Abstract Background Fasciola hepatica is an important zoonotic parasite that causes fasciolosis in a broad range of animals. No information is available about the prevalence of F. hepatica in Père David’s deer (Elaphurus davidianus), an endangered species in the world. Therefore, the purpose of the study was to evaluate the prevalence of fasciolosis in Père David’s deer in the Dafeng Elk National Natural Reserve, Jiangsu province, China. Results In this study, 142 fecal samples from Père David’s deer were analyzed for F. hepatica by microscopy and nest-PCR. Only one sample was positive for F. hepatica according to microscopy examination, while 18 of 142 (12.68, 95%CI: 2.841–22.45%) samples were positive for F. hepatica according to nest-PCR results. Conclusions This is the first report of prevalence of F. hepatica in Père David’s deer. The prevalence data indicated that F. hepatica was also present in this endangered animal, which may cause a potential threat to this precious species.