Objective To investigate mutations in the voltage-gated sodium channel gene (VGSC) responsible for knockdown resistance (kdr) in Aedes aegypti and analyze the prevalence characteristics of the VGSC gene in the key areas of Ae. aegypti distribution in Yunnan Province, China. Methods Wild adult or larval Ae. aegypti samples were collected from 7 counties/cities (Yingjiang County, Ruili City, Zhenkang County, Gengma Dai and Va Autonomous County, Menghai County, Jinghong Municipality, and Mengla County) in Yunnan Province. The gene VGSC was amplified by direct sequencing to analyze the point mutation rate and the composition of combined mutations. The Chi-square test was used to compare the differences in the kdr mutation rate of VGSC gene between different loci and between different sexes of Aedes albopictus. Results The DNA was successfully extracted and VGSC gene were sequenced for a total of 561 Ae. aegypti mosquitoes (276 females, 242 males, and 43 larvae) in 7 counties/cities. Four point mutations were found in the VGSC gene. S989P, V1016G, and F1534C mutations were present in every county/city. The S989P mutation rate ranged from 71.43% to 100%, with a mean of 92.51% (95% confidence interval [CI]: 90.33%- 94.70%). The V1016G mutation rate was 100%. The F1534C mutation rate was 55.81%-100%, with a mean of 85.38% (95%CI: 82.45%-88.32%). The Y1527F mutation was first detected in Ruili Municipality, Gengma and Zhenkang County, and the mutation rate ranged from 0 to 35.45%, with a mean of 8.73% (95%CI: 6.39%-11.08%). There were 4 types of co-occurring mutations: S989P+V1016G, V1016G+F1534C, S989P+V1016G+F1534C, and S989P+V1016G+Y1527F+F1534C. Single point mutations were not found. There were no significant differences in the mutation rates of 989, 1016, 1527, and 1534 loci and the composition ratios of combined mutations in female and male mosquitoes in six counties/cities except Yingjiang County. Conclusions The kdr mutation in the VGSC gene of Ae. aegypti in Yunnan Province is mainly characterized by multiple-point joint mutations, and a new Y1527F mutation appears. Kdr mutations surveillance in Ae. aegypti should be carried out regularly to grasp the changes in its resistance to insecticide targets in time.
[1] Beaty BJ,Black W,Eisen L,et al. The intensifying storm:Domestication of Aedes aegypti,urbanization of arboviruses,and emerging insecticide resistance[M]//Mack A. Global health impacts of vector-borne diseases:Workshop summary. Washington:National Academies Press,2016:64-66.
[2] 李春敏,董学书,杨明东. 云南省埃及伊蚊地理分布与季节消长[J]. 中国媒介生物学及控制杂志,2018,29(4):394-396,399. DOI:10.11853/j.issn.1003.8280.2018.04.019.Li CM,Dong XS,Yang MD. Geographical distribution and seasonal variations of Aedes aegypti in Yunnan province[J]. Chin J Vector Biol Control,2018,29(4):394-396,399. DOI:10.11853/j.issn.1003.8280.2018.04.019.(in Chinese)
[3] 刘慧. 东南亚国家加紧应对登革热[N]. 人民日报,2023-07-25(017). DOI:10.28655/n.cnki.nrmrb.2023.007493.Liu H. Southeast Asian countries are stepping up their response to dengue fever[N]. 2023-07-25(017). DOI:10.28655/n.cnki.nrmrb.2023.007493.(in Chinese)
[4] 张海林. 云南省登革热和基孔肯雅热跨境传播、本地流行、发展趋势及对策[J]. 中国媒介生物学及控制杂志,2021,32(1):12-20. DOI:10.11853/j.issn.1003.8280.2021.01.002.Zhang HL. Cross-border spread,indigenous transmission,development trend,and control strategy for dengue fever and chikungunya fever in Yunnan province,China[J]. Chin J Vector Biol Control,2021,32(1):12-20. DOI:10.11853/j.issn.1003. 8280.2021.01.002.(in Chinese)
