Multi-frequency BIA (mfBIA) equipment has been shown to be a non-invasive and reliable method to assess a muscle as a whole or at fibre level. In the equine world this may be the future method of assessment of training condition or of muscle injury. The aim of this study was to test if mfBIA reliably can be used to assess the condition of a horse's muscles in connection with health assessment, injury and both training and re-training. mfBIA measurements was carried out on 10 'hobby' horses and 5 selected cases with known anamnesis. Impedance, resistance, reactance, phase angle, centre frequency, membrane capacitance and both extracellular and intracellular resistance were measured. Platinum electrodes in connection with a conductance paste were used to accommodate the typical BIA frequencies and to facilitate accurate measurements. Use of mfBIA data to look into the effects of myofascial release treatment was also demonstrated. Our findings indicate that mfBIA provides a non-invasive, easily measurable and very precise assessment of the state of muscles in horses. This study also shows the potential of mfBIA as a diagnostic tool as well as a tool to monitor effects of treatment e.g. myofascial release therapy and metabolic diseases, respectively.