Use of Hoechst 33342 Staining To Detect Apoptotic Changes in Bovine Mononuclear Phagocytes Infected withMycobacterium aviumsubsp.paratuberculosis

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Abstract
Mycobacterium aviumsubsp.paratuberculosisis an intracellular pathogen of macrophages that causes a chronic enteritis (Johne's disease) in ruminants. The purpose of this study was to determine whetherM. aviumsubsp.paratuberculosisinfection causes apoptosis in bovine monocytes. Using Hoechst 33342 staining, we observed increased numbers of apoptotic monocytes within 6 h of infection withM. aviumsubsp.paratuberculosis, and these numbers increased further at 24 and 48 h. This effect appeared to require viable bacilli, because monocytes infected with heat-killedM. aviumsubsp.paratuberculosisdid not exhibit a significant increase in apoptosis. Preincubation of monocytes with bovine growth hormone prior to infection withM. aviumsubsp.paratuberculosisdid not significantly alter the number of apoptotic cells.

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