Mycoplasma bovis was identified by a specific lesion, conventional bacterial culture, immunohistochemistry, and polymerase chain reaction in 2 feedlot bison found dead with severe, chronic, caseonecrotic pneumonia; polyarthritis; and laryngitis. On microscopic examination, pulmonary lesions were characterized by prominent, well-defined areas of caseous necrosis and bronchiectasis. Immunohistochemical analysis of lung exhibited staining in bronchiolar epithelium and in random areas of caseous necrosis. On gross examination, the laryngeal lesion observed in 1 animal was typical of changes seen in cases of calf diphtheria. Nasal swabs taken from 6 clinically ill bison from the same feedlot revealed 1 animal shedding M. bovis by the nasal route. No other pathogens were recovered from the pulmonary or laryngeal lesions; however, Mannheimia haemolytica was cultured from the nasal swabs of 2 clinically ill bison, although not from the animal found to be shedding M. bovis. Several other affected bison had swollen joints and exhibited lameness and a reluctance to move. Changes observed in dead and clinically ill bison from this feedlot are similar to what has been described in the literature as chronic pneumonia and polyarthritis syndrome in feedlot cattle caused by M. bovis. Based on the severity of the lesions, and the number of dead and affected animals, bison in a feedlot setting appear to exhibit sensitivity to infection with M. bovis.