Rowers pull a handle to generate propelling force while moving their bodies back and forth. In this study, we considered a method for estimating the energy expenditure for the work done by a moving body, which was defined as internal work in ergometer rowing. Ten collegiate rowers participated in this study. They increased only the stroke rate while keeping the power output constant in three power output conditions. During the exercise, we measured the oxygen uptake (V O 2) and physical activity level, which was assessed by using a triaxial acceleration sensor. The power output conditions were 90% and 100% of power output at the lactate threshold (90%LT and 100%LT) and no load condition (NL). V O 2 and physical activity level increased with increasing stroke rate, and they indicated a strong linear relationship in each condition. The slope of the regression line was significantly larger under the NL condition than under the 90%LT and 100%LT conditions; however, 90%LT and 100%LT did not differ significantly. These results suggest that the relationship between physical activity level and V O 2 , which was assessed under 90% LT or 100%LT condition, will be useful to estimate the energy expenditure for the internal work done in ergometer rowing.