Early-life stage (ELS) toxicity tests are recognized as an advancement over current testing methodologies in terms of cost, animal use, and biological relevance. However, standardized ELS tests are not presently available for some vertebrate taxa, including birds. The present study describes a Japanese quail (Coturnix japonica) ELS test that is a promising candidate for standardization and applies it to test 8 environmental chemicals (ethinylestradiol, benzo[a]pyrene, chlorpyrifos, fluoxetine, lead(II)nitrate, trenbolone, seleno-L-methionine, hexabromocyclododecane). Individual chemicals were injected into the air cell of unincubated Japanese quail eggs at 3 concentrations, all predicted to cause ≤20% mortality. Survival to embryonic day 16 was consistently high (>90%) among the vehicle-injected controls. All chemicals, except ethinylestradiol, were detected in liver tissue, most at concentrations suggestive of embryonic clearance. Adverse effects were observed for 5 of the 8 chemicals; chlorpyrifos (41.1 µg/g) significantly increased developmental abnormalities and decreased embryo and gallbladder mass. Ethinylestradiol (54.2 µg/g) and hexabromocyclododecane (0.02 µg/g) decreased embryo mass and tarsus length, respectively. Benzo[a]pyrene (0.83 µg/g) and fluoxetine hydrochloride (32.7 µg/g) exceeded the 20% mortality cutoff. No effects were observed following lead(II)nitrate, seleno-L-methionine, or trenbolone exposure up to 10.7, 0.07, and 4.4 µg/g, respectively. Overall, our ELS approach was time- and cost-effective, caused minimal mortality in controls, effectively delivered diverse chemicals to the embryo, and permitted identification of apical outcomes, all of which provide support toward standardization. Environ Toxicol Chem 2019;39:141-154. © 2019 SETAC.

中文翻译：

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一种生命早期阶段的替代测试策略，用于评估环境化学物质对鸟类的影响。

在成本，动物使用和生物学相关性方面，生命早期阶段（ELS）毒性测试被认为是对当前测试方法的改进。但是，目前尚不适用于某些脊椎动物类群（包括鸟类）的标准化ELS测试。本研究描述了日本鹌鹑（Coturnix japonica）ELS测试，该测试有望成为标准化的候选方法，并将其用于测试8种环境化学品（炔雌醇，苯并[a] py，毒死rif，氟西汀，硝酸铅（II），群勃龙，硒代） -L-蛋氨酸，六溴环十二烷）。将单独的化学药品以3种浓度注入未孵化的日本鹌鹑蛋的气室中，所有这些化学成分预计会导致≤20％的死亡率。在注射媒介物的对照中，到胚胎第16天的存活率一直很高（> 90％）。除乙炔雌二醇外的所有化学药品，在肝组织中检测到了大部分的浓度，这些浓度提示存在胚胎清除的可能。观察到了8种化学药品中有5种的不良反应。毒死rif（41.1 µg / g）显着增加了发育异常并减少了胚胎和胆囊的质量。乙炔雌二醇（54.2 µg / g）和六溴环十二烷（0.02 µg / g）分别降低了胚胎质量和架长度。苯并[a] py（0.83 µg / g）和盐酸氟西汀（32.7 µg / g）超过了20％的死亡率临界值。硝酸铅（II），硒代L-蛋氨酸或群勃龙分别暴露至10.7、0.07和4.4 µg / g时未观察到影响。总体而言，我们的ELS方法既节省时间又具有成本效益，在对照组中造成的死亡率极低，有效地将多种化学物质传递至胚胎，并可以确定根尖结局，所有这些都为标准化提供了支持。Environ Toxicol Chem 2019; 39：141-154。©2019 SETAC。