Objective To evaluate the safety and efficacy of unilateral topical application of rocuronium bromide in scops owls. Animals studied Ten healthy adult scops owls. Procedures Birds weighting between 82‐111 g were enrolled. Complete physical and ophthalmic examinations were performed. Each animal received a single dose of 0.15 mg of rocuronium bromide (30 µL) in a randomly selected eye. Static pupillometric evaluations were performed before and after drug instillation at 0, 30, 60, 90, and 120 minutes, in a room with fixed light intensity. Physical and ophthalmic examinations were carried out to evaluate possible adverse effects. Results Median pupil (95% CI) size at t0 was 7.10 mm (5.51‐7.41) for placebo eyes and 7.22 mm (6.93‐7.48) for treated eyes, showing no statistical differences (P > .05). When compared to the placebo eye, significant mydriasis was achieved at t30 [8.18 mm (7.22‐9.00)] (P = .014) and lasting until t90 [7.35 mm (6.20‐9.52)] (P = .004). Maximal mydriasis was obtained at t60 [8.63 mm (7.72‐9.81)] (P = .001). During this period, the treated eye no longer responded to direct light stimulation. Complete mydriasis was observed in 5/10 birds (mean weight 97.4 g). Pupil size at t90 and t120 did not differ from baseline (P > .05) in treated eyes. No adverse effects were seen during the study period. Conclusion Single‐dose topical rocuronium bromide (0.15 mg) is a safe and effective medium duration mydriatic agent in scops owls. Further studies are needed to evaluate bilateral topical application and standardize the mydriatic protocol.