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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.

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