Waterbird die-offs caused by Clostridium botulinum neurotoxin type E (BoNT/E) have occurred sporadically in the Great Lakes since the late 1960s, with a recent pulse starting in the late 1990s. In recent die-offs, round gobies (Neogobius melanostomus) have been implicated as vectors for the transfer of BoNT/E to fish-eating birds due to the round goby invasion history and their importance as prey. Dreissenid mussels (Dreissena spp.) are also potentially involved in BoNT/E transmission to birds and round gobies. We examined gut contents of waterbirds collected in Lake Michigan during die-offs in 2010 -2012, and the gut contents of culled, presumably BoNT/E-free double-crested cormorants (Phalacrocorax auritus). Round gobies were found in 86% of the BoNT/E-positive individuals, 84% of the BoNT/E-negative birds, and 94% of the BoNT/E-free cormorants examined. Double-crested cormorants, ring-billed gulls (Larus delewarensis), and common loons (Gavia immer) consumed larger-sized round gobies than horned and red-necked grebes (Podiceps auritus and P. grisegena), white-winged scoters (Melanitta deglandi), and long-tailed ducks (Clangula hymealis). Other common prey included dreissenid mussels, terrestrial insects, and alewives (Alosa pseudoharengus). Our data emphasize the importance of round gobies and mussels in diets of Lake Michigan waterbirds and suggest they may play a role in the transfer of BoNT/E to waterbirds; however, round gobies and mussels were found in BoNT/E-positive, -negative, and -free individuals, suggesting that other factors, such as alternative trophic pathways for toxin transfer, bird migratory timing and feeding locations, prey behavior, and individual physiological differences across birds, may affect the likelihood that a bird will succumb to BoNT/E intoxication.