9 first-lactation dairy cows in a closed dairy herd had swelling in the forelimbs and forelimb lameness. Mycoplasmal arthritis and mastitis were diagnosed. Swelling of the carpal joint, diffuse subcutaneous edema from the carpal to metacarpophalangeal joints, and forelimb lameness were evident in 9 first-lactation cows 7 to 21 days after parturition. Diagnostic testing revealed that 3 of 3 bulk-tank milk samples, 3 milk samples from cows with clinical mastitis, 2 fluid samples obtained from arthritic joints, and samples from the lungs and spleen of a cow that had died yielded positive results for Mycoplasma spp. Nucleic acid sequence analysis performed by use of a PCR assay on the joint fluid and lung tissues confirmed infection with Mycoplasma bovis. Affected cows were treated by IM administration of flunixin meglumine and dexamethasone for 3 days. All cows were nonresponsive to treatment (3 cows died, and the other 6 were culled). Follow-up culture for Mycoplasma spp of milk samples from the bulk tank and from all lactating cows was recommended to screen for chronic subclinical carriers. Mycoplasmal infections may cause unusual initial clinical signs or an atypical history. When dairy cattle, including those residing in closed herds, have lameness, swelling of the carpal or metacarpophalangeal joints, edema of the distal portions of the forelimbs, or polyarthritis, infection with Mycoplasma spp should be investigated. Delay in diagnosis of mycoplasmal infections in dairy herds can result in substantial financial loss and the establishment of chronic subclinical carriers.