Seasonal investigation of virus dynamics by flow cytometry was conducted in the Yangtze river estuarine area in April, August, November 2002 and February 2003, and a supplemental investigation in the inner estuary and downstream of the river was conducted in October 2005. The majority of the total viral abundance was bacteriophage and only 5.4% of the total was algal virus. Total viral abundance varied with season and location, ranging from 6.75×105–1.68×107 particles/ml, and the virus:bacterium ratio (VBR) ranged from 1.52 to 72.02 with a mean of 8.7. In the present study, viral abundance peaked in both the summer and the winter, unlike the typical seasonal pattern reported in the literature, in which viral abundance peaks in the summer when bacterial hosts are also at their most abundant. However, the driving forces for the two peaks reported here were totally different, the summer viral abundance peak coupled with the development of bacterial hosts which were controlled largely by temperature year-round and by trophic state occasionally, while the winter one seemed to be multi-factor controlled. The host-phage interaction was no longer predominant in control of the winter viral abundance as bacterial abundance was lowest in this season. The winter low temperature would help maintain a high viral abundance as high temperatures might increase viral inactivation and viral decay; the VBR peak values actually occurred in the winter. More importantly, the high virus-containing freshwater discharge in winter due to a higher proportion of anthropogenic sewage relative to low natural flooding in winter run-off, turned out to be the first factor contributing to the high winter viral abundance and VBR values. In addition, the variation of intrusion of warm and relatively oligotrophic water from oceanic currents played a role alternating the distribution patterns of temperature, salinity and trophic conditions and consequently the distribution patterns of virus and bacteria seasonally and spatially. Dynamics of virus in the Yangtze river estuarine area is thus characterized by distinct seasonal and spatial variations due to natural forcing and by pronounced alternation of the regular patterns due to anthropogenic impacts.