Graduate Thesis Or Dissertation
 

Development of a new vaccine delivery system for immunizing fish and investigation of the protective antigens in Vibrio anguillarum

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  • Phenol-water and trichloroacetic acid extracts of Vibrio anguillarum were both antigenic and immunogenic when injected into coho salmon (Oncorhynchus kisutch). Cross-protection against virulent Vibrio anguillarum was observed with whole-cell bacterins and phenol-water extracts prepared from two serologically distinct groups. Gel diffusion studies indicated that intact somatic antigens were serotype specific. Breakdown products or impurities in serotype specific phenol-water extracts displayed antigenic cross-reactivity Generation times for Vibrio anguillarum LS 1-74 were shorter than for MAN 1669. Both isolates reached higher levels of growth at 18 than at 30 C. Spheroplasting and lysis was more characteristic of Vibrio anguillarum MAN 1669. An artificial, waterborn challenge was developed for Vibrio anguillarum LS 1-74. Similar challenges with members of the serologically unrelated group (Vibrio anguillarum MAN 1669 and MSC 2-75) were unsuccessful. Studies with motile and non-motile strains of Vibrio anguillarum indicated that the flagellar antigen is not required for immunogenicity. An economical, efficacious vaccine delivery system for immunizing fish has been developed which employs a high-pressure liquid spray apparatus operated at 6.3 to 7. 0 kilograms per square centimeter (90 to 100 pounds per square inch). Bacterin consisting of formalin-killed Vibrio anguillarum culture plus 0. 15 percent bentonite was both antigenic and immunogenic when sprayed on various size coho salmon. The technique, referred to as spray vaccination, was found to confer higher levels of immunity against virulent Vibrio anguillarum than oral vaccination. Immune serum raised by the technique was protective in passively immunized coho. Bacterins containing bentonite were found to be most effective when adjusted to pH 3. 0. Spray vaccinated fish possessed detectable agglutinating antibody after 112 days and were protected against challenge after 125 days. Aeromonas salmonicida and Bacterial Kidney Disease bacterins induced increased humoral antibody production in spray vaccinated fish.
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