Isolation,Identification Of Aeromonas Hydrophila From Pelodiscus Sisnensis And Immunogenicity Analysis Major Outer Membrane Proteins Of The Bacteria

Objective Using Koch postulates to isolate and identificate pathogenic bacteria from pelodiscus sisnensis; Vaccination efficiency of surface antigens of Aeromonas hydrophila; With appropriate way to immunize pelodiscus sisnensis detecting the change of immunological factors and disease resistance, to select the commom prptective antigen of Aeromonas hydrophila.Methods(1)Through physiological and biochemical characteristics, virulent factors, 16 S r RNA and animal experiment to identificate the isolated bacterias;(2)Design and synthesis four pairs specific primers to PCR, and construction the outer membrane protein expression strains;(3)In the present study, the four main outer membrane proteins of Aeromonas hydrophila wereprepared and the pelodiscus sisnesis, on post-immunization day 28, serum antibody titers of the pelodiscus sisnesis were evaluated; The efficiency of vaccination was tested by artificial challenge via intraperitoneal injection of live bacteria on post-immunization day 28 and 35, respectively.Results(1) The isolated bacteria from pelodiscus sisnesis were proved to be Aeromonas hydrophila,which oxidase(+), can use glucose, maltose and other glycosides, the ability of mobility, with the virulent factors of hemolysin and aerolysin;(2)The four recombinant outer membrane proteins were expressed accurately, the SDS-PAGE showed the protein bands were consistent with the expected proteins;(3)Although significant humoral and innate immune responses were elicited in all vaccination groups, the challenge produced effective protection in Omp TS and Omp38, poor protection in both Omp F and Aha1.Conclusion(1)The physiological and biochemical characteristics, virulent factors, 16 S r RNA show that the isolated bacteria is Aeromonas hydrophila, the animal experiment show that the isolated bacteria has strong pathogenicity;(2)The four recombinant outer membrane proteins were expressed in E.coli accurately;(3)The challenge produced effective protection in Omp TS and Omp38, suggests the possibility of Omp TS and Omp38 as vaccine candidate antigens of the vaccine development.