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To evaluate the feasibility and safety of a transvaginal approach for laparoscopic ovariohysterectomy (OVH) in dogs and to compare it with conventional laparoscopic OVH.Prospective study.Adult healthy female mixed breed dogs (n = 24).Dogs (weighing 14–17 kg) were anesthetized and positioned in dorsal recumbency for ovariohysterectomy. Dogs were prepared for either conventional (n = 12) or transvaginal (n = 12) laparoscopic OVH. For conventional laparoscopic OVH, 3 midline abdominal portals were used and for the transvaginal approach, 2 midline abdominal portals and one vaginal portal were used. The transected ovarian pedicles, broad ligament, and uterus were removed through the umbilical region in the conventional method and through the vagina in the transvaginal method. Mean surgical time, intraoperative and postoperative complications, clinical and hematologic findings, and wound complications were compared.OVH was successfully performed without complications using both methods. Mean ± SD surgical times were similar between conventional (34.2 ± 4.03 min) and transvaginal (37.0 ± 3.56 min) methods. No significant differences, hematologic and clinical variables, were found between groups. The vaginal port could limit surgical maneuvers ergonomically during manipulation of the uterine body.Transvaginal approach for laparoscopic OVH is a feasible technique with the advantage of requiring one less abdominal portal.