Thornton Jonathan, Gomes Bruno, Ayres Constância, Reimer Lisa
Vector Biology Department, Liverpool School of Tropical Medicine, Liverpool, L35QA, UK.
Laboratório de Bioquímica e Fisiologia de Insetos, Oswaldo Cruz Institute (IOC-FIOCRUZ), Rio de Janeiro, 21040-360, Brazil.
Wellcome Open Res. 2020 Dec 16;5:183. doi: 10.12688/wellcomeopenres.15974.2. eCollection 2020.
: Laboratory reared mosquito colonies are essential tools to understand insecticide action. However, they differ considerably from wild populations and from each other depending on their origin and rearing conditions, which makes studying the effects of specific resistance mechanisms difficult. This paper describes our methods for establishing multiple resistant strains of from two colonies as a new resource for further research on metabolic and target site resistance. : Two resistant colonies of , from Cayman and Recife, were selected through 10 generations of exposure to insecticides including permethrin, malathion and temephos, to yield eight strains with different profiles of resistance due to either target site or metabolic resistance. Resistance ratios for each insecticide were calculated for the selected and unselected strains. The frequency of alleles (F1534C and V1016I) in the Cayman strains was determined using TaqMan assays. A comparative gene expression analysis among Recife strains was conducted using qPCR in larvae (CCae3A, CYP6N12, CYP6F3, CYP9M9) and adults (CCae3A, CYP6N12, CYP6BB2, CYP9J28a). : In the selected strain of Cayman, mortality against permethrin reduced almost to 0% and became fixated by 5 generations. A similar phenotype was seen in the unselected homozygous resistant colony, whilst mortality in the susceptible homozygous colony rose to 82.9%. The Recife strains showed different responses between exposure to adulticide and larvicide, with detoxification genes in the temephos selected strain staying similar to the baseline, but a reduction in detoxification genes displayed in the other strains. : These selected strains, with a range of insecticide resistance phenotypes and genotypes, will support further research on the effects of target-site and/or metabolic resistance mechanisms on various life-history traits, behaviours and vector competence of this important arbovirus vector.
实验室饲养的蚊虫群体是了解杀虫剂作用的重要工具。然而,它们与野生种群有很大不同,并且彼此之间也因起源和饲养条件而存在差异,这使得研究特定抗性机制的影响变得困难。本文描述了我们从两个群体建立多个抗性品系的方法,作为进一步研究代谢抗性和靶标位点抗性的新资源。:从开曼群岛和累西腓选取了两个抗性群体,通过对包括氯菊酯、马拉硫磷和双硫磷在内的杀虫剂进行10代暴露,产生了8个因靶标位点或代谢抗性而具有不同抗性谱的品系。计算了所选品系和未选品系对每种杀虫剂的抗性比率。使用TaqMan分析确定了开曼品系中kdr等位基因(F1534C和V1016I)的频率。使用qPCR对累西腓品系的幼虫(CCae3A、CYP6N12、CYP6F3、CYP9M9)和成虫(CCae3A、CYP6N12、CYP6BB2、CYP9J28a)进行了比较基因表达分析。:在开曼的所选品系中,对氯菊酯的死亡率几乎降至0%,并且在5代后kdr等位基因固定下来。在未选的纯合抗性群体中也观察到了类似的表型,而敏感纯合群体中的死亡率上升到了82.9%。累西腓品系在接触成蚊杀虫剂和幼虫杀虫剂时表现出不同的反应,双硫磷所选品系中的解毒基因与基线相似,但其他品系中的解毒基因有所减少。:这些具有一系列杀虫剂抗性表型和基因型的所选品系,将支持进一步研究靶标位点和/或代谢抗性机制对这种重要虫媒病毒载体的各种生活史特征、行为和媒介能力的影响。