A historical collection of hesperornithiform fossils from the Gammon Ferruginous, Pembina, and Millwood members of the Pierre Shale (Campanian, Upper Cretaceous) in southern Manitoba, Canada, was examined to revise their taxonomy. Only two species of Hesperornis have been recognized in previous studies on the Pierre Shale in Manitoba, but our study recognizes six species of two genera, including H. lumgairi sp. nov. H. regalis is the most common species but absent in the uppermost unit within the studied sequence. The result of this study supports the paleobiogeographic subdivision of the Campanian vertebrate fauna within the Western Interior Seaway, but not the faunal boundary that distinguishes the avian fauna of Manitoba from that of South Dakota and Kansas.