The effects of intramuscular (IM) alfaxalone were evaluated in three species of Australian frogs: Australian green tree frogs, (Litoria caerulea); green and golden bell frogs (Litoria aurea); and booroolong frogs (Litoria booroolongensis). Heart rate, respiration rate, responses, reflexes, spontaneous movement, time to effect, and duration of effect were measured. Other observations, including change in color and pupil size, were also noted. Alfaxalone administered IM at 30 mg/kg in Australian green tree frogs and 20 mg/kg in green and golden bell frogs achieved sedation and caused bradypnea. In booroolong frogs, a light plane of anesthesia and bradypnea occurred at 20 mg/kg IM. Apnea was not observed in any of the frogs, and there was no statistically significant bradycardia. No mortalities or adverse effects were noted.