The physical and gynecological examination of a Holstein heifer single-born with a disorder of sexual development showed anatomical abnormalities such as the presence of a scrotum and 2 vulvae and an anal sphincter that was positioned on the right side of the body. Also, an early pregnancy was diagnosed. Cytogenetic and hormone analysis was requested, and the animal showed normal female metaphases (60,XX) and hormonal profiles. However, in gross anatomy and histological examinations, a structure compatible with a penis, the absence of a uterine body, 2 exophytic structures, and a septum in the vagina were detected.