Since 1978, surveys of Lake Ontario preyfish communities have provided information on the status and trends of the benthic preyfish community related to Fish Community Objectives that includes understanding preyfish population dynamics and community diversity. Beginning in 2015, the benthic preyfish survey expanded from US-only to incorporate Canadian sites, increasing the survey’s spatial coverage to a lake-wide scale. Additionally, sampling in eastern US 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) with survey techniques, gear and timing comparable to Lake Michigan. This alignment provides a necessary biological reference point for measuring the success of Lake Ontario Bloater reintroduction. In 2022, the collaborative benthic preyfish survey completed 171 bottom trawl tows across main lake and embayment sites at depths from 6 to 222 m. In total, the 2022 survey sampled 141,552 fish from 34 species. Round Goby was the most numerically abundant species comprising 36% of the total catch, followed by Alewife (Alosa pseudoharengus) and Deepwater Sculpin (Myoxocephalus thompsonii), at 20% and 16%, respectively. Alewife accounted for most (623 kg) of the fish biomass sampled during the 2022 survey (total=2,197 kg), followed by Deepwater Sculpin (547 kg), and Round Goby (262 kg). Slimy Sculpin (Cottus cognatus) lake-wide biomass density (0.03 kg/ha) remained low relative to historical observations from US waters during the 1980-1990s and was similar to the average from the previous three survey years (2019-2021 average 0.04 ± 0.02 kg/ha). Lake-wide Deepwater Sculpin biomass density reached a new high (4.4 kg/ha) in 2022. Embayment catches continue to have unique species assemblages compared to main lake habitat. Historically common native benthic preyfish species like Trout-perch (Percopsis omiscomaycus), Spottail Shiner (Notropis hudsonius), and darters (Etheostoma spp.), that are now rare at main lake trawl sites, still occur in some embayment trawl sites.