Peek-A-Boo test: a simple test for assessing the effect of diazepam on medaka (Oryzias latipes) behavior

Abstract: The potential of pharmaceuticals and personal care products to alter the behavior of aquatic organisms is a growing concern. To assess the actual effect of PPCPs on aquatic organisms, the method of a simple but effective behavior test has been required. In this study, we devised a simple behavior test (Peek-A-Boo test) to assess the effect of anxiolytics on model fish behavior (medaka, Oryzias latipes). In the Peek-A-Boo test, we investigated the response of medaka to the picture of a predator fish (donko fish, Odontobutis obscura). The diazepam exposure test (0.8, 4, 20, 100 µg/L) revealed that the time taken for test medaka to approach the picture was 0.22–0.65 fold shortened and the time spent by test medaka in the close area to the picture was 1.8–2.7 fold elongated in all of the diazepam exposure groups compared with the solvent control group (P < 0.05). Hence, it was confirmed that the Peek-A-Boo test could detect the medaka behavioral alteration caused by diazepam with high sensitivity. The Peek-A-Boo test devised in this study is a simple behavior test with high sensitivity for fish behavioral alteration. Files: coord_images.zip includes the csv files of fish swimming coordinates in the experiment using five other fish images. coord_diazepam.zip includes the csv files of fish swimming coordinates in the diazepam exposure experiment.

摘要：药物及个人护理品（pharmaceuticals and personal care products，PPCPs）改变水生生物行为的潜在风险正日益引发学界关注。为评估PPCPs对水生生物的实际影响，亟需开发一种简便高效的行为学检测方法。本研究开发了一种简便的行为学检测方案——躲猫猫试验（Peek-A-Boo test），用于评估抗焦虑药物对模式生物青鳉（Oryzias latipes）行为的影响。在该躲猫猫试验中，我们检测了青鳉对捕食性鱼类沙塘鳢（Odontobutis obscura）图像的响应情况。地西泮暴露试验设置0.8、4、20、100 μg/L四个浓度梯度，结果显示：与溶剂对照组相比，各暴露组受试青鳉抵达目标图像区域的时长缩短至对照组的0.22~0.65倍，且受试青鳉在图像近区域停留的时长延长至对照组的1.8~2.7倍（P < 0.05）。由此证实，躲猫猫试验可高灵敏度地检测出由地西泮诱导的青鳉行为改变。本研究开发的躲猫猫试验是一种简便且可高灵敏度检测鱼类行为改变的行为学检测方法。数据集文件说明：coord_images.zip 包含使用另外5种鱼类图像开展的试验中鱼类游泳轨迹坐标的csv文件；coord_diazepam.zip 包含地西泮暴露试验中鱼类游泳轨迹坐标的csv文件。