A vaginal septum was diagnosed in an English bulldog bitch during routine estrous cycle staging. The septum extended 16 cm from the vestibulo-vaginal junction to just caudal to the cervix. The septum was removed by two applications of an Nd:YAG laser via a flexible fiberoptic endoscope. Complete healing occurred over a 5-week period and was uneventful. The bitch was bred and subsequently delivered four puppies vaginally. Transendoscopic laser ablation provided a noninvasive approach to surgically remove a vaginal septum in the dog.