To gain insight into the adaptive mechanisms to tripedal locomotion and increase understanding of the biomechanical consequences of limb amputation, this study investigated kinetic and temporal gait parameters in dogs before and after the loss of a hindlimb was simulated. Nine clinically sound Beagle dogs trotted on an instrumented treadmill and the ground reaction forces as well as the footfall patterns were compared between quadrupedal and tripedal locomotion. Stride and stance durations decreased significantly in all limbs when the dogs ambulated tripedally, while relative stance duration increased. Both vertical and craniocaudal forces were significantly different in the remaining hindlimb. In the forelimbs, propulsive force increased in the contralateral and decreased in the ipsilateral limb, while the vertical forces were unchanged (except for mean force in the contralateral limb). Bodyweight was shifted to the contralateral and cranial body side so that each limb bore ~33% of the dog's bodyweight. The observed changes in the craniocaudal forces and the vertical impulse ratio between the fore- and hindlimbs suggest that a nose-up pitching moment occurs during the affected limb pair's functional step. To regain pitch balance for a given stride cycle, a nose-down pitching moment is exerted when the intact limb pair supports the body. These kinetic changes indicate a compensatory mechanism in which the unaffected diagonal limb pair is involved. Therefore, the intact support pair of limbs should be monitored closely in canine hindlimb amputees.