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Cardiac effects of the β2-adrenergic agent clenbuterol have been the focus of many studies, but effects on myocardial velocities and myocardial deformation parameters have not yet been evaluated in horses affected with recurrent airway obstruction (RAO) using tissue Doppler imaging (TDI) and two-dimensional speckle tracking (2DST). In our study, 7 horses affected by RAO were treated over 14 days with clenbuterol, 0.8 μg/kg every 12 hours. Standard echocardiographic, TDI (pulsed wave and color TDI), and 2DST examinations were performed before and after the treatment period. Myocardial function was recorded in the right parasternal short-axis view. Percent of fractional shortening and two-dimensional echocardiography (2DE) measurements did not show any significant changes after 2 weeks of treatment. Early diastolic velocity, E, increased significantly after clenbuterol in the left ventricular free wall (LVFW; P = .001). The E/late diastolic velocity (A) quotient (P = .003) and the isovolumetric contractility (P = .035) also increased significantly after treatment. Time parameters, particularly the time interval between the Q-wave in the echocardiograph and atrial release, the time of diastole and Tei index (parameter of global ventricular function), decreased significantly after clenbuterol administration in the LVFW (P = .014/P = .028/P = .015, respectively). The 2DE speckle tracking revealed a significant increase of the early diastolic systolic strain rate (P = .01) in the LVFW after therapy. In conclusion, 2 weeks of treatment with clenbuterol at a dosage of 0.8 μg/kg every 12 hours led to improved cardiac function in severely RAO-affected horses. This could be a sign of myocardial restoration (re-remodeling) after therapy.