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Separating the individual effects of climate variability and human activities on streamflow is more important than just knowing their combined effects. In this paper, using a scenario-based hydrological simulation approach, the streamflow changes caused by climate variability and two different types of human activities (i.e. land-use change and large reservoirs operations) as well as the contribution rates of these three factors over 272 sub-basins in the Yangtze river basin were quantified and compared among 5 different periods (i.e. 1988–1992 (P1), 1993–1997 (P2), 1998–2002 (P3), 2003–2007 (P4) and 2008–2012 (P5)). Results demonstrate that, at the annual scale, climate variability played a leading role in the change in outflow of most sub-basins. With regard to the seasonal variations in discharge at Datong station, climate factors played a predominant role during P1-P2 and P2-P3. Since the Three Gorges Reservoir began operating in 2003, the discharge was enhanced by reservoirs in January-May and reduced by reservoirs in July-December. Reservoir and climate factors codetermined seasonal streamflow change during P3-P4 and P4-P5. Land-use change made the smallest contribution to seasonal discharge fluctuations. This study can support decision-making in regional water resources planning and management. [ABSTRACT FROM AUTHOR]