[5] 兰学梅,魏春,朱进,等. 中国-老挝登革热重点地区埃及伊蚊孳生习性及杀虫剂抗性水平比较研究[J]. 中国媒介生物学及控制杂志,2023,34(2):238-243. DOI:10.11853/j.issn.1003. 8280.2023.02.016.Lan XM,Wei C,Zhu J,et al. A comparative study of breeding habits and insecticide resistance of Aedes aegypti in key areas of dengue fever in China and Laos[J]. Chin J Vector Biol Control,2023,34(2):238-243. DOI:10.11853/j.issn.1003.8280.2023. 02.016.(in Chinese)
[6] 郑宇婷,杨春梅,杨明东,等. 云南边境地区登革热媒介伊蚊生态学及抗药性监测[J]. 中国媒介生物学及控制杂志,2022,33(1):38-43. DOI:10.11853/j.issn.1003.8280.2022.01.007.Zheng YT,Yang CM,Yang MD,et al. Ecological and insecticide resistance surveillance of dengue vector Aedes in Yunnan border region of China[J]. Chin J Vector Biol Control,2022,33(1):38-43. DOI:10.11853/j.issn.1003.8280.2022.01.007.(in Chinese)
[7] 宋晓,史琦琪,程鹏,等. 病媒昆虫的抗药性分子机制研究进展[J]. 中国媒介生物学及控制杂志,2018,29(6):657-661,665. DOI:10.11853/j.issn.1003.8280.2018.06.029.Song X,Shi QQ,Cheng P,et al. Research progress in molecular mechanisms of vector insect's resistance to insecticides[J]. Chin J Vector Biol Control,2018,29(6):657-661,665. DOI:10.11853/j.issn.1003.8280.2018.06.029.(in Chinese)
[8] Dang K,Doggett SL,Veera Singham G,et al. Insecticide resistance and resistance mechanisms in bed bugs,Cimex spp. (Hemiptera:Cimicidae)[J]. Parasit Vectors,2017,10(1):318. DOI:10.1186/s13071-017-2232-3.
[9] Chen ML,Du YZ,Nomura Y,et al. Chronology of sodium channel mutations associated with pyrethroid resistance in Aedes aegypti[J]. Arch Insect Biochem Physiol,2020,104(2):E21686. DOI:10.1002/arch.21686.
[10] Li CX,Kaufman PE,Xue RD,et al. Relationship between insecticide resistance and kdr mutations in the dengue vector Aedes aegypti in Southern China[J]. Parasit Vectors,2015,8:325. DOI:10.1186/s13071-015-0933-z.
[11] 师灿南. 景洪市登革热媒介伊蚊对常用杀虫剂的抗药性及机制初步研究[D]. 北京:中国疾病预防控制中心,2017.Shi CN. Insecticides resistance and its underlying mechanisms for dengue vectors Aedes albopictus and Ae. aegypti in Jinghong city,Yunnan province[D]. Beijing:Chinese Center for Disease Control and Prevention,2017. (in Chinese)
[12] 石清明. 云南省埃及伊蚊种群遗传特征研究[D]. 北京:中国人民解放军军事医学科学院,2017.Shi QM. Genetic characteristics of Aedes aegypti (Diptera:Culicidae) in Yunnan province[D]. Beijing:Academy of Military Sciences of Chinese Peopleʼs Liberation Army,2017. (in Chinese)
[13] 王戈. 云南省中老和中缅边境地区埃及伊蚊种群遗传特征研究[D]. 北京:中国人民解放军军事医学科学院,2023. DOI:10.27193/d.cnki.gjsky.2022.000140.Wang G. Study on population genetic characteristics of Aedes aegypti in China-Laos and China-Myanmar border areas of Yunnan province[D]. Beijing:Military Academy of Sciences,2023. DOI:10.27193/d.cnki.gjsky.2022.000140.(in Chinese)
[14] 兰学梅,杨明东,杨锐,等. 云南省埃及伊蚊对拟除虫菊酯类抗性群体的击倒抗性基因突变分析[J]. 中国人兽共患病学报,2020,36(12):993-999. DOI:10.3969/j.issn.1002-2694. 2020.00.163.Lan XM,Yang MD,Yang R,et al. Analysis of knockdown resistance gene mutations in pyrethroid insecticide resistant populations of Aedes aegypti in Yunnan province[J]. Chin J Zoonoses,2020,36(12):993-999. DOI:10.3969/j.issn.1002-2694.2020.00.163.(in Chinese)
[15] 陆宝麟. 中国动物志昆虫纲第八卷双翅目:蚊科(上卷)[M]. 北京:科学技出版社,1997:221-223.Lu BL. Fauna sinica. Insecta,Volume 8. Diptera:Culicidae Ⅰ[M]. Beijing:Science Press,1997:221-223.(in Chinese)
[16] Endersby-Harshman NM,Ali A,Alhumrani B,et al. Voltage-sensitive sodium channel (Vssc) mutations associated with pyrethroid insecticide resistance in Aedes aegypti (L.) from two districts of Jeddah,Kingdom of Saudi Arabia:Baseline information for a Wolbachia release program[J]. Parasit Vectors,2021,14(1):361. DOI:10.1186/s13071-021-04867-3.
