While estimating the prevalence of the Dermacentor variabilis (Say) symbiont (DVS) in dog ticks on Martha‘s Vineyard, MA, we identified DNA that may represent a heretofore unrecognized Francisella sp. Polymerase chain reaction targeting a portion of the 16S rDNA specific for DVS yielded an amplicon that was only 96.6% similar to that of DVS accessioned in GenBank. Phylogenetic analysis of the 16S and 23S rDNA genes suggests the presence of a distinct bacterium closely related to the other endosymbionts of Dermacentor spp. Fifty-five percent of dog ticks tested from three sites in Massachusetts showed evidence of infection with this new agent, called Dermacentor variabilis francisella (DVF), whereas 100% tested positive for DVS. All larval progeny of dog ticks known to contain DVF also showed evidence of colonization, demonstrating that this agent may be maintained by transovarial transmission. Coinfection of ticks with both Francisella species did not seem to interfere with transmission.