Preparation and Purification of Pathogenic Bacterial Immune Serum from Japanese Flounder(Paralichthys olivaceus) with Ascites

The methods used in preparation of high-titer anti-Edwardsieua tarda(Et) serum by immunized rabbits with E.trada were described as the following:Firstly,the antigen of Et was prepared by the rabbits vaccinated by an ear vein.The agglutination efficiency of the immune serum was found to be 1︰2560 with micro-reaction plate.Then,the antibody was purified by Millipore Montage antibody purified kit,and the purified antibodies by SDS-PAGE electrophoresis with less miscellaneous focus had the main strip of about 50 kD.E.tarda was positive in the Dot-ELISA while Aeromonas sobria,A.hydrophila,Vibrio fluvialis,V.alginolyticus,V.anguillarum,Shewanella putrefaciens,Providencia alcalifaciens,Enterobacter sakazakii and Escherichia coli were negative in the assay.The specificity and high titer of immune serum were produced in this study,leading to a strategy to detect Bifidobacteriumin dairy products serologically.The specificity and high titer E.tarda immune serum provide the foundation for the rapid detection of pathogenic bacteria in Japanese flounder(Paralichthys olivaceus)with ascites.