Combining biomarkers from salmonmetabolomics

Combining biomarkers from salmonmetabonomics, genomics and proteomics with high-field NMR profiling of fish feed nutrients The organization of this project involved using lab platforms which yield large datasets (so called "-omics" for metabolomics (molecules), genomics (genes) and proteomics (proteins)) to develop aquafeed from raw materials to feed to fish.

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Project
Combining biomarkers from salmonmetabonomics, genomics and proteomics with high-field NMR profiling of fish feed nutrients The organization of this project involved using lab platforms which yield large datasets (so called "-omics" for metabolomics (molecules), genomics (genes) and proteomics (proteins)) to develop aquafeed from raw materials to feed to fish. The project was managed as a joint HAVBRUK and FUGE program with the raw material and feed work in the HAVBRUK program. Therefore, two objectives were made: 1) Develop new technology and procedures for performing feed trials designed for using new biotechnology derived biomarkers and metabonomics. 2) Use high field NMR to characterize the feed chain of salmon: from feed ingredients to processed feed and fish flesh.

Back ground 
The salmon feed industry has relied primarily on growth trials to assess new raw materials and feeds. Depending on the starting weight of the fish, they are grown for two to four months in land based tanks, fish are weighed, and feed properties are measured physically and chemically. The first objective of this program was to determine the best way to perform trials, measure performance, and combine this data with feed physical and chemical information.
Traditionally the development of feed by the aquaculture industry relied on proximate analysis (protein, fat, water, ash) of feed and sometimes amino acid composition, related to feed hardness and fish performance. The second objective of this program was to develop high-field NMR (HF-NMR) as a rapid nutrient profiling tool for feed and fish evaluation. This concept - metabolic feed profiling - is new to the animal feed industry, and also new to Norway.

Results
Related to new trial methods, individual fish were monitored for growth and feed intake and compared to traditional tank group performance. The results were similar, the individual fish growing within the mean or slightly better than groups, and the potential of repeated measures making individual fish a good model. Related to this statistics was
developed for expressing growth as repeated measures, rather than weight gain or specific growth rate. This technique has become our standard tool, and it was presented at an international conference to disseminate the information.
Raw material and feed metabolite profiling was done using HF-NMR. The data was then analyzed with bio-informatics tools, allowing us to relate large data sets from raw materials to feed to fish.

Figure 1. PCA plot based on metabolomics of soy products analyzed using HF-NMR. Figure 1 is an example of etabolomics (nutrient profiling based on measuring 38 nutrients in various soy products using HF-NMR) of soy roducts which EWOS uses in commercial salmon feed. The three different types of products (color coded by product type) are distinct based on nutrients which vary according to product origin and processing technique.

Application and utility value
Based on the work of this program trials procedures were changed to give more repeated measures of fish erformance, and more chemical analysis of feed and feed ingredients is done routinely. The significance of this work is added precision and knowledge, and in addition increased efficiency in work and reduced cost. This information has been
communicated in an international conference as a poster and keynote speech. Because of the usefulness of these techniques the industry partner is now the sole leasing agent for the HF-NMR instrument used for this project. Standard protocols are done for many ingredients and feed on a routine basis.

Addtional information
The industry partner on this project intends to create high-throughput individual fish facilities related to feed development. New statistical models with larger data sets are being made and are equally useful.

174222/S40 Ansvarlig: EWOS Innovation
Prosjektleder: Richard Taylor
Kontaktperson: Jan Vidar Jakobsen, EWOS Innovation Adresse: 4335

Telefon: 55 69 78 11 E-mail: jan.vidar.jakobsen@ewos.com
Lenker: http://www.ewos.com/portal/wps/portal/EwosInnovation/