The U.S. Geological Survey’s Great Lakes Science Center (GLSC), Eastern Ecological Science Center, and Columbia Environmental Research Center (CERC), and the State University of New York (SUNY) Brockport have conducted in collaboration with partner agencies a cooperative program to monitor thiamine concentrations in lake trout eggs since the late 1990s. In 2021, egg thiamine concentrations were highly variable at each sampling site. No eggs samples with thiamine concentrations less than the 4 nmol/g threshold recommended for successful lake trout reproduction were collected in Lakes Superior, Huron, Erie, and Champlain. In contrast, every site in Lakes Michigan and Ontario and Cayuga Lake had some lake trout eggs below 4 nmol/g. Time series of mean lake trout egg thiamine concentrations showed high temporal and spatial variability within the Great Lakes region.