Graduate Thesis Or Dissertation
 

Properties of anti- Aeromonas salmonicida antibodies from juvenile coho salmon (Oncorhynchus kisutch)

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/6m311s412

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  • After parenteral immunization with Aeromonas salmonicida cells, the agglutination activity in juvenile coho salmon (Oncorhynchus kisutch) serum was associated with the macroglobulin serum fraction. Antibody activity was diminished by 30 minutes exposure to 50 C but not 45 C. Reduction with 2-mercaptoethanol and alkylation markedly lowered but did not completely destroy agglutination activity. The residual activity appeared to be maintained by noncovalent forces. The minimum antibody concentration required for agglutination of A. salmonicida cells was approximately 0.04 μgm protein/ml. The extreme sensitivity of this bacterial agglutination system is an important consideration in the interpretation of coho salmon anti- A. salmonicida titers. Specific antibodies were isolated from pooled coho salmon serum by dissociation of bacterial cell wall immune complexes with 15 percent NaCI. The procedure resulted in the recovery of about 30 percent of the total anti- A. salmonicida antibody. The purified antibodies had a sedimentation coefficient of approximately 17S. They migrated with the slowest moving serum component in alkaline polyacrylamide gel electrophoresis, and were restricted to the anodic portion of the serum immunoglobulin arc in immunoelectrophoretic separations. In spite of the restriction in electrophoretic mobility, the antibodies were quite heterogeneous and a minimum of 11 electrophoretic subpopulations of light chains were observed. Electron microscopy and sodium dodecyl sulfate-acrylamide gel electrophoresis revealed that the coho salmon anti- A. salmonicida rnacroglobulin was composed of four IgM-like subunits. No evidence for a low molecular weight antibody was obtained.
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