Environmental isolates P1SW and V3SW as a bivalent vaccine induce effective cross-protection against Edwardsiella tarda and Vibrio anguillarum

Abstract

Edwardsiella tarda and Vibrio anguillarum are severe fish pathogens. In this study, we aimed at selecting avirulent environmental isolates with application potential in the prevention of E. tarda- and V. anguillarum-associated diseases. For this purpose, we selected and analyzed 2 seawater isolates, P1SW and V3SW, belonging to the genera Pseudomonas and Vibrio, respectively. When administered to turbot Scophthalmus maximus via immersion and oral feeding, P1SW and V3SW at a dose of 2 × 108 colony-forming units caused no mortality, but both strains were able to disseminate into internal organs in a transient, time-dependent manner. When turbot were immunized with P1SW, V3SW, or P1SW plus V3SW (named P1V3) via immersion plus oral routes, the latter with vaccines embedded in sodium alginate microspheres, moderate protection against E. tarda and V. anguillarum was induced by V3SW, and moderate protection against E. tarda was induced by P1SW. Compared to P1SW and V3SW, P1V3 elicited a significantly stronger protection against both E. tarda and V. anguillarum. Immunological analysis showed that (1) P1SW, V3SW, and especially P1V3 activated head kidney macrophages, (2) P1V3 induced significantly higher levels of serum antibodies against E. tarda and V. anguillarum than P1SW and V3SW, and (3) P1V3-induced antibodies were able to bind E. tarda and V. anguillarum and enhance serum bactericidal activity. These results indicate that P1V3 as a naturally delivered vaccine elicited a humoral immune response against both E. tarda and V. anguillarum and, as a result, was cross-protective against E. tarda and V. anguillarum infection.