Introduction: The objective of this study was to describe a laparoscopic abomasal cannulation (LAC) technique, and compare the extent of the surgical trauma after LAC and open abomasal cannulation (OAC) by examining postoperative visual analog scale (VAS) pain scores and serum values of interleukin-6 and tumour necrosis factor-α in sheep. Material and Methods: Twelve healthy ewes, weighing 38-43 kg, were used. Three-portal laparoscopic techniques were used for LAC procedures. OAC was performed by a right flank laparotomy. Results: Abomasal cannulation was accomplished in all sheep without major intraoperative and postoperative complications. The abomasal contents were collected easily in both groups. Comparative studies found that open procedures exhibit a more pronounced short-term increase in cytokines and significantly higher VAS pain scores than the corresponding laparoscopic procedures. Conclusion: The laparoscopic technique proved to be less traumatic than the conventional open technique.