The recent recognition of streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) in dogs caused by Streptococcus canis highlights our lack of knowledge regarding the mechanisms of virulence of this organism. Fifteen isolates of S. canis from cases of canine STSS and/or NF were examined for the presence of 10 Streptococcus pyogenes-associated virulence genes by Southern hybridizations using gene probes generated by PCR. The isolates lacked DNA with homology to eight of the 10 gene probes (speA, speB, speC, mf, ssa, scp, hasA, ska) under low stringency conditions. Thirteen and 15 of 15 isolates hybridized with streptolysin O and M protein gene probes, respectively. Twelve of 15 S. canis isolates were resistant to phagocytosis in canine blood. Electron microscopy revealed the presence of proteinaceous cell surface fibrillae. These results suggest that S. canis possesses M proteins and encodes streptolysin O, but lacks some of the other recognized virulence genes with significant homology to those in S. pyogenes.