Protocols to determine metabolic resistance in ticks were mainly derived from reports published using mosquitoes and agriculturally important insects without prior standardization. In the present study, biochemical assays were standardized to quantify acaricide metabolizing enzymes in tick homogenates. Three variables viz., age, number of larvae and reaction time were optimized using reference susceptible IVRI-I and deltamethrin resistant IVRI-IV (Resistance Factor = 194) tick strains. The optimum conditions for estimation of general esterases were 10-15 day old 40 larvae with 15 mins reaction time, 15-20 day old 40 larvae with 20 mins reaction time for Glutathione S- transferase, while 10-15 day old 80 larvae with 5 mins reaction time for monooxygenase. The standardized protocols were further validated in multi acaricide resistant strain (IVRI-V) and in nine field isolates having variable resistant factors to different acaricides. In all the nine heterogeneous field isolates, a significant correlation (p < .05) between resistance to synthetic pyrethroids and over-expression of esterases and monooxygenase was noticed. Similarly, esterases and GST activities were significantly correlated with resistance to organophosphates. The details of the assay protocol are explained for adoption in different laboratories.

中文翻译：

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用于估计酯酶、单加氧酶和谷胱甘肽 S-转移酶以表征杀螨剂抗性的蜱特异性生化工具的标准化

确定蜱代谢抗性的协议主要来自使用蚊子和农业上重要的昆虫发表的报告，未经事先标准化。在本研究中，生化测定被标准化以量化蜱匀浆中的杀螨剂代谢酶。使用参考易感 IVRI-I 和溴氰菊酯抗性 IVRI-IV（抗性因子 = 194）蜱菌株优化三个变量，即年龄、幼虫数量和反应时间。估计一般酯酶的最佳条件是10-15日龄40条幼虫与15分钟反应时间，15-20日龄40条幼虫与谷胱甘肽S-转移酶反应时间20分钟，而10-15日龄80条幼虫与5单加氧酶的反应时间为分钟。标准化方案在多杀螨剂抗性菌株 (IVRI-V) 和对不同杀螨剂具有不同抗性因素的九个野外分离株中得到进一步验证。在所有九个异源田间分离株中，注意到对合成拟除虫菊酯的抗性与酯酶和单加氧酶的过度表达之间存在显着相关性 (p < .05)。同样，酯酶和 GST 活性与对有机磷的抗性显着相关。分析协议的细节被解释为在不同的实验室采用。酯酶和 GST 活性与对有机磷的抗性显着相关。分析协议的细节被解释为在不同的实验室采用。酯酶和 GST 活性与对有机磷的抗性显着相关。对检测方案的细节进行了解释，以供不同实验室采用。