Tebuconazole, a widely used fungicide, can severely disrupt the reproductive process of various organisms. In this study, we investigated the subacute effects of tebuconazole on the earthworm to fully understand its toxic implications. Herein, untargeted metabolomics, mRNA assay and biochemical approaches were adopted to evaluate the subacute effects of Eisenia fetida earthworms, when exposed to tebuconazole at three different concentrations (0.5, 5 and 50 mg/kg) for seven days. SOD enzyme activity test displayed that tebuconazole exposure interfered with the earthworms' ROS. ANN mRNA expression was down-regulated after tebuconazole exposure. ¹H nuclear magnetic resonance (¹H-NMR)-based untargeted metabolomics study showed that 5 mg/kg tebuconazole exposure interfered with earthworms' metabolism. Twelve significantly changed metabolites were identified. The pathway analyses indicate that tebuconazole can disrupt the earthworm's metabolism, particularly in the AMP pathway, which impact the reproduction. This may explain the tebuconazole's mechanism of action behind the down-regulation of the expression of ANN mRNA, which is related to the earthworm's reproductive process. We comprehensively evaluated the mRNA expression, enzyme activity, and metabolomics, and acquired sufficient information for evaluating the toxicity of tebuconazole.

戊唑醇（一种广泛使用的杀菌剂）会严重破坏各种生物的繁殖过程。在这项研究中，我们调查了戊唑醇对worm的亚急性作用，以充分了解其毒性含义。在本文中，当暴露于三种不同浓度（0.5、5和50 mg / kg）的戊唑醇7天时，采用非靶向代谢组学，mRNA分析和生化方法来评价艾氏nia的亚急性效应。SOD酶活性测试表明，戊唑醇暴露会干扰worm的ROS。戊唑醇暴露后，ANN mRNA表达下调。¹ H核磁共振（¹基于1 H-NMR的非靶向代谢组学研究表明，暴露于5 mg / kg的戊唑醇会干扰worm的代谢。确定了十二种显着变化的代谢物。途径分析表明戊唑醇可以破坏worm的代谢，特别是在AMP途径中，这会影响繁殖。这可能解释了戊丁唑在下调ANN mRNA表达后的作用机理，这与worm的繁殖过程有关。我们全面评估了mRNA的表达，酶活性和代谢组学，并获得了足够的信息来评估戊唑醇的毒性。