The aim of this study was to investigate the effect of combined atropine low-dose dobutamine stress test on left ventricular parameters in adult warmblood horses, to establish a potential protocol for pharmacological stress echocardiography. Seven healthy untrained warmblood horses aged 9 to 22 years were used. Heart rate (HR) and left ventricular B- and M-mode dimensions were recorded at baseline and during stress testing with 35 μg/kg atropine IV followed by incremental dobutamine infusion of 2 to 6 μg/kg/min. HR increased significantly (P < .05) during the pharmacological challenge, and a maximal HR of 156.6 ± 12.5 bpm was reached at maximal dobutamine infusion rate. Systolic and diastolic interventricular septum thickness, systolic and diastolic left ventricular free wall thickness, and fractional shortening increased significantly and reached a maximum at the highest infusion rate (mean ± SD: 4.51 ± 0.27 versus 5.65 ± 0.31 cm, 2.89 ± 0.19 versus 3.78 ± 0.10 cm, 3.72 ± 0.34 versus 4.77 ± 0.18 cm, 2.44 ± 0.28 versus 3.11 ± 0.34 cm, 34.98 ± 3.82 versus 50.56 ± 3.42%, respectively). Systolic and diastolic left ventricular internal diameter decreased significantly during dobutamine infusion. Left ventricular external and internal area were significantly lower at a dobutamine infusion rate of 2 μg/kg/min but no further decrease was observed during the subsequent steps. Systolic and diastolic myocardial area was significantly lower after the administration of dobutamine but not significantly different during dobutamine infusion, when compared to baseline values. This pharmacological stress test induced significant changes in left ventricular echocardiographic parameters in adult warmblood horses. Additional research should evaluate the value of this stress test in horses suffering from cardiac disease.