Abstract: As a top predator, the Eurasian otter (Lutra lutra) is an indicator of healthy freshwater ecosystems and a flagship species for conservation. Once widespread in China, the species is now distributed mainly in the upper reaches of the great rivers of western China. However, a lack of knowledge on local otter populations continues to hinder their conservation in China. Here, we conducted a detailed study on habitat use of Eurasian otters in Yushu City and Tangjiahe National Nature Reserve in western China using transect surveys. At both study sites, otters preferred to defecate on large rocks close to or protruding from the river and about 50 cm above the waterline. In Yushu, no spraints were found along the 5 km river bank section in the downtown area, with otters preferring sprainting sites with natural banks, riparian zones, and lower human population density. However, this pattern was not obvious at Tangjiahe, where river transformation and human disturbance are minor. Otter river use intensity was negatively correlated with elevation and human population density in Yushu in both seasons. In Tangjiahe, otter river use intensity was positively correlated with prey mass and flow rate and negatively correlated with human population in spring, but positively correlated with human population and negatively correlated with flow rate in autumn. These results reflect the flexible habitat use strategies of otters at different sites, underlining the necessity to study otters living in different regions and habitat types. We provide suggestions for river modification and call for more site-specific studies to promote otter conservation in China.