Research news 2016 by Havforskingsinstituttet / NIFES

2016

RESEARCH NEWS

N AT I O N A L I N S T I T U T E O F N U T R I T I O N A N D S E A F O O D R E S E A R C H

Dear reader Does the food children eat affect their ability to learn? Do children who eat fish do better at school than children who eat meat? Or is it the opposite? These are just a few of the questions that scientists at NIFES are attempting to find answers to. In order to learn more about whether different diets actually affect children's concentration and their ability to learn, NIFES has conducted two extensive food trials comprising 713 kindergarten children and lower secondary school pupils from Bergen. Whatever the outcome, there is no question that children should eat seafood. Norwegian seafood is safe, and it contains many important nutrients that are important for children, among others. Iodine and the omega-3 fatty acid DHA are known to be important for children's development. Another population group that should eat more seafood is pregnant women. In Norway and the rest of the world, pregnant women are advised to increase their seafood intake for the sake of their babies. The Norwegian health authorities previously advised pregnant women not to eat more than two meals of fatty fish a week. Now we know better, and there is no longer any reason for pregnant women to limit their intake of fatty fish to two meals a week. We know this because NIFES annually monitors undesirable and illegal substances in farmed salmon. The analyses show that the levels of undesirable substances in farmed salmon are well below the maximum limit,

and that the levels of some of these substances have decreased further in recent years. Monitoring wild fish, farmed fish and its feed is one of NIFES's main tasks. We do this to establish whether Norwegian seafood is safe for consumers to eat. Contaminant residues are found in most foods. This makes it crucial to have proper programmes in place to document the state of play and make findings available to consumers and the authorities. This is what NIFES does. Thorough monitoring of undesirable substances in stocks of Greenland halibut along the continental shelf slope over the last three years is also the background for reopening the previously closed fishing grounds. NIFES conducts research in a food chain perspective. This means that we research the whole chain, from the fish feed to the food that ends up on our plates, and finally, to the effect seafood has on our bodies. One of our key areas of research is therefore fish feed. Today, there is limited access to marine ingredients for fish feed, and more and more plant ingredients are being used in the feed. It is therefore important to find out what effect the new feed ingredients have on fish health and welfare, as well as on the food that consumers serve at their tables. The change in feed ingredients can also result in changes in the undesirable substances consumed by the fish. NIFES also conducts research on what the fish feed of the future can contain. Insect meal, mussel meal and animal offcuts are a few of the alternatives scientists are working on. There are also interesting research

developments in the last link of the chain, i.e. how seafood affects our bodies. A lot of the health benefits of eating seafood are often linked to the marine omega-3 fatty acids in fatty fish. New trials with mice and people suggest however that lean seafood can also reduce the risk of developing both cardiovascular disease and type 2 diabetes. There have also been major organisational changes at NIFES over the last year. In February, I had the pleasure of taking over from Ă&#x2DC;yvind Lie who had been at the helm of NIFES for the past 18 years. With a background in both fisheries research and the university system, I look forward to being involved in further developing NIFES over the coming years. NIFES is also involved in the concept study for the localisation of the marine institutions in Bergen. This work aims to find solutions that will put the city of Bergen in an even better position to deliver world-class marine research. And NIFES will continue to play an important role in this context. In this year's Research News, you will get an overview of the research that is currently being conducted at NIFES. Happy reading!

Ole Arve Misund Director General

LEADER

Innhold

Leader

THIS IS NIFES

RESEARCH PROGRAMMES AND SECTIONS

LABORATORIES

ADMINISTRATION AND COMMUNICATION

SUSTAINABLE DEVELOPMENT OF FISH FARMING

NIFES MONITORS FISH FEED: Few maximum limits exceeded in fish feed

16 16

NEW FISH FEED INGREDIENTS MEAN NEW RISKS: Mussel meal in feed? Different interaction between nutrients and undesirable substances Maximum limits for selenium in fish feed

18 18

CAN NUTRITIONAL EFFECTS BE TRANSFERRED TO THE NEXT GENERATION? High levels of omega-6 affect the next generation Vitamin status affects the next generation Pollutants affect fish genes FISH NUTRITION FOR ROBUST FISH New analysis method for improved fish health Cell models to counteract stress Sterile salmon with cataracts By-product counteracting cell death Development of cataracts in salmon Composition of fatty acids and salmon's health Redox status measuring fish welfare How robust are farmed salmon to unfavourable environmental conditions? The time salmon spawn can affect the nutritional content of their eggs Can salmon tolerate plant sterols? 4

CONTENTS

19 20

22 22 23 24 25 25 25 26 28 29 30 31 32 33 34

THE PRACTICAL NUTRITIONAL NEEDS OF FARMED SALMON: 35 Minerals in modern feed 35 More plant-based feed creates new needs 36 Little marine omega-3 in feed for salmon 37 The nutritional needs of sterile salmon 38 Testing large-scale salmon production 39 FEED FOR FARMED MARINE SPECIES 40 Difficult to make the change to dry feed for halibut larvae 40 NEW FISH FEED INGREDIENTS Offcuts as an alternative feed ingredient Waste as resource Insects as fish feed Insects as a sterol source for Atlantic salmon

42 42 43 43 44

THE RIGHT FEED FOR FARMING LOUSE EATERS The genome of the ballan wrasse has been mapped Feed for the fish without a stomach Abundance of nutrients in commercial fish feed

45 45 46 47

PHARMACEUTICALS FROM FISH FARMING Delousing agents affect the genes of lobsters

48 48

ARE FISH PRODUCTS WHAT THEY CLAIM TO BE? 49 New DNA methods can reveal fish fraud 49 High levels of undesirable substances in certain marine oils 49 SAFE SEAFOOD IS BASED ON KNOWLEDGE

MONITORING: BASELINE STUDIES Mapping undesirable substances in haddock Analyses of undesirable substances in Atlantic halibut Monitoring our most important fish species

52 52 53 54

MONITORING OF SEAFOOD Monitoring of undesirable substances in farmed fish No Anisakis in Norwegian farmed salmon Studying the occurrence of Anisakis in wild fish in the European market

56 56 57 58

Salt tolerance in Anisakis Monitoring of parasites and hygiene in the wild fish sector General management plans of the seas Examining mercury in tusk Safe seafood near submarine wreck Did mercury from the submarine off Fedje spread to the environment? The maximum limit for mercury was not exceeded in cod caught in the Oslofjord Good status for Norwegian bivalves Spot check-based monitoring of new species and farmed salmon Monitoring of undesirable substances in seafood from the Vatsfjord Monitoring imported seafood Undesirable substances and microorganisms in cultivated kelp Good quality of frozen and defrosted cod fillets NIFES supports the Norwegian Food Safety Authorityt THE EFFECT OF UNDESIRABLE SUBSTANCES IN THE EARLY STAGES OF LIFE Undesirable substances affect bone metabolism DEVELOPMENT OF METHODS Analysis methods for organic contaminants Analysis methods for metals New standard methylmercury method soon to be available Method for the determination of pharmaceuticals in seafood Developing better cell models Micro plastics in seafood Joint PCB determination of whale blubber

59 60 61 62 63 64 65 66

Seafood, pollutants and obesity How are cell cultures and mice affected by undesirable substances? Interactive effects of methylmercury and various nutrients SEAFOOD AND LEARNING ABILITIES IN CHILDREN Is a child's health affected by its mother's diet? A trial of meals with meat or fish given to kindergarten children Diet and concentration at school Salmon to German children

88 90 92 95 96 97 98 99

68 HEALTH EFFECTS ON PEOPLE Lean seafood counteracts lifestyle diseases Children's iodine status better than their mothers'

100 100 101

72 73 74

NUTRIENTS IN SEAFOOD Salmon is still a good source of omega-3 fatty acids. More knowledge about the population's intake of iodine Analyses of nutrients in pizza and fish products

102 102 103 105

75 75

COOPERATION ACROSS THE WORLD Assistance on seafood safety and a good monitoring system in Angola Marine farming of Cobia on Cuba Aim to ensure iodine intake in Europe

107 108 109 110

SEMINARS Nordic iodine meeting Fish nutrition seminar

111 111 112

AQUACULTURE NUTRITION NIFES is chief editor of international aquaculture nutrition journal

113 113

COLLABORATORS AND FUNDING SOURCES

114

70 71

77 78 79 80 81 81 82 83

HOW CAN YOUR HEALTH BENEFIT FROM EATING SEAFOOD?85 HEALTH EFFECTS OF SEAFOOD, STUDIED THROUGH MODELS 86 The positive health effects of seafood are also affected by the food it is eaten with 86

CONTENTS

THE NATIONAL INSTITUTE OF NUTRITION AND SEAFOOD RESEARCH

The National Institute of Nutrition and Seafood Research (NIFES) is a research institute performing tasks in relation to governmental administration. We are affiliated to the Ministry of Trade, Industry and Fisheries. NIFESâ&#x20AC;&#x2122; area of research is both nutrition and welfare for farmed fish, and the effects of fish and seafood consumation on human health. We provide scientific documentation and advice to national authorities, industry and the public sector as a basis to ensure that seafood is safe and healthy to eat. NIFES is a supplier of knowledge, and has a responsibility to make its research known and available. We offer courses in human and fish nutrition, educating master and PhD students in collaboration with the University of Bergen and the University of Copenhagen. NIFES has also got the editorial responsibility for the international scientific journal Aquaculture Nutrition.

THIS IS NIFES

RESEARCH PROGRAMMES AND SECTIONS Our research is organised in two programmes that are divided into five sections, which each work with a large network of partners. Our overriding focus is on research and dissemination. In order to make our research known, we frequently attend conferences, seminars and trade fairs both in Norway and abroad.

FISH NUTRITION: Acting director of RESEARCH FOR FISH NUTRITION: Livar Frøyland

Head of Research: Rune Waagbø

13 scientists and 3 PhD students*

Changes in the composition of feed can cause nutrient based fish welfare challenges. Our research generates knowledge about the suitability of new fish feed ingredients, such as vegetable oils, insect meal and animal byproducts to meet the nutritional needs of farmed fish. This includes research on the fish's nutritional requirements for amino acids, fatty acids, vitamins and minerals to ensure that farmed fish achieve optimal growth

THIS IS NIFES

Fish feed must be healthy and safe for the fish, and produce a seafood product that is healthy and safe for people. The fish nutrition programme aims to increase knowledge about the nutritional needs of farmed fish and its tolerance of undesirable substances, which ensures the production of safe and nutritious food and robust fish in today's aquaculture industry. Our expertise also contributes to risk assessments that form the basis for the development of legislation relating to fish nutrition. The programme has editorial responsibility for the international journal Aquaculture Nutrition (Wiley, UK). Livar Frøyland also represents NIFES in different cooperation forums and working groups, both in Norway and abroad.

without production disorders, and are robust in relation to stress, infection pressure and climate change. We also study the fish's nutritional physiology during the reproduction phase and in early stages of life, and we try to understand how changes in the levels of both nutrients and undesirable substances in the fish's diet can alter normal development (from egg to juvenile fish). This includes nutrition-driven epigenetic impacts across generations.

Head of Research: Robin Ørnsrud

11 scientists and 1 PhD student*

We carry our research on the undesirable substances in feed that can affect fish health and food safety, in order to gain knowledge about the safe maximum limits for undesirable

substances in fish feed now and in the future. We generate knowledge regarding the extent contaminants and additives are transferred from the feed to the fillet. We also research interaction effects, also known as the cocktail effect, of several undesirable substances and of several undesirable substances in combination with nutrients. Our work increases knowledge of how the level of nutrients and undesirable substances in the feed can change the metabolism in fish, and whether the changes are inherited (epigenetics).

SAFE AND HEALTHY SEAFOOD Director of Research: Ingvild Eide Graff

The research and monitoring activities of the Safe and Healthy Seafood programme aim to increase our knowledge of the content of nutrients and undesirable substances in seafood. With the aid of animal and cellular models we also study how different substances interact. It is important for consumers, industry and the authorities to understand the effects on health of consuming seafood. In addition; we perform human intervention and observational studies. Our knowledge and research data contribute to both risk-benefit assessments and recommendations of seafood intake.

Head of Research: Amund MĂĽge

Head of Research: Lise Madsen

Head of Research: Marian Kjellevold

A large proportion of Norwegian seafood comes from catches of wild stocks. In order to improve our knowledge of the content of undesirable substances in species such as herring, mackerel, cod and saithe, we monitor and analyse these species through comprehensive monitoring programmes termed baseline studies. We also contribute to knowledge about parasites, human pathogenic bacteria and bacteria which compromise quality.

In order to identify the mechanisms that underlie the effects on health of seafood consumption, we study what happens to rodents when we give them seafood. We focus in particular on the effects of nutrients and undesirable substances on obesity and type 2 diabetes in mice. This section also does research on the content of undesirable substances in farmed salmon and other seafood.

We study the effects of seafood on human beings. We can do this by means of dietary intervention studies, in which participants consume a defined quantity of seafood in a short space of time, and we measure a number of health markers before and after the study. An alternative method is to use a food questionnaire to register how much seafood people eat, and to relate this to their state of health.

10 scientists and 2 PhD students*

10 scientists and 1 PhD student*

6 scientists and 1 PhD student*

*as of 31.12.15

THIS IS NIFES

LABORATORIES NIFES has four modern laboratories, and is the national reference laboratory for several analytical methods for seafood. NIFES analyses nutrients, undesirable substances and a number of quality parameters, mainly in fish and other types of seafood. On the basis of our results and research on fish nutrition and safe seafood, the institute provides advice to the authorities, industry and the public sector documenting that seafood is safe and healthy to eat. The overarching aim of all our laboratories is to develop and establish reliable and efficient analytical methods to meet the demands of the future. The laboratories are involved in student courses together with our research staff, and education of apprentices. NIFES is accredited according to standard NS-EN ISO 17025, and we have about 60 accredited methods. The results of our analyses are available via the Sjømatdata (Seafood Data) open-source database, which can be found on our website. We also provide analytical data to the Norwegian

Director of Research LABORATORIES:

Gro-Ingunn Hemre

THIS IS NIFES

Foodstuff Table, the Norwegian Scientific Committee for Food Safety, EFSA and FAO/WHO. Under the terms of the EEA Agreement, NIFES has been appointed national reference laboratory for a number of parameters. As the national reference laboratory, NIFES takes part in ringtests organised by the European Reference Laboratory (EU-RL). We also participate in working meetings with other EURLs, in order to keep abreast of methodological developments and new regulations, which are reported back to the Norwegian authorities. The Nutrients Laboratory analyses vitamins, amino acids and fats, as well as additives and energy content. We are working on the automation of a number of methods, such as Vitamin D and fatty acids, in order to increase our capacity.

The Element Analysis Laboratory analyses elements, total fats and proteins. Our methods

Head of Department NUTRIENTS LABORATORY:

Annbjørg Bøkevoll

Head of Department ELEMENTS LABORATORY:

Marita Kristoffersen

cover both heavy metals such as lead and mercury and essential trace elements such as iodine and selenium. The laboratory also has the responsibility for sample reception at NIFES. The sample reception unit plays a central role in that it registers and stores samples until they are sent to the appropriate laboratories at NIFES. The Contaminants Laboratory analyses a wide range of undesirable substances, including dioxins, pesticides and medical residues. Requirements regarding the preparation and analysis of samples are stringent, due to our range of extremely sensitive instruments.

The Molecular Biology Laboratory carries out analyses in microbiology, biochemistry, parasitology and molecular biology. The laboratory also manages the experimental animal department, which houses both rodents and zebrafish.

Head of Department CONTAMINANTS LABORATORY:

Bergitte Reiersen

Head of Department MOLECULAR BIOLOGY LABORATORY:

Anette Kausland

THIS IS NIFES

ADMINISTRATION AND COMMUNICATION NIFES is served by a support team that both ensures that we are able to perform our research in an optimal manner, and that our results are made known and available to the general public.

Deputy Director General Terje Digranes

The administration provides support for our research activities, and is responsible for office functions such as accounting, purchasing, personnel and archiving.

THIS IS NIFES

Head of Department Kathrin Gjerdevik

General Services offer services such as user support, maintenance and development. We are responsible for IT, the service office, and general operation and maintenance of buildings. HSE is another of our areas of responsibility. We also have general responsibility for the Laboratory Information Management System (LIMS), which is a database for the storage of analytical results, etc., which in turn forms the basis of our Internet-based Seafood Data service.

Director of Communications:: Anne Dorthea MĂŚland

We support the information dissemination work of our research staff, and are in constant dialogue with the authorities, the media and the general public concerning the work of NIFES. We are responsible for NIFESâ&#x20AC;&#x2122; interface with the rest of the world; via the Internet, in print, through our own publications and in the media, and we focus on offering high-quality written and visual communications services.

Photo 1: A German TV-team records sample preparation. Photo 2: French jouralists visit the Sample reception unit. Photo 3: Historical photo of the quayside of Nordnesboder 4. Photo 4: NIFES viewed from the sea.

THIS IS NIFES

SUSTAINABLE DEVELOPMENT OF FISH FARMING Healthy and safe food for the growing global population

Salmon

Trout

Fish feed

The sustainable production of seafood through aquaculture plays an important role in ensuring enough nutritious food to feed the world's growing population. But growth in this sector is contingent on a corresponding growth in access to feed. At NIFES, we research, among other things, alternative feed ingredients that are not suitable to use as food. Household waste cannot be used directly in fish feed because it contains too much carbohydrates. However, it can be used to feed insects, which then convert the carbohydrates (food waste) into good sources of protein and fat. In initial trials with salmon, in which insect meal was the only source of protein, the salmon grew as much as those fed on fishmealbased feed. However, the fat from these insects contains very little omega-3 fatty acids (EPA and DHA). We are therefore studying whether this changes if the food waste is enriched with, or replaced by, seaweed and kelp containing EPA and DHA. Another option in aquaculture is to use offcuts from chicken and pork directly in fish feed. In a feed trial using salmon, in which plant feed was compared with feed based on animal by-products from chicken and pork, the results showed that the animal offcuts resulted in just as good growth as the plant-based feed. This indicates a potential for converting an unexploited resource into good, healthy food. Research is in its infancy in this area, but it does suggest a potential feed for the future. An additional means of increasing food production is to exploit more of the resources in the sea, such as harvesting at a lower trophic level (mesopelagic species) for example or cultivating marine algae. Such

resources can potentially be used directly in food production or as a feed resource in farming both on dry land and in the sea. Knowledge of the fish's minimum nutritional requirements is therefore a basic precondition, so that the raw ingredients can be chosen on the basis of how much they contain of the nutrients that farmed fish need. The minimum nutritional requirements shall ensure the production of robust fish that can tackle the challenges posed by modern aquaculture. All of the ingredients in fish feed also contain different levels of undesirable substances, and it is vital to determine the tolerance level of fish for these substances, and how much is transferred from the feed to the edible part of the fish. Nutrients and undesirable substances (cocktail) in feed affect fish health, metabolism and the transfer of substances from the feed to the fillet, and we therefore need to know more about the cocktail effect in farmed fish. More aquaculture and better exploitation of the resources from the 'blue field', combined with better cooperation with the 'green field' to develop the feed of the future, will make an important contribution to securing enough safe and healthy food.

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NIFES MONITORS FISH FEED Few maximum limits exceeded in fish feed Funding: The Norwegian Food Safety Authority

In 2014, NIFES found no maximum limits were exceeded for heavy metals and organic undesirable substances in complete feeding stuffs or feed ingredients. The exception was one complete feedingstuff that contained residues of the pesticide hexachlorobenzene that exceeded the maximum limit. Fish feed ingredients and fish feed from the feed industry are analysed every year to identify any undesirable substances and certain nutrients, and NIFES conducts these analyses on behalf of the Norwegian Food Safety Authority, which is notified of any levels that exceed the maximum limit. In the feed report for 2015, which contains analysis from 2014, a total of 126 different samples were analysed: 78 complete feedingstuffs, 10 fishmeals, 10 vegetable feed ingredients, 12 vegetable oils, 7 fish oils and 9 premixes. Hexachlorobenzene (HCB) was found to slightly exceed the maximum limit in one complete feedingstuff. The maximum limit for HCB in a complete feedingstuff is 10 micrograms per kilo, and 11 micrograms per 16

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kilo was found in this one feed. Many of the feeds also contained vitamin D3 and selenium in excess of the maximum limits, but it is generally difficult to know whether nutrients have been added to the feed or whether they naturally occur in the ingredients. No Salmonella bacteria were found in any of the samples. This year, the programme was expanded to include analyses of several new pesticides. The pesticide pirimiphos-methyl was found in 55 per cent of the analysed rapeseed oils, and the levels were generally low. Rapeseed oil is the oil that currently replaces fish oil in Norwegian fish feed for farmed salmon. Analyses of the fatty acid profile of the 78 complete feedingstuffs showed that the levels of the marine omega-3 fatty acids EPA and DHA are similar to last year's analyses. There was great variation, however, in the analysed feeds. This is primarily a result of the fact that NIFES analyses feed from all the growth phases in the fish's life cycle, and that they have a varying content. As feed ingredients are a global commodity, the list of pesticides in use expands and changes. NIFES plans to include more types of pesticides in its monitoring programme. In

addition, the programme will also study fatsoluble mycotoxins that can accumulate in fish, and that may thereby affect both fish health and human health. NIFES is also planning to map vitamins in its next monitoring report.

