Graduate Thesis Or Dissertation
 

Development of monoclonal antibodies against the myxosporean, ceratomyxa shasta, and characterization of the salmonid host response

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  • Monoclonal antibodies (Mabs) were produced against the myxosporean parasite, Ceratomyxa shasta, and were characterized by indirect immunofluorescent techniques, western blot analysis, and immunoelectron microscopy. The resulting Mabs reacted specifically with the trophozoite and sporoblast stages of the parasite, but not with the mature spore, indicating that certain antigens are stage-specific. Two of the Mabs produced also reacted with trout white blood cells as evidenced by indirect fluorescent antibody techniques (IFAT) and Western blot analysis. Further characterization of these cross-reacting antibodies showed that they were directed against carbohydrate epitopes on both the parasite and trout immunoglobulin heavy chain. One hybridoma produced antibodies of high specificity for prespore stages of C. shasta. Ultrastructural analysis of the parasite using immunogold labeling showed that the Mab specifically bound antigen located in the cytoplasm of the trophozoites. The C. shasta-specific antibodies produced by monoclonal technology, standard histological methods, and scanning electron microscopy were used to investigate the host response of Sa lmo gairdneri against infection by the parasite. In infected fish the parasite was first observed in the mucosal epithelium of the posterior intestine using IFAT. As the trophozoites proliferated, the infection spread anteriorally and into the submucosa, muscularis, and serosa of the intestine. Hepatic tissues were also infected early in the disease process. In the terminal stages of the infection, trophozoites had penetrated the stomach, pyloric caeca, pancreas, and adipose tissues and were also observed in the blood and kidney. In fish refractory to infection, the parasite was observed in the lumen of the intestine. Examination of infection in a moribound fish by scanning electron microscopy showed extensive destruction of the mucosal folds of the posterior intestine. Although a vigorous tissue response was evident in infected tissues, trout antibodies specific for the parasite were not detected by either Western blot analysis or IFAT. The distribution of the infective stage of C. shasta in the Columbia River basin was also investigated. The range of the parasite was extended in the Columbia River to its confluence with the Snake River and in the Snake River to Oxbow Dam.
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