PT - JOURNAL ARTICLE AU - Dominic Mason AU - Susanne Zajitschek AU - Hamza Anwer AU - Rose E O’Dea AU - Daniel Hesselson AU - Shinichi Nakagawa TI - Low Repeatability of Aversive Learning in Zebrafish (<em>Danio rerio</em>) AID - 10.1101/2020.11.16.385930 DP - 2020 Jan 01 TA - bioRxiv PG - 2020.11.16.385930 4099 - http://biorxiv.org/content/early/2020/11/17/2020.11.16.385930.short 4100 - http://biorxiv.org/content/early/2020/11/17/2020.11.16.385930.full AB - Aversive learning – avoiding certain situations based on negative experiences – can profoundly increase fitness in animal species. The extent to which this cognitive mechanism could evolve depends upon individual differences in aversive learning being stable through time, and heritable across generations, yet no published study has quantified the stability of individual differences in aversive learning using the repeatability statistic, R (also known as the intra-class correlation). We assessed the repeatability of aversive learning by conditioning approximately 100 zebrafish (Danio rerio) to avoid a colour cue associated with a mild electric shock. Across eight different colour conditions zebrafish did not show consistent individual differences in aversive learning (R = 0.04). Within conditions, when zebrafish were twice conditioned to the same colour, blue conditioning was more repeatable than green conditioning (R = 0.15 and R = 0.02). In contrast to the low repeatability estimates for aversive learning, zebrafish showed moderately consistent individual differences in colour preference during the baseline period (i.e. prior to aversive conditioning; R ~ 0.45). Overall, aversive learning responses of zebrafish were weak and variable (difference in time spent near the aversive cue &lt;6 seconds per minute), but individual differences in learning ability did not explain substantial variability. We speculate that either the effect of aversive learning was too weak to quantify consistent individual differences, or directional selection might have eroded additive genetic variance. Finally, we discuss how confounded repeatability assays and publication bias could have inflated average estimates of repeatability in animal behaviour publications.Summary Statement Zebrafish exhibit low repeatability (intra-class correlation) in an aversive learning assay possibly due to past selection pressure exhausting genetic variance in this learning trait.