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通过酯酶和谷胱甘肽 S-转移酶的相互作用阐明两种内栖蚯蚓物种对农药的敏感性。

Elucidating pesticide sensitivity of two endogeic earthworm species through the interplay between esterases and glutathione S-transferases.

机构信息

Univ Avignon, Aix Marseille Univ, CNRS, IRD, IMBE, Pôle Agrosciences, 301 Rue Baruch de Spinoza, BP 21239, 84916, Avignon, France.

INRAE, UMR 1114 EMMAH Domaine Saint Paul, 84914, Avignon Cedex 09, France.

出版信息

Chemosphere. 2021 Jan;262:127724. doi: 10.1016/j.chemosphere.2020.127724. Epub 2020 Jul 18.

DOI:10.1016/j.chemosphere.2020.127724
PMID:32805653
Abstract

Earthworms are common organisms in soil toxicity-testing framework, and endogeic species are currently recommended due to their ecological role in agroecosystem. However, little is known on their pesticide metabolic capacities. We firstly compared the baseline activity of B-esterases and glutathione-S-transferase in Allolobophora chlorotica and Aporrectodea caliginosa. Secondly, vulnerability of these species to pesticide exposure was assessed by in vitro trials using the organophosphate (OP) chlorpyrifos-ethyl-oxon (CPOx) and ethyl-paraoxon (POx), and by short-term (7 days) in vivo metabolic responses in soil contaminated with pesticides. Among B-esterases, acetylcholinesterase (AChE) activity was abundant in the microsomal fraction (80% and 70% of total activity for A. caliginosa and A. chlorotica, respectively). Carboxylesterase (CbE) activities were measured using three substrates to examine species differences in isoenzyme and sensitivity to both in vitro and in vivo exposure. CbEs were mainly found in the cytosolic fraction (80% and 60% for A. caliginosa and A. chlorotica respectively). GST was exclusively found in the soluble fraction for both species. Both OPs inhibited B-esterases in a concentration-dependent manner. In vitro trials revealed a pesticide-specific response, being A. chlorotica AChE more sensitive to CPOx compared to POx. CbE activity was inhibited at the same extent in both species. The 7-d exposure showed A. chlorotica less sensitive to both OPs, which contrasted with outcomes from in vitro experiments. This non-related functional between both approaches for assessing pesticide toxicity suggests that other mechanisms linked with in vivo OP bioactivation and excretion could have a significant role in the OP toxicity in endogeic earthworms.

摘要

蚯蚓是土壤毒性测试框架中常见的生物,由于其在农业生态系统中的生态作用,目前推荐使用内栖物种。然而,人们对它们的农药代谢能力知之甚少。我们首先比较了沟金针虫和暗褐异唇蚓中 B-酯酶和谷胱甘肽 S-转移酶的基础活性。其次,通过使用有机磷(OP)毒死蜱-氧(CPOx)和对氧磷(POx)的体外试验以及受污染土壤中短期(7 天)体内代谢反应来评估这些物种对农药暴露的易感性。在 B-酯酶中,乙酰胆碱酯酶(AChE)活性在微粒体部分丰富(A. caliginosa 和 A. chlorotica 的总活性分别为 80%和 70%)。使用三种底物测量羧酸酯酶(CbE)活性,以检查物种间同工酶的差异以及对体外和体内暴露的敏感性。CbEs 主要存在于细胞质部分(A. caliginosa 和 A. chlorotica 分别为 80%和 60%)。GST 仅在两种物种的可溶性部分发现。两种 OP 均以浓度依赖的方式抑制 B-酯酶。体外试验显示出一种农药特异性反应,即与 POx 相比,A. chlorotica AChE 对 CPOx 更敏感。CbE 活性在两种物种中受到相同程度的抑制。7 天的暴露表明 A. chlorotica 对两种 OP 的敏感性较低,这与体外实验的结果相反。这两种评估农药毒性的方法之间没有相关性,这表明与体内 OP 生物活化和排泄相关的其他机制可能在内栖蚯蚓的 OP 毒性中发挥重要作用。

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