OBJECTIVE: To evaluate the effect of treadmill incline on muscle activity and joint range of motion (ROM) in hind limbs of dogs. ANIMALS: 8 purpose-bred healthy adult hounds. PROCEDURES: Activities of the hamstring (semimembranosus, semitendinosus, and biceps femoris muscles), gluteal (superficial, middle, and deep gluteal muscles), and quadriceps (femoris, vastus lateralis, vastus intermedius, and vastus medialis muscles) muscle groups and hip and stifle joint ROM were measured with surface electrogoniometric and myographic sensors in hounds walking on a treadmill at 0.54 m/s at inclines of 5%, 0%, and -5% in random order. Mean electromyographic activities and mean ROMs at each inclination were compared for swing and stance phases. RESULTS: Treadmill inclination did not affect duration of the stance and swing phases or the whole stride. When treadmill inclination was increased from -5% to 5%, hip joint ROM increased and the degree of stifle joint extension decreased significantly. In the beginning of the stance phase, activity of the hamstring muscle group was significantly increased when walking at a 5% incline versus a 5% decline. In the end of the stance phase, that activity was significantly increased when walking at a 5% incline versus at a 5% decline or on a flat surface. Activity of the gluteal and quadriceps muscle groups was not affected when treadmill inclination changed. CONCLUSIONS AND CLINICAL RELEVANCE: Treadmill inclination affected joint kinematics only slightly. Walking on a treadmill at a 5% incline had more potential to strengthen the hamstring muscle group than walking on a treadmill with a flat or declined surface.