The construction of an estuary barrage, an instream structure in the lower reaches of a river, causes significant physical changes in water flow patterns and river morphology, and results in altered environmental conditions. Here, we examined the impact of the Geum River estuary barrage, completed in 1990, on fish assemblages by using a literature search and fresh surveys of fishways in the barrage. We found that fish assemblages upstream and downstream of the barrage were altered following its completion. After construction, more species were found in the freshwater area, with a particularly great increase in freshwater species. Conversely, estuarine and marine species were only consistently caught in the downstream salt-water area, although the number of species increased. In total, 15,829 fish from 47 species and 20 families were identified at the three types (pool and weir, rubble type, and boat passage) of fishways in the barrage. The dominant species were Chelon haematocheilus, an estuarine species, Coilia nasus, a diadromous species, and Erythroculter erythropterus, a freshwater species. The mean total length of fish (101.9 ± 76.0 mm) in the boat passage fishway was approximately 100 mm lesser than those in the pool and weir (207.2 ± 112.8 mm) and rubble type (205.8 ± 112.7 mm) fishways. The boat passage fishway was the most efficient for fish movements. The current fishway system is not sufficient for fish migration, and thus additional ways are required to improve the system such as the boat passage. Few estuarine or diadromous species were found in both freshwater and salt-water areas, but freshwater fishes that accidently moved to salt-water area actively used fishways. Therefore, fishway management in the Geum River estuary barrage has to focus on freshwater fish; however, this may need to change to a focus on migratory fishes depending on ecological life cycles of migratory fish.