Fear and anxiety in fish are generally evaluated by video-based behavioral analysis. However, it is difficult to distinguish the psychological state of fish exclusively through video analysis, particularly whether the fish are freezing, which represents typical fear behavior, or merely resting. We propose a system that can measure bioelectrical signals called ventilatory signals and simultaneously analyze swimming behavior in real time. Experimental results comparing the behavioral analysis of the proposed system and the camera system showed a low error level with an average absolute position error of 9.75 ± 3.12 mm (about one-third of the body length) and a correlation between swimming speeds of r = 0.93 ± 0.07 (p < 0.01). We also exposed the fish to zebrafish skin extracts containing alarm substances that induce fear and anxiety responses to evaluate their emotional changes. The results confirmed that this solution significantly changed all behavioral and ventilatory signal indices obtained by the proposed system (p < 0.01). By combining the behavioral and ventilatory signal indices, we could detect fear and anxiety with a discrimination rate of 83.3% ± 16.7%. Furthermore, we found that the decreasing fear and anxiety over time could be detected according to the peak frequency of the ventilatory signals, which cannot be measured through video analysis.
Keywords: fear and anxiety; real-time measurement; ventilatory signals.