The efficacy and safety of the combination of medetomidine and ketamine was examined in order to establish an adequate chemical immobilization protocol in the Eurasian otter (Lutra lutra) for use during translocation projects in Spain. Thirty-eight Eurasian otters ranging in body mass from 3 to 8.7 kg (mean 5.3 kg) were successfully anesthetized on 82 occasions. The dosage of ketamine was 5.1 ± 0.8 (3.4–6.6) mg/kg (mean ± SD; range) combined with mede-tomidine at a dosage of 51 ± 8 μg/kg (34–66 μg/kg). In most cases anaesthetic effect occurred within 3 min and the mean induction time was 5.5 ± 3.2 min. The mean pulse rate was 95 beats/min. The mean respiratory rate was 32 respirations/min while the relative oxyhemoglobin saturation was 93%. According to these results, this anesthetic protocol is considered safe and can be recommended in wild caught Eurasian otters for immobilization during translocation projects. It is safe, rapid and can be reversed when needed with atipamezole. However caution is required as heart depression resulting in bradychardia may occur.