Behavioral analysis of fish activity by a computer image processing system

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We developed a computer image processing system to acquire positional coordinates of goldfish freely moving in an aquarium. The system was composed of two CCD cameras and two interface I/O boards. The output was fed into an IBM PC/AT compatible computer. By using this image processing system, we could get a trace map of moving goldfish on real time for a long period. Furthermore, we constructed various analytical histograms (moving velocity, right or left turning, inclination of body axis) to quantitatively score fish behavior. We applied these analyzes for the goldfish behavior after treatment of hemisection of the optic nerve. The experimental procedure induced abnormal turning and inclinating behaviors toward the intact side. This abnormality continued for 7-10 days and then were gradually recovered to control fish until 30-40 days after optic nerve injury. To confirm regeneration of the optic nerve, a tracer HRP was injected to one eye, in which the optic nerve had been hemisectioned, and optic nerve terminals in the optic tectum were followed after treatment. The retino-tectal connection of treated goldfish was completed like control by 30 days after hemisection. The regenerating optic nerve was also quantified by ^3H-proline labeling method. These results indicate complete regeneration of the goldfish optic nerve after hemisection at a cellular and a behavioral level. The all different methods described above were good correlated each other. Therefore, the computer image processing system is a very useful quantitation tool for fish behavior and is applicable to the other field of fish science.