The available database comprises research projects in Fisheries, Aquaculture, Seafood Processing and Marine Biotechnology active in the time period 2003-2022.
BlueBio is an ERA-NET COFUND created to directly identify new and improve existing ways of bringing bio-based products and services to the market and find new ways of creating value from in the blue bioeconomy.

More information on the BlueBio project and participating funding organizations is available on the BlueBio website: www.bluebioeconomy.eu

Last Update: 2024/06/19

NA
Marine Biotechnology
Aquaculture
Modern tools for the study of host-agent interactions of emerging fish diseases
National Programme
National
Atle Lillehaug
atle.lillehaug@vetinst.no
NVI - Norwegian Veterinary Institute (Norway)
NA - Columbia University (United States of America),  ITU - Instambul Technical University (Turkey),  NVH - Norwegian School of Veterinary Science (Norway),  NA - University of Glasgow (United Kingdom),  UV - University of Valencia (Spain), 
2008
2013
€ 1,572,487
https://prosjektbanken.forskningsradet.no/en/project/FORISS/186907?Kilde=FORISS&distribution=Ar&chart=bar&calcType=funding&Sprak=no&sortBy=date&sortOrder=desc&resultCount=30&offset=0&ProgAkt.3=SIP-FKD-SIP+finansiert+av+Fiskeri-+og+Kystdepartementet&source=FORISS&projectId=173523
The formidable growth of Norwegian aquaculture during the last decades, dominated by the production of Atlantic salmon, also has the appearance of new diseases as a consequence. There is also a continuous research based development of new scientific methods related to microbiology, pathology and epidemiology, and particularly within the area of molecular genetics there has been remarkable innovative activity in the scientific communities. Diagnostics, control, prevention and treatment of new fish diseases must be based on knowledge of the disease causes and pathogenesis, and there was a great need for strengthened implementation of modern methods to study such mechanisms. Due to the great importance of Norwegian seafood production, potential food safety and food quality aspects of fish diseases should also be emphasized. The primary objective of the SIP "Modern tools for the study of host/agent interactions of emerging fish diseases" was to improve scientific tools in order to elucidate disease etiology, pathogenesis, and epidemiology of emerging fish diseases, as well as to comply with the responsibilities NVI has as a national and international reference laboratory". II. Scientific achievements An overview of results and activities will be given relating to aims for the different work packages under the SIP, including the involvement of research groups and institutions: (WP 1) Investigations of disease pathogenesis Aim: Develop and test in situ techniques on tissue sections for use in disease investigations and studies of pathogenesis Primary cell cultures are established based on gill epithelium from Atlantic salmon, and these are utilized in the study of disease development. In situ-hybridization is applied in cell culture for the examination of the pathogenesis of infectious salmon anaemia (ISA), together with other methods like immunohistochemistry. A new method for the identification of the cell receptor for ISA-virus is developed, and it has been demonstrated that existence of this receptor in a cell is a prerequisite for the infection with ISA. These studies have also made use of bath challenge trials with live fish, representing the first successful bath challenges with ISA-virus. Hybridization methods have also been used to identify the microsporidium Paranucleospora theridion in situ in tissue sections. The parasite has also been detected by use of synthetic fluorescein-marked oligoprobes, in house produced DIG-marked DNA-probes and RNA-probes. The results are hold together with histo-pathological examinations and a general staining technique. The gill epithelial cell cultures have also been used for the demonstration of apoptosis ('cell death') as a response marker. Other methods and reagents developed include peptide antibodies used to demonstrate hypoxia in tissues, and a monoclonal antibody, which binds both to salmon endothelium and erythrocytes, is produced. In Atlantic cod, cellular reactions during inflammatory responses to agents like Francisella noatunensis and nodavirus have been described. Probes and other methods have been used to demonstrate the presence of interferon, IL1, IL8 and IL10-expressing cells. These studies have been carried out in close cooperation with other research project; i.e. the strategic institute program 'Improving preparedness through research' (NRC-funding), and the industry funded project "Mitigation of ISA" (Novartis Animal Health Canada). The most significant collaborating partner in this work package has been the group of Professor Erling Olav