The importance of Dermacentor spp. in the transmission of tick-borne pathogens is not well recognized in Europe. To investigate the role of Dermacentor spp. in the transmission of tick-borne pathogens, questing ticks were collected in 9 sites from southern to northwestern France (Camargue Delta to Eastern Brittany) where Dermacentor spp. exist and tick-borne diseases had occurred previously. Three tick species were collected during the spring and autumn of 2009. Collected ticks (both males and females) included D. marginatus (n = 377), D. reticulatus (n = 74), and I. ricinus (n = 45). All ticks were analyzed by PCR or reverse line blot for the presence of pathogens' DNA. Pathogens analyzed were based on veterinarian reports and included Anaplasma phagocytophilum, Coxiella burnetii, Anaplasma marginale, Borrelia burgdorferi, Bartonella spp., Babesia spp., Theileria spp., and Francisella sp. Francisella tularensis was not detected in any of the analyzed ticks. In D. marginatus, infection prevalence for A. phagocytophilum (3%) was similar to that found in I. ricinus in Europe. Other pathogens present in D. marginatus included A. marginale (0.5%), Bartonella spp. (9%), C. burnetii (12%), F. philomiragia (1.3%), and Theileria annulata/Babesia bovis (0.3%), which were detected for the first time in France. Pathogens detected in D. reticulatus included A. marginale (1%), Bartonella spp. (12%), C. burnetii (16%), Borrelia spp. (1.5%), and F. philomiragia (19%). Pathogens detected in I. ricinus included A. phagocytophilum (41%), Bartonella spp. (9%), C. burnetii (18%), A. marginale (1%), Borrelia spp. (4.5%), and Babesia sp. (7%). This study represents the first epidemiological approach to characterize tick-borne pathogens infecting Dermacentor spp. in France and that may be transmitted by ticks from this genus. Further experiments using experimental infections and transmission may be now conducted to analyze vector competency of Dermacentor spp. for these pathogens and to validate such hypothesis.