Suryanto Michael Edbert, Luong Cao Thang, Vasquez Ross D, Roldan Marri Jmelou M, Hung Chih-Hsin, Ger Tzong-Rong, Hsiao Chung-Der
Department of Chemistry, Chung Yuan Christian University, Taoyuan 320314, Taiwan; Department of Bioscience Technology, Chung Yuan Christian University, Taoyuan 320314, Taiwan.
Department of Chemical Engineering & Institute of Biotechnology and Chemical Engineering, I-Shou University, Da-Shu, Kaohsiung City 84001, Taiwan.
Ecotoxicol Environ Saf. 2023 Oct 15;265:115507. doi: 10.1016/j.ecoenv.2023.115507. Epub 2023 Sep 22.
The freshwater crayfish, Procambarus clarkii is an excellent aquatic animal model that is highly adaptable and tolerant. P. clarkii is widely used as a toxicity model to study various pharmaceutical exposure. This animal model has complex behavioral traits and is considered sensitive to environmental changes, making it an excellent candidate to study psychoactive drugs based on a behavioral approach. However, up to now, most behavioral studies on crayfish use manual observation and scoring that require panelists. In this study, we aim to develop an automation pipeline to analyze crayfish behavior automatically. We use a deep-learning approach to label body parts in multiple crayfish, and based on the trajectory results, the intra- or inter-individual crayfish were calculated. Reliable and fast results of several behavior endpoints in multiple crayfish were retrieved. We then validated the detection performance of numerous crayfish in specific gender groups (male-male and female-female). Based on the result, the male crayfish displayed significantly higher aggression than females. We also tested the antidepressant exposure on this animal model to evaluate the psychoactive effects of this drug. As male crayfish display more distinct agonistic behavior than females, we exposed them to sertraline (SRT) 1 ppb for 7 and 14 days. It was revealed that sertraline was able to alter several behavioral endpoints in crayfish. Significant increases in extend claw ratio, total distance moved, average speed, and rapid movement were displayed in sertraline-exposed crayfish but decreased interaction time and longest interaction time. In addition, SRT 14 days exposure could atler the aggressiveness and bold behavior In the present method, DeepLabCut (DLC) has been utilized to analyze the locomotion behavior of multiple crayfish. This established method provides rapid and accurate ecotoxicity measurements using freshwater crayfish, which beneficient and applicable for environmental research.
克氏原螯虾是一种适应性强且耐受性高的优秀水生动物模型。克氏原螯虾被广泛用作毒性模型来研究各种药物暴露。这种动物模型具有复杂的行为特征，被认为对环境变化敏感，使其成为基于行为方法研究精神活性药物的优秀候选对象。然而，到目前为止，大多数关于小龙虾的行为研究都采用需要专家小组的人工观察和评分。在本研究中，我们旨在开发一种自动化流程来自动分析小龙虾的行为。我们使用深度学习方法对多只小龙虾的身体部位进行标记，并根据轨迹结果计算小龙虾个体内或个体间的情况。检索到了多只小龙虾几个行为终点的可靠且快速的结果。然后，我们验证了特定性别组（雄性 - 雄性和雌性 - 雌性）中多只小龙虾的检测性能。基于结果，雄性小龙虾表现出比雌性更高的攻击性。我们还在这个动物模型上测试了抗抑郁药暴露，以评估这种药物的精神活性作用。由于雄性小龙虾比雌性表现出更明显的争斗行为，我们将它们暴露于1 ppb的舍曲林（SRT）中7天和14天。结果表明，舍曲林能够改变小龙虾的几个行为终点。暴露于舍曲林的小龙虾显示出伸展螯比例、移动总距离、平均速度和快速移动显著增加，但相互作用时间和最长相互作用时间减少。此外，SRT暴露14天可改变攻击性和大胆行为。在本方法中，已利用DeepLabCut（DLC）来分析多只小龙虾的运动行为。这种既定方法使用淡水小龙虾提供了快速准确的生态毒性测量，这对环境研究有益且适用。