Koppang, Norwegian School of Veterinary Science; (WP 2) Perform advanced studies of host-pathogen interplay Aim: Evaluate functional importance of immune system components, with emphasis on bridging the knowledge-gap between 'descriptive' genomics and functional data Characterization of cells from the immune system, as well as signal substances, in Atlantic salmon has included the production of polyclonal antisera against salmon CD3? and IL2. The methods are based on a combination of immunizing with DNA and protein, respectively. DNA-immunizing is made by injection of DNA-plasmid, which is taken up in fish cells (electroporation). Further characterization of the immune response is based on the principles of pyro-sequencing. Several assays have been developed to identify immunoglobulin classes T, M and D in salmon. Molecular biological methods have been used to analyse variation in complement factor 3 in different fish species. Recombinant interleukin-2 has been produced in mammalian cells, and has been characterized. This r-IL-2 has been used as stimulants in salmon leucocytes, and the effects include proliferation and prolonged persistence of T-cells. Activation of genes coding for variable sites in salmon B- and T-cell receptors has been demonstrated. Other recombinant proteins produced include CD4, Pax5, IFN' and TNF'. RT-PCR methods for interleukin-2 receptor, interleukin-21, interleukin-22 and ki67, all assumed to be T-cell markers, has been established and tested in assays. A quantitative real-time PCR for analysis of mRNA from single cells have been developed. From cell suspensions, cells were isolated by micromanipulation devices. Pyro-sequencing have been used to characterize expression profiles of cell populations, e.g. r-IL-2 stimulated leukocytes. These sequencing techniques have also been applied for analysis of the variable part of the T-cell receptor ('-chain). The primary collaboration partners have been Professors Espen Rimstad and Anne Storset at the Norwegian School of Veterinary Science, as well as Professor Tony Raga and PhD student Neus Garcia-Sanchez at the University of Valencia. Spain; (WP 3) Modern tools for identification of emerging agents Establish fish "viromes", develop strategies for generic amplification of viruses from fish, and develop microarray-based diagnostics for rapid screening of fish populations Two new fish viruses are identified and described, piscine reovirus (PRV), which is associated with heart- and skeletal muscle inflammation (HSMI), and piscine myocarditis virus (PMCV), found in farmed salmon suffering from cardiomyopathy syndrome (CMS). Pioneer technology within pyro-sequencing has been applied, based on large-scale sequencing of nucleic acids, together with bio-informational processing of results, aiming at the identification of unknown infectious agents. The two new viruses PRV and PMCV have been studied further, by help of PCR-methods for the detection of the specific viruses in fish material, in situ-hybridization for detection in tissue sections, and virus antigens are constructed by use of recombinant technique for the production of specific antibodies against the viruses, which are very useful in diagnostic purposes. Distribution of the viruses, relating both to geographical regions and fish hosts, has been studied, and prevalence's of the agents in both farmed and wild populations. PRV is widespread in farmed salmon, both in healthy fish and in fish with HSMI, however, virus titters are higher in sick fish. Only a limited part of the wild salmon population seems to be infected. For PMCV, the association between infection and disease seems to be more directly connected; virus detection seems to be restricted to fish in disease outbreaks. A study based on reverse genetics has been performed to implement the methodology, and human astrovirus was used as a model. A reverse genetic construct has been obtained, and some modifications have been made to adapt to our laboratory conditions. Development of microarray-based diagnostic methods was not prioritized during the project period. As a consequence of the path-breaking results with the pyro-sequencing method, the scientific application of this approach was prioritized. The principal collaboration partners in this work package were the group of Professor W. Ian Lipkin at Columbia University, New York, and the group of Professor Matthew L. Meyerson, Harvard Medical School; (WP 4) Develop dynamic fish disease models in order to improve the epidemiological understanding of disease dynamics in aquaculture Aim: Develop dynamic fish disease models in order to improve the epidemiological understanding of disease dynamics in aquaculture In order to establish models for the spread of infectious diseases within and between populations, pancreas disease (PD) caused by salmonids alphavirus has been