Conspecific Identity Determines Interactive Space Area in Zebrafish Shoal

[Image: see text] Sex ratio of shoals has been shown to influence shoaling behavior in many fishes. This study tests whether the conspecific identity influences shoal performance (shoal area, interactive distances, distance traveled, and thigmotaxis) of zebrafish (Danio rerio) via group tracking. We conducted a two-dimensional analysis of shoals with different sex ratios (male only, female only, male rich, and female rich) of a five-membered shoal. Parameters describing the shoal structure and individual behavior were derived using video tracking and a custom-made program. We found that mixed-sex shoals had significantly lesser shoal area and interactive distance compared to single-sex shoals (approximate difference of 80% for shoal area and 50% for interactive distance). Our findings shed light on complex interactive behaviors of zebrafish in a shoal that are affected by differences in sex ratios of interacting individuals. The outcomes from this study can be used to design better zebrafish shoaling experiments for clinically relevant research like human nerve disorders and social deficits.