This prospective echocardiographic study investigated the respective impacts of left ventricular (LV) pacing and simultaneous and sequential biventricular pacing (BVP) on ventricular dyssynchrony during exercise in 23 patients with compensated heart failure and ventricular conduction delays. During exercise, LV pacing and BVP significantly (p <0.05) improved mitral regurgitation and LV dyssynchrony compared with spontaneous activation. LV segmental electromechanical delays were significantly prolonged during LV pacing, leading to increased systolic time (p <0.05), decreased LV filling time (p <0.05), and decreased stroke volume (p <0.05) compared with BVP. After optimization of the interventricular delay with sequential BVP, additional benefit was obtained during exercise in terms of stroke volume and mitral regurgitation (p <0.05). The optimal interventricular delay was different at rest and during exercise in 57% of the patients. Changes from at rest to exercise in LV dyssynchrony were correlated with changes in stroke volume (r = -0.61, p <0.01) and changes in mitral regurgitation (r = 0.60, p <0.01).