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To determine the feasibility of hand-assisted laparoscopic ovariectomy via colpotomy for bilateral ovariectomy in standing mares.Case series.Mares (n = 21) aged 2–21 years, weighing 312–582 kg.After a pilot study in 6 mares, the technique was used in 15 mares. Ovaries were identified and manipulated using 2 laparoscopic portals in the left paralumbar fossa. Transection and hemostasis of the ovarian pedicle was achieved using a LigaSure Atlas™ or chain écraseur. Ovaries were removed through a colpotomy by an assistant.In 19 mares, both ovaries were successfully identified from the left portals whereas in 2 mares, laparoscopic portals in the right flank were needed to access the right ovary because the working length of the equipment was insufficient to reach the right side from the left portals.Bilateral laparoscopic ovariectomy by means of 2 same-sided portals in combination with hand assistance via colpotomy is a technically feasible, safe, and effective technique in small- to average-sized mares.