Thiamine deficiency complex (TDC) causes mortality and sublethal effects in Great Lakes salmonines and results from low concentrations of egg thiamine that are thought to be caused by thiaminolytic enzymes (i.e., thiaminase) present in the diet. This complex has the potential to undermine efforts to restore lake trout Salvelinus namaycush and severely restrict salmonid production in the Great Lakes. Although thiaminase has been found in a variety of Great Lakes fishes, the ultimate source of thiaminase in Great Lakes fishes is currently unknown. We used logistic regression analysis to investigate relationships between thiaminase activity and phylogenetic or ecological characteristics of 39 Great Lakes fish species. The taxonomically more ancestral species were more likely to show thiaminase activity than the more derived species. Species that feed at lower trophic levels and occupy benthic habitats also appeared to be more likely to show thiaminase activity; these variables were correlated with taxonomy, which was the most important predictor of thiaminase activity. Further analyses of the relationship between quantitative measures of thiaminase activity and ecological characteristics of Great Lakes fish species would provide greater insight into potential sources and pathways of thiaminase in Great Lakes food webs. ?? Copyright by the American Fisheries Society 2008.