Testing experimental subunit furunculosis vaccines for rainbow trout

Abstract

11 Aeromonas salmonicida subsp. salmonicida (AS) is the etiological agent of typical furunculosis in salmonid 12 fish. The disease causes bacterial septicemia and is a major fish health problem in salmonid aquaculture 13 worldwide, inducing high morbidity and mortality. In this study we vaccinated rainbow trout with subunit 14 vaccines containing protein antigens that were selected based on an in silico antigen discovery approach. 15 Thus, the proteome of AS strain A449 was analyzed by an antigen discovery platform and its proteins 16 consequently ranked by their predicted ability to evoke protective immune response against AS. Fourteen 17 proteins were prepared in 3 different experimental subunit vaccine combinations and used to vaccinate 18 rainbow trout by intraperitoneal (i.p.) injection. We tested the proteins for their ability to elicit antibody 19 production and protection. Thus, fish were exposed to virulent AS 7 weeks post-vaccination by applying a 20 novel, multi-puncture challenge method. The immune response in fish was evaluated following vaccination 21 and challenge by measuring antibody levels and recording survival. The control group showed 56 % 22 mortality whereas the groups of fish vaccinated with experimental subunit vaccines exhibited significantly 23 lower mortalities (17-30 %). These results imply that in silico-predicted protective protein antigens of AS 24 have significant protective properties and should be considered for further validation as potential candidates 25 for a subunit vaccine against furunculosis. 26 27