Recording the ventilation activity of free-swimming zebrafish and its application to novel tank tests.

Bioelectric signals related to ventilatory movements in fish can be detected via externally located electrodes. In this study, a technique to continuously monitor the electric ventilatory signals in free-swimming zebrafish was developed. This technique was applied to monitoring ventilation activity as a physiological measure in conjunction with various behavioral measures in a novel tank environment. It was found that in addition to ventilation rate, time domain analysis of changes in ventilation rate is useful for evaluating the emotional state of zebrafish. By integrating the physiological and behavioral measures in analyses, a 1 h novel tank test trial revealed that the habituation process involves two phases. The first phase, which lasted 10 min, involved rapid attenuation of the initial fear/anxiety response to encountering a novel environment. The second phase lasted 20 min and involved further attenuation of anxiety and an increase in exploration behavior. These data suggest that combining ventilation-related physiological measures with conventional behavioral measures enables multidimensional examination of the habituation process in a novel tank environment with more precision than is possible when relying on behavioral responses alone.