In this study, dual energy X-ray absorptiometry (DXA) was performed on the calcaneus (CAL) and accessory carpal bone (ACB) of early-age neutered (n = 11) and sham-operated puppies (n = 10) to investigate changes in bone mineral density (BMD) and bone mineral content (BMC) over time. Prepubertal ovariohysterectomy and sham surgery were performed at 10 weeks, while BMD and BMC were measured at 12, 16, 20 and 24 weeks of age. BMD and BMC of CAL and ACB of ovariohysterectomised puppies were found to be higher. The differences between the groups were statistically unimportant; however, the fourth measurement of the ACB showed a significant (P < 0.05) difference. Sexually intact puppies weighed less than gonadectomised puppies in all measurements. Because of the positive correlations between body weight and bone mineral density, BMD and BMC of early-age neutered puppies were found to be higher. These results suggest that BMD and BMC are not affected by prepubertal ovariohysterectomy until six months of age in mixed-breed puppies.