Linking behavioural effects of thyroid active chemicals with molecular biomarkers and their application towards the evaluation of sewage treatment technologies

城市废水将各种药物和个人护理产品引入水生环境。这些化学品有可能通过干扰激素功能来扰乱各种生物过程，但二级和三级处理并不是专门设计来去除这些内分泌干扰化合物（EDCs）的。此外，缺乏用于检测EDC活性的有效的环境和生态相关的方法，特别是对于甲状腺激素（TH）干扰活性。TH对所有脊椎动物的正常生长、行为、发育和代谢至关重要。TH作用的一个最显著的例子是它们触发青蛙蝌蚪蜕变成小青蛙的绝对要求。在蛙科物种，这代表了人口最多的青蛙家庭，蝌蚪从水生过渡到陆地生活方式的方式具有相当大的相似之处围产期人类发展。拟议的项目结合了学术和政府科学家的专业知识，建立基于本地两栖动物哨兵物种，牛蛙，和定量真实的时间聚合酶链反应（QPCR）的标准化测定，以确定TH-破坏活动锚定的行为结果。拟议的项目将使用RNA-Seq发现牛蛙蝌蚪肝脏和大脑嗅叶中TH作用的最敏感和最强大的指标基因转录本，并将其与选择迷宫和电嗅觉评估中嗅觉线索反应测量的神经功能变化联系起来。这些信息将指导QPCR阵列的开发，以评估去除效果。 的TH-干扰活动的国家的最先进的和新兴的城市污水处理技术产生的废水。这项工作的结果将1）形成生物锚定的基础，用户友好的检测方法用于评估活性物质，2）提供有关EDC去除效果的关键信息，以影响未来的设计和处理厂的工程，3）指导监管机构将基因表达终点作为水质指南的一部分。

Municipal wastewater effluents introduce a variety of pharmaceuticals and personal care products into the aquatic environment. These chemicals have the potential to perturb diverse biological processes by disrupting hormone function yet secondary & tertiary treatments are not specifically designed to remove these endocrine disrupting compounds (EDCs). Moreover, effective environmentally- and ecologically-relevant methods for detecting EDC activity are lacking, particularly for thyroid hormone (TH)-disrupting activities. THs are essential for normal growth, behaviour, development, and metabolism in all vertebrates. One of the most striking examples of TH action is their absolute requirement for triggering frog tadpole metamorphosis into a froglet. In Ranid species, which represent the most populous of the frog families, the tadpole transitions from an aquatic to a terrestrial lifestyle in a manner possessing considerable parallels to perinatal human development. The proposed project combines the expertise of academic and government scientists to establish standardized assays based upon a native amphibian sentinel species, Rana catesbeiana, and quantitative real time polymerase chain reaction (QPCR) to identify TH-disrupting activities anchored to behavioural outcomes. The proposed project will discover gene transcripts that are the most sensitive and robust indicators of TH action in the bullfrog tadpole liver and brain olfactory lobe using RNA-Seq and link these to changes neurological function as measured by olfactory cue responses in choice maze and electro-olfactory assessments. This information will guide the development of QPCR arrays to evaluate the efficacy of removal of TH-disrupting activity in effluents produced by state-of-the-art and emerging municipal wastewater treatment technologies. The results of this work will 1) form the foundation of biologically-anchored, user-friendly assays for assessing hormone-active substances, 2) provide crucial information regarding the efficacy of EDC removal to influence future design and engineering of treatment plants, and 3) guide regulators on the incorporation of gene expression endpoints as part of water quality guidelines.