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NEW FISH FEED INGREDIENTS MEAN NEW RISKS Mussel meal in feed? Cooperation: Ocean Forest EWOS Innovation Funding: Ocean Forest, VestMarin

NIFES will study how much diarrhetic shellfish poisoning, if any, is transferred to salmon from fish feed containing mussel meal. Scientists will also study possible toxic effects in the fish. In today's open cages, considerable

quantities of nitrogen and phosphorus are released into the marine environment from the fish's metabolism and excrement. These nutrient salts are eaten by microalgae, which are then eaten by filter feeders such as mussels. By cultivating mussels in and around fish farms, some of these nutrient salts are absorbed. Mussels can be harvested and

used as a marine ingredient in fish feed. NIFES participates in a project that is studying the use of mussel meal in fish feed. An analysis previously carried out by NIFES shows that mussel meal contains many of the vital nutrients fish need, and is therefore suitable for use in fish feed. It is also a very good source of the omega-3 fatty acids EPA and DHA. However, mussels can also contain algal toxins, such as diarrhetic shellfish poisoning (DSP toxins) and paralytic shellfish poisoning (PSP toxins). NIFES's role in the project is to study the toxins in the mussel meal. The presence of these toxins in the mussel meal may be a challenge for the fish. NIFES will therefore study the transfer of diarrhetic toxins from the feed to the fish fillets, and the possible toxic effects they can have on salmon.

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Different interaction between nutrients and undesirable substances Cooperation: Norwegian School of Veterinary Science (NVH), Norwegian University of Life Sciences (NMBU), Radboud University Nijmegen (RuN), the Netherlands, The Norwegian Institute for Water Research (NIVA), University of Oslo (UiO), SINTEF Funding: The Research Council of Norway

Marine nutrients reduced the negative effects of undesirable substances and pesticides, while vegetable nutrients strengthened the negative effects. The increased use of vegetable oils in fish feed has resulted in a strong reduction of organic pollutants in farmed salmon. Fish feed in which marine feed ingredients have been replaced by vegetable ingredients, however, also poses new challenges to fish health and seafood safety. This is due, among other things, to the fact that the vegetable ingredients contain other undesirable substances than the marine ingredients, such as the, benzopyrene and phenanthrene, and the pesticides chlorpyrifos and endosulfan. However, some nutrients are known to affect, even protect against, or strengthen the effect of pollutants and pesticides. The introduction of new vegetable feed ingredients has increased the need for more knowledge on how this interaction between nutrients and undesirable substances affects the fish. In this project, scientists have therefore studied the interaction between the 'new' undesirable substances benzopyrene, phenanthrene,

chlorpyrifos and endosulfan and four nutrients. Scientists studied the interaction between the nutrients and the undesirable substances in several different ways, looking at, among other things, absorption in the intestines and interaction in the cells in cell models.

negative effects of the pollutants and pesticides. The cell trials also showed that the vegetable nutrients gamma-tocopherol, vitamin E from plants, and the omega-6 fatty acid arachidonic acid reinforced the negative effects.

Trials show that the undesirable substances benzopyrene and phenanthrene are absorbed in all parts of the intestines. Absorption is however slower when plant oil is used instead of fish oil. The scientists also found that there is higher absorption of phenanthrene than benzopyrene. The scientists used cell models in order to learn more about the interaction between undesirable substances and nutrients. The undesirable substances generally had a normal, additive effect on each other, but the pesticides chlorpyrifos and endosulfan reinforced the effect of each other. Cell trials also showed that marine nutrients reduced the toxicity of the undesirable substances more than the vegetable nutrients. The marine nutrients EPA, an omega-3 fatty acid, and alpha-tocopherol, or vitamin E, reduced the

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Maximum limits for selenium in fish feed Cooperation: National Food Institute, Technical University of Denmark, BioMar AS, Ewos Innovation, Skretting AS, The Norwegian Seafood Federation Funding: The Norwegian Seafood Research Fund

NIFES is researching the effects of increased selenium intake in salmon. The findings can be used to establish a maximum limit for selenium in salmon feed. Marine ingredients are the natural source of the mineral selenium in salmon feed. The levels of fishmeal in salmon feed were previously relatively high, but, in the development of new sustainable salmon feed, fishmeal is replaced by alternative plantbased feed ingredients that can reduce the content of selenium in salmon feed. In order to ensure strong and robust salmon, it is advantageous to add selenium to the natural levels found in marine salmon feed. However, it is not currently permitted to add selenium to salmon feed because of the maximum limit for selenium in animal feed. The current maximum limit for selenium in feed therefore limits the scope of action for replacing marine feed ingredients with alternative ingredients, which have lower, natural levels of selenium. The maximum limits were established in the EU as a result of risk

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assessments, among other things. This project will therefore provide knowledge that is needed in connection with the risk assessment of selenium in salmon feed by establishing the upper tolerance level for selenium in salmon. In 2015, one fish trial was conducted in which the scientists wished to find markers for the early effects of increased selenium intake in salmon. One feed trial with Atlantic salmon, which were given feed containing selenite or selenomethionine, was conducted and markers for the toxic effect of selenium were determined using screening methods. The most important finding was that the antioxidant selenium becomes a pro-oxidant at high dosage, i.e. it has the opposite effect. Too high concentrations of selenium can thereby cause oxidative damage to fish. The biomarkers that have been identified will be used in a forthcoming trial to establish threshold values for the undesirable effects of selenium in salmon.

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CAN NUTRITIONAL EFFECTS BE TRANSFERRED TO THE NEXT GENERATION? High levels of omega-6 affect the next generation Cooperation: University of Nordland (UiN) Funding: The Research Council of Norway

Trials with zebrafish show that if the parent generation eats an omega-6rich diet, their offspring may be affected. More plant ingredients in today's fish feed has considerably reduced the proportion of marine omega-3 fatty acids in the feed, which are partly replaced by plant-based oils containing varying amounts of omega-6 fatty acids. But how are the farmed fish affected by this change in the composition of fatty acids? In this project, NIFES scientists have used zebrafish as a model for studying how dietary changes affect the methylation pattern of DNA. This is a way in which the body can switch certain genes on and off without

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altering the DNA sequence itself. This property can lead to cells, organs and organisms changing their physiological qualities. As this methylation can, in some cases, be transferred from parents to offspring through the generations, the scientists wished to see whether any changes in the methylation pattern could be passed from one generation of zebrafish to the next. Analyses show that high levels of the omega6 fatty acid arachidonic acid in the diet of the parent generation affects both the methylation pattern and the gene expression of the offspring at maturity. The studies also showed signs of increased inflammation and a change in the ability to metabolise fat in the

parent generation. The scientists have previously found in the project that the composition of fatty acids affects vitamin A metabolism in both the parent and offspring generation. The diet with a high omega-6 content resulted in no differences in growth.

Vitamin status affects the next generation Cooperation: Oslo University Hospital, Stirling University, UK, Nord University, Medical University of Vienna, Austria, The Institute of Marine Research Funding: The Research Council of Norway

A low vitamin B level in parents' fish feed did not affect the growth of the next generation. Feed trials with zebrafish did however show epigenetic effects in the offspring. A significant proportion of the marine ingredients in fish feed are now replaced by plant-based ingredients. If extra vitamins are not added to the plant-based feed, it can have negative consequences for the fish that eat the feed, but can it also have consequences for subsequent generations?

This project takes a closer look at how a lack of vitamin B affects fish. Plant-based feed has a lower vitamin B content. These vitamins are important, among other things, for enabling the body to regulate the expression of genes, which, in other organisms, can be passed on to the next generation. This is called epigenetics, and little is known about the regulation of gene expression in fish. Epigenetics is a discipline that describes how the environment affects which genes are switched on or off, just like a light switch. The manner in which the genes are switched on and off can be passed on from the mother and father to the next generation, but this has not yet been documented for fish. This makes it important to determine whether broodstock feed has the potential to improve the health of the offspring. The hypothesis in this project is that the quantity of vitamin B added to the plant-based feed of one generation affects how the DNA can be used in the next generation.

In this project, scientists have conducted a long-term feed trial with zebrafish, from the parent generation to the offspring generation. The zebrafish in the parent generation were either given too low or too high levels of vitamin B in the feed. The difference in vitamin B levels resulted in differences in growth in the parent generation. All of the fish in the next generation were then given enough vitamin B, and the scientists found no differences in growth. They did however find major differences in the genes that were expressed in the offspring both at the embryo stage and later in life. The scientists also found differences in DNA methylation, which is one of the ways the body switches certain genes on and off. The scientists will now study which cells and signal pathways between the cells are regulated by changed DNA methylation patterns in the offspring generation.

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Pollutants affect fish genes Cooperation: Norwegian Institute for Water Research (NIVA) Funding: The Research Council of Norway

How can pollutants change the DNA of fish, even through several generations? That is what scientists at NIFES are trying to find out. Epigenetics is a field that studies how genes can be switched on or off without changing the gene itself. These changes can affect several generations, also in fish. Undesirable substances are one of the things that can result in epigenetic changes. When mature animals are exposed to certain pollutants, several generations of offspring, without any exposure to the pollutant, can nonetheless be negatively affected. This can result in more disease in fish â&#x20AC;&#x201C; generations after the fish that was actually exposed to the pollutant in question. Similarly, environmental pollutants in early stages of life can affect fish later in life through epigenetic mechanisms. In order to obtain more knowledge in this field, scientists in the project conducted trials using zebrafish, in addition to other tools. This species is a useful model in epigenetic research, as it has a short generation time compared with species like salmon and cod. In 2015, scientists therefore conducted trials in which the fish were exposed to the substances bisphenol A, genistein and DDE. These substances were chosen because they 24

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are found in the environment, they are endocrine disruptors and they are thought to damage reproductive ability and affect epigenetic mechanisms in animals. Bisphenol A, for example, is in many products and has been found in mussels and cod liver along the Norwegian coast. Trials with zebrafish embryos showed that very low concentrations of bisphenol A affected genes that are important for DNA methylation, i.e. one of the mechanisms that switch genes on and off. Trials with liver cells

from male salmon also showed endocrine disruptor effects of bisphenol A, genistein and DDE at relatively low concentrations. Analyses of samples from these trials will continue, and new exposure trials with zebrafish embryos will be conducted in 2016. This research will provide useful information about how environmental pollutants that fish are exposed to can lead to diseases in fish, through hereditary changes that are not caused by changes in the gene sequence.

FISH NUTRITION FOR ROBUST FISH New analysis method for improved fish health

Cell models to counteract stress

Funding:

Cooperation: Ewos Innovation AS Funding: The Research Council of Norway, Ewos Innovation AS

The Ministry of Trade, Industry and Fisheries

The proportion of omega-6/omega-3 fatty acids in fish feed is crucial to fish health. How omega-6 and omega-3 fatty acids will affect the health of salmon can be determined based on the type and level of the pheromones eicosanoids. Roughly speaking, we can say that the eicosanoids that are derived from omega-6 fatty acids increase inflammatory reactions, while eicosanoids derived from omega-3 reduce inflammation. All cell membranes contain fatty acids in what are known as phospholipids. The eicosanoids derive from the phospholipids in the cell membrane. However, studies published about eicosanoids are linked to exposure to pollutants, bacterial substances or nutrients, and do not look at how these nutrients affect the fatty acid composition of the phospholipids in the cell membrane. NIFES therefore wishes to develop a simple, analytical strategy for the simultaneous determination of phospholipids and eicosanoids. This will enable us to understand the dynamics of the activation and synthesis of eicosanoids during inflammation in living organisms.

Cell models are good tools for studying stress in fish cells. These models can thereby be used to learn more about how to counteract stress in fish. In recent years, scientists have started studying how well the nutrients in feed can be exploited, including whether they have additional effects on top of supporting growth and deposition in the fish. Scientists at NIFES are set to study this using specific amino acid modulation, in which amino acids or their metabolites are added to cell models and then compared to cells without added amino acids/metabolites before and after using different stress models.

The scientists will determine whether different amino acids can counteract the stress response. They will do this by isolating liver and head kidney cells from salmon, then cultivate them in the laboratory before finally conducting trials on them by stressing the cells. In doing so, the scientists at NIFES can study whether and how specific amino acids can counteract oxidation or inflammatory reactions. They will thereby have a functional effect in the cells in addition to contributing to growth and deposition (functional qualities). No final results have been reached to date, and scientists are initially working on developing suitable cell models for the project objective.

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Sterile salmon with cataracts Cooperation: University of Bergen, the Institute of Marine Research Funding : The Research Council of Norway

There is a higher incidence of cataracts in triploid salmon. Scientists at NIFES are trying to find out why sterile salmon experience this problem. Triploid salmon are salmon with three sets of chromosomes, as opposed to two sets of chromosomes which is the norm. This makes triploid salmon sterile. It is therefore possible with triploid farmed salmon to avoid the negative consequences to the environment of fish escaping and reproducing with wild salmon. However, triploid salmon demonstrate periodic differences in growth, physiology, behaviour and appearance in relation to diploid salmon, which only have two sets of chromosomes. Among other things, there is a higher incidence of cataracts in triploid fish. This is an eye condition in which the lens becomes unclear, due in part to damage to the lens proteins. This thereby reduces sight. The condition is a welfare issue in salmon farming. One of the reasons for cataracts is that the fish have a problem with water balance in 26

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the lens, which in turn can lead to damage to the lens structure. Incorrect regulation of the tear ducts, or aquaporins which is the technical term, in the lens may be directly linked to the development of cataracts, according to studies of mammals and zebrafish. In this project, the scientists wished to study whether there were any differences in the genes that were expressed in triploid and diploid salmon, and whether this could explain why triploid salmon are more prone to cataracts than diploid salmon. Scientists did this by studying the patterns in the gene expression in lenses from triploid and diploid salmon with and without cataracts. In general, there were only minor differences in the gene expression between diploid and triploid salmon. In total, there were 165 genes that were expressed differently in the lenses from diploid and triploid salmon with cataracts. Many of these genes

had a lower expression in triploid salmon than in diploid salmon. Some of the genes with a lower expression in triploid salmon with cataracts code proteins that contribute to the maintenance and configuration of proteins, which can suggest a reduced ability to repair damaged proteins. This can help to explain why triploid salmon are more prone to develop cataracts. The scientists will also identify specific markers that can be linked to robustness.

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By-product counteracting cell death Cooperation: HordafĂ´r AS, the International Research Institute of Stavanger (IRIS) Funding: The Research Council of Norway

Hydrolysed fish protein, a by-product from the fisheries industry, can counteract cell death. The fish farming industry wants ingredients that are not too expensive and that contain nutrients that contribute to maximum growth, without jeopardising fish health and welfare. Documentation is therefore needed of the qualities of the by-products from the fisheries industry, such as hydrolysed fish protein. This can have a practical effect during periods of increased oxidative stress. If a protein is hydrolysed, it means that it is broken down into smaller fragments.

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Hydrolysed fish protein is a by-product from fisheries that can be used in fish feed. Trials conducted on liver cells and head kidney cells from salmon show that hydrolysed fish proteins counteract cells committing 'suicide' (apoptosis) after being exposed to oxidative stress. When the cells were exposed to oxidative stress without the addition of hydrolysed fish proteins, apoptosis was activated and the cells died. The main objective of this project is to document any functional qualities of the low molecular weight water-soluble part of hydrolysed proteins. Trials have also been

conducted to study whether hydrolysed proteins can counteract inflammatory stress by exposing the cells to bacterial lipopolysaccharide (LPS). The trial shows that hydrolysed proteins protect against LPS-initiated inflammatory responses in the cells. Contingent on the functional properties of hydrolysed proteins that are found in the cells also being found in trials with animals, they can, in the feed, have a preventive effect during periods of increased metabolic stress in farmed fish.

Development of cataracts in salmon Cooperation: Nofima Funding: The Norwegian Seafood Research Fund

Salmon smolt that have been fed feed consisting of plant meal and plant oils develop serious cataracts compared with salmon fed with fishmeal and fish oil, despite the addition of EPA and DHA. Cataracts are an eye condition resulting from damage to the lens and the gradual loss of sight for the fish. This is a production-related disease, and it is an important ethical and welfare challenge in salmon farming. The objective of this project was to find out whether low levels of the fatty acids EPA and DHA in the feed in the early stages of life increase the risk of developing cataracts later

in life. We also wanted to see whether different salmon genera have different susceptibility for developing cataracts. The results suggest that the composition of the feed when the fish are released into the sea is crucial to their susceptibility for developing cataracts, also later in life. Salmon smolt that were given experimental feed without fish oil and fishmeal, with a varying EPA, DHA and EPA+DHA content (02 per cent) during the period where their bodyweight was between 40 grams and 400 grams, all developed serious cataracts at 1.3 kilos. Very few of the fish that were given a control feed based on fish oil and fishmeal (2.2 per cent EPA + DHA) developed cataracts during the period. The feed the fish were given from they weighed 400 grams to 1.3 kilos was not crucial to the development of cataracts, and the fish that changed to commercial feed at 400 grams, had the same cataract

score as those given experimental feed throughout the period. Salmon that were given the experimental feeds to which high concentrations of EPA and DHA were added, had as high cataract scores as the salmon that were fed feed with a low EPA and DHA content, but their lens metabolism (metabolomics) was affected by the EPA and DHA content of the feed. The development of cataracts in genera that were selected on the basis of their ability to make EPA and DHA from shorter omega-3 fatty acids from plants, showed that there were genetic differences as to how susceptible the different salmon genera were to cataracts. It also indicated that a low content of fish oil during the freshwater phase could be advantageous. In order to support the production of robust salmon with good sight under good welfare conditions, it is important to carry out an evaluation of what the use of alternative ingredients and the EPA and DHA level in the feed means for the development of cataracts.

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Composition of fatty acids and salmon's health Cooperation: Nofima, Norway Skretting ARC Funding: The Norwegian Seafood Research Fund

Plant oils in the feed can affect the health of fish and their resistance to viral diseases. As a result of limited access to fish oil, and a steadily growing aquaculture industry, plant oils are used in the production of commercial feed for salmon. It is therefore important to find out whether this has consequences for the fish's health and how it affects welfare and robustness. The objective of this project was to increase knowledge about how the composition of fatty acids in the feed affects fish health and their resistance to viral diseases. This was initially studied by evaluating the state of health of salmon in a feeding trial. We wanted to find out whether omega-6, monounsaturated and saturated fatty acids in the feed affected the conversion of alphalinolenic acid (ALA, 18:3 n-3; an omega-3 fatty acid from plants) into the marine omega3 fatty acids EPA (20:5 n-3) and DHA (22:6 n-3). The fish were fed eight different diets, composed of different amounts of plant oils that are each characterised by containing a lot of one type of fatty acid: Saturated fat 30

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(palm oil), unsaturated fat (rapeseed oil) and omega-6 (soybean oil) in addition to fish oil. The diets had the same amounts of ALA, EPA and DHA, but different levels of saturated, monounsaturated and omega-6 fatty acids, and thereby different omega-6/omega-3 ratios. The results showed no differences in growth and welfare between fish fed on the different diets. The plant fatty acid 18:2n-6 in the feed affected the level of the long-chained fatty acids, with a higher content of the omega-6 fatty acid arachidonic acid (ARA), and a lower EPA and DHA content in the cell membranes of the liver. The levels of proinflammatory eicosanoids (PGE2) in the blood of the omega-6 groups (soya) were also higher. In order to determine whether the different fatty acids in the feed had any significance for resistance to viral diseases, fish from four diet groups previously dominated by saturated fat (palm oil), unsaturated fat (rapeseed oil), omega-6 (soya oil) and a high omega-6 content (high soya content) were infected with SAV3 (the virus that causes the pancreas disease, PD). Inflammatory and immune responses were also studied in vitro

(in the laboratory), as a model for studying the mechanisms of infection. The results of the infection trials showed that there was no difference in the amount of virus in the hearts of fish from the different diet groups, but that there was a difference in how the fish responded to the viral infection. Fish previously fed on a feed with a high content of omega-6 fatty acids had a stronger inflammatory response and immune response, while fish fed on the saturated fat diet (palm

Redox status measuring fish welfare Cooperation: Heinrich Heine University, Germany Funding: The Ministry of Trade, Industry and Fisheries

oil) appeared to have a better outcome, with less inflammation and faster repair of diseased tissue. Head kidney cells isolated from salmon were stimulated with LPS in vitro, as a model for inflammation caused by bacterial infection. Cells from fish fed omega-6 diets (soya and high soya content) produced more proinflammatory eicosanoids (PGE2, inflammatory signaling) measured by the cell culture medium, while cells from fish fed on saturated fat (palm oil) had a higher concentration of anti-inflammatory eicosanoids (PGE3 and LTB5). The trials show that plant oils in the feed can affect the health of fish and their resistance to infectious diseases.

This project aims to establish a zebrafish model as a tool for researching robust fish. Changes in an organism, e.g. in the event of illness, malnutrition or poisoning can lead to a change in the so-called redox status. This means that the fish's redox status can be used as a template for how it is doing, e.g. in connection with the introduction of new feed ingredients. The objective of this project is to increase understanding about redox regulation and how redox status templates can be used to assess fish welfare. The objective was to develop a range of tools in order to be able to evaluate redox status.

We have started charaterisation of these lines, and will focus in 2016 on developing methods that can be used to measure redox status in all organisms, and conduct a study on salmon where we will look at the development of redox status through its life cycle. This project will expand the toolbox we use to measure whether fish are robust and in good health. The project implements some of the most recent biological developments in research on fish. This knowledge is already being used in different projects at NIFES.

