This study was designed to describe and clarify muscle activities which occur while walking in water. Surface electromyography (EMG) was used to evaluate muscle activities in six healthy subjects (mean age, 23.3±1.4 years) while they walked on a treadmill in water (with or without a water current) immersed to the level of the xiphoid process, and while they walked on a treadmill on dry land. The trials in water utilized the Flowmill which has a treadmill at the base of a water flume. Integrated EMG analysis was conducted for the quantification of muscle activities. In order to calculate the %MVC, the measurement of maximal voluntary contraction (MVC) of each muscle was made before the gait analysis, thus facilitating a comparison of muscle activities while walking in water with those on dry land. The %MVCs obtained from each of the tested muscles while walking in water, both with and without a water current, were all found to be lower than those obtained while walking on dry land at a level of heart rate response similar to that used when walking on dry land. Furthermore, the %MVCs while walking in water with a water current tended to be greater when compared to those while walking in water without a water current. In conclusion, the present study demonstrated that muscle activities while walking in water were significantly decreased when compared to muscle activities while walking on dry land, that muscle activities while walking in water tended to be greater with a water current than without, and that the magnitude of the muscle activity in water was relatively small in healthy humans. This information is important to design water-based exercise programs that can be safely applied for rehabilitative and recreational purposes.