Nodavirus has become a serious pathogen for a wide range of cultured marine fish species. In the present work, the expression of genes related to immune and inflammatory responses of sea bream (Sparus aurata L.), considered as non susceptible species, was studied both in vitro and in vivo. No replication of the virus was observed in head kidney macrophages and blood leukocytes. Moreover, the enhancement of expression of several immune genes (tumor necrosis factor alpha (TNFalpha), interleukin-1-beta (IL-1beta), interferon-induced Mx protein) was not detected in both head kidney macrophages and blood leucocytes in response to an in vitro infection with nodavirus. However, in vivo, nodavirus was detected 1 day post-infection (p.i.) by a reverse transcription-polymerase chain reaction (RT-PCR) in blood, liver, head kidney and brain of experimentally infected sea bream, while its presence clearly decreased in blood after 3 days p.i. Also, a transitory increment of the expression of TNFalpha and IL-1beta was detected in the brain of intramuscular (i.m.) infected sea bream 3 days p.i. In head kidney, the over expression of TNFalpha was only observed 1 day p.i. The expression of Mx, an interferon induced gene, was increased in brain and head kidney of infected sea bream, reaching values of 1300-fold compared to controls in brain three days post-infection. For comparative purposes, we analyzed the expression of the same genes on a susceptible species, such as sea bass (Dicentrarchus labrax) and, although the same pattern of expression was observed both in brain and kidney, the magnitude was different mainly in the case of brain, the key organ of the infection, where higher expression of TNFalpha and lower expression of Mx compared with control was observed.

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The transmission of viral encephalopathy and retinopathy (VER) was investigated in juvenile sea bream, Sparus aurata L. Two different infection routes [intraperitoneal (i.p.) and intramuscular injection (i.m.)] were tested at two different temperatures (20 and 26 °C) using sea bream of mean weight 0.7, 2 and 4 g, as well as an immersion challenge performed at 26 °C with sea bream of 0.7 g. Successful transmission of the disease was only achieved by i.m. injection. Mortalities of 100% occurred in sea bream of 0.7 g at day 15 post‐infection and 47% in sea bream of both 2 and 4 g at day 30 post‐infection in all the experimental infections at 26 °C. No mortalities were ever observed with infections at 20 °C. When mortalities were observed, the virus was detected by immunoperoxidase staining in the SSN‐1 cell line inoculated with tissues from infected fish. Histological examination of both normal and infected fish showed a vacuolization in the bipolar and granular layers of the retina of the infected sea bream. This is the first experimentally induced transmission of VER in sea bream. Differences were observed at the time of disease onset depending on water temperature, the route of infection and the age of the juvenile fish.

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The marine environment includes diverse microeukaryotic organisms that play important functional roles in the ecosystem. With molecular approaches, eukaryotic taxonomy has been improved, complementing classical analysis. In this study, DNA metabarcoding was performed to describe putative pathogenic eukaryotic microorganisms in sediment and marine water fractions collected in Galicia (NW Spain) from 2016 to 2018. The composition of eukaryotic communities was distinct between sediment and water fractions. Protists were the most diverse group, with the clade TSAR (Stramenopiles, Alveolata, Rhizaria, and Telonemida) as the primary representative organisms in the environment. Harmful algae and invasive species were frequently detected. Potential pathogens, invasive pathogenic organisms as well as the causative agents of harmful phytoplanktonic blooms were identified in this marine ecosystem. Most of the identified pathogens have a crucial impact on the aquacultural sector or affect to relevant species in the marine ecosystem, such as diatoms. Moreover, pathogens with medical and veterinary importance worldwide were also found, as well as pathogens that affect diatoms. The evaluation of the health of a marine ecosystem that directly affects the aquacultural sector with a zoonotic concern was performed with the metabarcoding assay.