Experiments on the natural history of Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis (HME), would be facilitated by the availability of a laboratory animal model for transmission to vector ticks. Five strains of mice were evaluated for their susceptibility to infection with E. chaffeensis and transmission competence: C57BL/6 mice, inducible nitric oxide synthase (iNOS) deficient mice, MHC I deficient (beta2m /) mice, MHC II deficient mice (Abb /), and B and T cell deficient (Rag1 /) mice. Mice were inoculated with a low passage isolate of E. chaffeensis, and infection and morbidity were monitored for 57 days. Three xenodiagnostic infestations with A. americanum nymphs were performed 1, 8, and 15 days following inoculation. C57BL/6 mice cleared the organism in less than 17 days, with no indication of morbidity, and mounted a rapid, strong antibody response. Transmission to feeding A. americanum nymphs was seen in 1/30 nymphs fed on C57BL/6 mice immediately after inoculation. In MHC I and iNOS deficient mice, pathogen DNA was detected up to 17 or 24 days, respectively, after inoculation. Persistent infection for the duration of the experiment (57 days) was observed in MHC II deficient mice. However, E. chaffeensis was not detected in ticks fed on iNOS, MHC I, or MHC II deficient mice. Susceptibility to infection was greatest in Rag1 knockout mice, with significant morbidity and mortality within 24 days after inoculation. E. chaffeensis DNA was detected in up to 55% of replete nymphs that fed on Rag1 mice. However, E. chaffeensis was not detected in molted adult ticks from the same cohorts.