In 2015, we gained access to two zebrafish lines that express redox-sensitive green fluorescent protein (roGFP), whose fluorescence changes when the cells' redox status changes. This will make it possible to measure changes in redox status directly, such as changes due to the effect of nutrients or contaminants.

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How robust are farmed salmon to unfavourable environmental conditions? Cooperation: Institute of Marine Research, University of Bergen, University of Arkansas, USA, Hokkaido University, Japan, University of Victoria, Canada, University of Ghent, Belgium, Bergen University College, Skretting ARC Funding: The Research Council of Norway

High water temperatures and CO2concentrations affect growth, metabolism and antioxidant defence mechanisms in salmon. A changing climate means that it is important to study how robust farmed fish are to high temperatures and higher concentrations of CO2 in seawater. This will enable us to adapt production strategies and tailor feed to increase farmed fish's mastery and performance. The objective of this project is to map the fish's reactions to unfavourable environmental conditions. We focus in particular on how temperatures above the optimal temperature for growth affects appetite, growth regulation, metabolism and antioxidant defence mechanisms. A number of short-term and long-term experiments have been conducted on salmon of different sizes during the course of the

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project. The results show that high temperatures affect big salmon to a greater extent than small salmon. This means that the appetite and growth of big salmon are greatly reduced compared with big salmon at an optimal temperature, while small salmon are less affected. Analyses of growth hormones found there to be lower levels and thereby lower stimulation of growth in salmon of all sizes. High temperatures also led to the fish consuming more of the antioxidant vitamins E and C, because of increased stress (oxidative pressure). The results from this project also show that periods of high temperatures and low oxygen levels in the water can result in less nutritious salmon. The amino acid histidine and histidine compounds are found in a number of tissue types. They perform the important functions of supporting antioxidant defence mechanisms, water balance and pH regulation. The results

of this project show that histidine and the compound anserine in the muscle and Nacetyl-L-histidine in the heart are not affected by temperature in salmon and trout. The same conclusion was found for histidine compounds in the muscle tissue of carp and tilapia. This shows that histidine compounds serve as important and stable buffers at high temperatures. On the other hand, histidine compounds in the salmon's heart are reduced if it is exposed to high CO2 levels, which can have unfortunate consequences for the salmon. Unfavourable environmental conditions for farmed fish can arise during periods of production, and it is therefore important to know what happens to the fish and what is required to maintain their robustness.

The time salmon spawn can affect the nutritional content of their eggs Cooperation: SalmoBreed AS, Benchmark holding plc Funding: Salmobreed AS

Salmon can spawn during large parts of the year, but the time they spawn can affect the quality of their eggs. Smolt are no longer only released at certain times of the year in salmon farming, and roe suppliers are therefore required to extend the season, with roe being stripped from the salmon for as long as possible at both ends of the season. Stripping the salmon means collecting the eggs of sexually mature fish. For the groups that are stripped last, it may take a long time from feeding stops until the roe is stripped, which may affect egg quality. The objective of the project is to find out whether roe from fish that spawn late have a less optimal nutritional status, and when such a reduction in quality takes place. The results showed that the time the salmon spawn clearly affects the nutritional content of the eggs. This can be due to both the feeding regime and the nutritional content of the broodstock feed.

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Can salmon tolerate plant sterols? Cooperation: Nofima, Skretting ARC Funding: The Norwegian Seafood Research Fund

In this project, we wish to clarify whether changes in the level of plant sterols in the salmon's feed has negative effects on its health. Many trials in which fish oil in the feed has been replaced by vegetable oil have shown an increased accumulation of fat in the liver of salmon, often combined with an increased level of fat in the blood. This is often seen in connection with the use of rapeseed oil, which contains a high level of plant sterols. Plant sterols consist of molecules that resemble cholesterol in humans. We wished to find out whether plant sterols were responsible for the accumulation of fat in fish. In humans, plant sterols are best known for reducing the absorption of cholesterol in the intestines. We therefore wished to see whether the ratio between plant sterols and cholesterol was of significance, and whether the addition of cholesterol could potentially counteract the negative effects of plant sterols on the health of salmon.

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In 2015, cell trials were conducted on liver cells from salmon that were exposed to different plant sterols and a combination of plant sterols and cholesterol. The latter showed certain effects on genes relating to fat and cholesterol metabolism in the liver. Samples of rapeseed oils have also been collected for plant sterol analysis from different suppliers and geographical areas. There was a lot less variation in plant sterol content than expected. This is good news for the reliability of plant sterol content in commercial feed. A big salmon feeding trial started in September. The trial includes a total of nine feed groups that are fed different levels of plant sterols and feeds with different ratios of plant sterols and cholesterol. The trial is being conducted at two temperatures, at 6Â°C and 12Â°C. The fish are eating and growing well, and an intermediate sampling at 12Â°C

was conducted in December. The trial will conclude in 2016. When all the analyses have been completed, we will be able to define a safe level of plant sterols in feed for salmon, and whether the addition of cholesterol and the ratio of plant sterols and cholesterol has any significance. This is important knowledge with respect to flexibility in choosing feed ingredients, and for ensuring fish health and welfare.

THE PRACTICAL NUTRITIONAL NEEDS OF FARMED SALMON Minerals in modern feed Cooperation: King's College London, UK, National Food Institute DTU, Denmark, Institute of Marine Research, Skretting ARC Funding: The Research Council of Norway and Skretting ARC

The chemical form determines the need for minerals in fish feed Previous projects have shown that farmed salmon now need a higher content of minerals added to plant-based feed. The APREMIA project looks at the chemical forms of the minerals, and to what extent they determine the availability of, and thereby the requirement, in the fish. There are limited marine ingredients for salmon feed. If we replace them with other ingredients, however, the levels of minerals in the feed changes, as does their overall composition.

minerals needed to ensure fish health and robustness, and it will increase the environmental impact of minerals lost via excrement. High mineral supplements in fish feed today can also affect each other with respect to absorption in the intestines and in their biological functions. The chemical form (e.g. inorganic or organic) of mineral supplements can have a bearing on their bioavailability, but the absorption paths and tissue break-down of the different chemical forms has not been established in fish.

chemical forms of minerals in the feed ingredients and finished feeds, in addition to contributory factors that may increase the requirement. Speciation is distinguishing chemical forms from each other. In this project, we will develop a chemical 'speciation' method for certain minerals, in order to predict mineral digestibility, based on the breakdown of the chemical forms in the feed. The work on developing analytical methods to determine zinc compounds in the feed and fish is under way.

The determination of mineral requirements in Atlantic salmon strongly depends on the

Replacing them with plant ingredients also reduces the bioavailability (how well different substances are absorbed) of minerals. This is because of the presence of plant fibre and phytate. The lower bioavailability of minerals in modern feeds will therefore affect the level of

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More plant-based feed creates new needs Cooperation: Institut National de la Recherche Agronomique INRA, France (coordinator), University of Stirling, UK, Consejo Superior de Investigaciones Científicas CSIC, Spain, Hellenic Centre for Marine Research (HCMR), Greece, Universidad de Las Palmas de Gran Canaria (ULPGC), Spain, Research institute for fisheries, aquaculture and irrigation (HAKI), Hungary, Wageningen University (WU), the Netherlands, Università dell’Insubria (USI), Italy, Centre of Marine Sciences (CCMAR), Portugal, BIOMAR, Denmark, SPAROS Lda, Portugal, Viviers de Sarrance VDS, France, NOREL S.A., Spain, Aranykarasz Mezogazdasagi Halaszaties Szaktanacsadoi Szolgaltato Bt (KARAS), Poland, Alevines y Doradas, S.A ADSA, Spain, AquaTT, Ireland Funding: ARRAINA is a EU FP7 project, The Ministry of Trade, Industry and Fisheries

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More nutrients should be added to plant-based feed for salmon to meet the fish's needs. This is one of the recommendations of the EU project ARRAINA. The European aquaculture industry increasingly uses feed based on feed ingredients other than fishmeal and fish oil. In the EU project ARRAINA, scientists at NIFES, among others, are looking at plant-based feed and how we can ensure that the fish are getting all the nutrients they need to ensure good growth and health. As fishmeal and fish oil have provided many nutrients, including many of the B vitamins, the transition from using these ingredients to a plant-based feed could lead to a lack of or imbalance in nutrients. Based on two doseresponse trials, one in freshwater and one in seawater, the scientists have discovered that more nutrients should be added in greater amounts to plant-based feed for salmon to meet its needs. When more nutrients are added, the salmon demonstrate better growth, without any increase in the size of their liver and intestines. When the liver and intestines increase in relative size, this can be the first symptom of an imbalance in the feed. Increasing the content of certain specific B vitamins is also recommended to maintain enzyme activity. Enzymes are proteins that are

responsible for the chemical processes in living organisms. The project also makes new recommendations for the B vitamins niacin, cobalamin, folacin and pyridoxine in fish feeds, while the applicable 2011 recommendations still meet the salmon's need for biotin, riboflavin, thiamin and pantothenic acid. In this project, the scientists have also looked at the absorption and metabolism of the nutrients that have traditionally been regarded as antioxidants: vitamin C, vitamin E and selenium. In one feeding trial, the plant-based feed ingredients appeared to contain sufficient quantities of vitamin E and selenium to prevent clinical deficiency diseases in the fish. Based on the results from this part of the project, the current practice of adding 100200 milligrams of vitamin E per kilogram and a similar level of vitamin C to salmon feed, but a slightly higher level of vitamin C than vitamin E is still advised. Concentrations higher than the 20-50 milligrams per kilogram that prevent clinical deficiency diseases will protect the fish against episodes of oxidative stress.

Little marine omega-3 in feed for salmon Cooperation: Skretting ARC, Centre for Aquaculture Competence (CAC) Funding: The Research Council of Norway

NIFES has proposed a minimum limit for marine omega-3 fatty acids in salmon feed. The marine ingredients that salmon feed traditionally contained have ensured that the fish have consumed plenty of long-chained marine omega-3 fatty acids (EPA and DHA). The main source of EPA and DHA is fish oil, and, to some extent, fishmeal. Apart from these, there are currently no alternative sources of the volumes required in the aquaculture industry. The growing industry and limited access to fish oil have made it impossible to maintain the same high EPA and DHA content in the feed as previously. Atlantic salmon have been shown to both store and produce marine omega-3 fatty acids (EPA and, in particular, DHA) when it gets little through the feed. The overriding goal of this project is to determine how much EPA and DHA salmon actually need, and the consequences reducing the content in the feeds can have for growth and health. We also look at how the feeds can be optimised to ensure that salmon 'save' as much EPA and

DHA as possible (by burning other fatty acids, but storing these in the body), and thus make more of these fatty acids themselves. Atlantic salmon were fed low levels of EPA and DHA from 1.3 to 7.4 per cent of the fatty acids (4 - 24 mg kg-1 feed) in two longterm trials run at 6Â°C and 12Â°C. The growth of the fish was negatively affected by low EPA and DHA. The trial also showed that salmon can be a net producer of DHA when the feed contains low levels, with a retention of 120 to 200 per cent. This means that we measured up to twice as much DHA in the fish at the end of the trial as it had received from the feed. EPA retention was lower, and there were no differences in the EPA tissue levels between the different feeding groups. We found reduced DHA tissue levels when there were low levels in the feed, but the greatest effect was seen in the red blood cells. In the retina and brain, only low temperature was seen to have an effect. The results show that salmon have a specific EPA+DHA requirement of >2.7 per cent of the fatty acids for optimal

growth in the sea and to maintain the DHA level in the tissue. We also analysed samples from one largescale commercial production facility, where 5 and 8 per cent EPA+DHA was used. These results supported the findings of the smaller scale feeding trials. During the growth phase, this fish was exposed to stress in the form of repeated delousing treatments and an outbreak of pancreas disease at the site. There were no significant differences in growth and mortality rates between the diet groups, which indicated that reducing EPA+DHA from 8 to 5 per cent of the fatty acids does not reduce the fish's robustness. The goal of the last feeding trial was to determine whether omega-3 fatty acid retention was affected by other classes of fatty acids in the diet, such as the level of monounsaturated, saturated and omega-6 fatty acids. The trial was based on a mix design, with rapeseed oil, palm oil and soya oil, which, respectively, provided high contents of monounsaturated, saturated and omega-6 fatty acids. Linseed (rich in linolenic

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The nutritional needs of sterile salmon Cooperation: The Institute of Marine Research, Marine Harvest, Aqua Gen AS, Ewos, Alfred Wegner Institute, Germany Funding: The Research Council of Norway, the Norwegian Seafood Research Fund

acid, 18:3n-3) and fish oil (rich in EPA and DHA) were used to balance the diets to equivalent levels of short-chained omega-3 fatty acids from plants (18:3n-3) and EPA and DHA. There were no differences in growth, but the palm oil diet resulted in lower growth per kilo feed consumed and reduced fatty acid metabolism. Retention data showed that a high content of omega-6 in the feed, up to 43 per cent of the fatty acids, had no negative effect on the production of longchained omega-3 fatty acids, but a high omega-6 content had a negative effect on the incorporation of omega-3 fatty acids in the cell membranes.

By adding the amino acid histidine to feed, we can reduce the development of cataracts in sterile salmon smolt at different freshwater and seawater temperatures. Good production characteristics have been cultivated in farmed salmon over several decades, making it genetically distinct from wild salmon. If farmed salmon escape, they can spawn with wild salmon and genetically affect the wild salmon. To minimise the consequences of fish escaping, it is possible to produce triploid salmon, which are sterile. This is done by pressure-treating the eggs just after fertilisation, so that the salmon end up with two sets of genetic material from the mother and one from the father, rather than one set from each of the parents as in diploid salmon. Triploid trout are often farmed in other countries, while triploid salmon farming is less common as they do not perform as well as diploid salmon when farmed. The main reasons for this seem to be a lower tolerance for high seawater temperatures and that they are more prone to developing welfare diseases such as bone deformities and cataracts. This is most likely due to triploid

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salmon having different nutritional needs to diploid salmon. This also means that these welfare diseases can be reduced by appropriately adjusting the composition of the feed. If we solve these problems, triploid salmon has the potential to become a more environmentally friendly species of farmed fish. In this project, a feeding trial with diploid and triploid salmon smolt was conducted in freshwater and seawater at two temperatures (10 and 16Â°C). In addition, two different feeds were used containing different amounts of the amino acid histidine (low and normal level). The results show little development of cataracts at 10Â°C in freshwater, regardless of the level of histidine, while at 16Â°C, there was severe cataract development in both diploid and triploid salmon. The severity was highest in triploid salmon, but was somewhat reduced when using a normal level of histidine in the feed. After transfer to seawater, both the diploid and triploid salmon that were given a lowhistidine feed developed more severe cataracts than those that were given the normal level of histidine, and this was again more pronounced in triploid salmon. Diploid

Testing large-scale salmon production Cooperation: Marine Harvest, Skretting AS, AKVAgroup Funding: CAC (Centre of Aquaculture Competence)

smolt that were kept at a low temperature and that were given the normal histidine feed did not develop cataracts in the trial. We found a lower content of the histidine compound N-acetyl-L-histidine (NAH) in the lenses of triploid salmon than in diploids, which is related to the severity of cataracts. Since increased levels of histidine in the feed did not reduce cataracts as much in triploids as in diploids, we assume that triploid salmon have a greater need for histidine to maintain eye health after being transferred to seawater. The final project report was published in 2015.

At the Centre of Aquaculture Competence (CAC), NIFES is testing full-scale-production use of feed with reduced dependence on fishmeal and fish oil. The Centre of Aquaculture Competence (CAC) in Rogaland holds a trial licence to test feed and feeding conditions on a full-scale premise, from releasing to harvesting the salmon. This has proven to provide valuable information, which gives research, the industry and its administration a so-called 'proof of concept'. Over several generations of full-scale production, we have gained confirmation that plant ingredients can increasingly be used to replace marine ingredients. The studies have covered production, fish health, product quality, seafood safety and sustainability assessments of the feed and environment. In a large-scale trial in which we reduced the content of the fatty acids EPA and DHA in the feed from 8% (standard feed) to 5% (low omega-3) of the fatty acids (from 26 to 16

grams per kilo feed) throughout the production cycle in the sea, the performance, health and robustness of the salmon were not affected. This was despite practical matters such as several delousing rounds, an outbreak of pancreas disease (PD) and fatalities in connection with gill infection. As expected, the fatty acids in the fillet varied, with 7.5 per cent of the fatty acids (13 grams per kilo fillet) in salmon given standard feed and 5.44 per cent EPA and DHA of the fatty acids (9 grams per kilo fillet) in salmon given low omega-3 feed. Pigment spots can occur in the fish fillet during largescale production, which is not seen in smallscale feeding trials. We cannot say whether it was the reduced content of EPA and DHA or the increased ratio between omega-6 and omega-3 fatty acids that caused the increased incidence of pigment spots in the fish fillet. These spots mean the product is of poorer quality, and they are occasionally seen in commercial production. The feed was also observed to have possible effects on gill infection and pigment deposits, but further studies are required to reach a final conclusion.

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FEED FOR FARMED MARINE SPECIES Difficult to make the change to dry feed for halibut larvae

Making the change from live feed to dry feed for halibut larvae will simplify halibut farming. This is, however, easier said than done. The DIVERSIFY project's strategy is to both improve live feed and to attempt to get the larvae to eat dry feed as early as possible.

European aquaculture is a modern industry that employs 190,000 people and has an annual turnover of over seven billion euros. The EU project DIVERSIFY has identified some species of fish that are thought to have great farming potential and that can contribute to increasing the diversity of farmed fish in Europe. The project focuses on overcoming known bottlenecks.

Norway has focused on halibut, which, though farmed for a long time, still has some bottlenecks that need to be overcome before farming can become profitable. This applies not least to the species' dependence on live feed. NIFES is participating in DIVERSIFY, by conducting research on nutrition for halibut larvae and fry. In 2015, the scientists concentrated on the start feeding of larvae. Halibut larvae need live feed organisms during their first period of feeding. When farmed, they are fed the crustacean Artemia for the first 45 to 50 days. This is normally not as nutritious as the larvae's natural prey, copepod, and this affects the quality of fry. Since copepod are difficult to cultivate in large enough quantities, the scientists have tested out different ways of improving the quality of Artemia. Artemia were cultivated in a nutritious environment for

Halibut fry. 40

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Halibut larvae.

three days rather than just being hatched. This increased the content of a number of nutrients in the prey. In our halibut larvae feeding trial, where newly hatched Artemia were compared with on-grown Artemia, the scientists could not find differences in growth or the quality of fry, because the normal Artemia gave unusually good results. When halibut fry is commercially produced, the quality of fry varies greatly and the industry

Halibut yolk sac larvae.

therefore wants to improve its results by using on-grown Artemia. Since this project attempts to eliminate the dependency on live feed, it is also important to try dry feed early on during the larvae phase. Many trials have been done on this previously, but getting the halibut larvae to make the change to dry feed has proved difficult. In 2015, the scientists set up a tank

Cooperation: Hellenic Center For Marine Research (Hcmr), Greece, Institut De Recerca I Tecnologia Agroalimentaries (Irta), Spain, Fundacion Canaria Parque Cientifico Tecnologico De La Universidad De Las Palamas De Gran Canaria (Fcpct), Spain, Israel Oceanographic And Limnological Research Limited (Iolr), Israel, The University Court Of The University Of Aberdeen (Uniabdn), Uk, Stichting Dienst Landbouwkundig Onderzoek (Dlo), Belgium, The Institute Of Marine Research (Imr), Norway, Instituto Español De Oceanografia (Ieo), Spain, Université De Lorraine (Ul), France , Technische Universiteit Eindhoven (Tu/E), The Netherlands, Aarhus Universitet (Au), Denmark, Asociación Empresarial De Productores De Cultivos Marinos (Apromar), Spain , Universitá Degli Studi Di Bari Aldo Moro (Uniba), Italy, Institut Francais De Recherche Pour L'exploitation De La Mer (Ifremer), France, Universidad De La Laguna (Ull), Spain, Facultes Universitaires Notre-Dame De La Paix De Namur (Fundp), France , The National Institute Of Nutrition And Seafood Research (Nifes), Norway, Fundacion Centro Tecnologico Acuicultura De Andalucia (Ctaqua), Spain, Conselleria Do Mar - Xunta De Galicia (Cmrm), Spain, Skretting Aquaculture Research Centre As (Sarc), Norway , Danmarks Tekniske

system in which the conditions were especially suitable for feeding with dry feed. Following this, several commercial types of feed were tried and one feed was found to give a good feed intake from day 27 after start feeding. In 2016, research will be conducted on how early in the larvae phase this feed can be used.

