Date Awarded

2004

Document Type

Dissertation

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Virginia Institute of Marine Science

Advisor

Wolfgang K. Vogelbein

Abstract

Striped bass (Morone saxatilis) in Chesapeake Bay, USA, are experiencing an epizootic of mycobacteriosis. This disease, caused by bacteria in the genus Mycobacterium, causes granulomatous lesions of the skin and viscera. Diseased fish are often emaciated, and fish with skin lesions may be significantly disfigured. The overall goal of this work was to examine aspects of the pathobiology of mycobacteria in striped bass via laboratory exposure studies and cellular assays. Striped bass were injected intraperitoneally with a sublethal dose of Mycobacterium marinum, M. shottsii, or M. gordonae and sampled for histology and bacteriology at regular intervals to 45 weeks post-injection (p.i.). Fish injected with M. marinum developed granulomas in the mesenteries, spleen and anterior kidney. Acid-fast bacilli (AFB) were rare in initial stages of disease whereas granulomas at 8 weeks p.i. and later frequently contained large numbers of AFB. Secondary disease was observed in some fish between 26 and 45 weeks p.i., with granuloma disintegration, severe inflammation, and elevated splenic bacterial densities. Relative to fish injected with M. marinum, fish injected with M. shottsii or M. gordonae did not develop severe pathology. Granulomas were observed in the mesenteries, but were not observed in the spleen or, with one exception, anterior kidney. M. shottsii and M. gordonae both established persistent splenic infections. The ultrastructure of developing M. marinum granulomas in experimentally infected bass was examined. Formation of large macrophage aggregations containing intracellular bacilli was observed within the peritoneal cavity shortly after injection. M. marinum were always contained within phagosomes, and apparent phagolysosomal fusion was frequently observed. Epithelioid transformation of macrophages was observed. Ultrastructural observation of bacilli within granulomas agreed with histologic findings. The in vitro interaction between macrophages and intracellular M. marinum was examined ultrastructurally and with a quantitative bactericidal assay. Phagosomes containing M. marinum were fused at high rates by pre-labeled lysosomes. No differences in lysosomal fusion rates were observed between phagosomes containing live or heat-killed M. marinum. Intracellular M. marinum remained largely viable for the duration of the assay (72 hours). Heat-killed M marinum were resistant to lysis within phagolysosomes.

DOI

https://dx.doi.org/doi:10.25773/v5-5a8q-ap90

Rights

© The Author

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