Fascioliasis is a common foodborne and waterborne zoonotic trematode parasitic disease infection worldwide. However, yak’s fascioliasis is rarely reported on Tibetan Plateau. In this study, serological survey and associated risk factors of Fasciola hepatica (F. hepatica) infection in Tibetan yaks was investigated using an ELISA assay. The results showed that the 305/849 (35.92%) studied animals were sero-positive for F. hepatica with the further distribution of 24.20% (38/157), 38.02% (92/242) and 38.89% (175/450) from Bayi, Mainling, and Gongbo’gyamda areas, respectively. The seroprevalence of F. hepatica increased from 20.75% to 48.8% with age of yaks, with the further distribution of 20.75%, 30.23%, 39.55%, 37.70% and 48.8% for age groups: <0.5 year, 0.5-1 year, >1-2 year, >2-4 years and >4 years, respectively. The prevalence in the male and female yaks was 32.51% and 39.05%, respectively. The physical examination showed a poor body condition with emaciation, weakness, loss of appetite, anemia, jaundice, and abdominal edema in infected yaks, while on dissection, many prominent nodules of parasitic migration were present on the liver surface. Our results provided a baseline data for the exposure of yaks to F. hepatica infection in the study area. Therefore, some important strategies and surveillance should be adopted to control F. hepatica on Tibetan Plateau, China.