Universitet (Dtu), Denmark, Sterling White Halibut As (Swh), Norway, Ichthyokalliergeies Argosaronikou Anonymi Etairia (Argo), Greece , Azienda Agricola Ittica Caldoli (Ittical), Italy, Dor Dgey Yam Ltd (Dor), Israel, Vas. Geitonas & Co Ltd Ee (Gei), Greece, Aquaculture Forkys Ae, (Forkys), Greece, Canarias Explotaciones Marinas Sl (Canexmar), Spain, Asialor Sarl (Asialor), France, Culmarex S.A.U. (Culmarex), Spain, Irida Ae-Products For Animal ProductionServices (Irida), Greece, Ayuntamiento De A Coruna (Mc2), Spain, Syndesmos Ellhnikon Thalassokalliergeion Somateo (Fgm), Greece, Bundesverband Der Deutschen Fischindustrie Und Des Fischgrosshandels E.V. (Bvfi), Germany , Hungarian Aquaculture Association (Masz), Hungary, Asociacion Nacional De Fabricantes De Conservas De Pescados Y Mariscos-Centro Tecnico Nacional De Conservacion De Productos De La Pesca, European Food Information Council Aisbl (Eufic), Belgium, Kentro Meleton Agoras Kai Koinis Gnomis Anonimi Emporiki Etairia (Hrh), Greece Funding: EU, FP72

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NEW FISH FEED INGREDIENTS Offcuts as an alternative feed ingredient Cooperation: CRAW (Centre wallon de Recherches agronomiques), Belgium , RIVM (Rijks instituut voor volksgezondheid en milieu), the Netherlands , IUPA (Institut Universitari de Plaguicides i AigĂźes. Universitat Jaume I Avda), Spain, EWOS AS, Nutricontrol, the Netherlands Funding: The Research Council of Norway, the BIONĂ&#x2020;R programme; 227387/E40

By-products from meat production from land animals can be a good ingredient in fish feed. It is, however, essential to avoid beef products. The parts of a slaughtered animal that do not end up on the consumer's table can become processed animal proteins (PAP). This can include blood meal, feather meal or meat and bone meal, much of which is a good source of protein and fat and is very suitable for use in fish feed. Using these animal by-products, however, was banned in Europe following the outbreaks of mad cow disease a few decades ago. The ban on using PAP in fish feed was lifted in 2013, following an EU risk analysis. The exception is PAP from beef, which is still banned. This project assesses the risk associated with

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using PAP from land animals, mainly pork and chicken, in feed for Atlantic salmon. The scientists will screen the by-products for undesirable substances, such as medicine residues from land animals. The scientists will also study the transfer of these undesirable substances from the feed to the farmed fish. They will also use different techniques to examine the possibilities of identifying PAP from beef. In 2015, a total of 19 commercially produced by-products were tested for beef. At least ten per cent of the by-products analysed had traces of beef in them and are therefore illegal to use as fish feed ingredients. In addition, new methods were developed to identify which species the animal by-products come from. The scientists are also working to develop methods to identify which species and which tissue the animal by-products

come from. This will, among other things, be useful for identifying the source of beef contamination in feed. In 2015, medicine residues that are banned or not registered for use in salmon farming were also found. The scientists will conduct trials to determine whether these substances are transferred from the feed to the edible parts of the farmed fish.

Waste as resource

Insects as fish feed

Cooperation: NorInsect AS, Vekst i Stordal Funding: The Research Council of Norway

Cooperation: University of Stirling (UoS), Skottland, Norwegian Institute of Bioeconomy Research (NIBIO), Norwegian University of Life Sciences (NMBU), University of Bergen (UiB), Uni Research, GildeskĂĽl forskningsstasjon (GIFAS), EWOS Innovation, Protix Biosystems BV, Netherlands, National University of Ireland Galway (NUI), Ireland, Universitetet of Barcelona, Spain Funding: Research Council of Norway

Can insect meal and fat become important ingredients in the feed industry of the future?

Meal and fat from insects can become an important ingredient for the feed industry

In industrialised countries, vegetable byproducts are produced from the food industry that are not fit for human consumption. These waste products can be used in several different ways. Some can be used for livestock feed, while others can be composted or used, for example, as biofuel. It has now been found that these waste products can also be used as feed for insects, which can in turn be used as fish feed.

When searching for new raw materials for fish feed, it is important to establish that the ingredients are safe and healthy for the fish. At the same time, they should be sustainable and not be in competition with food for human consumption. It now seems that insects may be a good alternative. In the project AquaFly we are studying how the nutritional composition of insects may vary with different feeding regimes, and whether insects can be used as a nutritious feed ingredient for salmon.

Insects have the ability to efficiently transform vegetable waste with a high fibre content into fat and protein. Insect fat and protein can in turn be used as a feed resource in the salmon farming industry, as shown in the NIFES project AquaFly. In the Stordal project, we will study which local waste products are available as insect feed in Norway and how the nutritional composition of insects changes when they are fed these products.

We have studied how composition of the diet can influence both the nutrients composition and undesirable substances in the insects. We found that the insects to some degree reflected the kind of fat, minerals and vitamins they had received, such as iodine and vitamin-E. This suggests that it is possible to tailor nutrient composition in the insects to be optimal for salmon. In addition, we found that the transfer of heavy metals and arsenic from diet to insects can limit the use of certain ingredients. Feeding trials with salmon will be carried out during 2016, using insect meal and insect fat.

In this part of the project, we are investigating the use of plant ingredients from the marine environment, for example seaweed, which exist in large quantities along the Norwegian coast. By using marine raw material to feed the insects, nutrients like marine omega-3 and iodine are introduced. However, undesirable substances such as heavy metals and arsenic may also be transferred to the insects through these ingredients.

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Insects as a sterol source for Atlantic salmon Cooperation: University of Stirling, UK and Protix, the Netherlands Funding: Research Council of Norway

Feed trials show that the composition of sterols in insect larvae is affected by what they eat. Cholesterol is essential to many physiological processes. Vertebrates, like humans and fish, produce cholesterol and do not have to get it from their food. Phytosterol, the plant world's answer to cholesterol, lowers the cholesterol level in people's blood. In Atlantic salmon, however, high levels of phytosterol in the diet are linked to increased accumulations of fat in the liver and in the blood. Plant ingredients, which have become increasingly important as a feed resource in recent years, is the salmon's source of phytosterols, while fish oil and other animal oils provide cholesterol. Protein and fat from insects is a potential new source of feed ingredients for farmed salmon and will also change the sterol composition in the feed. In this project, scientists have studied how what the insects eat can affect their sterol composition. The effect of different combinations of sterols on the fish's cholesterol absorption and metabolism has also been studied. 44

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The scientists let three species of insect larvae grow on different types of seaweed. When black soldier flies grew on a standard diet, i.e. a vegetable diet, the insects contained a lot of phytosterols and very little cholesterol. Insects cannot produce cholesterol on their own, and need a sterol source in their diet. Planteating insects do not get much cholesterol from the food they eat, and some of these insects have therefore developed the ability to turn phytosterol into cholesterol. It is thought that other insects are able to use phytosterols rather than cholesterol in many biological processes. However, the scientists saw no sign of the black soldier flies being able to produce cholesterol, and the cholesterol probably only came from what the larvae had eaten. When ground seaweed was mixed into the diet, the amount of cholesterol in the insects increased. The amount and type of sterols in black soldier flies that had eaten seaweed was quite similar to that in the seaweed flies. Most of the sterol in the insects that had eaten seaweed was fucosterol, which there is a lot of some types of seaweed.

The effect of different phytosterols on fish has been studied using a cell model of liver cells from salmon. The cell trial has indicated that proteins that are important to the regulation of the gene expression in the cells were affected by phytosterol, and that this may mean that different sterols are capable of affecting the lipid metabolism in the cells. In a new feed trial with salmon, fat and protein from black soldier flies will be used in the fish feed. The results of this trial will provide new knowledge about insects as a source of sterols for Atlantic salmon.

THE RIGHT FEED FOR FARMING LOUSE EATERS The genome of the ballan wrasse has been mapped Cooperation: Centre for Ecological and Evolutionary Synthesis (CEES), University of Oslo Funding: The Norwegian Seafood Research Fund

Scientists at NIFES have now mapped the DNA of the louse-eating ballan wrasse. They combined two different methods to solve this great puzzle. Ballan wrasse is a relatively new species in fish farming and is used as a louse eater to combat the problem of salmon lice. Scientists at NIFES have now also sequenced the fish's genome. This means that they have mapped the total DNA of the species. The starting point for the genome sequencing was a lucky ballan wrasse selected from a fish farming facility in Ă&#x2DC;ygarden. It was sequenced by combining two technologies. This was done because the two technologies cover each other's weaknesses. The first method of sequencing (Illumina) can take readings of many, but short chains of nucleotides. Nucleotides is the appellative for the four types of molecule that code a genome. Illumina sequences are also very reliable since they contain very few mistakes. The other method (PacBio) produces very long readings of

the genome's nucleotides, but it is more often incorrect. It is assumed that between 10 to 15 per cent of the nucleotide readings taken will be incorrect. When all these pieces of a genome are in place, the work on solving the puzzle begins. The pieces have to be put together. To do this, the genome has to be sequenced many times. In this project, the scientists sequenced the whole genome 111 times using the Illumina method and 33 times using the PacBio method. In this way, the pieces of the sequence overlap many times. It is then possible to use computer programs to put the pieces together and pick out the nucleotides that have been incorrectly read. The completed map of the ballan

wrasse genome is 805 megabases, a commonly used unit of length for DNA. This is in the same magnitude as the genomes of cod, medaka and tilapia. For the sake of comparison, the genome of salmon is assumed to be around 3,000 megabases. The project has also demonstrated that it is possible to produce a complete, complex and commented genome for a 'new' species of fish in a relatively short period of time and at an increasingly lower cost. Now that we have knowledge about this genome, we can identify the genes and variations in genes that control the fish's physiology, immunology and metabolism and use these to study the fish's nutrition and welfare.

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Feed for the fish without a stomach Cooperation: Nofima, the Norwegian University of Life Sciences (NMBU), the University of Bergen, Department of Biology, Marine Harvest AS Funding: The Research Council of Norway

Farming ballan wrasse to combat lice can be a challenge since it does not have a stomach. Despite ballan wrasse being the most sought after louse eater in salmon farming, it is a relatively new species in the fish farming industry. We have therefore known little about the intestine's structure and how the various nutrients are digested and absorbed. This knowledge is important if we are to find the optimal feed. Norway has broad experience of farming a number of different species of fish. They share many common traits and

their digestive systems tend to be relatively similar. Ballan wrasse, however, is different from other farmed species in that it has no stomach and only a simple intestine without blind pouches. The intestine is also quite short, only two thirds of the length of the fish. We are currently studying how a digestive system of this type works. The first factor we studied was how quickly food passed through the intestine and where the food was in the intestine during its journey. The first part of the intestine is wider than the rest, so one theory was that this part

Intestine of the ballan wrasse.

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worked as a storage space for food to be digested. We marked the feed with an inert mineral and took samples from the fish's intestine at two-hour intervals, starting four hours after feeding. Surprisingly, we found that the feed was almost at the very end of the intestine after four hours. Despite this 'sprint' to the end of the intestine, it took between ten and fourteen hours before the feed left the intestine in the form of excrement. The next stage is to find out which chemical signals in the feed and hormones in the body control the peristalsis.

Abundance of nutrients in commercial fish feed Cooperation: Akvaplan-niva, Nofima Funding: The Norwegian Seafood Research Fund

In order to study whether the nutritional requirements of lumpfish in captivity are met, we have compared the nutritional content of their eggs with eggs from wild fish. We found that farmed fish had an abundance of nutrients. Broodstock are sexually mature fish that produce roe and milt and are the parents of the next generation. It is important that it is given nutritious and balanced feed, because the broodstock feed forms the basis for the nutritional content of the eggs, which is the foundation for foetal and larval development and growth. Good treatment of broodstock forms the basis for good growth and development in offspring up until full maturity. In this project, we have studied whether all the nutritional needs of lumpfish broodstock are met when they are fed commercial fish feed. We compared the levels of nutrients in eggs from fish that have been kept in captivity and fed, with the levels in eggs from newly caught wild fish, which we assume to have a good

nutritional profile. We have also analysed feed samples. There was a big difference between the nutritional content of eggs from wild and farmed lumpfish. The eggs from farmed fish contained the same amount of protein but more fat. They also had more of a number of other nutrients, such as free amino acids, vitamins and minerals than the eggs of wild lumpfish. The result was surprising, but may be related to the fact that wild lumpfish eat a lot of jellyfish, which are assumed to be quite low in nutrients.

viability of eggs and fry. More research is needed in this area. The results represent a step towards developing feed adapted to lumpfish. This will make lumpfish farming more efficient and subsequently, salmon farming, which uses lumpfish as louse eaters.

It is hard to say how an abundance of nutrients affects the

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PHARMACEUTICALS FROM FISH FARMING Delousing agents affect the genes of lobsters Cooperation: The Institute of Marine Research Funding: The Ministry of Trade, Industry and Fisheries

A study reveals that non-fatal doses of teflubenzuron affect genes that are important for detoxification, stress handling and moulting in lobsters. Salmon lice are a common parasite in farmed salmon and are a major problem for the industry. Many different chemicals are used to treat salmon lice. In recent years, the use of so called flubenzurons, such as diflubenzuron and teflubenzuron, has increased, since salmon lice have become resistant to other substances. In collaboration with the Institute of Marine Research, NIFES has previously shown that these substances can be found in organisms living near fish farming facilities after the salmon have been medicated. Flubenzurons are particularly harmful to crustaceans, since the substances have a negative effect on moulting. In connection with a trial carried out by the Institute of Marine Research, NIFES examined the gene expression in claws as a result of teflubenzuron treatment. Through its feed, the lobster was exposed to concentrations of teflubenzuron that 48

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can be found in the areas around fish farming facilities. The study focused on how the delousing agent affects the expression of genes that are important for stress handling, detoxification and moulting. As a result of exposure, a small number of lobsters had damage to their claws and legs, such as deformities and stiff legs, while mortality was generally low. The lobsters received a concentrationdependent accumulation of teflubenzuron, with a 2.7 times higher concentration in the high-dose group than in the low-dose group. Significant changes in gene expression were found in 21 of the 39 genes examined. The study therefore showed that non-fatal doses of teflubenzuron affect genes that are important for detoxification, cellular stress and moulting. The trial indicates that flubenzurons, even at the low concentrations that can be found in the

environment, can have a negative effect on crustaceans that have habitats around the fish farming facilities. In 2015, the scientists also conducted follow-up studies on teflubenzuron and shrimps, which will continue in 2016. This research will improve our knowledge about the negative effects of using delousing agents in the aquaculture industry and give us useful information about biomarkers for exposure to flubenzurons in crustaceans which can be used in subsequent monitoring.

ARE FISH PRODUCTS WHAT THEY CLAIM TO BE? New DNA methods can reveal fish fraud

High levels of undesirable substances in certain marine oils

Cooperation: Centre for Ecological and Evolutionary Synthesis (CEES), Department of Biosciences, University of Oslo Funding: The Ministry of Trade, Industry and Fisheries

Cooperation: The Institute of Marine Research Funding The Norwegian Food Safety Authority

Are the correct species of fish named on the fish products you buy at the shop? Scientists at NIFES are developing new analysis methods that will be able to do exactly that.

Dioxins, dioxin-like PCBs and PCBs were found in levels that far exceeded the maximum limits in two chimera oils and one shark liver oil.

Studies from several countries have shown that fish products sold in shops can contain fillets from species other than those listed in the product description. In order to determine which types of fish are used in fish products on the Norwegian market, NIFES scientists are attempting to develop methods based on DNA identification. Such methods will provide the authorities with a tool that enables them to determine whether fish that is imported and sold in Norway is correctly labelled and is what it is supposed to be. In 2014 and 2015, NIFES established a new method for identifying the most frequently sold types of fish in Norway. The method can identify which species the ingredients in pure fish products (products made from only one species) come from.

NIFES is also working on a method to determine both the species and the amount of each species in mixed products, such as fish cakes and fish balls, where the composition of species is given as a percentage or is unknown. This has proved to be more of a challenge since the fish products sold in Norway can potentially be made up of fillets from tens of species. A method based on accumulation of a mitochondrial gene, followed by nextgeneration sequencing, was tested, but did not produce the desired result. The method could, to a certain extent, identify individual species in the products, but did not provide a good estimate of the proportions of the different species. In 2015, so-called shotgun sequencing was also tested. This is the quickest method of mapping genes and the work continues on analysing these data.

Every year, NIFES studies the content of undesirable substances in ten different marine oils sold as human food supplements. The results for the oils that were purchased in 2014 showed that the levels of undesirable substances varied a great deal. Three of the oils, two chimera oils and one shark liver oil had extremely high levels of dioxins, dioxin-like PCBs and non-dioxin-like PCBs, with levels that far exceeded the maximum applicable limits for these undesirable substances in the EU and Norway. One salmon oil also had high levels close to, but not exceeding, the maximum limits. As found in previous years, the other oils had low levels of contaminants well below the maximum limits for food safety. The results were reported to the Norwegian Food Safety Authority, which took action against the producers of the oils that exceeded the limits.

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SAFE SEAFOOD IS BASED ON KNOWLEDGE What are we actually eating?

Salmon

Herring

Scallop

The seafood industry is important for Norway, and we need to know that seafood is safe, both what we eat ourselves and what we export. We therefore need indepth knowledge about the levels of contaminants and infective agents in seafood. We analyse and report the real levels to be able to determine whether they breach Norwegian and international regulations, and, if relevant, pose a health hazard. This is part of the risk assessment carried out for seafood. The experience Norway and NIFES has acquired through research and management has great national and international significance, also for developing countries that wish to increase their exploitation of marine resources.

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MONITORING: BASELINE STUDIES Mapping undesirable substances in haddock Cooperation: The Institute of Marine Research Funding: The Ministry of Trade, Industry and Fisheries

Haddock from Norwegian waters contained little mercury, but a relatively large amount of arsenic. The first arsenic studies suggest that this concerns organic arsenic compounds, which are non-toxic. There are clear differences between geographical locations, the size of the fish and the seasons.

makes haddock the fifth most important commercial species of wild seafood, after cod, herring, mackerel and saithe. Haddock is caught in all Norwegian waters, but mainly in the north, between Troms/Finnmark and Svalbard. Because haddock is largely a bottom dweller, NIFES will compare the results of the analyses with sediment analyses to look for possible correlations.

A thorough mapping of substances that are harmful to health, such as heavy metals, arsenic, PCBs, dioxins, furans, flame retardants, polyaromatic hydrocarbons, perfluorinated substances and pesticides, was carried out on fillets and livers from more than 1,000 haddock in 23 different areas of the North Sea, the Norwegian Sea and the Barents Sea. Fish were collected from all of the stations in autumn, but they were also collected earlier in the year in ten of the areas, before and after spawning.

Since 2006, NIFES has carried out a series of baseline studies on the commercially important species, cod, herring, mackerel, saithe and Greenland halibut, and as a rule, each study includes more than 1,000 fish. The studies are unique in their scope and form a sound basis for assessing food safety. In this way, we can tailor further follow-up of areas of particular concern over time and in connection with emissions. Baseline studies are therefore of interest to consumers, fisheries management, the fisheries industry and environmental monitoring.

date is 0.15 mg/kg sampled off Lofoten in summer, and the lowest is 0.02 sampled off Finnmark in winter. These levels are well below the maximum limit for mercury, which is 0.5 mg/kg. Relatively high levels of arsenic have been found in both liver and fillets. The arsenic levels in the fillets was significantly higher during winter than summer. The first arsenic studies suggest that this concerns organic arsenic compounds, which are non-toxic. Cadmium levels were low in fillets, but somewhat higher in liver. Lead levels were low in the whole fish. So far, all the levels of heavy metals are well below the EU's maximum limits, where they exist, for food.

Haddock is a lean, white fish related to cod, and it is a common ingredient in fish cakes, fish balls and fish and chips. Today, haddock makes up about 1/13 of the total revenues from Norwegian fisheries, with over 100,000 tonnes of catch annually. This

More than 300 haddock from the Norwegian Sea and the Barents Sea have been analysed so far. Mercury levels were consistently low and were significantly lower during winter than summer, both in fillets and liver. The highest average level in fillets to

PFAS was found in haddock fillets, but in very few fish, which were from the Lofoten area. Other organic contaminants have not yet been studied. During 2016 and 2017, NIFES will analyse all of the fish and the final results will be reported in 2017.

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Analyses of undesirable substances in Atlantic halibut Cooperation: The Institute of Marine Research Funding: The Norwegian Food Safety Authority

The preliminary results of the mapping of Atlantic halibut indicate that the level of undesirable substances is generally low in this species, but levels above the maximum limit have nonetheless been found in some fish. In the period 2013-2016, NIFES will be carrying out a comprehensive baseline study to determine the levels of contaminants in Atlantic halibut. A total of 400 Atlantic halibut from various areas along the entire

Norwegian coast will be analysed for metals, perfluorinated alkyl substances (PFAS) and slowly degradable organic undesirable substances such as dioxins, PCBs and brominated flame retardants (PBDE). At the end of 2015, samples have been collected from around 390 halibut of all sizes weighing from 1 kg to 225 kg. Analyses of fillet samples from the halibut are still in progress, but the preliminary result indicates that both heavy metals and organic undesirable substances in the halibut fillets are well below the maximum limits for these

undesirable substances. However, a few fish, especially some of the biggest, have been found to contain levels of mercury and/or organic undesirable substances that exceed the maximum limits for fish fillets for human consumption. Halibut has a higher maximum limit for mercury than most other fish. It is the thickest part of the halibut fillet that has the highest level of organic undesirable substances. Around 15 per cent of the analysed fish had levels of dioxins and dioxin-like PCBs in the thick part of the fillet that were above the maximum limit for food safety, while only 1.8 per cent of the fish had levels above the maximum limit in the thin part of the fillet. The study of Atlantic halibut is part of a comprehensive baseline study that maps undesirable substances in several fish species that are important to Norwegian fisheries management. The goal is to establish basic levels for undesirable substances in the most important commercial fish species in Norwegian waters, as a basis for risk analyses and an adequate monitoring regime for Norwegian seafood.

