AIMS OBJECTIVES AND ACTIVITIES: Salmon farming has achieved major growth and success over a short period, but global demands mean that aquaculture will need to play a vital role in supplying animal protein to a fast-growing human population (World Bank 2014). Two overlapping bottlenecks currently limit progress in salmon aquaculture: (1) the ability to realise consistently high levels of fertility through artificial in vitro fertilisation, and (2) the ability to reliably store sperm both within and between spawning seasons. Fortunately, a very large body of research knowledge exists on reproduction, fertilisation, and sperm storage and cryopreservation in salmonids, which we will translate through this innovation project into practices that meet the specific needs of salmon aquaculture for improved breeding and stock enhancement. PROJECT PARTNERS AND AQUACULTURE CHALLENGES: Alastair Hamilton and his team at Landcatch Natural Selection Ltd will work with Matt Gage (UEA), combining practical expertise from industry with scientific knowledge and skills. Breeding and stock enhancements within salmon aquaculture have so far focused (successfully) on improvements in growth, feed conversion and disease resistance. However, it is essential at this early stage in salmon farming that we do not neglect challenges from fundamental reproductive biology, which must be carefully managed through the domestication process as new strains and processes are developed. Broodstock fertility already presents significant constraints on progress in salmon aquaculture, and this should be guarded against into the future. Fertility that is often below 50% currently limits productivity into the farm stage by reducing the number of eyed eggs and fry that can be generated for the grow-on stages to market. Low fertility also significantly constrains the ability to replicate cross-strain breeding selection and stock improvements. In addition to fertility limitations, ineffective sperm storage solutions place major breeding barriers on stock enhancement. Short-term (days to weeks) sperm storage using extenders and chilling can be implemented on the farm that researchers have used to achieve fertility up to 33 days from strip. Longer-term cryo-storage has a detailed history of investigation in fish, especially salmon, and methods are now developed that should allow reliable storage over years. Translating established knowledge and technology into farm salmon sperm storage solutions will transform aquaculture's ability to select for farm-friendly traits and enhance stocks, especially with the availability of modern genomic selection. DELIVERABLES AND OUTPUTS: The project will deliver better fertility and sperm storage solutions, immediately benefiting farm productivity, and transforming the ability to select for and diversify optimal aquaculture traits. The outputs will be technical reports and demonstrations of work practice that detail how improvements in fertility and sperm storage can be applied on the farm, disseminated direct to Landcatch through Dr Alastair Hamilton and his team. The reports will be evolved and optimised under discussion between the team members so that the farm practicality perspective is prioritised, allowing successful implementation of change among farm processes and workers directly as a result of successful Knowledge Exchange. Ultimately, reports will be published in peer-reviewed international impact journals, and disseminated at the European Aquaculture Society annual meeting in Croatia '17. Peer-reviewed outputs will be produced in rapid-publication journals with relevance to the aquaculture industry; we anticipate four papers: two describing improvements in sperm recovery, handling, analysis and fertilization, and two on short-term and long-term sperm storage solutions. PROJECT DURATION: 18 months, to encompass two spawning seasons and to allow 6-12 month testing of cryopreservation. TOTAL COST: £185k (80% FEC)

Vaccines for chronic viral pathogens in salmon- generation of interferon attenuated cell lines. Acroym: SalVacCell Project goal / key aims. There is a major requirement to generate new vaccines for viral pathogens in the salmon aquaculture industry. Viruses represent one of the major economic losses to the salmon industry, which is a direct reflection of the lack of highly protective vaccines. In order to improve vaccine design and testing high quantities of viruses are required, but at present this is not possible. This project will use cell lines that have been edited by CRISPR/cas9 to be highly permissive for growth of difficult to grow viruses. We will knockout the type I interferon pathway which is the major antiviral mechanism in animals. Specifically we will target key genes that regulate interferon induced cellular responses to viral infection. These cell lines will additionally be used to explain the underlying function of genes associated with natural viral resistance / susceptibility in salmon that is of direct importance to breeding programs. We have secured Industrial partners Benchmark (vaccine company) and Landcatch (fish breeding). Consortium agreements will clearly define roles and confidentiality of IP. OBJECTIVES 1. Development of IFN-deficient fish cell lines by knock-out of IFN function 2. Evaluation KO cell lines to mount an antiviral response to confirm phenotype 3. Use the cell lines to explain naturally occurring resistance / susceptibility to fish viruses 4. Comparison of yield for viral particle production and viral diagnostic turn-over time between traditional cell lines and newly developed cell lines KEY CHALLENGES The overarching challenge is to be able to produce high titres of virus for vaccine companies to be able to improve design and protection to economically important chronic viral pathogens. Many labs have attempted to knock down genes in salmonid cells, but to date this has not been achieved. To our knowledge we are the first to create such cell lines and as such our unfunded preliminary work has already overcome a major hurdle. The challenge of this project is to fully exploit our cell line technology for both disease management and also to greatly improve the basis of genetic selection. Deliverables- to be used by partners a. Engineered cell lines that are deficient in antiviral responses. b. Cells lines that can be used to produce high titre of viruses for vaccines c. Capacity to upscale viral production in industrial environment. d. Improved precision of selection for disease resistance in salmon breeding Project duration: 24 months Total project cost: £354622 Contribution requested from (100%) BBSRC/NERC £249,622 (£199697.60 80%) Contribution from MSS: secured £65K Contribution from Benchmark secured £20K Contribution from Landcatch secured £20K