ObjectiveTo evaluate the effect of hyaluronidase on uptake, duration and speed of elimination of xylazine–tiletamine–zolazepam administered in the subcutaneous fat over the dorsal lumbar region of swine. Study designBlinded, randomized, crossover study. AnimalsSix healthy Landrace/Large White pigs weighing 132±24 kg (mean±standard deviation). MethodsAnimals were administered xylazine (1 mg kg−1) and tiletamine–zolazepam (8 mg kg−1) (control treatment, CON), or xylazine–tiletamine–zolazepam at the same doses with hyaluronidase (400 IU) (treatment HYA). The treatments were administered into the dorsal lumbar adipose tissue, 2.5–3.0 cm laterally from the spinous process of the second lumbar vertebra. The latency, anesthesia and recovery periods were measured. Heart rate, noninvasive systolic, diastolic, and mean arterial pressures, respiratory rate, hemoglobin oxygen saturation and rectal temperature were recorded every 10 minutes for up to 50 minutes. ResultsOne animal in CON and one animal in HYA were responsive to stimulation and did not allow safe handling. No significant difference was found between treatments for latency (CON 11.3±5.9 minutes, HYA 7.4±5.1 minutes) and anesthesia (CON 53±53 minutes, HYA 49±38 minutes) periods. Recovery period was shorter in HYA (9±6 minutes) than in CON (32±16 minutes) (p < 0.05). Physiological variables were not significantly changed over time and were within accepted normal clinical limits for the species in both treatments. Conclusion and clinical relevanceHyaluronidase (400 IU) administered into adipose tissue in pigs did not reduce the latency and duration of dissociative anesthesia, but was associated with faster recovery.