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Monitoring our most important fish species Cooperation: The Institute of Marine Research Funding: The Ministry of Trade, Industry and Fisheries

The levels of undesirable substances in fillets of cod, saithe, mackerel, Norwegian spring-spawning herring and North Sea herring are low, while the levels of organic pollutants in cod and saithe liver can be high. Lower levels of organic pollutants were found in Greenland halibut than previously. The results are from the 2015 annual monitoring of a number of fish species from Norwegian waters. Since 2006, NIFES has carried out six baseline studies to map the content of undesirable substances in our most important species of fish. To date, extensive baseline studies have been conducted for Norwegian spring-spawning herring, North Sea herring, Greenland halibut, mackerel, cod and saithe. The results of these studies have formed the basis for risk analyses and more focused monitoring. Further monitoring of undesirable substances will be conducted for all species for which a baseline study has been carried out. The baseline study for Greenland halibut found challenges relating to the content of dioxins and dioxin-like PCBs in certain areas. As a result of this, two areas off the coast of 54

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Nordland have been closed to fishing Greenland halibut since 2012. Monitoring is therefore carried out every year to obtain new and updated knowledge about the levels in Greenland halibut in these areas in particular. In 2013, 2014 and 2015, Greenland halibut was sampled from 15 stations along the continental shelf slope between 65°30’N and 68°30’N. This includes three stations in the two fishing grounds closed to Greenland halibut fishing. Fillet samples from the fish were analysed for heavy metals and the organic pollutants dioxins, dioxin-like PCBs, PCB7 and brominated flame retardants. The results from each of the three years showed that the levels of dioxins and dioxin-like PCBs in Greenland halibut were clearly lower than in previous years. Despite great variation in the levels from year to year in previous studies, NIFES believes, based on the past three years of monitoring, that the level is now at a stable level below the maximum limit for food safety. NIFES therefore believes that there is no longer a scientific basis for maintaining the closure of the areas that have been closed since 2012 and has reported its conclusion to the Norwegian Food Safety Authority and the Directorate of Fisheries.

In 2014, samples of cod were collected from the Barents Sea, the Norwegian Sea and the North Sea, and samples of saithe were collected from the Norwegian Sea and the North Sea. The analyses of these samples was completed in 2015 and the results showed, as previously, that the content of heavy metals and arsenic was at a similar level to the baseline studies for these species. As before, the level of heavy metals in the fillets was low, and the average levels were well below the maximum limits for food safety for these contaminants in fish fillets. Three of the 125 saithe analysed had mercury levels that exceeded the maximum limit, but no cod had levels of heavy metals above the maximum limits for food safety. The content of organic pollutants in cod liver from the Barents Sea was somewhat lower than in the baseline study, while the level in saithe liver was somewhat higher than in the baseline study, both in the North Sea and the Norwegian Sea. As in the past, it was found that the levels of dioxins and dioxin-like PCBs in cod liver were relatively high, and somewhat higher than in saithe liver. In the Barents Sea, only 3 per cent of the cod liver samples had levels of dioxins and dioxin-like PDBs that exceeded the maximum limit, while in the North Sea, 48 per cent of cod and 42

per cent of saithe had levels of dioxins and dioxin-like PCBs in the liver that exceeded the maximum limit for food safety. The monitoring continues and in 2015, cod and saithe from the Barents Sea, the Norwegian Sea and the North Sea were collected. The analysis of these samples will be completed in 2016. Mackerel is monitored every year in Skagerrak and every third year in the North Sea. In 2015, analyses were conducted of metals and organic pollutants in mackerel samples collected in 2014 from Skagerrak. The results showed that the levels of both heavy metals and organic pollutants tallied well with the results of the baseline study. The level of heavy metals was very low and much

lower than the applicable maximum limit for fish fillets intended for human consumption, while the level of organic pollutants in mackerel fillets from Skagerrak was somewhat higher and 8 per cent of the samples had levels of dioxins and dioxin-like PCBs over the maximum limits. In autumn 2015, new mackerel samples were collected from Skagerrak, which will be analysed in 2016. Norwegian spring-spawning herring from the Norwegian Sea and North Sea herring are only monitored every third year, since the baseline studies of these species concluded that annual monitoring was not necessary. In 2014, Norwegian spring-spawning herring was collected from two positions in the

Norwegian Sea and North Sea herring from three positions in the North Sea, which were analysed in 2015. The results tallied well with the results of the baseline studies for these species. The level of heavy metals was very low in both Norwegian spring-spawning herring and North Sea herring and the level of organic pollutants was very low and under the maximum limits for food safety for Norwegian spring-spawning herring. Analyses of organic undesirable substances in North Sea herring will be completed in 2016.

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MONITORING OF SEAFOOD Monitoring of undesirable substances in farmed fish Funding: The Norwegian Food Safety Authority

The analyses of Norwegian farmed fish in 2015 showed the applicable maximum limits were not exceeded for pharmaceuticals and undesirable substances. Food safety was assessed as good. In order to check that farmed fish intended for human consumption does not contain residues of undesirable substances or legal pharmaceuticals in concentrations that are harmful to health, or that illegal pharmaceuticals are not used to treat farmed fish, Norway has implemented a control system in accordance with the EU's guidelines (Directive 96/23). The Norwegian Food Safety Authority is responsible for the Norwegian system and NIFES conducts the analyses of the farmed fish. The samples analysed for illegal substances are collected at different stages of the fish's life cycle, whereas samples analysed for residues of legal pharmaceuticals and undesirable substances are collected from harvesting facilities and are representative of fish ready for human consumption. In 2015, a total of 2,457 pooled samples were taken. Each pooled sample comprises five fish from the same cage, and the result is deemed to be

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representative of that cage. Levels above the applicable maximum limits for undesirable substances were not found. Several of the substances, such as dioxins, dioxin-like PCBs, PCB-6, mercury, arsenic, DDT and certain brominated flame retardants have showed a downward trend over the past 12 to 14 years. This is mainly due to a change in the feed, whereby traditional ingredients are increasingly being replaced by vegetable ingredients that contain less undesirable substances than that normally found in fish oil and fish meal. A total of 944 pooled samples were analysed for legal pharmaceuticals.

Antibiotics or pharmaceuticals used to treat intestinal parasites were not found in any of the samples. Residues of the delousing agents emamectin, cypermethrin and diflubenzuron were found in a number of samples. All of the results were below the applicable maximum limits and there is therefore no risk to food safety. Forty-four samples were analysed for cypermethrin, 133 for emamectin and 128 for diflubenzuron. Cypermethrin was found in seven pooled samples. The highest measured level was 21 ng/g, while the applicable maximum limit is 50 ng/g. Emamectin was found in eight pooled samples. The highest measured level was 32 ng/g, while the maximum limit is 100 ng/g. Diflubenzuron residues were found in six of the analysed

No Anisakis in Norwegian farmed salmon Cooperation: The Norwegian Seafood Research Fund, the project's respective reference groups, the Norwegian Food Safety Authority Funding: The Norwegian Seafood Research Fund

pooled samples. The highest measured level was 14 ng/g, while the applicable maximum limit is 1,000 ng/g. A total of 883 pooled samples were analysed for illegal substances. Crystal violet, a substance that inhibits the growth of bacteria and fungi, was found in six pooled samples, all of which came from the same facility. One sample was analysed as a routine analysis, while the others were analysed due to crystal violet being found in the first sample. When crystal violet is used to treat salmon, the substance is quickly converted into leuco crystal violet in the fish, but this was only found in very low levels in the samples analysed. Crystal violet was not found in the samples collected at the harvesting facility three months later. The findings of crystal violet were reported to the Norwegian Food Safety Authority, which concluded that the samples were contaminated during the sampling process at the fish farming facility. Based on these results from 2015, food safety is deemed to be good for farmed fish. This is in line with the results from previous years.

The risk of salmon produced in Norway containing Anisakis is very low. Anisakis is the common name for the larvae of a group of parasitic roundworm very commonly found in wild saltwater fish. If a fish that has not been properly prepared (heated/frozen) is eaten, live Anisakis from the fish can cause acute gastrointestinal illness. The current EU and EEA regulations conclude that wild fish to be consumed raw must be frozen at a temperature of minus 20Â°C for 24 hours prior to consumption. For a number of years, Norwegian farmed salmon has been exempt from the freezing requirement. The grounds for the exemption was that the fish were only fed heat-treated dry feed that cannot contain live parasites. The need for a nationwide study of the Anisakis situation in both salmon and rainbow trout was identified following findings of Anisakis in so-called 'loser fish' (salmon) from a farming facility. 'Loser fish' are fish with very poor growth, which are therefore small and thin, and they are filtered out early in the harvesting process.

in 4,184 farmed salmon. The facilities from which samples were collected are representative of all of the salmon-producing counties. All of the fish were tested for Anisakis using a method known as the UVpress method. No Anisakis were found in salmon intended for human consumption. Five Anisakis were found in three loser fish from facilities in Southern or Western Norway. A corresponding study of rainbow trout is also underway, and 345 fish have been tested so far. In 2016, a further 690 rainbow trout will be analysed, with a ratio of 80 per cent fish intended for consumption and 20 per cent loser fish. The study concludes that the probability of salmon produced in Norway containing Anisakis is very low.

In an extensive nationwide study of salmon, we have mapped the occurrence of Anisakis Loser fish at the bottom. SEAFOOD SAFETY

Studying the occurrence of Anisakis in wild fish in the European market Cooperation: PARASITE project consortium and WP2 partners Funding: The EU

NIFES has led an extensive study on the occurrence of parasites that may be harmful to health, in 15 of the commercially most important fish species from the biggest European fishing grounds. NIFES was one of the main institutions that took part in a recently completed EU project, 'Parasite risk assessment with integrated tools in EU fish production value chains', or 'PARASITE'. Certain common parasites found in wild marine fish can affect food safety and the aesthetic quality of a product. An example is the larvae of roundworms called Anisakis. The main aim of the project was to collect data supporting the development of efficient administration and control measures linked to the occurrence of parasites that are harmful to health in fishery products sold in Europe. The project involved 15 R&D partners and six European fishery companies, including representatives from Norway. NIFES led the work on mapping the occurrence of parasites in 15 of the commercially most important fish species from

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Fish meat samples are flattened and lit by UV-light searching for anisakis.

the big European fishing grounds, including the North Sea and the Barents Sea. Within the framework of the work package, we also took a closer look at the occurrence of parasites in several of the most common species of fish for sale in Europe. These included farmed catfish, or Pangasius, from Vietnam and tuna from the Philippines. We have collected large quantities of data as a part of the work package that NIFES was responsible for. The preliminary results suggest great variation in parasite loads between the

different species of fish, but also between different groups of sizes of the same species of fish. This means that larger and older fish, as a rule, have a greater Anisakis load than smaller, younger fish. The fishing ground however appears to be of less significance to the Anisakis load. We only found minor differences, for example, in mackerel of approximately the same size, which were fished in the North Sea and around the Faroe Islands.

Salt tolerance in Anisakis Cooperation: The Norwegian Seafood Research Fund, the project’s consultancy group, SINTEF Fisheries and Aquaculture Funding: The Norwegian Seafood Research Fund

Reducing the quantity of salt increases Anisakis' survival in marinated herring products. Anisakis is a genus of parasites that we find in wild fish in the sea. The most common type of Anisakis found in saithe, herring and mackerel etc. is Anisakis simplex. The survival of Anisakis larvae in the production of marinated herring has proven to depend most on the concentration of salt in the marinade. More knowledge is therefore needed about the salt tolerance of Anisakis. This is important information if a recommendation is to be issued for less salt to be used in marinated herring. The goal of this project is to determine and document whether a reduced or altered salt content in the marinade affects the survival of Anisakis in marinated herring products. Two types of trials were set up. One was a petri dish trial with Anisakis larvae in different concentrations of sodium chloride (NaCl) and potassium chloride (KCI). The second was a trial with a known quantity of Anisakis larvae in bits of herring marinated in marinades with

different concentrations of NaCl and KCI, and at different temperatures (-2°C, 0°C and 4°C). The main finding from the trials was that Anisakis larvae survive longer when the salt concentration is reduced. Potassium chloride was also found to cause greater Anisakis mortality than sodium Anisakis proven in UV-lit samples. chloride. In the trial using bits of for quite a while at high salt concentrations, marinated herring, the temperature had a but the question is whether they can still greater effect on the survival/mortality of spread infection. Further trials are needed to Anisakis larvae than the salt concentration. At determine this with certainty, in which 4°C, more Anisakis survived over a longer infectivity is tested using mouse trials, for period than at -2°C, irrespective of the salt example. concentration. New trials are under way, in which we test five different marinades stored at different Anisakis larvae have been shown to survive

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Monitoring of parasites and hygiene in the wild fish sector Cooperation: The Norwegian Seafood Research Fund, the Norwegian Food Safety Authority Funding: The Ministry of Trade, Industry and Fisheries

Monitoring helps to maintain a focus on good hygienic standards and the food safety of Norwegian pelagic fisheries products. Every year, NIFES studies the microbiological quality of and potentially harmful infective agents in the most commercially important types of fish caught by the Norwegian fisheries fleet and sold for consumption. The studies are organised as a national monitoring programme. In general, samples are collected and fish are prepared for subsequent analysis, on board the fishing vessels. This means that the original catch and storage conditions are reflected in the samples. In some cases, we follow the fish through the entire production chain, from catch and temporary storage on board, to bringing it ashore and further processing at

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the relevant fish processing facilities. The main objective of the monitoring programme is to obtain epidemiological basic data for the incidence of parasites and microorganisms. Particular emphasis is placed on parasitic roundworm (Anisakis) and potentially harmful bacteria such as Listeria monocytogenes, in the commercially most important types of fish for the Norwegian fisheries. On the background of this basic data, we will be able to identify possible changes or tendencies in the incidence of the aforementioned infective agents from year to year, which will in turn form a basis for providing advice to the public administration and the industry. One of the activities is investigating whether, or to what extent, Anisakis can migrate from the intestines to the flesh of herring and mackerel during storage

in the refrigerated tanks on board vessels prior to delivery of the catch. Initial trials suggest that Anisakis can migrate in the fish after it is dead (known as post-mortem migration), and that this is widespread in both types of fish. Our findings may form a basis for issuing a recommendation to the industry that fish should be stored for as short a period as possible between catch and further processing to reduce the risk of Anisakis in the end product. A sterile sample of Anisakis will be obtained for further work on the issue of bacterial contamination of fish due to Anisakis infections. The DNA will be isolated and sequenced using deep sequencing in order to characterise the bacterial community.

General management plans of the seas Cooperation: Sampling is organised by the Institute of Marine Research through their own research cruises or fishermen in the Norwegian Reference Fleet. A number of public administration institutions and research institutes are involved in following up the management plans. The monitoring group is chaired by the Institute of Marine Research, and The Management Forum for Norwegian Sea Areas is chaired by the Norwegian Environment Agency. Funding: The Ministry of Trade, Industry and Fisheries

NIFES is monitoring undesirable substances in seafood from the Barents Sea, the Norwegian Sea and the North Sea, as part of the work on management plans for Norwegian waters. The management plan work is a linchpin in Norway's work on the marine environment. In Spring 2016, the Marine Ecosystem Monitoring Group (The Monitoring Group) published a status report on the environment in the North Sea and Skagerrak, in which NIFES had a special responsibility for seafood safety. The monitoring of a number of indicators in the North Sea and Skagerrak showed that seafood from these areas is generally safe to eat. The exception is cod liver, which in a great many cases, and particularly in Skagerrak, has levels of dioxins and dioxin-like PCBs that exceed the maximum limits for food safety.

basis for the continuous assessment of the condition of the ecosystem. NIFES participates in the monitoring group, which coordinates the monitoring work, and in the Management Forum for Norwegian Sea Areas, which is tasked with assessing the condition of the ecosystem and whether the goals of the management plans have been achieved. Each year, the monitoring group publishes a status report for one of the main areas, so that each sea area is reported on every third year. The status reports published by the monitoring group are largely based on monitoring a range of indicators that, together, provide as much information as possible about the condition of the ecosystem. NIFES is

monitoring the following indicator species for undesirable substances: prawn, cod, capelin, Arctic cod, herring, blue whiting, sand lance, Greenland halibut, tusk and European plaice. The results of this monitoring are published on www.miljĂ¸status.no, where they are updated every year or every third year. In 2015, The Management Forum for Norwegian Sea Areas worked a great deal on considering different factors that impact on the sea areas, and finding a common map basis for all of the bodies involved. This was conducted as a project led by Barentswatch and the Norwegian Mapping Authority.

General management plans have been prepared for the Barents Sea and the area around Lofoten (2006), the Norwegian Sea (2009) and the North Sea and Skagerrak (2013). The management plans are followed up through extensive monitoring that forms the

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Examining mercury in tusk Cooperation: The Institute of marine research (IMR) coordinated the sampling, which mainly was performed by fishermen of the reference fleet. Funding: The Norwegian food safety authority.

Tusk and other deep sea species of fish from coastal and fjord regions has higher levels of mercury and persistent organic pollutants than deep sea species from open waters. Previous investigations have shown that the deep sea species of tusk may have relatively high levels of mercury and mercury levels that exceeded the EUsâ&#x20AC;&#x2122; upper limit in some areas. This is especially the case in the fjords and coastal areas in Southern and Western Norway. We know little about other territories where tusk are caught. Therefore, it has been a need for a baseline study to identify levels of mercury and other pollutants in tusk along the Norwegian coast, in addition to some fjords and in the sea. At the same time, it has also been a need of acquiring more knowledge about other species that live in the same areas. This particularly applies to common ling, but other deep sea species that are caught together with tusk and common ling are also interesting. The collection of samples took place from 2013 to 2015, with the majority of the collections being conducted in 2014. In total, 1181 tusks from 52 locations were collected. 796 samples of common ling was 62

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caught in 38 locations. Furthermore, 549 samples of bycatch were also collected. This included species such as haddock, catfish, greater forkbeard, whiting (merlangius), European Pollock, blue ling, European hake, rose fish, rat fish and also some more. Most of the analysis were completed in 2015. Filet of individual fish and pooled samples of liver were analysed for metal, and pooled samples of fillets and liver were analyzed for POPs. The preliminary results shows that tusk in general has higher mercury levels than the others species, except blue ling, rat fish and blackmouth catshark. The results also show that the mercury levels in tusk decreases from south to north and from the fjords to the sea, with concentrations above the maximum limit in some areas. Common ling and other deep sea species

from the same areas mostly have lower levels of mercury. When it comes to organic pollutants, the levels in fillet was very low, whereas not surprisingly quite high levels were found in the liver among most of the species. Species like blue ling, pollack, whiting and common ling had higher levels of organic pollutants in the liver than tusk. The levels of organic pollutants also decreased from south to north and from the fjords to the ocean. This project forms a solid data basis for undesirable substances in tusk and other deep sea species. The government may use this knowledge in order to give the right advice to the whole population or to specific groups in society. Hopefully more thorough analyzes of data can also give us some answers to why the levels differ as much as they do.

Safe seafood near submarine wreck Cooperation: The Institute of Marine Research (sampling coordination) Funding: The Norwegian Coastal Administration

New results suggest that tusk absorb little mercury from the seabed around the wreck off the coast of Fedje. Since 2004, NIFES has monitored mercury in seafood caught in the area around the submarine wreck off Fedje. The results show year after year that mercury concentrations in tusk and crab in the area do not exceed the maximum limit for food safety. In 2015, the Norwegian Food Safety Authority lifted its warning for pregnant and breastfeeding women to avoid seafood from the area. At the end of WWII, the German submarine U-864 was on its way to Japan loaded with mercury, among other things, when it was torpedoed and sunk in February 1945. Split in two with parts of the wreck and some of the load spread over the seabed, the submarine has remained at a depth of 150 metres about three kilometres west of Fedje. High concentrations of mercury were found in sediments around the submarine wreck and NIFES has monitored the mercury content of seafood from the area every year, on assignment for the Norwegian Coastal Administration. In summer 2015, new samples of tusk and

crab were collected from the area near the wreck, and from four nautical miles north and four nautical miles south of the wreck. Tusk fillets and liver and crabs' claw meat and brown meat were analysed for mercury. The NIFES report concludes, as in previous years, that the concentrations of mercury in tusk and crab from the area around the submarine wreck are largely below the EU and Norway's applicable maximum limits for food safety. Only one of the 75 analysed samples of tusk fillets had a higher mercury content than the maximum limit of 0.5 mg/kg, with a mercury concentration of 0.57 mg/kg. The mercury level in tusk caught near the wreck did not exceed the background level for the coast, but is somewhat higher than that found in the open sea. However, brown meat from crabs caught near Fedje show a slightly higher concentration of mercury compared with other areas along the coast. Before mercury is absorbed in organisms and bioaccumulated in the food chain, it is converted from metallic mercury or inorganic mercury into methylmercury. Research assigned by the Norwegian Coastal Administration shows that there is little

methylation in sediment from the area around the wreck. There has been a need, nonetheless, to find out whether the metallic mercury is absorbed by organisms in the area. In 2015, therefore, tusk fillets and liver, in addition to the total mercury, was also analysed for methylmercury. The results suggest that tusk caught in the area near the submarine wreck absorb little of the metallic mercury, both in the fillets and the liver. The previous year's methylmercury analyses results show that crabs probably absorb some of the metallic mercury from the sediment, but that this is not bioaccumulated in the claw meat and is probably gradually secreted.

