Simple SummaryDue to their significant size and aggressiveness, the capture of adult crocodiles carries significant risk, both in terms of stress and injuries to themselves and to operators. Neuromuscular blocking agents act by inducing flaccid muscle paralysis, thereby reducing the physical and chemical risks associated with transportation and anesthesia. Pancuronium bromide and its antagonist, neostigmine methylsulfate, have been successfully used in juvenile and subadult saltwater crocodiles (Crocodylus porosus), but their applications in larger animals (body weight > 230 kg or total length > 3.8 m) or in Nile crocodiles (Crocodylus niloticus) have never been described. We trialed a dose recommendation in nine Nile crocodiles using pancuronium bromide that was originally established for small- and medium-sized saltwater crocodiles, the effect of which can be reversed using neostigmine. We found that the recommended dose caused a prolonged recovery time in adult male Nile crocodiles, and we propose a weight-independent dose for Nile crocodiles with body weight ≥ 300 kg or total length ≥ 4.0 m, which we successfully trialed in 32 animals.Abstract(1) Background: Pancuronium bromide is a neuromuscular blocker used for immobilizing crocodiles that can be reversed with neostigmine. A recommended drug dose has only been established for saltwater crocodiles (Crocodylus porosus), mostly based on trials in juveniles and subadults. After trialing a dose recommendation in a small cohort of nine Nile crocodiles (Crocodylus niloticus), we developed and applied a new dose recommendation for large adult Nile crocodiles. (2) Methods: we trialed and adapted a pancuronium bromide (Pavulon 4 mg/2 mL) dose in Nile crocodiles originally established for saltwater crocodiles and applied the new dose for the immobilization of 32 Nile crocodiles destined for transport. Reversal was achieved with neostigmine (Stigmine 0.5 mg/mL). (3) Results: Nine crocodiles were included in the trial phase; the induction time was highly variable (average: 70 min; range: 20–143 min), and the recovery time was prolonged (average: 22 h; range: 50 min–5 days), especially in large animals after reversal with neostigmine. Based on these results, we established a dose-independent recommendation (3 mg pancuronium bromide and 2.5 mg neostigmine) for animals weighing ≥ 270 kg (TL ≥ ~3.8 m). When applied to 32 adult male crocodiles (BW range: 270–460 kg; TL range: 3.76–4.48 m), the shortest induction time was ~20 min and the longest ~45 min. (4) Conclusions: Pancuronium bromide and its antidote, neostigmine, are effective for the immobilization and reversal of adult male Nile crocodiles (TL ≥ 3.8 m or BW ≥ 270 kg) when given in a weight-independent fashion.