The aim of this case series was to describe a novel technique of single-incision laparoscopic ovariectomy in dogs using the SILS Port (Covidien), a single-port multiple-access device, in 40 client-owned dogs. A single 3 cm incision was made caudal to the umbilicus and the SILS Port device was bluntly introduced. Three cannulae were inserted in the SILS Port through the access channels. In the first 20 cases, a transabdominal suspension suture was used to transfix the ovaries. In all cases, ovariectomy was performed using a standard straight non-roticulated laparoscopic grasper and a vessel sealer/divider device. Mean (sd) duration of the ovarian resection was 25.1 (6.1) minutes (range 16 to 39 minutes). In five dogs (with transabdominal suspension suture), minor bleeding in the mesovarium or in the spleen was observed. Since the SILS Port allows simultaneous use of two instruments and a telescope through a single incision, the suspension suture is not mandatory. The lack of a transabdominal suspension suture increased collision between instruments and the telescope, but triangulation capabilities remained sufficient to achieve visualisation, sufficient manoeuvrability and safe vessel sealer/divider device application. The time to perform ovarian resection remained unaltered with or without suspension suture and regardless of the fat score of the ovarian ligament. Complications were less frequent without a suspension suture.