We recorded the electromyographic (EMG) activity of 7 skeletal muscles in the forequarters and 1 in the hindquarters of 6 Thoroughbred horses during overground walking, swimming in a circular pool, and walking and trotting in a water treadmill. Bipolar fine wire electrodes were inserted into the muscles and the EMG signals were recorded using a telemetric system. The splenius exhibited tonic EMG activity during swimming. The brachiocephalicus showed its highest intensity during swimming followed by the walk and trot in the water treadmill and then walking overground. The triceps brachii caput longum had a similar activity pattern to the brachiocephalicus. The brachialis showed only weak EMG activity in all 3 types of locomotion. The extensor digitorum communis had higher intensity of EMG activity in the walk in the water treadmill than in other kinds of locomotion. The flexor digitorum profundus exhibited the most intense EMG activity during swimming. These results indicated that swimming evoked strong EMG activity in the antigravity muscles in spite of reduced gravitational force. Walking in the water treadmill may require more intensified EMG activity of the forelimb than the trot in the same treadmill.