  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
