BackgroundSurgical approaches to the equine rectum and perirectal area are described in the literature. However, surgeries in this region can be challenging.ObjectiveTo describe the surgical anatomy of the presacral space and to evaluate its access using a retroperitoneoscopic approach.Study designEx vivo experiment.MethodsPreliminary dissections were performed in two cadavers to define the boundaries of the presacral space and to determine portal locations for the surgical approach. After that, nine cadavers were used for experimental presacral retroperitoneoscopic procedure in a standing position. Following retroperitoneoscopy, cadavers were dissected to confirm the anatomical structures observed during the endoscopic procedures, to control the location of each portal and to record iatrogenic trauma.ResultsThe presacral space was bordered by the vertebral column from the ventral aspect of lumbosacral promontorium to the first coccygeal vertebra dorsally and by the presacral fascia and peritoneum ventrally. Lateral limits were composed of the sacrosciatic ligament and transversalis fascia. Cranial and caudal borders were composed of the peritoneum and coccygeal and levator ani muscles respectively. Retroperitoneoscopic portals were placed between the external anal sphincter and semimembranosus muscles and between the base of the tail and the external anal sphincter muscle through the anococcygeal fascia to enter the space by its caudal border. The retroperitoneal space was reached in all cases and the dorsal and lateral aspects of the rectum were visualised after creation of a working space.Main limitationsUse of cadaver specimens do not permit to evaluate the tolerance in living animals and the surgical complications such as rectal damage, haemorrhage and infection.ConclusionThis study provides an anatomical description and surgical access of the presacral space with a minimal invasive approach. Retroperitoneoscopy allows access to the rectum and the dorsal aspect of the pelvis.