In this study, using a continuous behavior measurement technique, the short-term behavioral responses and tolerance limits of red seabream Pagrus major fingerlings to sudden exposure to low salinity in a controlled environment were observed. The activity of the fingerlings suddenly exposed to 21.4, 17.3, and 9.8 psu increased temporarily at the initial exposure to show irregular swimming behavior, but then recovered a stable activity pattern through rapid salinity adaptation. However, the organisms suddenly exposed to 7.3 and 4.3 psu could not withstand the salinity stress, and their swimming behavior was severely disturbed and all individuals died within 48 hours. The findings suggest that red seabream underwent a temporary salinity stress process at the beginning of the exposure to concentrations of 10.0 psu or higher. At these concentrations, osmotic control was possible within at least 11 hours, so stable metabolic activity was also possible. However, organisms suddenly exposed to concentrations below 5.0 psu exceeded the tolerance to low salinity and the sublethal limit. In red seabream exposed to this concentration range, severe behavioral and metabolic disturbances were observed, and death was observed due to osmotic control failure. In conclusion, a salinity range of 5.0 to 10.0 psu can be predicted to correspond to a concentration range in which the osmotic control ability of the red seabream fingerlings is lost, and sub-lethal reactions occur.