To determine whether the agent of Lyme disease disseminates in vertebrate hosts directly after deposition by an infecting tick, a 6-mm disk of skin was excised from the sites where nymphal Ixodes dammini ticks infected by Lyme disease spirochetes, Borrelia burgdorferi, had fed. Infection in each mouse was tested by examining xenodiagnostic ticks that had engorged on these mice 4 weeks later and by serologic testing. Generalized infection was aborted when the site of inoculation was excised within 2 days after the infecting tick detached but not after 2 weeks. In contrast, all mice became infected when the bite site remained intact. Spirochetes could be cultured from the tissues around the site of attachment solely when the sample was ablated within a week after infecting ticks detached. These observations suggest that infecting ticks deliver the agent of Lyme disease directly into the skin and that such spirochetes multiply locally for some days before disseminating to remote sites.