It has been proposed that manipulating proprioceptive signals of the equine distal limb as part of a rehabilitation process in cases of musculoskeletal pain or neurologic deficits can be used to correct postural control and restore normal motor programs. This trial has examined the effect of treatment with a light-weight and loose-fitting bell boot (82 g) on an imbalance of muscle gluteus superficialis function in horses as measured using acoustic myography (AMG). Eight horses were trained over a 60-minute period every 3 days for 6 weeks, a protocol based on preliminary findings. Acoustic myography measurements, recording the coordination, spatial and temporal summation of muscle contractions, were made at the start (baseline) and at the finish (week 6) after a warmup period and following a set procedure of physical activity. Walking, trotting, and cantering during a left-hand circle at the start of the trial revealed a slight but significant asymmetry between the left and right hind limb muscle, which improved successfully after 6 weeks of proprioception training. Data for the right-hand circle, which revealed no significant asymmetry, during walk, trot, and canter at the start, showed no change after 6 weeks of training at the walk and trot but developed an imbalance during cantering, the result of overcompensation. This study demonstrates that functional musculoskeletal asymmetry measured during periods of activity can not only be accurately detected using AMG but it also reveals an association between the program of proprioceptive training adopted and an improvement in muscular imbalance