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Fig. 5 Lighting conditions are crucial to reveal anxiety behaviours in fish. A study was conducted to examine the relationship between lighting conditions and anxiety-like behaviour in fish. The light?dark preference test was used to evaluate the effects of MDMA and oxytocin receptor agonist, WAY-267464. The results showed that MDMA administered at doses of 1.0 and 2.5??M increased the time spent in the dark zone (a). At a dose of 2.5??M, it also significantly reduced the time it took for the fish to cross into the dark zone (b). The compound had no effect on the frequency of crossing into the dark zone (c) or locomotion (d), indicating that the observed effects were not due to changes in overall movement. Previously shown to be anxiolytic ? WAY-267464 ? there was no change in the time spent in the light or dark zone in comparison with the control group (e), and even increased the time required to first move into the dark zone (f). The compound did not affect the frequency of crossing into the dark zone (g) or locomotion (h). Data were analysed using a one-way analysis of variance followed by Dunnett?s post hoc test (e, f). Kruskal?Wallis test with Dunn?s post hoc was used for datasets that failed normality testing or had significantly different variances (a?d, g, h). The confidence limit of *p?