The purpose of this study was to demonstrate whether acute vascular occlusion was safe and if it would result in changes to limb muscle size in horses. Six healthy, unfit Standardbred mares were used. Horses (standing at rest) wore an occlusion cuff at the most proximal position of the left forelimb. The right forelimb was used as control. An occlusion pressure of 200 mmHg was set for 5 min followed by a 2 min recovery. Three sets of occlusions were given to each horse. Muscle thickness was measured using B-mode ultrasound. The circumference of the forelimb and first phalanx was measured using a flexible tape measure. Pulsed-wave Doppler was performed on the radialis artery with a 5–10 MHz mechanical transducer at baseline and at each occlusion. Peak flow velocity (PFV) and the flow velocity integral (FVI) were measured each time. Mid-forelimb, but not first phalanx, girth was increased (P<0.05) in the occluded but not in the control leg following occlusion. Extensor and flexor muscle thickness was increased (P<0.05) in the occluded but not in the control leg. There were no changes (P>0.05) in PFV or FVI at any measurement time point. Acute vascular occlusion may be a suitable and safe model for studying muscle hypertrophy in horses.