Distribution of knockdown resistance genotypes in Aedes albopictus in Jinghong, Yunnan province, China, 2018-2019

doi:10.11853/j.issn.1003.8280.2020.01.002

Abstract

Abstract: Objective To investigate knockdown resistance (kdr) genotypes and their distribution in the field populations of the dengue vector Aedes albopictus in different areas of Jinghong, Yunnan province, China, and understand their insecticide resistance levels, and to provide a basis for scientific control of Ae. albopictus. Methods Collected adults of Ae. albopictus were identified by morphology, and then soaked in anhydrous ethanol and stored at -20℃. DNA was extracted from each mosquito, and PCR was used to amplify the partial fragment of the voltage-gated sodium channel (VGSC) on the nerve cell membrane. Then sequencing was performed to analyze single or multiple point mutations. Results A total of 160 Ae. albopictus mosquitoes were collected from the east, south, west, north, and middle of Jinghong in September 2018 and July 2019. Mutations were detected at the V1016, I1532, and F1534 loci of the VGSC gene. There were two alleles at the V1016 locus, namely wild-type GTA/V (239/74.69%) and mutant GGA/G (81/25.31%); three genotypes, i.e., the wild-type homozygote V/V (91/56.88%), wild/mutant heterozygote V/G (58/36.25%), and mutant homozygote G/G (11/6.88%). There were two alleles at the I1532 locus, namely wild-type ACC/I (309/96.56%) and mutant ATC/T (11/3.44%); two genotypes, i.e., the wild-type homozygote I/I (149/93.13%) and wild/mutant heterozygote I/T (11/6.87%). There were four alleles at the F1534 locus, namely wild-type TTC/F (92/28.75%), mutant TCC/S (221/69.06%), mutant TGC/C (6/1.88%), and mutant TTG/L (1/0.31%); five genotypes, i.e., the wild-type homozygote F/F (10/6.25%), wild/mutant heterozygotes F/S (69/43.13%) and F/L (1/0.63%), and mutant homozygotes S/S (77/48.13%) and C/C (3/1.88%). Conclusion The kdr gene mutation rate is high in Ae. albopictus in Jinghong, and shows a trend towards a high level and diversified development. This suggests that close attention should be paid to the insecticide resistance level, so as to guide scientific and rational use of insecticides.

Key words: Aedes albopictus, Insecticide resistance, Knockdown resistance gene, Gene mutation, Jinghong

摘要： 目的检测云南省景洪市登革热媒介白纹伊蚊现场种群的击倒抗性（kdr）基因在不同区域的基因型及分布特点，以了解其抗药性水平，为白纹伊蚊科学防控提供依据。方法采集白纹伊蚊成蚊，经形态学方法鉴定后，用无水乙醇浸泡，并于-20℃冻存备用。提取单只蚊虫的DNA，PCR扩增神经细胞膜上电压门控钠离子通道（VGSC）基因部分片段，测序后分析VGSC基因单位点及多位点联合突变情况。结果 2018年9月和2019年7月在景洪市东、西、南、北、中5个方位共采集白纹伊蚊160只，结果显示，其VGSC基因V1016、I1532及F1534位点均存在突变。V1016位点有2种等位基因，即野生型GTA/V（239/74.69%）和突变型GGA/G（81/25.31%）；3种基因型，即野生型纯合子V/V（91/56.88%）、野生/突变型杂合子V/G（58/36.25%）和突变型纯合子G/G（11/6.88%）。I1532位点有2种等位基因，即野生型ACC/I（309/96.56%）和突变型ATC/T（11/3.44%）；2种基因型，即野生型纯合子I/I（149/93.13%）和野生/突变型杂合子I/T（11/6.87%）。F1534位点有4种等位基因，即野生型TTC/F（92/28.75%）、突变型TCC/S（221/69.06%）、TGC/C（6/1.88%）和TTG/L（1/0.31%）；5种基因型分别为野生型纯合子F/F（10/6.25%）、野生/突变型杂合子F/S（69/43.13%）和F/L（1/0.63%），突变型纯合子S/S（77/48.13%）和C/C（3/1.88%）。结论景洪市kdr基因突变频率较高，且近年来呈现高水平、多样化发展趋势。提示应密切关注当地蚊虫抗药性水平，从而指导杀虫剂科学合理地使用。

