Used since the 1970s as an avian anesthetic, the neurosteroid alfaxalone has been reformulated to avoid side effects from its castor oil excipient. This case report describes the clinical use of a new alfaxalone formulation (Alfaxan) as an intravenous anesthetic induction agent in wild isoflurane-anesthetized rose flamingos (Phoenicopterus roseus). Twenty-five male and female rose flamingos underwent orthopedic surgery using isoflurane anesthesia. The animals were induced following one of two protocols: inhaled isoflurane by facemask (ISO; n = 9) or intravenous alfaxalone (2 mg/kg; ALF; n = 16). The time and quality of anesthetic induction (until first signs of muscle relaxation) and the time and quality of recovery (sternal recumbency) were recorded using a scoring system. Mild sedation was first observed at 18.4 ± 3.8 min and 1.7 ± 0.3 min, following isoflurane and alfaxalone administration, respectively (P < 0.001). Alfaxalone induction time was significantly shorter and induction quality was considered smoother than in the ISO group. Flamingos given alfaxalone induction required lower isoflurane concentrations for maintenance anesthesia than did flamingos induced with mask isoflurane (1.5–2 %vol vs. 4–5 %vol for ALF vs. ISO, respectively). Alfaxalone produced moderate cardiorespiratory effects not seen in the isoflurane induction group. Recovery times were similar with both protocols without significant differences in quality and length. The new alfaxalone formulation produces a safe and effective anesthetic induction in rose flamingos and has significant isoflurane-sparing effects during anesthesia.