Nervous necrosis virus (NNV) vaccination of carrier Senegalese sole (Solea senegalensis)

Viral encephalopathy and retinopathy (VER), caused by nervous necrosis virus (NNV), is one of the most threatening diseases affecting marine farmed fish, especially in early developmental stages. In addition, subclinical infections are commonly detected in both farmed and wild fish. In recent years numerous efforts have been made to achieve effective vaccines for the protection of different fish species against NNV infection. Vaccination experiments are always conducted on pathogen-free animals, but subclinically infected individuals may respond differently to immunization. In this study, we have observed a different response between experimentally-induced-NNV carrier Senegalese sole and NNV-free animals when immunized with a BEIinactivated vaccine, whereas mock-vaccinated carrier fish behaved like the NNV-free group. Analysis of the immune response after vaccination showed specific antibody production as well as up-regulation of the genes coding for the proinflammatory cytokines IL-6 and IL-8, the antiviral protein HERC4 and the cytotoxic T lymphocyte (CTL) and T helper markers CD8 and CD4 in vaccinated NNV-free and mock-vaccinated carrier individuals. However, no significant antibody production was detected and only cd4 up-regulation was recorded in the vaccinated carrier fish. After challenge, viral replication was similar in the vaccinated groups up to 15 days post-challenge (dpc), although thereafter the viral load in the vaccinated carrier fish reached values similar to the control fish, whereas in the vaccinated NNV-free sole remained 1.5-2 log lower. At 2 dpc 10% mortality was recorded in the vaccinated carrier group that did not correlate with a higher viral titre, but with an overexpression of inflammation-related genes. These results suggest that an acute inflammatory reaction could have been the cause of the fish death. Excepting the values recorded in this episode, poor modulation of the immune response was observed at the early days post-challenge. However, at 30 dpc the IgM levels in the vaccinated carrier fish were significantly higher than those of the NNV-free sole. Longer post-vaccination studies would be necessary to clarify if this antibody increase may be relevant for fish protection.