Long-term survival of Mycoplasma bovis in necrotic lesions and in phagocytic cells as demonstrated by transmission and immunogold electron microscopy in lung tissue from experimentally infected calves

In the lungs of cattle infected with Mycoplasma bovis persistence of the agent for several weeks after infection has been demonstrated by different methods, e.g. isolation of the organism, immunohistochemistry for antigens, and in situ hybridization. The presence of macrophages and neutrophils with cytoplasmic M. bovis antigen suggests that phagocytosis occurs in vivo. It is, however, unknown whether this intracellular immunolabeling detected residual antigen after phagocytosis of M. bovis or surviving organisms in macrophages that use the intracellular survival as a strategy for evasion of the immune response. The aim of this electron microscopic investigation was to study the distribution of M. bovis within caseonecrotic lung lesions and to examine the phagocytes for intracellular presence of the agent. In lung tissue sections from 9 experimentally infected calves originating from two different infection experiments large numbers of mycoplasmas were detected by transmission electron microscopy and by immunogold electron microscopy using M. bovis-specific polyclonal antibodies. M. bovis were found throughout caseonecrotic foci and within the lumen of bronchi containing exudate. The majority of mycoplasmas were located extracellularly within necrotic lung lesions and around neutrophilic granulocytes and macrophages, while fewer organisms were found within the cytoplasm of phagocytes. The results of this study show that there is long-time survival of numerous intact M. bovis in necrotic lung lesions even though large numbers of neutrophils and macrophages are present. These findings show that the phagocytes are not able to eliminate M. bovis from the lungs from necrotic and inflamed lung tissue and indicate that persistence of the agent is possibly due to its capacity to avoid phagocytosis.