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Did mercury from the submarine off Fedje spread to the environment? Cooperation: The University of Ghent, Belgium and the Norwegian Coastal Administration Funding: The Norwegian Coastal Administration

New methodology can tell us whether mercury from the wreck of the U-864 submarine off Fedje is spreading to the environment and to seafood. NIFES has monitored mercury in fish and crabs from the area around the wreck of the U-864 submarine off Fedje since 2004. The mercury levels on the seabed around the wreck of the submarine are high, but high levels have not been found in the fillets of tusk or crab claw meat, and only a slightly increased level of mercury has been found in brown crab meat. However, we have not been able to conclusively answer the question of whether the mercury from the submarine enters biological organisms and is spread to the environment. Mercury, like most elements, consists of several isotopes. By isotopes of an element is meant variations of the same element with different quantities of neutrons in the core that thereby result in different masses. The isotope composition in mercury varies according to where the mercury originates from and what types of processes the mercury has been exposed to previously. Using a very accurate measuring method, the distribution of the various isotopes in mercury can be 64

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determined with certainty. The more similar the conditions are between different mercury isotopes, the more likely it is that the mercury comes from the same source. By measuring the distribution of isotopes in different biological samples, in addition to mercury from the wreck, we can attempt to conclude whether the mercury in the biological sample, e.g. a crab, comes from the submarine or from other sources. In this project, NIFES is cooperating with Ghent University in Belgium on mapping the isotope distribution in both metallic mercury from the wreck, in sediments from the seabed around the wreck and in crabs and tusk. It has been necessary to develop a suitable method for analysing stable mercury isotopes in biological material. A PhD student in Ghent has worked on this aspect of the project. The results of the crab samples collected in 2014 are now ready, while work is under way on the analyses of tusk samples collected in 2015. The mercury in the brown meat of crabs near the wreck had a different isotope composition compared with crabs four nautical miles north and south of the wreck. The composition of mercury in crabs near the wreck site was more like the metallic mercury

from the wreck than the mercury in the crabs caught further south and north. The differences were less distinct in the claw meat. This means that analyses of stable mercury isotopes can trace some of the mercury in the brown crab meat caught four miles north and south of the wreck back to the submarine, and that the high level of mercury we find in the brown crab meat probably comes from the submarine. However, it is important to remember that the mercury level was nonetheless low in both claw meat and brown meat, and not an issue with respect to food safety.

The maximum limit for mercury was not exceeded in cod caught in the Oslofjord Cooperation: Sven Marius Hofgaard, fisherman in the Oslofjord Funding: The Norwegian Food Safety Authority

The maximum limit for mercury was not exceeded in any of the 99 cod tested from the inner Oslofjord, but the biggest cod from Frognerkilen/Bygdøy had concentrations above the expected levels. In 2015, NIFES tested 99 cod from five stations in the inner Oslofjord. The stations were situated on the border of the outer Oslofjord, by Håøya, and from there, up as far as Ingierstrand in the east. All the fish were divided into three weight groups per station: under 700 g, over 1,300 g, and those in between. The average concentration of mercury increased with the weight of the fish. Only the highest weight group, over 1.3 kg, and only fish from the area around Frognerkilen, had an average mercury level that just exceeded 0.2 mg/kg (0.24 mg/kg). A total of seven of the 99 fish had levels exceeding 0.2 mg/kg, and of those, five were from the station at Frognerkilen. The fish with the highest level, 0.31mg/kg, from Ingierstrand, weighed 3 kg and was thereby the biggest fish in the data set. The average for all the fish was 0.06 mg/kg mercury and 736 g in weight.

The levels of arsenic, cadmium and lead in fillets was consistently low. Levels of PCBs, dioxins and furans in cod liver was consistently high, and higher than the maximum limit at all stations and in all weight groups. The highest levels of these groups of substances were also found at the station at Frognerkilen, and the lowest levels were from near the outer Oslofjord. The concentrations were nonetheless within the levels otherwise found in the liver of coastal cod. In a previous analysis, cod that was found to have approximately three times as high concentrations as cod from the area around Frognerkilen, did not generally, however, have measurable concentrations in the fillet. There is a general food warning against eating cod liver from our coastal areas.

and PFOS were also found in liver, but with a decreasing tendency up the fjord. This suggests a source outside the inner Oslofjord, further out at sea. NIFES believes that there may be grounds for reconsidering the food warning against eating cod fillets from the Oslofjord. This programme will continue throughout 2016, but the focus will shift to the Årdalsfjord.

Cod liver also had high levels of the flame retardants PBDE and HBCD. These levels increased the further up the inner Oslofjord you went. Perfluorinated compounds, PFOSA

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Good status for Norwegian bivalves Cooperation: The bivalve industry Funding: The Norwegian Food Safety Authority

Norwegian bivalves have low levels of undesirable substances and a good microbiological status. Every year, NIFES checks and monitors microorganisms and undesirable substances in bivalves and other selected shellfish, on assignment for the Norwegian Food Safety Authority. In 2015, mussels, oysters, scallops, horse mussels, softshell clams and sea urchins were collected. Most of the samples were of cultivated mussels. The results for 2015 have yet to be finalised, but the results for 2014 were reported to the Norwegian Food Safety Authority in July 2015 and are mentioned here. The purpose of this monitoring is to document that bivalves harvested for human consumption are not produced in locations that are contaminated by pathogenic microorganisms or contain undesirable substances in levels above the maximum limits. Bivalves can absorb intestinal bacteria such as E. coli or Salmonella, where this is present in the water where they are growing. Analyses of these microorganisms can therefore reveal whether the waters the 66

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shellfish are harvested from are polluted by nearby sewers and whether the bivalves therefore pose a potential health hazard. The occurrence of these microorganisms can also indicate whether the samples contain pathogenic viruses, particularly norovirus. Different areas are classified by their microbiological quality, so it can be determined whether bivalves can be harvested directly for human consumption (A areas) or whether they must be either rereleased or heat-treated before they can be sold (B and C areas). Shellfish can also store undesirable substances to varying degrees. Through the national control programme, the samples were analysed for the content of undesirable substances that exceeded the applicable maximum limits. At the same time, we collect information about the levels of different substances that are normal in different species of shellfish. The samples collected and analysed for microorganism content in 2014 comprised 253 pooled samples of mussels, 27 of oysters, 27 of scallops, five of horse mussels and one sample of pullet carpet shells. Four samples of sea urchins were also submitted. A total of 92 per cent of the 317 analysed samples contained E. coli below

the EU requirement for areas where shellfish are harvested directly for consumption. Salmonella was not found in any of the 50 analysed samples. In addition to samples submitted by the Norwegian Food Safety Authority, a microbiological analysis was also conducted of 148 samples submitted by the industry. In 91 per cent of the samples, the content of E. coli was below the EU's general maximum limit for an A area (equal to or below 230/100 g sample material). In addition, 24 analyses of heavy metals and organic pollutants in mussels, three in oysters, six in scallops, six in pullet carpet shells and two in sea urchins were also conducted. None of the samples contained undesirable substances at levels that exceeded Norway and the EU's maximum limits. One sample of sea urchin submitted by the industry contained low levels of heavy metals. The results are in line with previous findings.

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Spot check-based monitoring of new species and farmed salmon Cooperation: The Institute of Marine Research Funding: The Ministry of Trade, Industry and Fisheries

NIFES has expanded its spot checkbased monitoring of wild fish to include four new species, in addition to more samples of farmed salmon. In this project, NIFES carries out spot checkbased monitoring of contaminants in wild fish and farmed salmon. This monitoring has been carried out for many years, and the results are published on NIFES's Seafood Data

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database website. In recent years, the priority has been to conduct baseline studies of the most common fish species, ahead of spot check-based monitoring. However, the extra funding allocated since 2014 makes it possible to expand the spot check-based monitoring of wild fish to include four new species, in addition to more samples of farmed salmon. The spot check-based monitoring in 2014 targeted pollock, European hake and Norwegian lobster, and two species of

lanternfish. Lanternfish can become an important resource as an industrial fish. Sampling was also planned of bluefin tuna, but this was dropped as no fish were caught in 2014. In 2015 five new species were included; snow crab, Pacific oysters, monkfish, argentine and turbot. Two samples of bluefin tuna caught as by-catch were also purchased from a shop. The analysis results from the spot checks in 2015 are not ready, but will be in 2016. The samples from 2014 have now largely been analysed. The levels of mercury in pollock and European hake are the same as

for cod from fjord areas, while the levels of organic undesirable substances in the hake fillet were low. The levels of cadmium, mercury and organic undesirable substances in whole lanternfish (Mueller's pearlside and Northern lanternfish) were low. Both raw and cooked Norwegian lobster had a low level

of cadmium in the tail meat. The mercury content was relatively high, on a par with lobster, exceeding the maximum limit of 0.5 mg/kg wet weight by 4 per cent. The brown meat of these species, like the brown meat of common crab, had high levels of both cadmium and organic undesirable

substances, but a maximum level has yet to be set for undesirable substances in the brown meat of crustaceans. Analyses of farmed salmon showed similarly low levels of heavy metals and organic undesirable substances as in previous years.

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Monitoring of undesirable substances in seafood from the Vatsfjord Cooperation: Raunes Fiskefarm (sampling) Funding: Jakob Hatteland Resources AS

The results from 2014 show that seafood from the Vatsfjord appears to be somewhat affected by mercury and PCBs, around the same level as numerous other fjords in Western Norway. The content of undesirable substances was the same in 2014 as in 2013. In the Vatsfjord, a small fjord innermost in the Boknafjord in Rogaland, there has been concern that the fjord has been polluted by industry. There has been particular concern about a plant that has dismantled decommissioned oil rigs since 2004. The results of this study show that seafood from the Vatsfjord appears to be somewhat affected by mercury and PCBs, but that the 2014 level is roughly the same as the 2013 level. In 2014, we studied, for the second time, the content of undesirable substances in seafood from the Vatsfjord. Samples of mussels, crab, cod and tusk that were collected in 2014 have now been analysed for various organic undesirable substances and heavy metals. Most of the samples of mussels, crab and

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cod fillets were below the maximum limits for metals and organic undesirable substances, where such limits apply. The average mercury level in the cod fillet was 0.22 mg per kilo. The average level was higher in 2014 than in 2013, which is probably due to more big fish and fewer small fish being analysed in 2014 than in 2013. Cod and tusk livers had overall concentrations of dioxins and dioxin-like PCBs and PCB6 that were above the maximum limits. These levels were, in part, high. On the basis of previous studies, the Norwegian Food Safety Authority has issued a general warning against eating liver from self-caught fish in coastal and fjord areas.

Two out of ten tusk had a mercury content that was above the maximum food safety limit, and the level in 2014 was the same as that measured the year before. It has been shown that tusk can contain a relatively high level of mercury in the muscle, and the concentration in the fish from the Vatsfjord was on a par with previous analyses of tusk from the coast of Western Norway. Crab claw meat had a high level of mercury compared with the background level for the coast, while mussels had low levels. It thus appears that seafood from the Vatsfjord is somewhat affected by mercury and PCBs, around the same level as numerous other fjords in Western Norway.

Monitoring imported seafood Funding: The Norwegian Food Safety Authority

On assignment for the Norwegian Food Safety Authority, NIFES annually studies samples of seafood imported to Norway from countries outside the EU/EEA area. The quality and safety of these products are good overall. In 2014, the Norwegian Food Safety Authority collected 133 samples of seafood imported from countries outside the EU/EEA area. The samples were studied at NIFES using a selection of chemical and microbiological analysis methods. The samples included fish, crustaceans, shellfish, squid, marine oils and composite products. Both species caught in the wild and farmed species were represented, and the majority of samples were from saltwater. With a few exceptions, the analyses of the products collected in 2014 showed that they were of a good quality and that the applicable maximum limits for infective agents and contaminants were not exceeded. One sample of swordfish (Xiphia gladius) and one sample of processed squid (Loligo sp.) had a high level of bacteria. One out of eleven samples contained residues of a

degradation product of the dye malachite green in low concentrations. It is not permitted to use malachite green in connection with fish farming or other food production. The sample in question concerned farmed catfish (Clarias macrocephalus) imported from Thailand. Two out of 52 samples analysed contained dioxins and dioxin-like PCBs that exceeded the applicable maximum limits. Both samples were of squid oil. Norway is obliged to carry out veterinary border controls of goods imported to the EU/EEA area from countries outside this area. The veterinary border control is meant to uncover goods of poor quality or that contain microorganisms, parasites or undesirable chemical components that pose a risk to food safety. On assignment for the Norwegian Food

Safety Authority, NIFES is responsible for carrying out analyses and professional assessments and for preparing annual reports for the part of this work that concerns seafood. If the results of the analyses show that the applicable maximum limits have been exceeded, this is reported to the Norwegian Food Safety Authority. The analysis activity is also summarised in annual reports. This monitoring programme continued in roughly the same scope in 2015, but the results have yet to be completed.

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Undesirable substances and microorganisms in cultivated kelp Cooperation: The Royal Norwegian Society for Development leads the project. In addition to NIFES, the other partners are SINTEF Fisheries and Aquaculture, NOFIMA AS, Værlandet Fiskeredskap AS, Nikøy Eigedom AS, Seaweed AS and Geir Agnar Landøy Havella Funding: SINTEF Fisheries and Aquaculture, NOFIMA AS, Værlandet Fiskeredskap AS, Nikøy Eigedom AS, Geir Agnar Landøy Havella, and the Royal Norwegian Society for Development

The project studies the risk factors in cultivated kelp in Western Norway. In 2015, samples collected from the first production of sugar kelp, winged kelp and sea girdle by the company Seaweed AS were analysed by NIFES for microorganisms, iodine and other minerals as well as heavy

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metals. The first results show no incidence of potentially pathogenic bacteria or pollution indicators in the form of excrement. There were also low levels of heavy metals, while the level of iodine in sugar kelp and sea girdle were high, on a par with that found in wild kelp. There were lower levels of iodine, however, in winged kelp.

Further analyses of cultivated kelp are planned in 2016. We are also conducting a rat trial, in cooperation with the Technical University of Denmark. The goal is to study the bioavailability of iodine and cadmium in mammals.

Good quality of frozen and defrosted cod fillets Cooperation: SINTEF Fisheries and Aquaculture (head of project), IRIS AS Stavanger dept., Matís (Iceland), Røe Communication, the Norwegian University of Science and Technology (NTNU), Faculty of Engineering Science and Technology Funding: The Research Council of Norway, the BIONÆR programme

Controlled methods for freezing and defrosting cod ensures good quality fillets and can help to meet the industry's wish for year-round access to high-quality ingredients. Stable access to high-quality ingredients throughout the year is a challenge for the land-based industry. This is an issue, in particular, for seasonal fisheries such as the spawning-cod fisheries. Optimised procedures that can be used to freeze the fish during the fishing season, which can then be defrosted at any time during the year, will help to address this challenge. In this project, studies have focused on optimising the freezing and defrosting of white fish. In order to optimise the defrosting processes, several groups of frozen fish were defrosted using different defrosting methods. Two such defrosting trials have been conducted to date. In the first trial, Atlantic cod (<1,000 grams), caught in the Barents Sea by commercial trawlers, were frozen in blocks before rigor mortis set in. While frozen, the fish go through rigor mortis, which is important for maintaining the quality of the fish to be defrosted. At -35°C, it took eight weeks for the fish to go through rigor mortis. At -20°C,

it took five to six weeks for the fish to go through rigor mortis. In this trial, three different defrosting methods were tested; in aerated water, in non-aerated water and in a plate defroster (converted plate freezer). After defrosting, the fish was filleted and stored at 0°C. The temperature was logged during defrosting, and a number of different quality parameters were studied after defrosting, including chemical analyses, microbiological analyses and the Quality Index Method. QIM is a method in which the quality of raw fish is assessed and graded in accordance with a number of criteria. The overall grade is the quality index score, and it gives an indication of the storage time and how well the fish will keep. The QIM parameters include texture (natural/soft), colour (white/yellowish), smell (fresh sea smell /fish smell/sour), gaping (no gaping/pronounced gaping and disrupted). Microbiological analyses of the fish included germ count (how many bacteria there were), the number of H2S producing bacteria (quality reducing bacteria), coliform bacteria and E. coli (sign of pollution linked to excrement) and Listeria monocytogenes (potentially pathogenic). Aerated water gave the best results, while the converted plate

freezer gave the poorest results. In the second trial, the goal was to study how two different temperatures during the defrosting process affected the quality of the cod fillets after defrosting and storage of up to six days. The aerated-water method was used to defrost the fish, and the temperature of the water was set at either 10°C or 0.5°C. When defrosted, the fish was filleted and analysed, as in the first trial, to check whether there were differences in the quality of the fish after using the two different defrosting methods. As expected, the fish defrosted quicker at 10°C than at -0,5°C, but no significant differences in quality were found between the two groups. In order to identify any differences, the fish was stored for 10 to 14 days, as in the first trial. No findings were made in terms of food safety parameters of Listeria monocytogenes in any of the analysed samples. Both defrosting methods show that controlled methods for freezing and defrosting cod ensures good quality fillets and can help to meet the industry's wish for year-round access to high-quality ingredients.

SEAFOOD SAFETY

NIFES supports the Norwegian Food Safety Authority

Cooperation: Bacteriological contaminants; EURL, CEFAS (Centre for Environment, Fisheries and Aquaculture Science), and LERQAP (Laboratoire d’études et de recherches sur la qualité des aliments et sur les procédés agroalimentaires), Parasites, EURL ISS (Istituto Superiore di Sanità), Animal proteins in feedingstuffs: EURL CRAW (Centre wallon de recherches agronomiques), Medical residues in food of animal origin, directive 96/23/EC: EURL ANSES (French Agency for Food, Environmental and Occupational Health Safety), and BVL (Bundesamt für Verbraucherschutz und Lebensmittelsicherheit), and ISS (Istituto Superiore di Sanità), Additives in feed, and heavy metals in food and feed. EURL Joint Research Centre, European Commission: Institute for Reference Materials and Measurements Standards for Food Bioscience Unit, Dioxins and PCBs in feed and food, and Residues of pesticides in food of animal origin. EURL CVUA (Chemisches und Veterinäruntersuchungsamt), Funding: The Ministry of Trade, Industry and Fisheries

NIFES provides expertise to the Norwegian Food Safety Authority.

NIFES provides expertise to the Norwegian Food Safety Authority in areas in which it requires research-based expertise that the organisation itself does not possess. Through this project, the Norwegian Food Safety Authority has access to the expertise of and results from the scientists and laboratories at NIFES. This can be everything from hourly assignments to bigger analysis assignments. A contingency function is also included in the cooperation. This means that NIFES' competence and laboratories are available 24-hours a day, all year, if need be.

SEAFOOD SAFETY

In 2015, NIFES reviewed all of the new food warnings issued by the Norwegian Food Safety Authority in the field of marine food. This included, in 2015, new food warnings for the Oslofjord. NIFES has also developed sampling guidelines for monitoring, with the aim of providing advice about food warnings in polluted fjords and harbours. In order to be able to provide the correct results in an efficient manner, the laboratories must meet the current requirements regarding precision and efficiency. The EU requires that the analysis data used in laboratories in connection with monitoring is based on accredited methods.

In support of the laboratories' work, and of the Norwegian Food Safety Authority's need for expertise in NIFES's field of research, the administrative support project was used to maintain the accreditation status, implement new methods where required, and further develop established methods. The administrative support also covers participation in meetings arranged by EURL, Codex, the Norwegian Food Safety Authority, etc. NIFES also prepares expert statements and risk analyses in the field of seafood. Extra analyses can also be covered where a need has been mapped.

THE EFFECT OF UNDESIRABLE SUBSTANCES IN THE EARLY STAGES OF LIFE Undesirable substances affect bone metabolism Cooperation: The Institute of Marine Research, Centre for Ecological and Evolutionary Synthesis (CEES), The Foundation for Scientific and Industrial Research (SINTEF) , The Norwegian Institute for Water Research (NIVA), National Oceanic and Atmospheric Administration NOAA, USA, The University of Bergen , The University of Plymouth, UK Funding: The Research Council of Norway

Polyaromatic hydrocarbons in crude oil can cause bone deformities in fish. They can also affect bone metabolism. As part of a larger project chaired by the Institute of Marine Research, scientists at NIFES have studied how toxins affect bone metabolism and vitamin A metabolism in haddock. In this project, scientists at NIFES want to find out whether polyaromatic hydrocarbons (PAHs), a group of toxic substances in crude oil, have any effect on bone development and vitamin A metabolism

in the early life stages of haddock. The toxicity of the recognised carcinogenic PAHs has already been established, but the toxicity of the lighter, non-carcinogenic PAHs, however, is less known. These are found in both crude oil and vegetable oils. Scientists are therefore interested in learning more about their effect on fish. New studies on haddock larvae show that chemicals from crude oil cause bone deformities, particularly of the jaw. The same jaw deformities were not observed in older fish. This suggests that the deformities at early

stages of life can be fatal for the fish, which is why it is not found in older fish. An analysis of haddock's gene expression and examination of its bone mineralisation also suggests that the oil components cause delayed mineralisation. The scientists believe that this may be due to oil components affecting genes that are important for the maturation and activity of bone cells. Previous results have also shown that PAHs in vegetable ingredients can affect key processes in bone metabolism, but these PAH concentrations were much higher than those actually found in vegetable oils.