[17] 陈丽,姜进勇. 澜沧江-湄公河流域埃及伊蚊电压门控钠离子通道击倒抗性基因研究进展[J]. 中国病原生物学杂志,2023,18(3):354-357,363. DOI:10.13350/j.cjpb.230322.Chen L,Jiang JY. A review of knockdown resistance in voltage-gated sodium channel of Aedes aegypti in Lancang-Mekong river Basin[J]. J Pathog Biol,2023,18(3):354-357,363. DOI:10.13350/j.cjpb.230322.(in Chinese)
[18] Naw H,Võ TC,Lê HG,et al. Knockdown resistance mutations in the voltage-gated sodium channel of Aedes aegypti (Diptera:Culicidae) in Myanmar[J]. Insects,2022,13(4):322. DOI:10.3390/insects13040322.
[19] Son-Un P,Choovattanapakorn N,Saingamsook J,et al. Effect of relaxation of deltamethrin pressure on metabolic resistance in a pyrethroid-resistant Aedes aegypti (Diptera:Culicidae) strain harboring fixed P989P and G1016G kdr alleles[J]. J Med Entomol,2018,55(4):975-981. DOI:10.1093/jme/tjy037.
[20] Plernsub S,Saingamsook J,Yanola J,et al. Additive effect of knockdown resistance mutations,S989P,V1016G and F1534C,in a heterozygous genotype conferring pyrethroid resistance in Aedes aegypti in Thailand[J]. Parasit Vectors,2016,9(1):417. DOI:10.1186/s13071-016-1713-0.
[21] Kawada H,Oo SZM,Thaung S,et al. Co-occurrence of point mutations in the voltage-gated sodium channel of pyrethroid-resistant Aedes aegypti populations in Myanmar[J]. PLoS Negl Trop Dis,2014,8(7):E3032. DOI:10.1371/journal.pntd.0003032.
[22] Naw H,Su MNC,Võ TC,et al. Overall prevalence and distribution of knockdown resistance (kdr) mutations in Aedes aegypti from Mandalay Region,Myanmar[J]. Korean J Parasitol,2020,58(6):709-714. DOI:10.3347/kjp.2020.58.6.709.
[23] Boyer S,Lopes S,Prasetyo D,et al. Resistance of Aedes aegypti (Diptera:Culicidae) populations to deltamethrin,permethrin,and temephos in Cambodia[J]. Asia Pac J Public Health,2018,30(2):158-166. DOI:10.1177/1010539517753876.
[24] Shimono T,Kanda S,Lamaningao P,et al. Phenotypic and haplotypic profiles of insecticide resistance in populations of Aedes aegypti larvae (Diptera:Culicidae) from central Lao PDR[J]. Trop Med Health,2021,49(1):32. DOI:10.1186/s41182-021-00321-3.
[25] Fan YJ,O'Grady P,Yoshimizu M,et al. Evidence for both sequential mutations and recombination in the evolution of kdr alleles in Aedes aegypti[J]. PLoS Negl Trop Dis,2020,14(4):E0008154. DOI:10.1371/journal.pntd.0008154.
[26] Del Fiol FS,Balcão VM,Barberato-Fillho S,et al. Obesity:A new adverse effect of antibiotics?[J]. Front Pharmacol,2018,9:1408. DOI:10.3389/fphar.2018.01408.
[27] 兰学梅,朱进,李华昌,等. 云南省登革热重点地区埃及伊蚊对11种杀虫剂的抗药性调查[J]. 中国媒介生物学及控制杂志,2019,30(5):582-585. DOI:10.11853/j.issn.1003.8280. 2019.05.025.Lan XM,Zhu J,Li HC,et al. An investigation of the resistance of Aedes aegypti to 11 insecticides in key areas of dengue fever in Yunnan province,China[J]. Chin J Vector Biol Control,2019,30(5):582-585. DOI:10.11853/j.issn.1003.8280.2019.05.025.(in Chinese)
