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The Tonle Sap Lake in Cambodia supports several species of threatened megafish and contains one of the largest networks of freshwater fish reserves in the world. Despite these traits, this system remains understudied in terms of its utility for endangered fish conservation and restoration. This study was the first of a series of planned fish releases designed to test the effectiveness of conservation supplementation programs in the Tonle Sap Lake. In March 2022 (during the dry season), 1582 captive-reared fishes, including 1538 striped catfish Pangasianodon hypopthalmus, 42 giant barb Catlocarpio siamensis, and two Mekong giant catfish Pangasianodon gigas, were tagged and released into a 986-hectare fish reserve to assess post-release dispersal and survival. Brightly colored external tags with unique identification numbers were used to facilitate tag returns. A high-profile release event was held to raise awareness about the activity, bringing attention to the importance of fish reserves and endangered species conservation, and disseminating information about the research and tag return and reward program. This, in concert with other efforts, served to be an important education and outreach tool and increased tag return rates. We found that mortality from fishing was rapid and very high. Nineteen percent of released fishes were recaptured in the first 2 days after the fish release, and 46% were recaptured by day 83 after the release, indicating intense fishing pressure on the Tonle Sap Lake fisheries. Eighty percent of recaptured fishes were caught in stationary gill nets, most within 10 km of the release site. Fishing mortality rates were independent of fish size or source (pond-reared or cage-reared). Environmental DNA (eDNA) was found to be capable of detecting each of these species’ presence in the water at the release site and could prove to be a useful tool for endangered species monitoring and restoration. Future research should explore alternative release timing, release location, and other methods of increasing post-release survival. Ultimately, underlying sources of mortality, especially fishing, will need to be addressed for conservation supplementation programs to succeed in the Tonle Sap Lake. Conservation supplementation should not be viewed as a substitute for more fundamental conservation measures, such as maintenance of environmental flows, preservation of ecological connectivity, and science-based fisheries management.

Abstract Species’ responses to seasonal environmental variation can influence trophic interactions and food web structure within an ecosystem. However, our ability to predict how species’ interactions will vary spatially and temporally in response to seasonal variation unfortunately remains inadequate within most ecosystems. Fish assemblages in the Tonle Sap Lake (TSL) of Cambodia—a dynamic flood‐pulse ecosystem—were studied for five years (2010–2014) using stable isotope and Bayesian statistical approaches to explore both within‐ and among‐species isotopic niche variation associated with seasonal flooding. Roughly 600 individual fish specimens were collected during 19 sampling events within the lake. We found that fishes within the same species tended to have a broader isotopic niche during the wet season, likely reflecting assimilation of resources from either a wider range of isotopically distinct prey items or a variety of habitats, or both. Furthermore, among‐species isotopic niches tended to overlap and range more broadly during the wet season, suggesting that floodplain inundation promotes exploitation of more diverse and similar resources by different species in the fish community. Our study highlights that the flood‐pulse dynamic that is typical of tropical aquatic ecosystems may be an essential element supporting freshwater fish community structure and the fish diversity that underpins the TSL food web. This flow regime is currently threatened by regional dam development, which may in turn impact the natural function and structure of the fishery food web.