关键词: 白纹伊蚊, 抗药性, 击倒抗性基因, 基因突变, 景洪市

CLC Number:

ZHU Cai-ying, ZHAO Chun-chun, LUN Xin-chang, ZHU Jin, LI Hong-bin, JIANG Jin-yong, YAN Dong-ming, SONG Xiu-ping, WANG Jun, MENG Feng-xia. Distribution of knockdown resistance genotypes in Aedes albopictus in Jinghong, Yunnan province, China, 2018-2019[J]. Chines Journal of Vector Biology and Control, 2020, 31(1): 7-11.

朱彩英, 赵春春, 伦辛畅, 朱进, 李洪斌, 姜进勇, 闫冬明, 宋秀平, 王君, 孟凤霞. 云南省景洪市2018－2019年白纹伊蚊击倒抗性基因型分布研究[J]. 中国媒介生物学及控制杂志, 2020, 31(1): 7-11.

References

[1] 吴凡. 中国白纹伊蚊的分布和影响因素及登革热的风险评估研究[D]. 北京:中国疾病预防控制中心,2009. Wu F. A study on the potential distribution of Aedes albopictus and risk forecasting for future epidemics of dengue in China[D]. Beijing:Chinese Center for Disease Control and Prevention,2009.
[2] 张海林,米竹青,张云智. 白纹伊蚊垂直传播乙型脑炎病毒的研究[J]. 病毒学报,1996(1):42-47. DOI:10.13242/j.cnki.bingduxuebao.000958. Zhang HL,Mi ZQ,Zhang YZ. Vertical transmission of Japanese encephalitis virus in Aedes albopictus mosquitoes[J]. Chin J Virol,1996(1):42-47. DOI:10.13242/j.cnki.bingduxuebao. 000958.
[3] 刘转转. 白纹伊蚊感染寨卡病毒和登革病毒的媒介能力研究[D]. 广州:南方医科大学,2017. Liu ZZ. Vector competence of Aedes albopictus infected with Zika and dengue virus[D]. Guangzhou:Southern Medical University,2017.
[4] 张海林,米竹青,张云智. 白纹伊蚊和埃及伊蚊对基孔肯雅病毒的易感性和传播性的研究[J]. 中国病毒学,1994,9(3):195-200. Zhang HL,Mi ZQ,Zhang YZ. Susceptibility and transmissibility of Aedes albopictus and Ae. aegypti mosquitoes to Chikungunya virus[J]. Virol Sin,1994,9(3):195-200.
[5] 黄慕嫦,盛钊君,区博文,等. 江门市白纹伊蚊幼虫的抗药性现状调查[J]. 中华卫生杀虫药械,2018,24(4):345-347. Huang MC,Sheng ZJ,Ou BW,et al. Investigation on insecticide resistance of Aedes albopictus in Jiangmen city[J]. Chin J Hyg Insect Equip,2018,24(4):345-347.
[6] 于辛酉. 蚊抗药性检测靶标基因的筛选、鉴定及现场初步应用[D]. 南京:南京医科大学,2012. Yu XY. Screening,identification and field application of drug resistance target genes in mosquitoes[D]. Nanjing:Nanjing Medical University,2012.
[7] Dang K,Doggett SL,Singham GV,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.
[8] Ismail BA,Kafy HT,Sulieman JE,et al. Temporal and spatial trends in insecticide resistance in Anopheles arabiensis in Sudan:outcomes from an evaluation of implications of insecticide resistance for malaria vector control[J]. Parasit Vectors,2018,11:122. DOI:10.1186/s13071-018-2732-9.
[9] Zoh DD,Alou LPA,Toure M,et al. The current insecticide resistance status of Anopheles gambiae (s.l.) (Culicidae) in rural and urban areas of Bouaké,Côte d'Ivoire[J]. Parasit Vectors,2018,11:118. DOI:10.1186/s13071-018-2702-2.
[10] Kasai S,Ng LC,Lam-Phua SG,et al. First detection of a putative knockdown resistance gene in major mosquito vector,Aedes albopictus[J]. Jpn J Infect Dis,2011,64(3):217-221.
