The stress metabolome provides a thorough insight into the signals and hence mechanisms of response of organisms. This is an excellent tool to advance the understanding of interactions, especially for substances like nanomaterials (NMs), for which there is an urgent need for alternative methods for hazard assessment. The metabolome of Enchytraeus crypticus was studied for the first time. The case study, CuO NM (and CuCl₂) covered exposure along a time frame [0–7–14 days (d)] and two reproduction effect concentrations (EC10 and EC50). High-performance liquid chromatography-mass spectrometry based method (HPLC-MS) was used, with reversed phase (RP) separation and mass spectrometric detection in positive and negative modes. Metabolite profiling of Cu materials yielded 155 and 382 metabolite features in positive and negative modes, respectively, showing an expression related with time, material, and ECx. The number of differentially expressed metabolites (DEMs) decreased with exposure time (14 d) for CuO NM, whereas for CuCl₂ EC50 it increased. Overall, almost all DEMs are down-regulated for CuO NM and up-regulated for CuCl₂ (both modes). Early effects were mainly related to amino acids and later to lysophospholipids (down-regulation). Furthermore, the underlying mechanisms of CuO NM toxicity (e.g. neurotransmission, nucleic acids generation, cellular energy, and immune defense) differ from CuCl₂, where later metabolomic responses are mostly linked to the metabolism of lipids and fewer to amino acids. This study reports a large scale metabolome profiling for E. crypticus and identifies potential markers of Cu materials, which can help to align intelligent testing strategies and safer-by-design materials.

中文翻译：

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该Enchytraeus crypticus应激代谢- CuO的NM案例研究

压力代谢组提供了对信号的深入了解，从而对生物体的反应机理进行了深入了解。这是一个很好的工具，可以增进对相互作用的理解，尤其是对于像纳米材料（NMs）这样的物质，迫切需要替代方法进行危害评估。首次对隐孢子虫的代谢组进行了研究。案例研究，CuO NM（和CuCl ₂）覆盖的时间范围为[0–7–14天（d）]和两个繁殖效应浓度（EC10和EC50）。使用基于高效液相色谱-质谱的方法（HPLC-MS），以反相和反相分离（RP）和质谱检测。Cu材料的代谢物谱图分别以正和负模式显示155和382代谢物特征，显示出与时间，材料和ECx相关的表达。对于CuO NM，差异表达代谢物（DEM）的数量随暴露时间（14 d）的减少而减少，而对于CuCl ₂ EC50的增加。总体而言，几乎所有DEM的CuO NM均下调，而CuCl _2则上调（两种模式）。早期影响主要与氨基酸有关，后来与溶血磷脂有关（下调）。此外，CuO NM毒性的潜在机制（例如神经传递，核酸生成，细胞能量和免疫防御）不同于CuCl ₂，后者后来的代谢组学反应主要与脂质的代谢有关，而与氨基酸的代谢有关。这项研究报告了关于隐孢子虫的大规模代谢组谱分析，并鉴定了铜材料的潜在标记，这可以帮助调整智能测试策略和设计更安全的材料。