albopictus and the standardized use of insecticides in the region. Methods From July to August 2023, adult mosquitoes and larvae were collected from breeding sites of Ae. albopictus in Dalian and Yingkou in Liaoning Province. After mosquitoes were identified by morphology as Ae. albopictus, their resistance to three pyrethroid insecticides (permethrin, deltamethrin, and beta-cypermethrin) was determined by the adult mosquito WHO tube assay. The genomic DNAs of single mosquito were extracted, and the partial fragment of voltage-gated sodium channel (VGSC) gene was amplified by polymerase chain reaction. Descriptive statistical analysis method was used for the analysis of knockdown resistance genes mutations in Ae. albopictus after sequencing. Results Adult Ae. albopictus mosquitoes in Dalian had have possible resistance to permethrin, deltamethrin, and beta-cypermethrin. Adult Ae. albopictus mosquitoes in Yingkou area had developed resistance to permethrin, deltamethrin, and beta-cypermethrin. A total of 110 Ae. albopictus from two field populations were detected, and 220 gene sequences were obtained, with the length of about 400 bp. Mutations were detected at 1016, 1532, and 1534 loci of Ae. albopictus. At the 1016 locus, there were two alleles, namely wild-type GTA/V (82.73%) and mutant GGA/G (17.27%); there were three genotypes, namely wild-type homozygote V/V (68.18%), wild-type/mutant heterozygote V/G (29.09%), and mutant homozygote G/G (2.73%). At the 1532 locus, there were two alleles, namely wild-type ATC/I (86.82%) and mutant ACC/T (13.18%); there were three genotypes, namely wild-type homozygote I/I (76.36%), wild-type/mutant heterozygote I/T (20.91%), and mutant homozygote T/T (2.73%). At the 1534 locus, there were four alleles, namely wild-type TTC/F (50.46%), mutant TCC/S (23.64%), mutant TGC/C (15.45%), and mutant CTC/L (10.45%); there were eight genotypes, namely wild-type homozygote F/F (24.55%), wild-type/mutant heterozygotes F/S (24.55%), F/C (11.82%), and F/L (15.45%), mutant homozygotes S/S (9.09%) , C/C (7.27%), and L/L (2.73%), and mutant heterozygote S/C (4.54%). Conclusions Adult Ae. albopictus mosquitoes in Dalian of Liaoning Province have possible resistance to the three pyrethroid insecticides, and adult Ae. albopictus mosquitoes in Yingkou are resistant to the three pyrethroid insecticides. The knockdown resistance gene of Ae. albopictus is mutated in Dalian and Yingkou areas, with a high mutation rate and complex and diverse mutations at the 1534 locus. Sustained attention should be paid to the resistance level of Ae. albopictus and reduce the use of pyrethroid insecticides, so as to delay the emergence and development of insecticide resistance.
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