Thiaminase activity of alewife (Alosa pseudoharengus) is variable across Lake Michigan, yet factors that contribute to the variability in alewife thiaminase activity are unknown. The fatty acid content of Lake Michigan alewife has not been previously reported. Analysis of 53 Lake Michigan alewives found a positive correlation between thiaminase activity and the following fatty acid: C22:ln9, sum of omega-6 fatty acids (Sw6), and sum of the polyunsaturated fatty acids. Thiaminase activity was negatively correlated with C15:0, C16:0, C17:0, C18:0, C20:0, C22:0, C24:0, C18:ln9t, C20:3n3, C22:2, and the sum of all saturated fatty acids (SAFA). Multi-variant regression analysis resulted in three variables (C18:ln9t, Sw6, SAFA) that explained 71% (R2=0.71, P<0.0001) of the variation in thiaminase activity. Because the fatty acid content of an organism is related is food source, diet may be an important factor modulating alewife thiaminase activity. These data suggest there is an association between fatty acids and thiaminase activity in Lake Michigan alewife.