Background: Equine small intestinal resection and anastomosis is a procedure where optimizing speed, without compromising integrity, is advantageous. There is a range of different needle holders available, but little is published on their impact on surgical technique in veterinary medicine. The objectives of this study were to investigate if the needle holder type influences the anastomosis construction time, and the anastomosis quality (estimated by bursting pressure), and whether the bursting pressure is influenced by the anastomosis construction time. Single layer end-to-end jejunojejunal anastomoses were performed on jejunal segments harvested from equine cadavers. These segments were randomly allocated to four groups. Three groups based on the needle holder type that was used: 16.5 cm Frimand (Group 1), 16 cm Mayo-Hegar (Group 2) or 20.5 cm Mayo-Hegar (Group 3) needle holders. One (Group 4) as control without anastomoses. Anastomosis construction time was recorded. Bursting pressure was determined by pumping green coloured fluid progressively into the lumen whilst recording intraluminal pressures. Maximum pressure reached prior to failure was recorded as bursting pressure. Construction times and bursting pressures were compared between needle holder, and the correlation between bursting pressure and construction time was estimated.Results: Construction times were not statistically different between groups. Yet, the mean construction time was shortest in Group 2 followed by Group 3, with Group 1 having the longest mean construction time but showing most consistency. Segments from Group 2 and Group 3 burst at a statistically significantly lower pressure than those from Group 4. Group 4 and Group 1 were not different. The mean bursting pressure was highest in Group 4, followed by Group 1 and Group 2, with Group 3 having the lowest mean bursting pressure. Anastomosis construction time and bursting pressure were not correlated.Conclusions: The tested needle holders had a significant effect on bursting pressure, but not on anastomosis construction time. The Frimand needle holder shows promise to be superior for anastomoses in a clinical setting. However, future studies need to confirm these findings.