The food habits of 67 fish species collected from the mangrove estuary of the Urauchi River, Iriomote Island, southern Japan were investigated using gut content analysis. Ontogenetic changes in food preference were recognized in nine species, including mugilids, gerreids, mullids, gobiids and tetraodontids. In most cases, juveniles of these species fed mostly on small crustaceans (e.g. calanoid and cyclopoid copepods and gammaridean amphipods) or detritus. With their subsequent growth, larger prey items (e.g. crabs and polychaetes) became dominant. A cluster analysis based on dietary overlaps showed that the mangrove fish assemblage comprised eight trophic groups (zooplankton, small benthic crustacean, large benthic crustacean, polychaete, fish, detritus, plant and insect feeders). Of these, large and small benthic crustacean feeders, which consumed mainly crabs and gammaridean amphipods, respectively, were the most abundantly represented in terms of species, whereas polychaete and insect feeders were each represented by only two species.