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To understand the role of bighorn sheep (Ovis canadensis) in the epidemiology of anaplasmosis, we recovered a field isolate from a suspected enzootic area in southern California (USA). Whole blood was collected from three desert bighorn sheep (Ovis canadensis nelsoni) and inoculated into a susceptible splenectomized domestic sheep, calf and a susceptible spleen-intact bighorn sheep. No infection occurred in the calf, but a detectable infection developed in both the domestic sheep and bighorn sheep 24 days after inoculation. The infection in both domestic and bighorn sheep resulted in severe clinical disease but was resolved with the use of tetracycline. Using monoclonal antibodies and DNA probes, we confirmed that the isolate was Anaplasma ovis.