In the Great Lakes region, thiamine deficiency is considered a recruitment bottleneck for lake trout Salvelinus namaycush and has been correlated with the consumption of non-native alewife Alosa pseudoharengus. While alewife, the most abundant forage fish in Lake Ontario, are the predominant prey for lake trout, they also consume benthic prey such as round goby Neogobius melanostomus. Because variation in the proportion of alewife in lake trout diets is linked to variation in egg thiamine concentrations, understanding how factors such as region of capture and hatchery-strain of lake trout influence diet, are key to understanding the patterns of variation in egg thiamine concentrations observed in this species. With recent increases in natural recruitment of lake trout being observed in the western region of the lake, understanding if egg thiamine is a potential driver is crucial to the rehabilitation of lake trout. In this study, we evaluated egg thiamine concentrations in lake trout during 2019–2020. We found no significant difference in egg thiamine concentrations among regions. However, a stocked Lake Superior deepwater morphotype (Superior Klondike Wild – SKW) showed significantly higher egg thiamine concentrations compared to the lean morphotype including Seneca (SEN) and Lake Champlain Domestic (LCD) strains. An analysis of fatty acid signatures of each hatchery-strain suggested that the SKW strain consumed a higher proportion of round goby than lean strains. Overall, these results suggest that morphotypic differences in the feeding ecology of lake trout can result in biochemical changes which may influence the effectiveness of restoration efforts.