Thirteen healthy Labrador retrievers underwent a 5-stage incremental treadmill exercise test to assess its reliability. Blood lactate (BL), heart rate (HR), and body temperature (BT) were measured at rest, after each stage of exercise, and after a 20-min recovery. Reproducibility was assessed by repeating the test after 7 days. Two-way MANOVAs revealed significant differences between consecutive stages, and between values at rest and after recovery. There was also a significant reduction in physiological strain between the first and second trial (learning effect). Test reliability expressed as typical error (BL = 0.22 mmol/l, HR = 9.81 bpm, BT = 0.22 degrees C), coefficient of variation (BL = 19.3%, HR = 7.9% and BT = 0.6%) and test-retest correlation (BL = 0.89, HR = 0.96, BT = 0.95) was good. Results support test reproducibility although the learning effect needs to be controlled when investigating the exercise-related problems commonly observed in this breed.