Introduction and Historical SurveyOne of the more difficult problems encountered by investigators interested in the pathogenicity of the genus Mycoplasma is distinguishing between the mere association of these organisms with disease and a true cause-effect relationship. In this context, diseases that have been unequivocally shown to be due to mycoplasma infection alone assume particular significance, especially if such diseases occur naturally. An infectious mastitis of cattle is in this category.Mycoplasma infection of the bovine mammary gland was probably first observed by Alstrom (1955) in Sweden. He isolated pleomorphic organisms from bovine milk on serum-enriched media and considered them variants of bacteria present in some of the same milk samples. Retrospectively, it seems likely that these serum-dependent organisms were mycoplasma.The English investigators Davidson and Stuart (1960), in a brief report, described the isolation of mycoplasma from milk samples taken from cows with mastitis, but did not clearly establish an etiological relationship.Hale et al. ( 1962) at the University of Connecticut cultured mycoplasma from the milk of cattle suffering from severe mastitis. Similar results were reported by Carmichael et al. (1963), who isolated mycoplasma from severe mastitis cases in IS New York dairy herds. Both groups showed that the disease could be reproduced readily with pure cultures and described clinical and pathological changes that occurred in infected udders. In 1961, Langer isolated a hemadsorbing agent in tissue culture from the milk of a cow with bacteria-free mastitis. Langer initially considered this agent a virus, and designated it strain 56-R. Further study (Langer and Carmichael, 1963) showed that 56-R could be cultivated on artificial serum-enriched media, and that this isolate possessed characteristics of mycoplasma. Other milk samples drawn in 1961 from cows with mastitis had given negative results on conventional bacteriological media, but produced definite, though slight, changes in inoculated bovine kidney cell cultures. Although no microorganisms were identified, the milk samples were considered suspicious and were frozen for study at a later time. After the identification of 56-R as a mycoplasma, the stored samples were recultured on serum-enriched artificial media, and a portion of them yielded mycoplasma. These isolates, however, were different from strain 56-R. Severe, purulent bovine mastitis was produced experimentally with pure cultures of each strain.In 1963, Stuart et al. published the results of further studies on isolates from bovine mastitis in England, and presented proof that the mycoplasma which they had isolated were responsible for the outbreaks of severe mastitis that had been observed earlier (Davidson and Stuart, 1960).