Immobilization was studied in 202 free-ranging two-toed sloths (Choloepus didactylus). All the sloths were in good condition with a body weight > 2 kg, and were anesthetized for a variety of minor clinical procedures. Intramuscular anesthetic combinations included 0.1 mg/kg acepromazine + 10 mg/kg ketamine (A/K, n = 30), 1 mg/kg xylazine + 10 mg/kg ketamine (X/K, n = 89), 10 mg/kg tiletamine/zolazepam (T/Z, n = 37), and 0.04 mg/kg medetomidine + 3 mg/kg ketamine (M/K, n = 46) antagonized by 0.2 mg/kg atipamezole. The animals were quiet during the induction stage and complete recumbency was reached in (mean +/- SD) 2.5 +/- 2.0 min with A/K, 2.7 +/- 1.7 min with X/K, 1.8 +/- 0.6 min with T/Z, and 2.5 +/- 5 with M/K. Utilization of A/K was not satisfactory because of poor anesthetic level and lack of muscle relaxation. T/Z induced immobilization was characterized by deep anesthesia and good myorelaxation, but often was associated with irregular respiration and low relative oxyhemoglobin saturation values (SpO2). Ketamine in combination with alpha2-agonists, xylazine or medetomidine, provided suitable anesthesia, with good to excellent muscular relaxation, good analgesia, high SpO2 values, moderate bradycardia, but strong bradypnea with medetomidine. Anesthesia with M/K was reversed after 41.6 min of immobilization with atipamezole. Calm recoveries were obtained and the animals were able to hang up after 10.0 +/- 7.9 min. The first signs of arousal were observed within an average of 43 to 51 min after the injection of the three other combinations. Recoveries from X/K immobilization were quiet; sloths held on after 34 min. With T/Z, recovery duration was long and very irregular at 76.7 +/- 31.3 min, some animals required 3 hr before being able to hang up. Finally, ketamine in association with an alpha2-agonist appeared to give the best chemical immobilization in wild two-toed sloths for 40 min procedures including minor surgery.