Background: Long duration moderate-intensity aerobic exercise is better called an endurance exercise. The commonest machines used for indoor endurance exercise are treadmill and cycle ergometer. The preferred modality of endurance exercise should be the one that induces higher fat oxidation and lesser cardiovascular response. The aim of the present study is to compare treadmill walking with cycling on the stationary upright ergometer for cardiovascular responses and fat oxidation rate at similar energy expenditure.Methods: The present experimental cross-sectional study involved physically inactive but otherwise healthy males, aged 20.1±1.8 years having a normal body mass index. Twenty-one participants completed thirty-minutes of treadmill walking and stationary upright cycling on separate occasions to expend approximately 180 Kcal. Systolic blood pressure (SBP), diastolic blood pressure, and heart rate (HR) were recorded just before and immediately after the exercise. Rate pressure product (RPP), a linear correlate of myocardial oxygen uptake, was calculated as the product of SBP and HR divided by 100. Fat oxidation rate was calculated by an indirect calorimetric equation based on the respiratory gas exchange analysis. The paired t-test was applied for comparative analyses. P<0.05 was considered significant.Results: Treadmill walking caused a significantly lower RPP and HR while a significantly higher fat oxidation rate than cycling on the stationary upright ergometer.Conclusions: Treadmill endurance exercise could be preferred over cycling for young healthy males. However, further studies are required for the external validity of our results which are approximate rather than precise due to limited resources.