海南省埃及伊蚊抗药性测定及击倒抗性基因V1016G和F1534C突变

Determination of insecticide resistance in Aedes aegypti in Hainan and analysis of V1016G and F1534C knockdown resistance mutations

目的了解海南省埃及伊蚊抗性水平,为制定当地登革热媒介埃及伊蚊防控措施提供参考。方法2020年,在海南省儋州和昌江2地采集埃及伊蚊的幼虫和蛹,繁殖至子一代(F;1;)。采用成蚊接触筒法测定埃及伊蚊对常用杀虫剂的抗性,然后利用等位基因特异性聚合酶链反应(allele-specific polymerase chain reaction,AS-PCR)法测定埃及伊蚊击倒抗性(knockdown resistance,kdr)基因V1016G和F1534C突变情况。结果海南昌江与儋州埃及伊蚊接触溴氰菊酯(0.03%)、氯菊酯(0.40%)、高效氯氰菊酯(0.04%)和高效氯氟氰菊酯(0.02%)4种拟除虫菊酯类杀虫剂1 h的击倒率差异无统计学意义,24 h后的死亡率分别为95.56%、85.56%、81.11%、48.89%与94.44%、68.60%、61.11%、30.00%,除溴氰菊酯(0.03%)外,其他差异有统计学意义。昌江埃及伊蚊接触残杀威(0.03%)和噁虫威(0.20%)2种氨基甲酸酯类杀虫剂的死亡率均为100.00%,儋州埃及伊蚊分别为100.00%和98.89%;昌江与儋州埃及伊蚊接触马拉硫磷(1.50%)、毒死蜱(0.80%)2种有机磷类死亡率均为100.00%,接触杀螟硫磷(0.25%)死亡率分别为100.00%和98.89%。海南儋州和昌江埃及伊蚊检测到V1016G和F1534C 2种突变,其突变频率分别为12.50%(20/160)和97.50%(156/160),差异有统计学意义(χ^(2)=233.54,P<0.001),其中昌江埃及伊蚊同时存在V1016G和F1534C突变,其突变频率分别为20.00%和100.00%,而儋州埃及伊蚊仅F1534C发生突变,突变频率为98.75%。结论海南省埃及伊蚊对溴氰菊酯、氯菊酯、高效氯氰菊酯和高效氯氟氰菊酯4种杀虫剂均产生不同程度的抗性,但对残杀威、噁虫威、马拉硫磷、毒死蜱和螟硫磷均为敏感,其击倒抗性基因以F1534C突变为主。

Objective To understand the level of Aedes aegypti resistance in Hainan and to provide a reference for developing local preventive and control measures against the dengue vector Ae.aegypti.Methods In 2020,Aedes aegypti larvae and pupae were collected from Danzhou and Changjiang in Hainan Province and reared to the first filial generation(F1).The adult mosquito contact tube method was used to determine the resistance of Aedes aegypti to commonly used insecticides.The allele-specific PCR(AS-PCR)was used to detect the mutations in V1016G and F1534C knockdown resistance(kdr)gene in Aedes aegypti.Results There was no significant difference in the knockdown rate of Ae.aegypti from Changjiang and Danzhou after 1 hour of exposure to four pyrethroids insecticides,namely deltamethrin(0.03%),permethrin(0.40%),betacypermethrin(0.04%)and lambda-cyhalothrin(0.02%).However,the 24-hour mortality rates of Ae.aegypti from Changjiang and Danzhou city were 95.56%,85.56%,81.11%,48.89%and 94.44%,68.60%,61.11%,30.00%,respectively.Except for deltamethrin(0.03%),the other differences were statistically significant.Aedes aegypti in Changjiang showed 100.00%mortality to propoxur(0.03%)and bendiocarb(0.20%),while in Danzhou,these rates were 100.00%and 98.89%,respectively.Ae.aegypti both from Changjiang and Danzhou showed 100.00%mortality to malathion(1.50%)and chlorpyrifos(0.80%),and mortality rates to 0.25%fenthion were 100.00%and 98.89%,respectively.V1016G and F1534C mutations were found in Ae.aegypti from Danzhou and Changjiang with mutation frequencies of 12.50%(20/160)and 97.50%(156/160),showing significant differences(χ^(2)=233.54,P<0.001).In Changjiang,Aedes aegypti exhibited both V1016G and F1534C mutations with frequencies of 20.00%and 100.00%,respectively,whereas in Danzhou,only F1534C mutations occurred with a frequency of 98.75%.Conclusions Ae.aegypti in Hainan Province exhibited varying levels of resistance to four insecticides:deltamethrin,permethrin,beta-cypermethrin,and lambda-cyhalothrin,but showed sensitivity to propoxur,bendiocarb,malathion,and chlorpyrifos.The primary knockdown resistance gene mutation was F1534C.