There is a growing concern that chronic exposure to fungicides contributes to negative effects on honey bee development, life span, and behavior. Field and caged‐bee studies have helped to characterize the adverse outcomes (AOs) of environmentally relevant exposures, but linking AOs to molecular/cellular mechanisms of toxicity would benefit from the use of readily controllable, simplified host platforms like cell lines. Our objective was to develop and optimize an in vitro‐based mitochondrial toxicity assay suite using the honey bee as a model pollinator, and the electron transport chain (ETC) modulators boscalid and pyraclostrobin as model fungicides. We measured the effects of short (~30 min) and extended exposures (16–24 h) to boscalid and pyraclostrobin on AmE‐711 honey bee cell viability and mitochondrial function. Short exposure to pyraclostrobin did not affect cell viability, but extended exposure reduced viability in a concentration‐dependent manner (median lethal concentration = 4175 µg/L; ppb). Mitochondrial membrane potential (MMP) was affected by pyraclostrobin in both short (median effect concentration [EC50] = 515 µg/L) and extended exposure (EC50 = 982 µg/L) scenarios. Short exposure to 10 and 1000 µg/L pyraclostrobin resulted in a rapid decrease in the oxygen consumption rate (OCR), approximately 24% reduction by 10 µg/L relative to the baseline OCR, and 64% by 1000 µg/L. Extended exposure to 1000 µg/L pyraclostrobin reduced all respiratory parameters (e.g., spare capacity, coupling efficiency), whereas 1‐ and 10‐µg/L treatments had no significant effects. The viability of AmE‐711 cells, as well as the MMP and cellular respiration were unaffected by short and extended exposures to boscalid. The present study demonstrates that the AmE‐711‐based assessment of viability, MMP, and ETC functionality can provide a time‐ and cost‐effective platform for mitochondrial toxicity screening relevant to bees. Environ Toxicol Chem 2024;00:1–12. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

中文翻译：

---

AmE-711 蜜蜂细胞系线粒体功能的评估：啶酰菌胺和唑菌胺酯的作用

人们越来越担心，长期接触杀菌剂会对蜜蜂的发育、寿命和行为产生负面影响。现场和笼养蜜蜂研究有助于描述环境相关暴露的不良后果 (AO)，但将 AO 与毒性的分子/细胞机制联系起来将受益于使用易于控制、简化的宿主平台（如细胞系）。我们的目标是开发和优化基于体外的线粒体毒性测定套件，使用蜜蜂作为模型传粉者，使用电子传递链 (ETC) 调节剂啶酰菌胺和唑菌胺酯作为模型杀菌剂。我们测量了短时间（约 30 分钟）和长时间（16-24 小时）暴露于啶酰菌胺和唑菌胺酯对 AmE-711 蜜蜂细胞活力和线粒体功能的影响。短时间暴露于唑菌胺酯不会影响细胞活力，但长时间暴露会以浓度依赖性方式降低细胞活力（中位致死浓度 = 4175 µg/L；ppb）。在短期（中值效应浓度 [EC50] = 515 µg/L）和长期暴露（EC50 = 982 µg/L）情况下，线粒体膜电位 (MMP) 均受到唑菌胺酯的影响。短时间暴露于 10 和 1000 µg/L 吡唑醚菌酯会导致耗氧率 (OCR) 迅速下降，相对于基线 OCR，10 µg/L 降低约 24%，1000 µg/L 降低 64%。长期暴露于 1000 µg/L 吡唑醚菌酯会降低所有呼吸参数（例如，备用容量、耦合效率），而 1 µg/L 和 10 µg/L 治疗则没有显着影响。 AmE-711 细胞的活力以及 MMP 和细胞呼吸不受短期和长期接触啶酰菌胺的影响。本研究表明，基于 AmE-711 的活力、MMP 和 ETC 功能评估可以为与蜜蜂相关的线粒体毒性筛选提供一个时间和成本有效的平台。环境毒理化学2024；00：1–12。 © 2024 作者。环境毒理学和化学由 Wiley periodicals LLC 代表 SETAC 出版。本文由美国政府雇员撰写，他们的作品在美国属于公共领域。