Siamese fighting fish (Betta splendens) has been extensively exploited in the behavioral and physiological toxicology studies of drugs. Tacrolimus is an immunosuppressant drug largely used in liver and renal transplantations. Here we found that a 7-day exposure of male B. splendens to concentrations of 0.05 and 0.1 µg/mL Prograf® (tacrolimus) caused alterations in aggression and immunity indexes. Tacrolimus exposed fish presented lower opercular display in a mirror test which is indicative of reduced aggression. In addition, serum levels of lysozyme, IgM, alternative complement, and bactericidal activity of subjects exposed to 0.1 µg/mL tacrolimus were lower than those from the control treatment. These results showed the behavioral impairment and immunotoxic impacts of tacrolimus in a model of aquatic toxicology. The results suggest fishes provide a possible model for better understanding of the drug action in vertebrates, and possible consequences for the environment via its effects on non-target organisms in an ecotoxicology context.