1. Behavioral analysis based on video recording is becoming increasingly popular within research fields such as; ecology, medicine, ecotoxicology, and toxicology. However, the programs available to analyze the data, which are; free of cost, user-friendly, versatile, robust, fast and provide reliable statistics for different organisms (invertebrates, vertebrates and mammals) are significantly limited. 2. We present an automated open-source executable software (ToxTrac) for image-based tracking that can simultaneously handle several organisms monitored in a laboratory environment. We compare the performance of ToxTrac with current accessible programs on the web. 3. The main advantages of ToxTrac are: i) no specific knowledge of the geometry of the tracked bodies is needed; ii) processing speed, ToxTrac can operate at a rate >25 frames per second in HD videos using modern desktop computers; iii) simultaneous tracking of multiple organisms in multiple arenas; iv) integrated distortion correction and camera calibration; v) robust against false positives; vi) preservation of individual identification if crossing occurs; vii) useful statistics and heat maps in real scale are exported in: image, text and excel formats. 4. ToxTrac can be used for high speed tracking of insects, fish, rodents or other species, and provides useful locomotor information. We suggest using ToxTrac for future studies of animal behavior independent of research area. Download ToxTrac here: https://toxtrac.sourceforge.io

ToxTrac是一款用于图像跟踪的开源软件，可高速同时处理多个实验室环境中的生物，无需对被跟踪物体的几何知识，如果出现交叉则保留个体识别，并提供实用的运动信息和统计数据。

ToxTrac: 用于追踪生物的快速而强大的软件