SEAFOOD SAFETY

DEVELOPMENT OF METHODS

The food we eat consists of chemical compounds that can roughly be divided into nutrients and undesirable compounds. An international coordination of the maximum limits for undesirable compounds in food is necessary in order to ensure stable commercial conditions and secure access to safe food for the worldâ&#x20AC;&#x2122;s population. Standard analysis methods are invaluable tools to ensure identical use and enforcement of maximum limits for food and feed. Validated and accredited methods appropriate to the purpose are used to analyse the content of metals and other chemical elements in food and feed. Increasingly stringent requirements of the analyses methods as regards e.g. sensitivity, selectivity, correct data and robust methods mean that the methods must be continuously updated and improved. As the national reference laboratory for pesticides in fish feed and seafood, polyaromatic hydrocarbons, dioxins and PCBs in feed and food, and for metals in both feed and food, it is important that NIFES has extensive practical and scientific experience of the accredited analysis methods that are used. Contingency plans will thereby be available on short notice if a situation should arise where analysis is required.

METHODS

Analysis methods for organic contaminants Funding:

The Norwegian Food Safety Authority

The use of better and more sensitive analysis instruments means that we are constantly discovering new organic contaminants in the food we eat, and in the feed we give to fish and other livestock. The continuous development, updating, rationalisation and maintenance of such analysis methods is therefore important. In order to control the content of organic contaminants in feed and food, we use approved and accredited analysis methods.

METHODS

The analysis instruments are becoming more and more sensitive, which means that more and more substances, found in mere traces, can be analysed in the food we eat. Analytical method work is a continuous process and stricter and stricter requirements apply, including in relation to the methods' sensitivity, selectivity, accuracy and robustness. NIFES has been appointed the national reference laboratory for pesticides in fish feed and seafood, PAHs, dioxins and PCBs in

feed and food by the Norwegian Food Safety Authority. It is therefore important that NIFES has good practical and scientific experience of the analysis methods it applies, so that contingency plans are available at short notice if a situation arises. Due to more comprehensive requirements relating to the determination of pesticides in 2014, it was necessary to procure a new instrument for analysing such compounds. In 2015, the new instrument was installed and work began on developing the method for determining pesticides.

Analysis methods for metals Funding:

The Norwegian Food Safety Authority

To ensure that the methods used in connection with the control of food safety meet increasingly comprehensive requirements, the analytical methods must be developed and updated. When we analyse the content of metals and other chemical elements in food and feed, we use approved and accredited methods that are appropriate to the purpose. The EU's

requirements of the analysis methods' sensitivity, selectivity, accuracy and robustness are increasingly stringent. This means the methods must be continuously updated and improved. NIFES is the national reference laboratory for metals in feed and food, and it is therefore essential that we have up-to-date practical and scientific expertise about the methods applied, so that we can provide technical

and scientific assistance to the authorities as necessary. During the year, a new method for determining inorganic arsenic was approved in order to harmonise the provision with the new European standard method that is nearing completion.

METHODS

New standard methylmercury method soon to be available Funding:

The project is funded by the European Committee for Standardization (CEN) through funding allocated by the European Union (EU)

A new and precise standard method for the determination of methylmercury in seafood will soon be available. The food we eat consists of chemical compounds that can roughly be divided into nutrients and undesirable compounds. Standard methods are invaluable tools to ensure that different countries can analyse the levels of these substances in a given sample, and arrive at the same result. The use of such

METHODS

standard methods means the food authorities can ensure the identical application and enforcement of maximum limits in food and feed both at a national and international level. European maximum limits generally apply in Norway. Methylmercury is an undesirable compound that is found in food. It is the most poisonous form of mercury. Seafood is the main source of methylmercury for people, and it is therefore important to have good methods for

analysing this compound in seafood. A method for determining the content of methylmercury in seafood, developed by NIFES, has previously been tested in various laboratories in Europe. The results proved that the method was both robust and accurate. The method is expected to be available soon as a new European standard method (CEN method).

Method for the determination Developing better of pharmaceuticals in seafood cell models Funding:

The Ministry of Trade, Industry and Fisheries

NIFES analyses seafood products for residues of legal and illegal pharmaceuticals. NIFES's goal is to have good, robust and accredited methods. NIFES analyses fish fillets and other seafood products for residues of legal and illegal pharmaceuticals. Samples from the Norwegian Food Safety Authority's monitoring programme of farmed fish, conducted in accordance with EU Directive 96/23 make up the bulk of the analyses. Numerous requirements are made of the methods to ensure they are approved for analysis of official samples that are to be reported under the monitoring programme for farmed fish (EU 2002/657). At NIFES, we make continuous efforts to improve the methods, and it is NIFES' goal to have accredited methods that are sensitive, selective and robust. All of the analyses of pharmaceuticals at NIFES were conducted using LC-MS-QQQ. This is an analysis instrument with high sensitivity and selectivity.

Cooperation: The University of Bergen (UiB), East China Normal University (ECNU), China Funding: The Research Council of Norway

Scientists at NIFES are working on developing new, improved cell models, which can potentially reduce the number of fish that are used in current nutrition and toxicology research. When it comes to research on fish, trials based on whole fish are the 'gold standard'. This also applies to current research on nutrition and toxicology. In order to ensure healthy fish in the fish farming industry and to gain knowledge about the consequences of introducing any new undesirable substances on fish health, cell models can, however, be used instead of fish trials. They will give a quicker and more cost-efficient evaluation. Cell models also make it easier to test undesirable substances and the toxicity of different mixes on a greater scale than when using whole fish. However, in order for cell models to be as good as the 'gold standard', and to reduce the number of fish used in fish trials, it is important to develop better and more advanced cell models that mimic the organs of live fish. This will overcome the limitations of today's 2D models.

Scientists working on the 'In Vitro Fish' project are therefore developing new 3D cell models by using primary cells from the liver, fat tissue, head kidney and kidney â&#x20AC;&#x201C; all isolated from the same salmon. The new cell models enable the scientists to study the fish's physiology in a more organ-specific manner that resembles the whole fish to a greater extent. These new models can potentially replace and improve the existing methods and thereby reduce the number of fish that are used in current nutrition and toxicology research. To date in the project, the scientists have isolated adipocytes from salmon. Work is now under way on comparing different types of adipocytes. Primary kidney epithelium cells have also been isolated for use in co-cultures with liver cells. Scientists have also found that the established method for separating cells actually isolates kidney epithelium cells. Further work will focus on establishing 3D kidney epithelium cells in co-cultures with liver cells from salmon and evaluating the use of the co-culture in toxicological research.

METHODS

Micro plastics in seafood Cooperation: Led by Alfred-Wegener Institut, Bremerhaven, Tyskland, together with 25 partners from European countries Funding: Joint Programming Initiative (JPI) â&#x20AC;&#x201C; Oceans, Research council of Norway

NIFES is part of a European consortium that is going to investigate which method is best suited to analyse micro plastics in sea water, sediments and animals. Micro plastics is quite well dispersed in the marine environment, and is defined as plastic particles less than 5 millimetres. The particles are mainly residues from breaking down larger objects, raw material from plastic production, synthetic clothes, wear and tear of car tires and reminiscent from paint. They can also be small particles added to products which is supposed to have a polishing effect, for example cosmetics. Micro plastics has been identified globally in both water, sediments and biota, in several areas including the Northeast Atlantic and Artic waters. Therefore, it has the ability to affect the marine ecosystems on several levels, globally. Most types of plastic float on water, while microplastic eventually sink due to growth of organisms. The uptake and concentration of microplastic has been shown for many animals in the sea, like planktons, worms, bivalves, sea cucumbers, crabs, fish and 82

METHODS

whale. Micro plastic are concentrating organic undesirable substances such as polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB) and polybrominated flame retardants (PBDE) from the water, with a factor up to a million. It is likely that the acidic environment in the gut of animals and humans will release these substances. Furthermore, it has been shown that micro plastics in bivalves can migrate through tissues into the bloodstream and into the cells, where they cause inflammation. In mammals, it has been demonstrated that these substances is absorbed through the colon and transported to the placenta. Therefore, it is possible that micro plastic will affect both the seafood safety and food security. In order to understand how extensive plastic related pollution is in the ocean, it is important to be able to compare different studies of micro plastics. The methods used to isolate small parts of plastic from sediments and animals are quite advanced, and there are many different kinds of plastic. There are several methods in use to measure the level and type of micro plastics in animals, and the difference between the

methods are substantial. Thus, the results from different studies cannot be compared. Today, the difference in methods encompass sampling size, extraction method, filter size, analysis of type and level, in addition to data processing. NIFES is comparing the various methods available internationally, in order to develop a common system. This is the aim within the project BASEMAN, which is one of four JPI-Oceans-financed consortiums concerned with micro plastics. The participants in this project include 25 laboratories from eleven European countries. NIFESâ&#x20AC;&#x2122; role is to deliver identical samples of fish and bivalves to all participating laboratories, which has an added standardised type of micro plastics at a known level. The goal is to determine the best extraction protocol, and to compare methods for the identifying kind of plastic the samples consists of, and their levels. The methods are for example Raman/infrared spectroscopy/microscopy or pyrolysis-mass spectrometry. The project will last three years.

Joint PCB determination of whale blubber Funding:

The Ministry of Trade, Industry and Fisheries

In order to increase the reliability of exports of Norwegian blubber, Norway and Japan need to coordinate the determination of PCBs. Norwegian whale stock management is based on the UN's Convention on the Law of the Sea, and is based on the best scientific knowledge available and advice from the International Whaling Commission. Utilising the whole animal is important in whaling. A lot of the whale blubber is nonetheless thrown into the sea, while the

meat is mainly sold in Norwegian shops. However in Japan, the meat and the blubber are in demand. To ensure that the food is safe and to achieve reliability in international food trade, it is important to coordinate international cooperation on maximum limits and analytical methods. This will enable more of the food that is produced to go to consumption and waste to be kept at a minimum. Japan and Norway use very different methods to determine the compound PCB.

While the Japanese determine the total PCB (a total of the 209 theoretically possible PCB compounds), the EU and Norway have focused on determining the 12 dioxin-like PCBs separately, and six selected non-dioxinlike PCBs (PCB6), or the seven indicator PCBs (PCB7) separately. In this project, we have shown that there is a link between PCB7 and total PCB (PCB209). The documentation of this link will make it easier to export whale blubber to Japan in the future.

METHODS

AND HEALTH

HOW CAN YOUR HEALTH BENEFIT FROM EATING SEAFOOD? How does seafood affect your body?

Seafood provides many of the essential nutrients you need in your diet, but it can also contain undesirable substances and medicine residue. The content of environmental toxins in fish must be monitored to determine whether it is safe to eat seafood. This type of monitoring also ensures that research on undesirable substances in seafood focuses on the most relevant issues. In order to be able to provide advice on seafood intake, it is also important to understand how individual components in the seafood function together as well as separately. Knowledge about the interaction between individual components is important for understanding the basic mechanisms behind beneficial effects and the development of disease.

SEAFOOD AND HEALTH

HEALTH EFFECTS OF SEAFOOD, STUDIED THROUGH MODELS The positive health effects of seafood are also affected by the food it is eaten with Cooperation: Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic; the University of Copenhagen, Denmark; Skretting ARC; LerĂ¸y Seafood Group; Marine Harvest and the Norwegian Seafood Council Funding: The Norwegian Seafood Research Fund, the Ministry of Trade, Industry and Fisheries

The health effects of seafood also depend on what other ingredients make up the meal. Eating more seafood can potentially reduce the development of obesity and obesityrelated diseases. The nutrients in the food we eat have a combined effect. Seafood is generally not eaten on its own, but few studies on seafood have looked at it as part of a meal. This type of research is a useful basis for issuing advice on what foods to combine with seafood. The results of this project will contribute to an overall assessment of the significance of seafood as a part of a meal, in relation to the development of obesity and diabetes. In the project, we compare how whole meals of seafood or meat affect our metabolism and the feeling of fullness in both people and mice. Because carbohydrates are the most common accompaniment to fish, we have prepared meals for the research subjects in which salmon, cod and veal were combined with two types of carbohydrates. One of the carbohydrate sources, mashed potatoes, causes a rapid rise in blood sugar in participants, while the other source, 86

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wholegrain pasta, causes a moderate increase in blood sugar. These meals were eaten by healthy research subjects. After the meal, we measured the subjects' metabolism and the level of the hormone indicating hunger. When the research subjects ate veal, their metabolism was the same irrespective of whether the veal was combined with pasta or mashed potatoes. When the research subjects ate salmon with mashed potatoes, they had a higher diet-induced metabolism and heat production than when they ate pasta with the salmon. The research subjects who ate veal had a higher concentration of the hunger hormone ghrelin after the meal, than those who ate salmon. This may mean that salmon suppresses the secretion of ghrelin, and thereby has a more filling effect than veal. In parallel with the human trials, we have studied the effect of cod, chicken and pork in different diets for mice. The results of the studies show that cod mixed into the diet reduces the development of obesity and fatty liver, compared with mice given chicken and pork in their diet. We saw reduced development of obesity when cod was mixed

into the diet. This was the case both in a high-protein diet and in a Western diet, and can largely be attributed to more active brown fat cells, which increase heat release and energy consumption. We have also seen a reduction in the development of obesity in mice given a mix of lean fish (including cod) when compared with lean meat in a Western diet. The main reason for this appears to be reduced energy intake and appetite.

SEAFOOD AND HEALTH

Seafood, pollutants and obesity Cooperation: Copenhagen University (Denmark) Funding: Ministry of Trade, Industry and Fisheries

Mice that eat sugar in combination with a high fat diet become fat. This leads to a higher amount of fatsoluble pollutants being absorbed and stored in the fatty tissue. Many of the pollutants found in seafood are persistent organic compounds, and are therefore accumulated in fatty fish and in the fatty tissue of those who eat the fish. Several studies of the population have pointed towards that obese people with type 2 diabetes have more pollutants in their body than slim non-diabetics. Questions have therefore been raised whether these pollutants can be a contributing cause to development of the disease. Not all of the completed studies of the population show a connection between the level of pollutants, obesity and the development of type 2 diabetes, and a direct causality has so far not been shown. It is therefore unclear whether these pollutants is a direct contributing cause to disease development. The pollutants there has been a lot of attention towards in this context, are PCBâ&#x20AC;&#x2122;s

SEAFOOD AND HEALTH

(polychlorinated biphenyl) and DDE (dichlorodiphenyldichloroethylene), which is formed when the pesticide DDT (dichlorodiphenyltrichloroethane) is disintegrated. We have fed mice with fish oils that contain pollutants, and characterized weight development and storage of the pollutants in both fatty tissue and liver during a whole life span. Four pollutants (PCB 153, PCB 138, PCB 118 and DDE) were given in diets to mice, singular in different doses or in combination in another trial. The storage of the pollutants was measured in several tissues from the mice, and development of obesity and type 2 diabetes was registered. We found that pollutants had a dose dependent storage in both the fatty tissue and the liver when they were given in a high fat/high sugar diet. The different pollutants did on the other hand not lead to development of obesity, glucose intolerance or impaired insulin sensitivity. The composition of the diets did affect the total accumulation of pollutants, and increased levels of sugar in the high fat diet led to increased storage of the pollutants in the fatty tissue. The mice also

gained more weight. The effect was opposite when we increased the amount of protein in the high fat diet. Our results indicate that the pollutants which were examined in this project does not affect development of obesity or diabetes type 2, despite a considerable accumulation of pollutants in the fatty tissue of the mice. It nevertheless seems like development of obesity, provoked by fat and sugar, causes an increased accumulation of pollutants. Furthermore, an animal study with the persistent organic pollutant HBCD has also been done. Mice were exposed to a low or a high dose of HBCD in a high fat/high sugar diet. The fat source in the diet was either fish oil or soy oil. Mice fed a high fat diet based on the fish oil stored less of the pollutant in both the fatty tissue and the liver compared to mice that ate a diet with soy oil. This indicates that the fatty acid composition in the diet also could have an impact on the amount of pollutants that is stored in the body. Our trials show that pollutants are not necessarily a cause to obesity. However, the

results indicate that development of obesity can lead to higher levels of pollutants, such as PCBâ&#x20AC;&#x2122;s and DDE, being stored in the body. Intake of fish oil seems to be able to reduce the amount of certain types of pollutants being stored. This is an important contribution to a comprehensive assessment of the significance of seafood in the diet.

SEAFOOD AND HEALTH

How are cell cultures and mice affected by undesirable substances? Cooperation: University of Copenhagen, Denmark, Laval University, Canada, BGI Shenzhen, China Funding: The Research Council of Norway

Exposure to a mix of the undesirable substances we find in fatty fish has shown to contribute to cellular stress, which is an underlying mechanism in non-communicable diseases. We did not find the same response in mice. The theory that seafood protects against the development of non-communicable diseases has recently been the subject of debate. Most of the debate concerns whether the levels of undesirable substances in seafood increase the risk of non-communicable diseases. The growing obesity epidemic cannot be explained from an evolutionistic perspective, and one of the explanatory models is that increased exposure to organic undesirable substances over the last century has affected generations through genetic mechanisms that regulate how genes are expressed in the body. The effect of undesirable substances in seafood on animal models has largely been identified on the basis of short-term studies. A key part of this project is therefore to gain better insight into whether undesirable substances contribute to the development of lifestyle diseases across generations, and

SEAFOOD AND HEALTH

how different cells in the body are affected by these undesirable substances. In order to increase understanding of the cellular mechanisms affected by exposure to undesirable substances in seafood, liver cells have been exposed to the most common undesirable substances we find in fatty fish. The undesirable substances we studied had little individual effect on the liver cells. When we mixed several of the different substances into a cocktail, we observed an effect on the expression of genes that code protein which triggers stress in components of the cells, called endoplasmic reticulum stress. This resulted in an error in the protein production in the cells, which can lead to cell death, and is an underlying mechanism in lifestyle diseases. Further studies of protein expression in the cells confirmed the findings. Analyses of the livers of mice that had been exposed to the same cocktail through their feed, however, showed no sign of endoplasmic reticulum stress or sign of developing non-communicable diseases. We cannot yet explain this difference between cells and mice.

In a bigger trial that has run over more than a year, the effect of undesirable substances on gene expressions involved in metabolism are now being studied in an animal model over several generations. The weight development, fat deposits and ability to regulate blood sugar in the offspring of two generations of a male mouse exposed to a cocktail of undesirable substances we find in salmon, are being mapped, as will any changes in gene expressions. The results of this project will contribute to riskbenefit assessments for future advice and recommendations on seafood intake.

SEAFOOD AND HEALTH

Interactive effects of methylmercury and various nutrients Cooperation: Lund University, Sweden, University of California, Davis, USA, University of Copenhagen, Denmark, Flemish Institute for Technolology, Belgium, University of Bergen, National Research Institute of Fisheries Science, Japan, BGI-Shenzhen, China Funding: The Research Council of Norway

The goal of this project is to understand the interactive effects of methylmercury and various nutrients. Fish feed should contain a beneficial composition of nutrients that the fish need, but it also contains undesirable substances. Fishmeal, for example, may contain mercury, in the form of methylmercury, one of the most toxic forms of mercury. Methylmercury affects the nervous system and is particularly damaging to the development of the nervous system at early stages of foetal development in a number of species such as zebrafish and mice. Research indicates that the nutrient selenium neutralises the effects of mercury. So far however, we know little about how mercury causes such damage, and how mercury and selenium interact. NIFES is using zebrafish as a model system to study the effects of methylmercury, and to study the interaction between mercury and selenium. We have carried out a trial on five-day-old zebrafish larvae and found that changed behaviour can be used as a parameter to identify negative effects of methylmercury at doses far below the doses that lead to visible 92

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deformities. Adult zebrafish exposed to methylmercury absorbed and stored the substance in their brain. The accumulation in the brain was reduced when the zebrafish were simultaneously exposed to selenium. Zebrafish that were exposed to mercury, selenium and a combination of the two showed changes in levels of different proteins

in their brains. When the samples have been finally analysed, we will know more about the biological processes that are affected by these substances. In another part of the project, mice have been fed higher and higher concentrations of methylmercury to determine the effect of

storage in the different tissue, obesity, insulin secretion and glucose tolerance. The results of this study showed that the pancreas is an organ that stores a lot of methylmercury, and that the base level of insulin secretion was reduced somewhat, although a low effect of methylmercury was observed on obesity, insulin sensitivity and glucose sensitivity. In order to find out whether methylmercury affects beta cells, the insulin-producing cells in the pancreas, cell trials were conducted to study the toxicity of methylmercury on a beta cell line. The beta cell line proved to be particularly sensitive to methylmercury compared with other cell lines. However, methylmercury had little effect on the beta cell line's ability to secrete insulin. A trial with mice has been conducted to study how intestinal bacteria affect the absorption of methylmercury from the diet and its release from the body. In the marine food chain, bacteria probably play a part in the conversion (methylation) of inorganic mercury into the more toxic and easily absorbed methylmercury. Methylmercury is largely

released from the body through excrement, but efficient secretion depends on it being converted into inorganic mercury (demethylation). It is not known how intestinal bacteria in an organism can affect the methylation of mercury into methylmercury and, if relevant, the demethylation of methylmercury. In this animal trial, we therefore wish to study whether changes in intestinal bacteria in mice can affect the absorption, storage and secretion of methylmercury consumed through the diet.

counteracts these effects, is important to ensure good fish welfare. Trials with animals can also provide more information about how changes in the composition of intestinal bacteria affect the accumulation and secretion of mercury from the diet. The results of this project can increase our understanding of the interaction between methylmercury toxicity and how different nutrients can affect this relationship. That will give us a better basis for future risk-benefit assessments of seafood.

