This study aimed to determine, if there is a difference in energy expenditure during walking among athletes and non-athletes at two different speeds of walking. Ninety five healthy female students (47 athletes and 48 non-athletes) with a mean age of 22.4 (±1.6) years purposively participated in this study. Medical and sport participation history of the subjects was acquired through a questionnaire. Two experimental tests including anthropometric measurements,V O , and walking tests on treadmill at speeds of 3.00 and 3.5 mph were conducted. Results showed no difference in weight, height, body mass index, and leg length between both groups. The non-athletes expended a greater amount of energy than athletes (3.78±.1 and 2.95±.6 kcal.min -1 , respectively) at both speeds of 3.00mph and 3.5mph (4.89±1 and 3.94±.7 kcal.min -1 ). Based on energy requirements for walking at similar weights and speeds by ACSM's guideline, the female athletes walked at a slow, moderate and brisk pace. Most of the female non-athletes walked at a moderate, brisk and very brisk pace. This study revealed that regular exercise could improve walking efficiency, and the energy expenditure of walking would play an important role in the information processing for total energy requirement that progressively affects weight management and health.