부산대학교 도서관

Photobacteriosis caused by Photobacterium damselae subsp. piscicida (Pdp) remains one of the main infectious diseases affecting cultured fish in Mediterranean countries. Diverse vaccine formulations based in the use of inactivated bacterial cells have been used with unsatisfactory results, especially in newly cultured species like sole (Solea senegalensis). In this work, we describe the use of the outer membrane receptor (FrpA) of the siderophore piscibactin produced by Pdp as a novel subunit vaccine against photobacteriosis. FrpA has been cloned and expressed in Escherichia coli under an arabinose-inducible promoter. A recombinant protein (rFrpA) containing the pelB localization signal and a His tag was constructed to obtain a pure native form of the protein from E. coli outer membranes. The immunogenicity of rFrpA, and its protective effect against photobacteriosis, was tested by i.p. injection of 30 μg of the protein, mixed with Freund's adjuvant, in sole fingerlings with two immunizations separated by 30 days. Results showed that using either pure rFrpA or whole cells as immobilized antigens in ELISA assays, rFrpA induces the production of specific antibodies in sole. An experimental infection using fish vaccinated with rFrpA or formalin-killed whole cells of Pdp showed that both groups were protected against Pdp infection at similar levels, with no significant differences, reaching RPS values of 73% and 79%, respectively. Thus, FrpA constitutes a promising antigen candidate for the development of novel more effective vaccines against fish photobacteriosis. • The siderophore piscibactin of P. damselae subsp piscicida is transported into the cell by the outer membrane protein FrpA. • FrpA was cloned and expressed in E. coli under an arabinose-inducible promoter. • Native FrpA was obtained from the outer membranes of E. coli by addition of a pelB sequence and a His tag to frpA sequence. • FrpA was immunogenic in fish since it induced specific antibodies when it was injected in sole fingerlings. • Fish immunized with FrpA achieved a 73% (RPS) protection against experimental infections with P. damselae subsp piscicida. [ABSTRACT FROM AUTHOR]