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Tropical lakes and their associated floodplain habitats are dynamic habitat mosaics strongly influenced by seasonal variations in hydrologic conditions. In flood-pulse systems, water level oscillations directly influence the connectivity to floodplain habitats for fish. Here, we aimed to investigate whether seasonal changes in the water level of a flood-pulse system (the Tonle Sap Lake, Cambodia) differentially affect diet breadth and dietary overlap of three common and commercially important fish species (Anabas testudineus, Boesemania microplepis and Notopterus notopterus) presenting important differences in their life-cycle (e.g. seasonal migration). For this purpose, the three fish species were sampled at four locations spread over the lake and their stomach contents extracted for analyses. Dietary differences were investigated across seasons regarding the diet composition and diet breadth of each species as well as the amount of dietary overlap between species. We found that the proportion of empty stomachs changed similarly across seasons for the three species, thus suggesting that ecological differences between species are not sufficient to outweigh the effect of seasonal variations in resource abundance. In contrast, changes in diet composition were species-specific and can be explained by ecological and behavioral differences between species. Diet breadth differed between species in all seasons, except during the wet season, and tended to be higher during the dry season when dietary overlap was the lowest. These variations likely result from changes in the diversity and amount of resources and may lead to habitat use shifts with potential implications for competitive interactions. In particular, increasing connectivity to floodplain habitats may reduce the competitive pressure during the wet season, while resource scarcity during the dry season may constrain individuals to diversify their diet to avoid competition. Overall, our results suggest a considerable plasticity in the feeding behavior of the three species as demonstrated by seasonal variation in both diet breadth and dietary overlap. Such variations can be explained by a number of factors and processes, including changes in resource availability or competitive interactions between individuals for resources, whose relative influence might vary depending on the magnitude and the timing of the flood-pulse driving the connectivity to floodplain habitats. Gaining knowledge on the seasonal evolution of fish's diet is relevant for fisheries management and conservation and our result could be used to guide aquaculture development in Cambodia., Competing Interests: The authors have declared that no competing interests exist. The work has been performed within the TLSCC project in collaboration with the Inland Fisheries Research and Development Institute (IFREDI) and Fisheries Administration of Cambodia (FIA). This does not alter our adherence to PLOS ONE policies on sharing data and materials.