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Infection with Borrelia burgdorferi caused arthritis, myocarditis, glomerulonephritis, and pneumonitis in a cow. Spirochetes were detected by use of immunofluorescent staining in liver and lung specimens and were isolated from the liver. The carpal, stifle, and tarsal joints had marked synovial proliferation, and synovial fluid obtained from these joints had high antibody titers against B burgdorferi. The cow was from an area of Wisconsin that is not endemic for borreliosis.