Department of Ecosystem Analysis, Institute for Environmental Research, ABBt-Aachen Biology and Biotechnology, RWTH Aachen University, 52074, Aachen, Germany; Department Evolutionary Ecology and Environmental Toxicology, Faculty Biological Sciences, Goethe University Frankfurt, 60438, Frankfurt am Main, Germany.
Department of Ecosystem Analysis, Institute for Environmental Research, ABBt-Aachen Biology and Biotechnology, RWTH Aachen University, 52074, Aachen, Germany.
Chemosphere. 2022 May;295:133863. doi: 10.1016/j.chemosphere.2022.133863. Epub 2022 Feb 3.
In (eco-)toxicological studies the light/dark transition (LDT) test is one of the most frequently used behaviour assays with zebrafish eleutheroembryos. However, study results vary regarding data presentation and analysis and mostly focus on a limited amount of the recorded data. In this study, we investigated whether monitoring two behavioural outcomes (time and distance moved) together with analysing multiple parameters can improve test sensitivity and data interpretation. As a proof of principle 5-day old zebrafish (Danio rerio) eleutheroembryos exposed to either endocrine disruptors (EDs) or acetylcholine esterase (AChE) inhibitors were investigated. We analysed conventional parameters such as mean and sum and implemented additional endpoints such as minimum or maximum distance moved and new parameters assessing the bursting response of eleutheroembryos. Furthermore, changes in eleutheroembryonic behaviour during the moment of the light to dark transition were added. To improve data presentation control-normalised results were displayed in radar charts, enabling the simultaneous presentation of different parameters in relation to each other. This enabled us to identify parameters most relevant to a certain behavioural response. A cut off threshold using control data was applied to identify parameters that were altered in a biological relevant manner. Our approach was able to detect effects on different parameters that remained undetected when analysis was done using conventional bar graphs on - in most cases analysed - averaged, mean distance moved values. By combining the radar charts with additional parameters and by using control-based thresholds, we were able to increase the test sensitivity and promote a deeper understanding of the behaviour response of zebrafish eleutheroembryos in the LDT test and thereby increased its usability for behavioural toxicity studies.
在（生态）毒理学研究中，光/暗转换（LDT）试验是最常用于斑马鱼幼鱼的行为分析方法之一。然而，研究结果在数据呈现和分析方面存在差异，并且主要集中在有限数量的记录数据上。在这项研究中，我们研究了同时监测两种行为结果（移动时间和距离）以及分析多个参数是否可以提高测试灵敏度和数据解释能力。作为原理验证，研究了暴露于内分泌干扰物（EDs）或乙酰胆碱酯酶（AChE）抑制剂的 5 天大的斑马鱼幼鱼（Danio rerio）。我们分析了常规参数，如平均值和总和，并实施了其他终点，如移动的最小或最大距离以及评估幼鱼爆发反应的新参数。此外，还添加了幼鱼在光暗转换时的行为变化。为了改善数据呈现，控制归一化结果以雷达图形式显示，能够将不同参数彼此相关地同时呈现。这使我们能够确定与特定行为反应最相关的参数。使用控制数据的截止阈值用于识别以生物学相关方式改变的参数。我们的方法能够检测到对不同参数的影响，而当使用常规条形图分析时，这些影响在大多数情况下分析的平均移动距离值上仍然无法检测到。通过将雷达图与其他参数结合使用，并使用基于对照的阈值，我们能够提高测试灵敏度，并深入了解斑马鱼幼鱼在 LDT 试验中的行为反应，从而提高其在行为毒性研究中的可用性。
