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Pang H.-Y.,Guangdong Ocean University | Pang H.-Y.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Pang H.-Y.,Economic Animals of Guangdong Higher Education Institutes | Zhang X.-Z.,Guangdong Ocean University | And 14 more authors.
Aquaculture Research | Year: 2013

Vibrio alginolyticus is one of the most serious diseases in cultured marine and freshwater fish and shellfish. The absence of suitable vaccine or virulent marker can be the bottleneck to control V. alginolyticus infection. In this study, immunoproteomic approaches were undertaken to study the immunogenicity of the whole-cell protein of V. alginolyticus HY9901. The whole-cell proteins were analysed by two-dimensional gel electrophoresis and subsequent immunoblotting using the rabbit anti-V. alginolyticus HY9901 serum. A total of 55 immunogenic proteins were identified by immunoproteomic analysis. Of the 55 proteins, 51 are specific immunoreactive proteins and four are nonspecific immunoreactive proteins. Furthermore, outer membrane protein N (spot 2) was used as immunogens to immunize Epinephelus coioides for investigation of protective abilities and activities. The E. coioides immunized with OmpN has abilities to fight against infections caused by V. alginolyticus. The other novel immunogenic proteins may be developed as alternative antigens for further study of V. alginolyticus vaccine and diagnostics. These data show that immunoproteomics methods can be successfully applied in identifying immunogenic proteins of V. alginolyticus, which helps to search for the protective antigens in future. © 2012 Blackwell Publishing Ltd. Source


Cai S.H.,Guangdong Ocean University | Cai S.H.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Cai S.H.,Economic Animals of Guangdong Higher Education Institutes | Lu Y.S.,Guangdong Ocean University | And 8 more authors.
Journal of Fish Diseases | Year: 2013

The outer membrane proteins of the marine aquatic animal pathogen, Vibrio alginolyticus, play an important role in the virulence of the bacterium and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein-OmpU was cloned and expressed in Escherichia coli. Polyclonal antibodies were raised in rabbits against the purified recombinant OmpU, and the reaction of the antibody was confirmed by Western blotting using the isolated OmpU and the recombinant OmpU of V. alginolyticus. To analyze the immunogenicity of the recombinant OmpU, crimson snapper, Lutjanus erythropterus Bloch, were immunized by intraperitoneal injection, and antibody response was assessed by the enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the recombinant OmpU produced an observable antibody response in all sera of the vaccinated fish. The vaccinated fish were challenged by virulent V. alginolyticus and observed to have high resistance to infection. These results indicate that the recombinant OmpU is an effective vaccine candidate against V. alginolyticus in L. erythropterus. © 2013 Blackwell Publishing Ltd. Source

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