River regulation by the construction of reservoirs represents one of the greatest challenges to the natural flow regime and ecological health of riverine systems globally. The Danjiangkou (DJK) Reservoir is the largest reservoir on the Hangjiang River and commenced operations in 1967. The reservoir was upgraded in 2012 to provide water resource for the South–North water transfer project through central China. However, the effect of the reservoir operations on the downstream hydrological regime and ecological health of the Hanjiang River following the upgrade (increase in dam height and reservoir capacity) has not been examined thus far. The daily discharge series from four stations along the main stem of the Hanjiang River, including a site upstream, were examined from 1950 to 2017. The study series was divided into three periods based on the difference stages of the reservoir operation: (1) 1950–1966, (2) 1967–2012 and (3) 2013–2017. The nature of hydrological alteration, ecological flow requirement and potential ecological risk during the different periods were investigated. The results clearly indicate that the DJK reservoir has significantly modified the hydrological regime in the middle and downstream section of the Hanjiang River, with most significant modifications recorded immediately downstream of the reservoir. None of the observed ‘Range of Variability Approach’ hydrological indicators fell within the expected range at Huangjiagang following the increase in reservoir capacity. As a result, the ecological flow requirements could not be guaranteed, and the frequency and intensity of ecodeficit increased. The river ecosystem immediately downstream of the dam was observed to be at high risk of ecosystem degradation during the post-dam periods considered.