Swiss Centre for Applied Ecotoxicology, Dübendorf, Switzerland.
Air Pollution Control and Chemicals Division, Industrial Chemicals Section, Federal Office for the Environment, Bern, Switzerland.
Environ Toxicol Chem. 2022 Oct;41(10):2512-2526. doi: 10.1002/etc.5445. Epub 2022 Sep 13.
Estrogens are released to the aquatic environment by wastewater treatment plant (WWTP) effluents and can affect wildlife. In the last three decades, many in vitro assay platforms have been developed to detect and quantify estrogenicity in water. In 2018, the International Organization for Standardization (ISO) standardized protocols became available for three types of in vitro estrogen receptor transactivation assays (ERTAs) detecting estrogenicity in 96-well plates (ISO19040 1-3). Two ERTAs-lyticase Yeast Estrogen Screen (L-YES) and Arxula YES (A-YES)-use genetically modified yeast strains, whereas the third utilizes stably transfected human cells. One human cell based assay is ERα-CALUX, which is based on a genetically modified human bone osteosarcoma cell line. In the present study, we characterized the performance, comparability, and effectiveness of these three ERTAs, including an evaluation involving proposed water quality thresholds (effect-based trigger values [EBTs]). For a robust evaluation, we collected 52 effluent samples over three sampling campaigns at 15 different WWTPs in Switzerland. Estrogen receptor transactivation assay results were correlated and compared with results from chemical analysis targeting known estrogens. The three ERTAs showed comparable data over all campaigns. However, the selection of EBTs plays a significant role in the interpretation and comparison of bioassay results to distinguish between acceptable and unacceptable water quality. Applying a fixed cross-assay EBT for effluent of 4 ng L resulted in varying numbers of threshold exceedances ranging between zero and four samples depending on the ERTA used. Using assay-specific EBTs showed exceedances in eight samples (ERα-CALUX) and in one sample (A-YES), respectively. Thus, proposed EBTs do not produce similar risk profiles across samples and further refinement of assay-specific EBTs is needed to account for assay-specific differences and to enable the application of ERTAs as effect-based methods in environmental monitoring. Environ Toxicol Chem 2022;41:2512-2526. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
雌激素通过废水处理厂（WWTP）的废水排放到水生环境中，并可能影响野生动物。在过去的三十年中，已经开发了许多体外检测平台来检测和量化水中的雌激素。2018 年，国际标准化组织（ISO）发布了三种用于检测 96 孔板中雌激素的体外雌激素受体转录激活测定法（ERTAs）的标准化协议（ISO19040 1-3）。两种 ERTA-lyticase 酵母雌激素筛选（L-YES）和 Arxula YES（A-YES）使用基因改良的酵母菌株，而第三种则使用稳定转染的人类细胞。基于人细胞的 ERα-CALUX 测定法是基于基因改良的人骨肉瘤细胞系。在本研究中，我们对这三种 ERTA 的性能、可比性和有效性进行了表征，包括对提出的水质阈值（基于效应的触发值[EBT]）的评估。为了进行稳健的评估，我们在瑞士的 15 个不同 WWTP 进行了三次采样活动，共收集了 52 个污水样本。雌激素受体转录激活测定法的结果与针对已知雌激素的化学分析结果相关联和比较。在所有活动中，三种 ERTA 显示出可比的数据。然而，EBT 的选择在解释和比较生物测定结果以区分可接受和不可接受的水质方面起着重要作用。对于污水，应用固定的跨测定 EBT（4ng/L）会导致根据所使用的 ERTA，阈值超过的样本数量在零到四个之间变化。使用特定于测定的 EBT 显示出在八个样本（ERα-CALUX）和一个样本（A-YES）中超过。因此，所提出的 EBT 不会在样本之间产生相似的风险概况，需要进一步细化特定于测定的 EBT，以考虑测定特异性差异，并使 ERTA 能够作为环境监测中的基于效应的方法应用。Environ Toxicol Chem 2022;41:2512-2526。 版权所有 2022 年由 SETAC 代表 Wiley 期刊出版公司出版。