used as an example. A statistical model has been applied to estimate the probability for outbreaks of PD at farm and site level. The results can be illustrated with a map of risk for PD along the Norwegian coast. Risk factors for new PD outbreaks are the experience of PD at the same site or neighbouring site, local high biomass density, and density of production sites. Transmission dynamics for PD-virus within a population during early stages of disease outbreaks have been studied in challenge trials, together with data from field outbreaks. A framework for the quantitative measurement of a PD epidemic that could be useful for the evaluation of prevention methods has been presented. Effects of variables like fish density, water temperature, vaccination status have been included. A model for spread of PD between aquaculture sites within a fjord-system has demonstrated that new PD cases are connected to water currents, more than other risk factors. Using water contact between sites to model PD transmission and time of infection, it has been found that increasing sea temperatures is a triggering factor for PD outbreaks. The activities in this work package have been carried out in cooperation with dr. Hamdi Ogut, Karadeniz Technical University, Turkey, dr. Marianne Sandberg, Norwegian School of Veterinary Science, and dr. David Graham and his group, University of Glasgow, and has been in close collaboration with the PD-project TRISAV (NRC project nr. 190484). Water contact between farms sites were estimated based on a hydrodynamic model run by SINTEF. Seafood safety aspects of fish diseases. Aim: Investigate whether uptake, growth and release of human pathogenic bacteria during slaughter and processing are correlated to pathological changes in fish The purpose of seafood safety studies was to study indirect effects of established fish diseases on Listeria monocytogenes. The study material consisted of salmon suffering from winter ulcers or salmon lice infestation, or from populations with pancreas decease (PD). Salmon with sea-lice, PD or winter ulcers were included, and carcasses were inoculated with L. monocytogenes. There were no significant differences in bacterial growth between infected and symptom free fish, the tendency was even towards rapid growth of Listeria in fish with winter ulcers than in symptom free fish. Even the spread of Listeria between fish within a batch was the same for infected and symptom free fish. Salmon processed to be smoked was inoculated before salting and smoking, and a surprising and interesting observation was that the growth was nearly as rapid when the fish was inoculated by injection into and on the muscle surface. No difference in growth rates was seen, indicating that the studied fish diseases have no effect on the growth rate of Listeria. Comparison of expression of virulence genes in Listeria in fish with winter ulcers and symptom free fish gave the same results, i.e. no over- or under expression of virulence genes. Prevalence of Listeria was investigated; however, the bacterium was not detected in fish with or without sea lice or PD. Some positive samples were found in fish with sea ulcers, however, batches analysed in the winter ulcer season had lower prevalence's and lower levels of Listeria than symptom free fish analysed during the rest of the year, indicating that winter ulcers itself did not lead to contamination. Our studies indicate that fish with winter ulcers, PD and lice give no interfering effects with Listeria monocytogenes in fish for any of the parameters studied. The studies under the seafood safety aspects have been coordinated with the EU-project 'BASELINE', in order to utilize materials and methods optimally. Primary objective: Improve scientific tools in order to elucidate disease etiology, pathogenesis, and epidemiology of emerging fish diseases, as well as to comply with the responsibilities NVI has as a national and international reference laboratory. Secondary objectives: (1) Develop and test in situ techniques on tissue sections for use in disease investigations and studies of pathogenesis; (2) Evaluate functional importance of immune system components, with emphasis on bridging the knowledge-gap between descriptive genomics and functional data; (3) Establish fish viromes, develop strategies for generic amplification of viruses from fish, and develop diagnostics for rapid screening of fish populations, based on this approach; (4) Develop dynamic fish disease models in order to improve the epidemiological understanding of disease dynamics in aquaculture; (5) Investigate if uptake, growth and release of human pathogenic bacteria during slaughter and processing are correlated to pathological changes.
Disease; Salmon; Genetic; Fish; Fish biology;
Not associated to marine areas
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