Four-week-old northern bobwhite quail (Colinus virginianus) were inoculated subcutaneously with 10⁶ organisms from a low passage culture of Borrelia burgdorferi. Blood was collected weekly for culture, antibody detection, and immunoblot analysis. Three weeks postinoculation, viable spirochetes were isolated from the blood of one bird, but not from kidney, spleen, liver, or heart; all infected birds from which preinfection antibody titer had been established, had antibodies by the enzyme-linked immunosorbent assay (ELISA). The inoculated birds did not show clinical signs of disease and there were no detectable gross or histopathologic lesions. Borrelia burgdorferi was detected in sections of kidneys on fluorescent antibody tests. Using a polymerase chain reaction (PCR) analysis to detect Borrelia burgdorferi DNA in tissue samples, the expected PCR product (DNA) of 246 base pairs was visible on agarose gel stained with ethidium bromide. The identity of the PCR product was confirmed by slot blot hybridization with Borrelia burgdorferi specific DNA probe. Thus, these birds sustained infections for at least 3 weeks without clinical signs and may play a role in the transmission of Borrelia burgdorferi.