Aquatic Toxicology ( IF 4.5 ) Pub Date : 2021-05-19 , DOI: 10.1016/j.aquatox.2021.105864 Ruth Cooper 1 , Arthur David 2 , Tetsuhiro Kudoh 1 , Charles R Tyler 1
Effluents from wastewater treatment works (WwTW) exhibit both temporal and spatial variation in oestrogenicity, however few studies have attempted to quantify how this variation affects biological responses in fish. Here we used an oestrogen-responsive green fluorescent protein (ERE-GFP) transgenic zebrafish (Danio rerio) to quantify oestrogenic activity and health effects for exposure to three different WwTW effluents. Endpoints measured included survival/hatching rate, GFP induction (measured in target tissues or gfp mRNA induction in whole embryos) and vtg mRNA induction in whole embryos. Exposure to one of the study effluents (at 100%), resulted in some mortality, and exposure to all three effluents (at 50% and 100%) caused decreases in hatching rates. Higher levels of vtg mRNA corresponded with higher levels of steroidal oestrogens in the different effluents, with lowest-observed-effect concentrations (LOECs) between 31 ng/L and 39 ng/L oestradiol equivalents (EEQs). Tissue patterns of GFP expression for all three WwTWs effluents reflected the known targets for steroidal oestrogens and for some other oestrogenic chemicals likely present in those effluents (i.e. nonylphenol or bisphenolic compounds). GFP induction was similarly responsive to vtg mRNA induction (a well-established biomarker for oestrogen exposure). We thus demonstrate the ERE-GFP transgenic zebrafish as an effective model for monitoring the oestrogenic potency and health effects for exposure to complex mixtures of chemicals contained within WwTW effluents.
中文翻译:
使用 ERE-GFP 转基因斑马鱼胚胎幼虫评估的污水处理厂出水的雌激素效力和生物效应的季节性变化
废水处理厂 (WwTW) 的流出物在雌激素方面表现出时间和空间变化,但很少有研究试图量化这种变化如何影响鱼类的生物反应。在这里,我们使用雌激素响应绿色荧光蛋白 (ERE-GFP) 转基因斑马鱼 ( Danio rerio ) 来量化暴露于三种不同 WwTW 废水的雌激素活性和健康影响。测量的终点包括存活率/孵化率、GFP 诱导(在靶组织中测量或在整个胚胎中测量gfp mRNA 诱导)和vtg整个胚胎中的 mRNA 诱导。暴露于其中一种研究流出物(100%)会导致一些死亡率,而暴露于所有三种流出物(50% 和 100%)会导致孵化率下降。较高水平的vtg mRNA 对应于不同流出物中较高水平的甾体雌激素,最低观察效应浓度 (LOEC) 介于 31 ng/L 和 39 ng/L 雌二醇当量 (EEQ) 之间。所有三种 WwTW 流出物的 GFP 表达的组织模式反映了甾体雌激素和可能存在于这些流出物中的一些其他雌激素化学物质(即壬基酚或双酚化合物)的已知目标。GFP 诱导对vtg 的反应类似mRNA 诱导(一种成熟的雌激素暴露生物标志物)。因此,我们证明 ERE-GFP 转基因斑马鱼是一种有效模型,用于监测暴露于 WwTW 流出物中的复杂化学品混合物的雌激素效力和健康影响。