[11] 赵春春,朱彩英,开文龙,等. 海口市2018年白纹伊蚊击倒抗性基因型分布研究[J]. 中国媒介生物学及控制杂志,2019,30(1):7-11. DOI:10.11853/j.issn.1003.8280.2019.01.002. Zhao CC,Zhu CY,Kai WL,et al. Genotypes of knockdown resistance gene and their distribution in Aedes albopictus in Haikou,China,in 2018[J]. Chin J Vector Biol Control,2019,30(1):7-11. DOI:10.11853/j.issn.1003.8280.2019.01.002.
[12] 王晓花,陈辉莹,杨新艳,等. 海口市白纹伊蚊对菊酯类杀虫剂的抗药性及击倒抗性基因突变分析[J]. 第二军医大学学报,2015,36(8):832-838. DOI:10.3724/SP.J.1008.2015.00832. Wang XH,Chen HY,Yang XY,et al. Resistance to pyrethroid insecticides and analysis of knockdown resistance (kdr) gene mutations in Aedes albopictus from Haikou city[J]. Acad J Second Mil Med Univ,2015,36(8):832-838. DOI:10.3724/SP.J.1008.2015.00832.
[13] 陈翰明,高景鹏,姜进勇,等. 我国白纹伊蚊现场群体击倒抗性基因I1532和F1534突变检测及I1532T突变等位基因报告[J]. 中国媒介生物学及控制杂志,2018,29(2):120-125. DIO:10.11853/j.issn.1003.8280.2018.02.002. Chen HM,Gao JP,Jiang JY,et al. Detection of the I1532 and F1534 kdr mutations and a novel mutant allele I1532T in VGSC gene in the field populations of Aedes albopictus from China[J]. Chin J Vector Biol Control,2018,29(2):120-125. DIO:10.11853/j.issn.1003.8280.2018.02.002.
[14] Kasai S,Caputo B,Tsunoda T,et al. First detection of a Vssc allele V1016G conferring a high level of insecticide resistance in Aedes albopictus collected from Europe (Italy) and Asia (Vietnam),2016:a new emerging threat to controlling arboviral diseases[J]. Eurosurveillance,2019,24(5):1700847. DOI:10.2807/1560-7917.
[15] Zhou XJ,Yang C,Liu N,et al. Knockdown resistance (kdr) mutations within seventeen field populations of Aedes albopictus from Beijing China:first report of a novel V1016G mutation and evolutionary origins of kdr haplotypes[J]. Parasit Vectors,2019,12(1):180. DOI:10.1186/s13071-019-3423-x.
[16] 杨明东,姜进勇,郭晓芳,等. 2009-2014年云南省登革热流行病学调查与分析[J]. 中国病原生物学杂志,2015,10(8):738-742. DOI:10.13350/j.cjpb.150816. Yang MD,Jiang JY,Guo XF,et al. Epidemiological analysis and investigation of dengue fever in Yunnan province from 2009 to 2014[J]. J Pathog Biol,2015,10(8):738-742. DOI:10.13350/j.cjpb.150816.
[17] 孟凤霞,王义冠,冯磊,等. 我国登革热疫情防控与媒介伊蚊的综合治理[J]. 中国媒介生物学及控制杂志,2015,26(1):4-10. DOI:10.11853/j.issn.1003.4692.2015.01.002. Meng FX,Wang YG,Feng L,et al. Review on dengue prevention and control and integrated mosquito management in China[J]. Chin J Vector Biol Control,2015,26(1):4-10. DOI:10.11853/j.issn.1003.4692.2015.01.002.
[18] 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.
[19] Smith LB,Kasai S,Scott JG. Voltage-sensitive sodium channel mutations S989P+V1016G in Aedes aegypti confer variable resistance to pyrethroids,DDT and oxadiazines[J]. Pest Manag Sci,2018,74(3):737-745. DOI:10.1002/ps.4771. Epub 2017 Dec 8.
[20] 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.