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In order to understand the stress responses, we measured the concentration of stress hormones by using the fresh feces of wild otters in the Gwangju stream and Seomjin river during summer and winter in 2018. We compared seasonal and regional number of spraints (=otters’ feces), temperature, altitude, water width, land cover, and water quality with stress hormones. The stress hormone levels were higher in summer than in winter, and significantly different to season and region. In addition, the number of feces were different according to the season and region, and showed a negative correlation with stress hormone. This study suggested assessing stress hormones as another promising method for the analysis of status of wild otters and it will contribute to establish the management and conservation policy, especially in urban area.