Rizzuto Simone, Neri Franco Maria, Ercolano Valeria, Ippolito Alessio, Linguadoca Alberto, Villamar Bouza Laura, Arena Maria
European Food Safety Authority, Parma PR, Italy.
Environ Toxicol Chem. 2025 May 1;44(5):1435-1447. doi: 10.1093/etojnl/vgaf067.
Amphibians are the current model species for investigating the endocrine-disrupting (ED) properties through the thyroid modality in non-mammalian species. A recurrent question in the European Union (EU) regulatory endocrine assessment of pesticide active substances (2018/605) is whether the positive results from an in vivo screening test, that is, Amphibian Metamorphosis Assay (AMA), can be considered sufficient to conclude on the ED properties of a pesticide active substance or whether the larval amphibian growth and developmental assay (LAGDA) is a necessary step to further clarify the concerns identified in the AMA. Another one is the consideration of the extended AMA (EAMA). To further clarify some of the uncertainties around the use of the LAGDA and to help further consideration of the EAMA in a regulatory context, the statistical power of the three test designs was tested for all the parameters entailed to be measured in the respective study design (except for thyroid histopathology) by using data from real experimental studies. Our findings showed that the statistical power of the EAMA is in line with other Organisation for Economic Co-operation and Development standardized tests, that is, AMA, LAGDA. Our results also confirmed that the LAGDA is more powerful to detect effects on relevant parameters, that is, time to reach metamorphosis, compared to other in vivo tests. However, the difference in power was small, questioning its contribution to an overall weight of evidence already supporting the identification of a substance as an ED. These findings should be considered only in the context of hazard-based endocrine assessment of active substances (i.e., EU regulatory ED assessment of pesticide active substances, 2018/65), while they may not be fully applicable for a risk assessment-based approach.
两栖动物是目前用于通过甲状腺途径研究非哺乳动物内分泌干扰（ED）特性的模式物种。在欧盟对农药活性物质的监管内分泌评估（2018/605）中，一个反复出现的问题是，体内筛选试验（即两栖动物变态试验，AMA）的阳性结果是否足以得出农药活性物质具有ED特性的结论，或者幼体两栖动物生长发育试验（LAGDA）是否是进一步澄清AMA中发现的问题的必要步骤。另一个问题是对扩展AMA（EAMA）的考虑。为了进一步澄清围绕LAGDA使用的一些不确定性，并帮助在监管背景下进一步考虑EAMA，我们利用实际实验研究的数据，对各自研究设计中需要测量的所有参数（甲状腺组织病理学除外）的三种试验设计的统计效力进行了测试。我们的研究结果表明，EAMA的统计效力与经济合作与发展组织的其他标准化试验（即AMA、LAGDA）一致。我们的结果还证实，与其他体内试验相比，LAGDA在检测对相关参数（即达到变态的时间）的影响方面更具效力。然而，效力差异很小，这对其在已经支持将一种物质鉴定为内分泌干扰物的总体证据权重中的贡献提出了质疑。这些发现仅应在基于危害的活性物质内分泌评估（即欧盟对农药活性物质的监管内分泌评估，2018/65）的背景下考虑，而它们可能不完全适用于基于风险评估的方法。
