贵州省兴义市致倦库蚊对拟除虫菊酯类杀虫剂的抗性机制研究  

Mechanism of resistance to pyrethroid insecticides in Culex quinquefasciatus in Xingyi City,Guizhou Province

作　　者：孔雪雪 王丹[2] 周敬祝[2] 管毓威 罗小龙 胡勇[1] 梁文琴 KONG Xuexue;WANG Dan;ZHOU Jingzhu;GUAN Yuwei;LUO Xiaolong;HU Yong;LIANG Wenqin(School of Public Health,the Key Laboratory of Environmental Pollution Monitoring,and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang,Guizhou 561113,China;Vector Surveillance Section,Guizhou Center for Disease Control and Prevention,Key Laboratory of Microbiome and Infectious Disease Prevention and Control,Guiyang,Guizhou 550004,China)

机构地区：[1]贵州医科大学公共卫生与健康学院,环境污染与疾病监控教育部重点实验室,贵州贵阳561113 [2]贵州省疾病预防控制中心病媒生物监测科,贵州省微生物组与传染性疾病防控重点实验室,贵州贵阳550004

出　　处：《中国热带医学》2025年第3期323-327,共5页China Tropical Medicine

基　　金：贵州省科技计划项目(No.黔科合支撑(2022)一般178);贵州省传染病预防与控制人才基地科研团队-媒介生物及相关传染病监测预警中心(No.RCJD2107);贵州省微生物组与传染性疾病防控重点实验室资助项目(No.ZDSYS[2023]004)。

摘　　要：目的 了解贵州省兴义市致倦库蚊成蚊对3种常用拟除虫菊酯类杀虫剂的抗性水平,并检测代谢解毒酶活性变化及击倒抗性(kdr)基因突变情况,为当地致倦库蚊防制提供科学依据。方法 2024年在兴义市不同方位采集致倦库蚊幼虫,带回实验室饲养至成蚊,采用成蚊接触筒法测定其对拟除虫菊酯类杀虫剂抗药性。对抗药性测试后存活的试虫(抗性种群)为样本,采用酶标仪检测其多功能氧化酶(MFO)、谷胱甘肽-S-转移酶(GST)和非特异性酯酶(NSE)活性以及提取单只致倦库蚊成蚊基因组DNA,采用普通PCR扩增电压门控钠离子通道(VGSC)基因片段并测序,分析kdr基因突变情况。结果 兴义市致倦库蚊成蚊接触0.25%氯菊酯、0.025%溴氰菊酯和0.025%高效氯氰菊酯药膜24 h的死亡率分别为1.00%、0%和0.88%,均表现为抗性。抗氯菊酯种群MFO活力为正态分布,抗溴氰菊酯和高效氯氰菊酯种群MFO活力为偏态分布,3个种群的RR分别为1.17、1.03和1.07倍,野外种群MFO酶活力与敏感品系差异无统计学意义(P均>0.05);抗氯菊酯和高效氯氰菊酯野外种群的GST活力比敏感品系高,为敏感品系的1.06和1.45倍(P<0.05)。抗拟除虫菊酯类杀虫剂致倦库蚊成蚊的VGSC基因片段测序显示在1014位点存在突变,有2种等位基因,即野生型TTA(L)(1.26%)和突变型TTT(F)(98.74%);有2种基因型,即野生/突变型杂合子L/F(2.52%)和突变型纯合子F/F(97.48%)。3种抗拟除虫菊酯类杀虫剂致倦库蚊种群之间的抗性基因突变频率并无统计学意义(P>0.05)。结论 兴义市致倦库蚊成蚊对3种常用拟除虫菊酯类杀虫剂均已产生抗性,代谢解毒酶活性发生变化,且kdr基因发生突变。建议加强蚊媒抗药性监测,开展以清除孳生地为核心的环境治理工作,减少拟除虫菊酯类杀虫剂使用,考虑轮换使用其他类型的杀虫剂,以延缓其抗药性的发展。Objective To investigate the resistance levels of adult Culex quinquefasciatus mosquitoes to three commonly used pyrethroid insecticides in Xingyi City,Guizhou Province,as well as to examine the changes in metabolic detoxification enzyme activities and mutations in the knockdown resistance(kdr)gene thereby providing a scientific foundation for the prevention and control of Culex quinquefasciatus in the region.Methods In 2024,the larvae of Culex quinquefasciatus were collected from different locations in Xingyi and reared to adult mosquitoes in the laboratory,and their resistance to pyrethroid insecticides was determined using the adult mosquito contact tube method.Surviving samples(resistant population)from the resistance tests were examined for the activities of mixed function oxidase(MFO),glutathione-S-transferase(GST),and nonspecific esterase(NSE)using a microplate reader.The genomic DNA of individual adult Culex quinquefasciatus mosquitoes was extracted,and the voltage-gated sodium channel(VGSC)gene fragment was amplified via PCR and sequenced to analyze the mutations in the kdr gene.Results The 24-h mortality rates of adult Culex quinquefasciatus exposed to 0.25%permethrin,0.025%deltamethrin,and 0.025%beta-cypermethrin were 1.00%,0%,and 0.88%,respectively,indicating all populations of Culex quinquefasciatus exhibited resistance to the three pyrethroid insecticides.The activity of MFO in permethrin-resistant population followed a normal distribution,while deltamethrin-resistant and cypermethrin populations exhibited skewed distributions. The resistance ratios (RRs) for the three populations were 1.17, 1.03, and 1.07 times, respectively, with nostatistically significant differences in MFO activity between field populations and susceptible strains (all P>0.05). However, theGST activities of permethrin-resistant and beta-cypermethrin-resistant field populations were 1.06 and 1.45 times higher thanthose of sensitive strains (P<0.05). Sequencing of the VGSC gene fragments of adult Culex quinquefasciatus resistant

关 键 词：致倦库蚊 抗药性 代谢酶 击倒抗性基因 兴义市 

分 类 号：R384.1[医药卫生—医学寄生虫学]