Neuroactive chemicals are frequently detected in the environment. At sufficiently high concentrations or within mixtures, they could provoke neurotoxic effects and neurological diseases to organisms and humans. Fast identification of such neuroactive compounds in the environment could help in hazard assessment and risk mitigation. Behavior change is considered as an important endpoint and might be directly or indirectly connected to a neuroactive mode of action. For a fast evaluation of environmental samples and pure substances, we optimized the measurement of a behavioral endpoint in zebrafish embryos - the spontaneous tail coiling (STC). Evaluation of results is automated via the use of a workflow established with the KNIME® software. Analysis duration and developmental stage were optimized to 1 min and 25 ± 1 hpf respectively during measurement. Exposing the embryos in a group of 10 or 20 and acclimatizing for 30 min at room temperature proved to be reliable. The optimized method was used to investigate neurotoxic effects of 18 substances with different modes of action (MoA). The STC test accurately detected the effect of 8 out of 11 neuroactive substances (chlorpyrifos, chlorpyrifos-oxon, diazinon, paraoxon-methyl, abamectin, carbamazepine, propafenone and diazepam). Aldicarb and nicotine showed subtle effects which were considered to be conditional and imidacloprid showed no effect. For substances with unknown neuroactive MoA, 3 substances did not provoke any effect on the STC (pyraclostrobin, diuron and daunorubicin-hydrochloride) while 4 other substances provoked an increased STC (hexaconazole, aniline, dimethyl-sulfoxide and 3,4-dichloroaniline). Such unexpected effects indicate possible neuroactive side effects or unknown mechanisms of action that impact on the STC. In conclusion, the optimized STC parameters and the automated analysis in KNIME® indicate opportunities for the harmonization of the STC test and further development for prospective and diagnostic testing.

在环境中经常检测到神经活性化学物质。在足够高的浓度下或在混合物中，它们可能对生物体和人类引起神经毒性作用和神经系统疾病。快速识别环境中的此类神经活性化合物有助于进行危害评估和风险缓解。行为改变被认为是一个重要的终点，可能直接或间接地与神经活动模式有关。为了快速评估环境样品和纯物质，我们优化了斑马鱼胚胎行为终点的测量 - 自发尾卷曲 (STC)。通过使用由 KNIME® 软件建立的工作流程自动评估结果。在测量过程中，分析持续时间和发育阶段分别优化为 1 分钟和 25 ± 1 hpf。以 10 或 20 个为一组暴露胚胎并在室温下适应 30 分钟被证明是可靠的。优化后的方法用于研究 18 种不同作用模式 (MoA) 物质的神经毒性作用。STC 测试准确检测了 11 种神经活性物质中的 8 种（毒死蜱、毒死蜱-氧磷、二嗪农、甲基对氧磷、阿维菌素、卡马西平、普罗帕酮和地西泮）的作用。涕灭威和尼古丁表现出微妙的影响，被认为是有条件的，而吡虫啉没有表现出影响。对于具有未知神经活性 MoA 的物质，3 种物质对 STC（唑菌胺酯、敌草隆和柔红霉素盐酸盐）没有任何影响，而其他 4 种物质引起 STC 增加（己康唑、苯胺、二甲基亚砜和 3,4-二氯苯胺）。这种意想不到的效果表明可能影响 STC 的神经活性副作用或未知的作用机制。总之，优化的 STC 参数和 KNIME® 中的自动分析表明有机会协调 STC 测试以及进一步开发前瞻性和诊断测试。