Behavioral and biochemical effects of environmental concentrations of caffeine in zebrafish after long-term exposure

Caffeine (CAF) is widely detected in aquatic environments, serving as an indicator of anthropogenic contamination. Its high consumption, and persistence raise environmental concerns. This study was to evaluate the chronic effects in terms of growth rate, weight, behavior, and biochemical parameters of environmental concentrations of CAF on adult zebrafish. Adult zebrafish were exposed, for 30 d, to 0, 0.5, 1.5, and 300 µg L^-1 CAF, with behavior (feeding latency, exploration, aggression, sociability, sound response) and biochemical endpoints (acetylcholinesterase (AChE), lactate dehydrogenase (LDH), and cortisol levels) assessed at the end of the exposure. CAF 0.5 µg L^-1 increased feeding latency time, while 300 µg L^-1 reduced growth and weight. Exposure to CAF affect fish behavior in terms of vertical exploration, aggressiveness, shoaling, and sound responses although were concentration specific. All concentrations tested increased social behavior, with fish swimming closer to the shoal. At a biochemical level, CAF exposed showed reduced AChE activity, while LDH activity, and cortisol levels increased at 300 µg L^-1. Low concentrations of CAF caused neurotoxicity in zebrafish which may compromise their feeding behavior, and social interactions in the wild. These changes suggest potential ecological impacts of chronic exposure to CAF, such as impaired feeding and stress responses.