Background and Aim:Footrot is a contagious disease of ruminants leading to severe economic losses. This study aimed to estimate the prevalence, virulence, and serogroups ofDichelobacter nodosusand the prevalence ofFusobacterium necrophorumin footrot lesions of sheep and cattle.Materials and Methods:A total of 106 pathogenic lesion samples were taken from 74 sheep and 32 cattle exhibiting typical footrot lesions and were analyzed for the presence ofD. nodosusandF. necrophorumby real-time polymerase chain reaction (PCR). Both virulence and serogroup were estimated forD. nodosuspositive samples.Results:Among the 106 samples, 89 were positive by PCR forF. necrophorum,D. nodosus, or both.Dichelobacter nodosuswas detected at a rate of 78.3% versus 28.3% forF. necrophorum. VirulentD. nodosusstrains were detected in 67.5% of positive samples, with a higher rate in sheep (73.4%) than in cattle (47.4%). BenignD. nodosusstrains were detected in 57.8% of samples, with a lower prevalence rate in sheep (50%) than in cattle (84.2%). The positive samples ofD. nodosusrevealed the presence of three dominant serogroups (D, H, I) and three minor serogroups (G, C, A) by serogroup-specific multiplex PCR.Conclusion:The findings provided information on the prevalence ofD. nodosusandF. necrophorumstrains in footrot lesions of sheep and cattle in some regions of Morocco, which will be useful for developing an effective autovaccine for the prevention of this disease in cattle and sheep in these regions.