Vaccination with recombinant protein (r22C03), a putative attachment factor of Neoparamoeba perurans, against AGD in Atlantic salmon (Salmo salar) and implications of a co-infection with Yersinia ruckeri.

Amoebic gill disease (AGD) affects salmonids during the marine grow-out phase in the Tasmanian industry and in other major salmonid producing countries. During the period post-transfer to seawater, the bacterial condition yersiniosis can also cause high levels of mortality in Atlantic salmon grown in Tasmania, in addition to the hatchery outbreaks. The recombinant protein r22C03, a mannose-binding protein-like (MBP-like) similar to attachment factors of other amoebae, was tested as a vaccine candidate against AGD in a large scale challenge trial. Fish were immunised with r22C03 combined with FCA via intraperitoneal (i.p.) injection, and given a booster five weeks later by either i.p. injection (RP group) or by a dip-immersion (mRP). Fish were then challenged twice with Neoparamoeba perurans: the initial challenge 16 weeks after primary immunisation was terminated due to presence of ulcerative lesions in the skin of salmon; the second challenge was carried out after five weeks of treatment with oxytetracycline. These skin lesions might have been associated with a concurrent infection with Yersinia ruckeri, which was detected by real-time qPCR in serum of a large proportion of moribund and survivor fish after the AGD challenge. Before and during the N. perurans infection, levels of antibodies against r22C03 were measured by ELISA in serum, skin mucus and supernatant from skin and gill explants. For the second challenge, the average size of AGD lesions was recorded from histology sections and survival curves were obtained. Before AGD challenge, r22C03 induced antibody responses in serum and explants with both vaccination strategies. At the end of the challenge, levels of antibodies were lower than before challenge irrespective of treatment. Both vaccinated groups presented increased serum antibody responses, while only mRP presented antibody responses in skin mucus, and no significant antibody responses were measured in the explants. Antibodies did not confer protection to N. perurans infection, as no difference was observed in the survival curves of the vaccinated and control groups, and there was no effect on the gill lesion size. The concurrent yersiniosis infection probably represented more closely infection patterns observed in commercial settings. However, it could have interfered with the survival results and with the ability of the fish to respond to the amoebae infection.