Since 1978, bottom trawl surveys in Lake Ontario have provided information on the status and trends of the benthic prey fish community related to Fish Community Objectives that include understanding prey fish population dynamics and community diversity. Beginning in 2015, the benthic prey fish survey expanded from only U.S. sites to incorporate Canadian sites, increasing the survey’s spatial coverage to a lake-wide scale. Additionally, sampling in the eastern U.S. embayments (Black River, Chaumont, Guffin, and Henderson Bays), that were historically sampled during a September bottom trawl survey to index Yellow Perch (Perca flavescens; 1978-2007), resumed in 2015. The current survey provides abundance indices for sculpins, Round Goby (Neogobius melanostomus) and Bloater (Coregonus hoyi) using techniques, gear, and timing comparable to surveys on Lake Michigan. This alignment provides a necessary biological reference point for evaluating Lake Ontario Bloater reintroduction. In 2025, the benthic prey fish survey completed 100 bottom trawl sites across main lake and embayment habitats at depths from 6 to 168 m. Sampling in US waters was limited in 2025 compared to previous years. In total, the 2025 survey sampled 59,870 fish from 23 species. No Bloater were detected in the 2025 survey. Round Goby was the most common species comprising 46% of the total catch by number, followed by Deepwater Sculpin (Myoxocephalus thompsonii), White Perch (Morone americana), and Alewife (Alosa pseudoharengus) at 23%, 9%, and 9% respectively. Slimy Sculpin (Cottus cognatus) lake-wide biomass density continues to be lower than when lakewide sampling began in 2015; zero Slimy Sculpin were detected in US waters, however sampling in the main lake within US waters was limited to the southeastern area of Lake Ontario. Deepwater Sculpin biomass has remained high since population recovery began in 2010. Embayment sampling in 2025 was limited to only Chaumont Bay.