A significant limitation of behavioral ecotoxicology is the challenge of obtaining reproducible results due to a wide range of testing conditions. In particular, shoal size affects almost all aspects of fish behavior, but is rarely considered as a factor in ecotoxicological studies. In the present study, we compared the swimming and antipredator responses of different sized shoals of Arabian killifish (Aphaniops stoliczkanus) after exposure to environmentally realistic concentrations of the antidepressant medication sertraline. Groups of fish (1, 3 or 5 individuals) were exposed to either 5 or 50 ng/L sertraline. After 37 days, swimming behavior and responses to a predator alarm were measured. We found that the effects of group size were much stronger than the effects of sertraline on swimming. Group size was also the major factor influencing responses to the predator alarm, with single fish showing the strongest responses. Sertraline directly affected acceleration, turning speed and average distance to the arena wall. For all three parameters, there were significant interactions with shoal size, demonstrating that responses differed depending on the size of the group. We also found that effects of sertraline could still be observed 14 days after cessation of exposure. The study highlights the importance of considering social context and specifically shoal size when designing behavioral studies on chemicals. Failure to consider this may result in over- or under-estimation of risks.
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