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12. Chapter 5 - Aquaculture and conservation

Author

Guerra-García, José Manuel, Martínez-Pita, Inés, Šegvić-Bubić, Tanja, Manchado, Manuel, Arechavala-Lopez, Pablo, Calado, Ricardo, Marchio, Elizabeth, Gentry, Rebecca, Tlusty, Michael F., Rhyne, Andrew, Moreno, Patricia, Béjar, Julia, Alonso, María del Carmen, García-Rosado, Esther, and Jiménez-Prada, Pablo

Published
2023
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22. Transcriptomic profiles of Senegalese aole infected with nervous necrosis virus reassortants presenting different degree of virulence

Author

Universidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía, Universidade de Santiago de Compostela. Instituto de Acuicultura, Labella, Alejandro M., Garcia Rosado, Esther, Bandín Matos, María Isabel, Pereira Dopazo, Carlos, Castro, Dolores, Alonso, M. Carmen, Borrego García, Juan José, Universidade de Santiago de Compostela. Departamento de Microbioloxía e Parasitoloxía, Universidade de Santiago de Compostela. Instituto de Acuicultura, Labella, Alejandro M., Garcia Rosado, Esther, Bandín Matos, María Isabel, Pereira Dopazo, Carlos, Castro, Dolores, Alonso, M. Carmen, and Borrego García, Juan José

Abstract

Betanodaviruses [nervous necrosis virus (NNV)] are the causative agent of the viral encephalopathy and retinopathy, a disease that affects cultured Senegalese sole (Solea senegalensis). NNV reassortants, combining genomic segments from redspotted grouper nervous necrosis virus (RGNNV) and striped jack nervous necrosis virus (SJNNV) genotypes, have been previously isolated from several fish species. The wild-type reassortant wSs160.03, isolated from Senegalese sole, has been proven to be more virulent to sole than the parental genotypes (RGNNV and SJNNV), causing 100% mortality. Mutations at amino acids 247 (serine to alanine) and 270 (serine to asparagine) in the wSs160.03 capsid protein have allowed us to obtain a mutant reassortant (rSs160.03247+270), which provokes a 40% mortality decrease. In this study, the RNA-Seq technology has been used to comparatively analyze Senegalese sole transcriptomes in two organs (head kidney and eye/brain) after infection with wild-type and mutant strains. A total of 633 genes were differentially expressed (DEGs) in animals infected with the wild-type isolate (with higher virulence), whereas 393 genes were differentially expressed in animals infected with the mutant strain (37.9% decrease in the number of DEGs). To study the biological functions of detected DEGs involved in NNV infection, a gene ontology (GO) enrichment analysis was performed. Different GO profiles were obtained in the following subclasses: (i) biological process; (ii) cellular component; and (iii) molecular function, for each viral strain tested. Immune response and proteolysis have been the predominant biological process after the infection with the wild-type isolate, whereas the infection with the mutant strain induces proteolysis in head kidney and inhibition of vasculogenesis in nervous tissue. Regarding the immune response, genes coding for proteins acting as mediators of type I IFN expression (DHX58, IRF3, IRF7) and IFN-stimulated genes (ISG15, Mx, PKR, Gig1

Published
2018

24. Contributors

Author

Alonso, María del Carmen, Ambroso, Stefano, Arechavala-Lopez, Pablo, Ashton, Gail V., Bazairi, Hocein, Béjar, Julia, Boissery, Pierre, Brunet, Robert, Cabezas, M. Pilar, Cacabelos, Eva, Calado, Ricardo, Canning-Clode, João, Carlton, James T., Dissanayake, Awantha, El Asmi, Souha, El Ouamari, Najib, Espinosa, Free, Fa, Darren, Ferrario, Jasmine, Fonbonne, Sebastien, García-de-Lomas, Juan, García-Gómez, José Carlos, García-Gutiérrez, María Leonor, García-Rosado, Esther, Gentry, Rebecca, Gerovasileiou, Vasilis, Gestoso, Ignacio, Gómez-Gras, Daniel, Grinyó, Jordi, Gudefin, Anaïs, Guerra-García, José Manuel, Jiménez-Prada, Pablo, Lecaillon, Gilles, Lenfant, Philippe, León-Muez, David, Limam, Atef, Manchado, Manuel, Marchini, Agnese, Marchio, Elizabeth, Martínez-Laiz, Gemma, Martínez-Pita, Inés, Montseny, María, Moreno, Patricia, Muirhead-Davies, Natalie, Navarro-Barranco, Carlos, Peñalver-Duque, Patricio, Rhyne, Andrew, Ros, Macarena, Ruiz, Gregory M., Santín, Andreu, Šegvić-Bubić, Tanja, Selfati, Mohamed, Sempere-Valverde, Juan, Terrón-Sigler, Alexis, Tlusty, Michael F., and Warr, Stephen

Published
2023
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27. A combined RT-PCR and dot-blot hybridization method reveals the coexistence of SJNNV and RGNNV betanodavirus genotypes in wild meagre (Argyrosomus regius)

Author

Lopez Jimena, Benjamin, Cherif, Nadia, Garcia-Rosado, Esther, Infante, Carlos, Cano, Irene, Castro, Dolores, Hammami, Salah, Borrego, Juan J, and Alonso, Maria del Carmen

Subjects
SJNNV, betanodavirus, degenerate primers, RT-PCR, RGNNV, dot-blot hybridization
Abstract

Aims: To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. Methods and Results: The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46·87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90·62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56·25%). Conclusions: This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. Significance and Impact of the Study: This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time.

Published
2010

31. Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass(Dicentrarchus labrax) during the course of an experimental challenge.

Author

Lopez-Jimena B, Garcia-Rosado E, Thompson KD, Adams A, Infante C, Borrego JJ, and Alonso Mdel C

Subjects
Animal Fins metabolism, Animal Fins virology, Animals, Antibodies, Viral metabolism, Brain metabolism, Brain virology, Enzyme-Linked Immunosorbent Assay veterinary, Immunohistochemistry veterinary, In Situ Hybridization veterinary, Kidney metabolism, Kidney virology, Liver metabolism, Liver virology, Real-Time Polymerase Chain Reaction veterinary, Retina metabolism, Retina virology, Spleen metabolism, Spleen virology, Antigens, Viral metabolism, Bass, Fish Diseases virology, Nodaviridae, RNA Virus Infections veterinary
Abstract

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain(cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.

Published
2012
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