ISTAnt.zip

In this study, we analyzed grooming behavior in the ant <i>Lasius neglectus</i> in groups of three worker ants. The workers for the experiment were obtained from their laboratory stock colony, which had been collected from the field in 2022 in the Botanical Garden Jena, Germany. Ant collection and all experimental work were performed in compliance with international, national and institutional regulations and ethical guidelines.For the experiment, the body surface of one of the three ants was treated with a suspension of either of two microparticle types (diameter ~5 µm) to induce grooming by the two nestmates, which were individually color-coded by application of a dot of blue or orange paint, respectively. The three ants were housed in small plastic containers (diameter 28mm, height 30mm) with moistened, plastered ground and the interior walls covered with PTFE (polytetrafluoroethane) to hamper climbing by the ants. Filming occurred in a temperature- and humidity-controlled room at 23°C within a custom-made filming box with controlled lighting and ventilation conditions. We set up nine ant groups at a time (always containing both treatments) and placed them randomly on positions 1-9 marked on the floor in a 3x3 grid, about 3mm from each other. The experiment was performed on two consecutive days. Videos were acquired using a USB camera (FLIR blackfly S BFS-U3-120S4C, Teledyne FLIR) with a high-performance lens (HP Series 25mm Focal Length, Edmund optics 86-572) in OBS studio 29.0.0 \citep{bailey2017obs} at a framerate of 30 FPS and a resolution of 2500x2500 pixels. From each original video (105x105 mm), we generated nine individual videos .mkv (each ~32x32 mm, 770x770 pixels) by determining exact coordinates per container from one frame in GIMP 2.10.36 and cropping of the videos with FFmpeg 6.1.1. Annotation was performed over two consecutive days by three observers who had not been involved in the experimental setup or recording and were unaware of the treatment assignments to ensure bias-free behavioral annotation. They annotated the behavior of the ants during video observations, using custom-made software that saves the start and end frames of behaviors marked in a .csv file. In one of the videos, one of the nestmates' legs got inadvertently stuck to its body surface during the color-coding, interfering with its behavior, so the video was discarded. This left 44 videos from 5 independent setups (n=24 of treatment 1 and n=20 of treatment 2) of 10 minutes each for a total of 792 000 annotated frames. For each video, we provide the following information: the number of the set to which it belongs (1-5); the number of the position within the set reflecting the position of the ant group under the camera (1-9), for which we also provide ‘coordinates’ in the 3x3 grid (taking values -1/0/1 for both X and Y axis); treatment (1 or 2); the hour of the day when the recording was started (in 24h CEST); experimental day (A or B); the top left coordinate of the cropping square from the original video (CropX/CropY); the person annotating the video (given as A, B, C); the date of annotation (1: first day, 2: second day) and in which order the videos were annotated by each person (both reflecting a possible training effect of the person).

本研究以三只工蚁组成的蚁群为对象，分析了疏忽毛蚁（Lasius neglectus）的理毛行为。实验所用工蚁采自2022年于德国耶拿植物园野外采集并长期饲养的实验室种群。所有蚂蚁采集及实验操作均符合国际、国家及机构相关规定与伦理准则。

实验中，我们对三只蚂蚁中的一只体表施加两种微米颗粒（直径约5 µm）中的一种悬浮液，以诱导另外两只巢伴产生理毛行为；两只巢伴分别通过涂抹蓝色或橙色漆点进行单独颜色编码标识。三只蚂蚁被饲养于小型塑料容器（直径28 mm，高度30 mm）中，容器底部铺有湿润石膏，内壁涂有聚四氟乙烯（PTFE，polytetrafluoroethane）以阻碍蚂蚁攀爬。

实验在温湿度可控的房间内开展，室温设定为23°C，使用定制的光照与通风可控拍摄箱完成录制。我们单次同步设置9组蚁群（始终包含两种处理组），并将其随机放置在3×3网格标记的1-9号位置上，各组间距约3 mm。实验连续开展两天。视频采集使用USB摄像头（Teledyne FLIR公司FLIR Blackfly S BFS-U3-120S4C）搭配高性能镜头（Edmund Optics 86-572，HP系列25mm焦距），通过OBS Studio 29.0.0 citep{bailey2017obs} 完成，帧率为30 FPS，分辨率为2500×2500像素。

我们从每段原始视频（尺寸105×105 mm）中裁剪得到9段独立的.mkv格式视频（单段尺寸约32×32 mm，分辨率770×770像素）：首先通过GIMP 2.10.36在单帧图像中确定每个容器的精确坐标，再使用FFmpeg 6.1.1完成视频裁剪。

行为标注由三名未参与实验搭建与录制的观察者分两天完成，且他们对处理组分配不知情，以确保标注无偏倚。观察者使用定制软件标记视频中的蚂蚁行为，该软件会将标记行为的起始与结束帧保存为.csv文件。其中一段视频中，一只巢伴的腿部在颜色标识过程中意外粘在自身体表，干扰了其正常行为，因此该视频被剔除。最终有效数据集包含来自5个独立实验批次的44段视频（处理1组n=24，处理2组n=20），每段视频时长10分钟，总计792000个标注帧。

对于每段视频，我们提供以下元数据：所属实验批次编号（1-5）；组内位置编号（对应摄像头下蚁群的位置，1-9），同时提供其在3×3网格中的"坐标"（X轴与Y轴取值均为-1/0/1）；处理组编号（1或2）；录制开始的当日时刻（采用24小时制中欧夏令时（CEST））；实验天数（A或B）；原始视频中裁剪框的左上角坐标（CropX/CropY）；视频标注者（以A、B、C标识）；标注日期（1为第一天，2为第二天）；以及每位观察者对该视频的标注顺序（可反映观察者的训练效应）。