Simple SummaryDespite increasing interest in fish welfare, we still know very little about positive emotions in fish. This study had two aims: (1) to characterize a previously undescribed social behavior in zebrafish, “heightened-shoaling”, and (2) to evaluate whether heightened-shoaling may be a good candidate for future research into positive emotional behavior in zebrafish. Observing six groups of zebrafish housed in 110 L tanks furnished with a sloping gravel substrate, rocks, and artificial plants (10 fish/tank), we found that heightened-shoaling is marked by tight group cohesion, high behavioral synchrony (all fish engaging in the same behaviors at the same time), and low agonism. Episodes of heightened-shoaling occurred in all six groups, but rarely at the same time (only once out of 31 episodes), suggesting that the onset of heightened-shoaling is driven by internal group dynamics rather than external influences. Heightened-shoaling also appeared to be self-reinforcing as it had sustained durations—typically lasting for over 7 min and sometimes lasting for nearly half-an-hour. Collectively, these results are similar to the patterns that typify positive emotional behavior in other animals, for example, social grooming and social play. As the first description of heightened-shoaling, this research extends our knowledge of the range of zebrafish social dynamics and suggests a promising area for future research on positive emotions in fish.Zebrafish, a highly-social species of freshwater fish, are widely studied across many fields of laboratory science including developmental biology, neuroscience, and genomics. Nevertheless, as standard housing for zebrafish typically consists of small and simplistic environments, less is known about their social behavioral repertoire in more naturalistic settings. In particular, spontaneously occurring, socio-positive affiliative behaviors (e.g., social coordination and cohesion) that may be indicative of positive emotional experiences have rarely been reported or studied deliberately in zebrafish. Housing adult zebrafish (10 fish/tank) in large semi-natural tanks (110 L; n = 6) with sloping gravel substrate, rocks, and artificial plants, we observed a previously undescribed behavior: Distinct periods of spontaneous, synchronized, compact aggregations, what we call “heightened-shoaling”. This project aimed to quantify the characteristics of this distinctive behavior and compare parameters of heightened-shoaling to baseline periods (normal behavior) and pre-feed periods (feed-anticipatory behavior). First, across 4 days, we selected video-clips (100 s each) from within (i) instances of heightened-shoaling (n = 9), (ii) baseline periods (n = 18), and (iii) pre-feed periods (n = 18). For each of these video clips, we scan sampled every 10 s to determine fish orientations and location within the tank and agonistic behavior. Next, we used an all-occurrence sampling method to record the timing and duration of all episodes of heightened-shoaling behavior when tank-lights were on (8:00 h to 18:00 h) across 10 days. From the scan-sampling data, we found that compared to baseline periods, heightened-shoaling was characterized by increased shoal cohesion (p < 0.0001), increased adherence to the horizontal plane (p < 0.0001), decreased agonism (p < 0.0001), and no diving behavior (lower positions within the water column signal negative effect in zebrafish, p > 0.1). From the all-occurrence sampling data, we found 31 episodes of heightened-shoaling with instances observed in all six tanks and only a single case in which heightened-shoaling occurred in two tanks at the same time. The median episode duration was 7.6 min (Range 1.3–28.6). As the first systematic description of heightened-shoaling behavior, this research contributes to our knowledge of the range of zebrafish social dynamics living in naturalistic environments. Moreover, as a spontaneously occurring, protracted, affiliative behavior, heightened-shoaling appears to be a good candidate for future research into positive emotional behavior in zebrafish.