We investigated the experimental vector competence of Ixodes pacificus Cooley and Kohls and Ixodes spinipalpis Hadwen and Nuttall, and the reservoir competence of the dusky-footed woodrat (Neotoma fuscipes Baird) and the deer mouse (Peromyscus maniculatus [Wagner]), for Borrelia bissettii Postic, Marti Ras, Lane, Hendson, and Baranton. Both rodent species are capable reservoirs for B. bissettii; infection rates for I. pacificus or I. spinipalpis nymphs fed as larvae on infected animals ranged from 50 to 57%. Moreover, both I. pacificus and I. spinipalpis are efficient vectors of B. bissettii. Viable infections were recorded from all rodents known to be exposed to one or more infected nymphs of I. spinipalpis (seven deer mice, two white mice) or I. pacificus (seven deer mice). In contrast, none of four New Zealand white rabbits fed upon by 90 I. pacificus nymphs with a probable B. bissettii infection rate of >50% became infected or seroconverted. The attachment and feeding success of laboratory-reared nymphs similarly confined with deer mice in muslin-covered wire-mesh cages for 24 h ranged from 0% for I. pacificus to 17–73% for I. spinipalpis. Notably, the I. pacificus nymphs were physiologically capable of feeding; nymphs failing to attach to rodents fed readily when placed in feeding capsules upon rabbits. We conclude that the dusky-footed woodrat and the deer mouse are capable experimental reservoir hosts of B. bissettii, and that both I. spinipalpis and I. pacificus are efficient experimental vectors of B. bissettii. However, the reluctance of I. pacificus nymphs to feed on certain rodents may limit its importance as an enzootic vector of B. burgdorferi sensu lato spirochetes.