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We compared environmental effects on the macrobenthic community of the Han River Estuary in summer, when freshwater input from the Han River increased, and in spring, when freshwater input decreased. Field samples were taken from the upper region of the Shingok reservoir to the southern area of Ganghwado at 18 sampling sites after rainy (August 2006) and dry (March 2007) seasons. Macrobenthic fauna were collected using a Van Veen Grab (0.025 m2 and 0.1 m2) and environmental factors were measured simultaneously. Dominant species of macrobenthic fauna and the macrobenthic community were divided into two areas, the area of the Han River with no salinity (< 0.1 psu) and the southern part of Ganghwado with salinity (> 20 psu). The dominant species Byblis japonicus appeared at Junruri in the dry season. The distributions of two polychaetes, Hediste japonica and Nephtys caeca, were divided into the lower and upper areas of the Singok submerged weir. BIO-ENV (the matching of biotic to environmental patterns) analysis revealed that salinity was the most important factor affecting macrobenthic communities in the Han River Estuary, with other factors such as sediment grain size, bottom dissolved oxygen, and total organic carbon of sediment being secondary.