Knowledge about how methylmercury affects the nervous system, and how selenium

SEAFOOD AND HEALTH

SEAFOOD AND LEARNING ABILITIES IN CHILDREN

More and more attention is being devoted to how diet and nutritional status can promote concentration and learning in children. Observation studies have found a positive correlation between seafood intake and cognitive function, while intervention trials involving individual components such as omega-3 capsules show diverging results. Seafood contains several of the nutrients that can explain the positive effect, but no intervention studies involving seafood, children and young people have been carried out yet. In 2014, NIFES conducted a trial with salmon given to children in Germany, and in 2015 a trial was conducted with herring and mackerel given to kindergarten children in Bergen. In 2015, there was also conducted a trial with fatty fish given to lower secondary school pupils in Bergen. The analyses of these trials are ongoing and will result in several manuscripts that will be sent to different journals in 2016.

SEAFOOD AND HEALTH

Is a child's health affected by its mother's diet? Cooperation: The Centre for Child and Youth Mental Health and Child Welfare, Eastern and Southern Norway (RBUP Eastern and Southern Norway), Beijing Genome Institute (BGI), China Funding: The Research Council of Norway

Scientists at NIFES intend to answer this question by analysing saliva samples from infants. Through a collaboration between the projects 'Little in Norway' and 'Fish Intervention Studies', scientists will analyse saliva samples from infants. This is to find out whether the intake of fish, mercury status and level of the omega-3 fatty acid DHA in the mother affects the child's development through what is known in the field as epigenetic mechanisms. This means that the body can switch genes on and off without altering the DNA sequence itself. These epigenetic changes can be passed on and thereby affect further generations. Both nutrients and undesirable substances can lead to changes of this kind. Scientists have isolated the DNA from the saliva samples and these samples will be examined for epigenetic changes. The results will provide knowledge about the sequences of genetic material in which there are changes in gene activity.

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A trial of meals with meat or fish given to kindergarten children Cooperation: Regional Centre for Child and Youth Mental Health and Child Welfare (Uni Research Health), Ludvig-Maximillians University (LMU), Munich, Germany Funding: The Norwegian Seafood Research Fund, the Ministry of Trade, Industry and Fisheries

NIFES has conducted an intervention study on kindergarten children at 12 kindergartens in Bergen, where children were randomised to eat meals with either meat or fish. In this randomised intervention study conducted by NIFES, 232 children (aged four to six) from 12 kindergartens in Bergen were individually randomised to eat either meals of meat (lamb, beef, chicken, turkey) or meals of fish (herring, mackerel), served for lunch three times a week for a total of 16 weeks from January 2015. Research assistants weighed the meat and fish before and after the meals. Eleven children were

removed from the study and four did not take the cognitive tests. The final study sample therefore included 218 children. Tests conducted before and after the food trial included: The IQ test 'The Wechsler Preschool Primary Intelligence Scale (WPPSI-III)', blood tests (fatty acids, vitamin D, ferritin) and urine samples (iodine). Parents also answered a questionnaire about, among other things, diet and socioeconomics. We have processed a lot of the data, and the preliminary results show that 44 study meals were served and the children ate an average of 2,675 grams of meat-based lunch and 2,070 grams of fish-based lunch. The meat group's intake was greater than that of the children in the fish

group. As expected, there was an improvement in the children's cognitive tests from the start to the end of the intervention period in both intervention groups. In the work ahead to determine whether there are differences in the cognitive outcome measures between the fish and meat groups, we will take how much meat and fish the children ate into consideration, as well as the biological markers. The first article will be submitted for publication in the course of 2016.

SEAFOOD AND HEALTH

Diet and concentration at school Cooperation: Regional Centre for Child and Youth Mental Health and Child Welfare (Uni Research Health), The Artic University of Norway Funding: The Norwegian Seafood Research Fund, the Ministry of Trade, Industry and Fisheries

To what extent does diet affect young people's concentration and nutritional status? In 2015, NIFES has conducted a food trial among 481 Year 9 pupils from eight lower secondary schools in Bergen. The study concluded in June, after pupils had been given fatty fish, meat/cheese or omega-3 capsules for lunch at school for 12 weeks.

SEAFOOD AND HEALTH

How much each pupil ate, and the number of capsules taken, was registered by research assistants at each school throughout the food trial. At the start and at the end of the intervention, all the pupils took a concentration test and a reading and writing test. They also answered a questionnaire about diet, sleep habits, physical activity and their well-being (mental

health). We also collected blood, urine and hair samples from the pupils. The diet data showed that a third of the pupils ate fish for dinner two or three times a week, and that almost half of the pupils took an omega-3 supplement. The pupils rarely ate sweets at school and drank less fizzy drinks compared with that found in other studies of young people.

Salmon to German children Cooperation: The Regional Centre for Child and Youth Mental Health and Child Welfare (Uni Research Health), LudvigMaximillians University (LMU), Munich, Germany Funding: The Ministry of Trade, Industry and Fisheries

We have processed a lot of the data results, which show that young people generally eat too little fish, fruit and vegetables. The final results will be included in a PhD thesis towards the end of 2017.

This study shows that omega-3 status improved with the intake of salmon, even though the salmon had a low EPA and DHA content. In this study, 205 children aged four to six from Munich in Germany were randomly selected to eat three meals of either meat or salmon a week for a total of 16 weeks. The salmon used in the study had a slightly lower content of omega-3 fatty acids compared with commercially available salmon. The meals were eaten for dinner in the children's homes. The children were tested before and after the food trial. This included the IQ test 'The Wechsler Preschool Primary Intelligence Scale (WPPSI-III)', blood tests (fatty acids, vitamin D, ferritin), urine samples (iodine) and a questionnaire on, among other things, the children's diets, which the parents answered.

There were no significant differences between the children in the meat and salmon groups in the variables studied, WPPSI-III, fatty acids, iodine, ferritin, and nutritional status when the study started. During the trial, 34 of 48 study meals were consumed, and there were no significant differences in the number of study meals between children in the meat and salmon groups. The children in the salmon group saw a significant increase in the omega-3 fatty acids EPA and DHA during the trial, compared with the meat group, despite the salmon containing lower marine omega-3 levels than fish available to buy. NIFES is now working to determine whether the increase in EPA and DHA in children who ate salmon had any effect on the results of the tests the children took.

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HEALTH EFFECTS ON PEOPLE Lean seafood counteracts lifestyle diseases Cooperation: University of Copenhagen, Denmark, Laval University, Canada, Aarhus University, Denmark, University of Bergen, Helse Bergen health trust (Haukeland University Hospital), University of TromsĂ¸, Bergen University College Funding: The Research Council of Norway, the Ministry of Trade, Industry and Fisheries

The intake of lean seafood reduces obesity in mice and risk markers for the lifestyle diseases type 2 diabetes and cardiovascular diseases, both in mice and humans.

The right diet is an important factor for staying healthy. Both the World Health Organization and the Directorate of Health advise the general population to increase its intake of seafood. This is largely due to the content of marine omega-3 fatty acids in fatty fish. New research also indicates that the intake of lean seafood can have a beneficial effect on people's health â&#x20AC;&#x201C; even though it contains a lower level of omega-3 fatty acids than fatty fish.

intake of lean seafood protects mice against reduced insulin sensitivity, compared with mice that were only fed meat-based diets. Mice that were fed lean seafood had lower fasting insulin levels, less storage of fat in the liver and more glycogen in the liver. All of these findings are strongly associated with an improvement in insulin sensitivity in mice fed a diet of lean seafood. This means that the mice had a reduced risk of developing type 2 diabetes.

The goal of this project was to increase understanding of the intake of lean seafood, and to find out whether lean seafood in the diet can counteract the development of metabolic syndrome in model animals (mice), and reduce risk factors for metabolic syndrome in healthy humans.

Mice that were given lean seafood in their diet also generally had a better fat profile in the blood, which can be beneficial in counteracting the development of cardiovascular disease. We used a mouse model that is prone to developing cardiovascular disease, i.e. mice without apolipoprotein E (Apo E-/- mice). We were then able to show that seafood reduced the incidence of atherosclerotic plaque in the main artery (aorta) compared with mice given chicken fillet in their diet. This is a strong indication that lean seafood can reduce the development of cardiovascular disease in Apo E-/-mice.

We have shown through this project that the intake of lean seafood, compared with the intake of lean meat (chicken fillets or a mix of fillets from chicken, pork and beef) counteracts diet-induced obesity in mice fed a high-fat diet. There are also strong indications that the

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Children's iodine status better than their mothers' Cooperation: The Regional Centre for Child and Youth Mental Health and Child Welfare, Eastern and Southern Norway, the Norwegian Institute of Public Health Funding: The Norwegian Seafood Research Fund, the Ministry of Trade, Industry and Fisheries and the Research Council of Norway

In a randomised, controlled trial on healthy people, we showed that the intake of lean seafood (cod, pollock, saithe and scallops) for four weeks reduced the risk markers for cardiovascular disease, compared to a diet without seafood (meat, eggs and dairy products). We also showed that the intake of lean seafood changed how carbohydrates in food were metabolised after a meal, compared to after eating a meat-based diet. We believe that this is due to the improvement in insulin sensitivity after the lean seafood intervention, and that this is an underlying explanation for the reduction of cardiovascular disease risk factors. Based on data from the mouse trial and the controlled, randomised study of healthy people, we have shown that eating lean seafood affects the metabolism in a manner that, over time, can counteract the development of type 2 diabetes and cardiovascular disease.

Children have a satisfactory iodine status at the age of 18 months, despite their mothers having a mild iodine deficiency at the same time. The World Health Organization considers iodine deficiency to be the most important preventable cause of mental retardation globally. The effect of serious iodine deficiency is well-documented, and pregnant women are a particularly vulnerable group. The reason for this is that iodine is a key component in the thyroid hormones, which are critical to the normal development of the brain and nervous system in foetuses. There is little data about the effect or significance of the mother not having an optimal iodine status on the development of the child.

deficiency. The children, on the other hand, had a satisfactory iodine status. Milk and milk products are considered to be the most important source of iodine in the Norwegian diet, but lean fish is the food group with the highest iodine content. The participants in this study have a generally low seafood intake. Some baby food products are enriched with iodine, and this may be an important reason why the children's status was better than their mothers'.

In the project 'Little in Norway', we have followed a group of mothers from pregnancy until the children were 18 months. Preliminary results show that around 80 per cent of the mothers had a mild to moderate iodine deficiency during their pregnancy. Analyses of iodine in urine samples from the mothers and children when the children were 18 months old show that the mothers' iodine status had improved since the pregnancy, but that, as a group, they had a mild iodine SEAFOOD AND HEALTH 101

NUTRIENTS IN SEAFOOD Salmon is still a good source of omega-3 fatty acids. Funding:

The Ministry of Trade, Industry and Fisheries

Every year, NIFES analyses nutrients in farmed salmon. The analyses show that salmon is an important source of nutrients in the food we eat, and is still a very good source of the marine omega-3 fatty acids EPA and DHA. By analysing the nutrients in seafood, we can deduce the significance of seafood as a source of a number of important nutrients in our diet. The objective of this project is to obtain analyses of nutrients in fish and other seafood, and make these analyses available in the 'Seafood Data' on NIFES's website. In

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2015, more than 100 samples of farmed salmon were collected from ten different locations. The total fat content in the salmon was 16 grams per 100 grams, and the marine omega-3 fatty acids EPA and DHA constituted 7.3 per cent of this fat. On average, the salmon contained 0.45 grams of EPA and 0.69 grams of DHA. This indicates that a dinner portion of salmon, i.e. 150 grams, provides four to five times the daily EPA and DHA requirement.

There was on average 7 micrograms of vitamin D per 100 grams, and a dinner portion thereby provides the daily vitamin D requirement of 10 micrograms. The salmon contained an average of 4 micrograms of iodine per 100 grams, and the amount of iodine varied from 2 to 7 micrograms per 100 grams. Salmon has a low iodine content compared with lean fish species.

More knowledge about the population's intake of iodine Cooperation: Regional Centre for Child and Youth Mental Health and Child Welfare (Uni Research Health) Funding: The Norwegian Seafood Research Fund, the Ministry of Trade, Industry and Fisheries

In order to be able to assess the iodine intake of people in Norway, it is important to have knowledge about the iodine content of the foods that make up Norwegians' diet. In this project, we analyse the amount of iodine in different types of milk, in a selection of milk products and in different types of lean fish. These food groups are the most important sources of iodine in the Norwegian diet. Milk and milk products are the most important sources because we have a high

intake of these foods, while lean fish is the food group that contains the highest levels. The iodine levels in milk depend on what cows eat. Iodine is added to the feed, and there are therefore higher levels in the milk during winter. The previous figures for lean fish have been based on relatively few analyses with big standard deviations.

content of foods. In this project, many samples will therefore be analysed with the seasonal variations for milk and the catch area and size of the fish in mind. The objective is to gain more knowledge about the iodine levels in these food groups. This will enable us to take this into consideration when we are assessing iodine intake based on individual dietary data.

In order to be able to assess a person's iodine intake as accurately as possible, it is important to use good data on the iodine

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Analyses of nutrients in pizza and fish products Cooperation: The Norwegian Food Safety Authority Funding: The Norwegian Food Safety Authority

NIFES has studied the content of nutrients in pizza and fish products on assignment for the Norwegian Food Safety Authority. In this project, nutrient analyses were conducted on a selection of pizza and fish products on the Norwegian market. The results are available in the Norwegian Food Composition Table.

shows that the most sold pizzas contained a substantial amount of iodine. One 300-gram portion of pizza provided 45 micrograms of iodine, which corresponds to almost one third of the daily 150-gram requirement for adults. The iodine content of fish products varied from 11 to 110, with an average of 48 micrograms per 100 grams. One dinner portion thus provides around half of the daily requirement.

The total fat content was 8 grams per 100 grams in pizza products and 4.7 grams in the fish products. The results show that fish au gratin, fish fingers, breaded fish, fish cakes and fish burgers contained an average of 0.15 grams of EPA and DHA. This shows that dinner portions of these products provide the daily requirement for omega-3 fatty acids. The pizza products did not contain EPA and DHA.

A total of 24 pooled samples were included in the project, and each pooled sample consisted of three products from each producer. All of the pooled samples were analysed in the laboratories at NIFES for their content of water, proteins, beef, fat, fatty acids, cholesterol, nine vitamins (A, B1, B2, niacin, folacin, C, D, E, and K) five minerals (calcium, sodium, magnesium, potassium and phosphorus) and five trace elements (iron, zinc, selenium, copper and iodine). The determination of beta-Carotene, starch, fibre and saccharides was conducted by Eurofins. The iodine content in the eight pizza products was 15 micrograms per 100 grams. This

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COOPERATION ACROSS THE WORLD NIFES enjoys significant international cooperation in many of our research projects, and we participate in a range of international discipline forums and expert groups. A central aspect of our work is focusing on safe and healthy seafood in a food chain perspective. By means of competence transfer, we also actively help other countries to develop a knowledge-based and environmentally sustainable fisheries and aquaculture industry. In 2015, NIFES worked on projects in Cuba and Angola through the Center for Development Cooperation in Fisheries (CDCF), funded by the Ministry of Foreign Affairs. There is broad international cooperation in many of NIFES's projects, and, in connection with the NORAD work, NIFES has served as project manager for Norway's activity in Cuba related to cobia fish farming. Three of the institute's scientists were in Cuba in autumn 2015 to oversee the first ever harvesting of farmed cobia in Cuba. Ambassadors for the areas around Cuba also participated. In 2015, a project in Angola started to provide assistance in the work relating to food safety in the country. In the field of food safety, NIFES has contributed to putting fish on the agenda and has participated in several international projects on this topic. Proper nutrition is essential to maintaining people's health, and fish and knowledge about fish will play a key role in securing enough food for the growing population of the future. Fish provides nutrients that are important for good health, both in the west, with the increase in health problems such as cardiovascular disease and mental health, and for poorer populations who still suffer from deficiency diseases due to malnutrition. In this regard, the institute has actively contributed in delegations to the UN's Committee on World Food Security (CFS), where NIFES led a parallel arrangement under the CFS meeting in Rome, autumn 2015. NIFES also participated in a food security conference at Cornell University, USA.

In 2015, NIFES participated in several different EU projects and is represented in various reference groups appointed by the Research Council of Norway, for example in connection with Horisont 2020 and Joint Programming Initiatives (JPIs). Text contributions from NIFES have been used by both the JPIs and the commission in their work on drawing up new strategy documents and calls for applications. One of the senior scientists at the institute is a member of one of EFSA's expert panels, based on her expertise in risk analysis, nutrition and toxicology. NIFES is represented in the Norwegian Committee on Sea Mammals and provides research results on marine mammals, which is reported to the North Atlantic Marine Mammal Commission (NAMMCO) and International Whaling Commission (IWC). The institute is represented in several discipline forums related to analysis methods, such as the Nordic Committee on Food Analysis (NMKL), which approves and publishes methods for analysis of foodstuffs, and CEN, the European Committee for Standardization, which is an association consisting of the national standardisation organisations of 33 European countries. CEN forms a platform for the development of European standards and other technical documents related to different products, materials, services and procedures. The institution is also represented in the Codex Committee on Methods of Analysis and Sampling (CCMAS) which sets out standards and related texts organised by the UN's joint FAO/WHO 'Food Standards Programme'. These standards are intended to protect consumers' health and ensure fair practice in international food trade. CCMAS is a horizontal committee that develops general provisions, principles and guidelines for analysis and sampling in Codex.

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Assistance on seafood safety and a good monitoring system in Angola Cooperation: The Ministry of Fisheries (Ministerio des Pescas), Angola, The Institute of Marine Research (Centre for Development and Cooperation in Fisheries, CDCF) Funding: The Norwegian Agency for Development Cooperation (Norad), the Norwegian Embassy of Angola

Angola and Norway have entered into a cooperation agreement on seafood safety and a monitoring system. A subproject has been established through the Institute of Marine Research (CDCF) to ensure the safety of seafood and to monitor seafood, and by-products and bycatches for feed. NIFES has been involved in the project since 2015. Angola has a medium-sized fisheries industry, which is largely based on horse mackerel, but also on species of squid, shellfish and European hake. Harbour facilities and landing facilities are not developed to any extent, and a lot of fish is transported directly out of the country. To improve this situation, Angola wants Norwayâ&#x20AC;&#x2122;s help to improve inspections and sampling and to certify laboratories for the analyses of seafood, both in microbiology and chemical analyses. Angola has also expressed a wish to be able to use the waste products from fishing and

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gutting, either for fish or land animals. NIFES provides expertise about preservation and microbiology in this respect. In autumn 2015, NIFES conducted a project start-up visit to establish contacts and get under way with the project. NIFES had discussions with the Ministry of Fisheries in Luanda. A considerable amount of method work was also carried out at INIP Lab (the

public fisheries laboratory) in Luanda. NIFES helped to make improvements so the laboratory can be accredited for mercury analysis and for microbiological issues, among other things. Training is an important element of the work ahead, and Angolan inspectors and laboratory technicians will visit Norway in 2016.

Marine farming of Cobia on Cuba Cooperation: Centro Investigaciones Pesqueras, Cuba, The Institute of Marine Research (Centre for Development and Cooperation in Fisheries, CDCF), Frost Innovation, Aquaculture management & Consulting Funding: The Norwegian Agency for Development Cooperation (Norad) and the Ministry for Foreign Trade and Foreign Investment of Cuba (MINCEX), Cuba

In 2011, Cuba and Norway entered into an agreement to cooperate on developing sustainable marine fish farming. The first production round of the species Cobia has now concluded and the fish are on the market on Cuba. Cuba and Norway are cooperating on aquaculture. In 2011, a new five-year Norwegian-Cuban project was initiated. The goal of this project is to use Cuban and Norwegian expertise and technology to establish sustainable marine aquaculture activities in Cuba. The species Cobia (Rachycentron canadum) was selected for the project. This is an active predatory fish with lean, white meat. It is

considered one of the most important species for marine farming in tropical waters, for example in parts of Asia, the USA and the Caribbean. It is relatively robust, grows quickly, and the adult fish can weigh more than 50 kg and be up to two metres long. The Norwegian-Cuban cooperation made it possible to harvest the first Cobia at the end of August 2015, when the fish were between three and four kilos. The fish have now been sold to hotels in Cuba where it is served. Cuba already has freshwater fish farming operations, but the volume is only around 30,000 tonnes a year. The most commonly farmed species include carp, tilapia and catfish. The current project has had the goal of covering all components required for marine farming operations. This included

selecting a suitable species, finding a good location, procuring cages and other necessary aquaculture equipment, and building hatcheries and broodstock farms. Other necessary factors were procuring appropriate feed, information campaigns on Cobia, and organising harvesting, processing and distribution. Furthermore, to ensure that the project is the start of a lasting industry, it is very important to build broad marine aquaculture expertise. This includes teaching students at university level. The annual tropical cyclones that can hit Cuba represented a particular challenge in relation to the choice of location. After a review of relevant factors such as depth, wind and wave conditions, current patterns, oxygen saturation, infrastructure and competing activities, the Bay of Pigs (Bahia de Cochinos) was chosen. This bay is approximately 25 kilometres long and 8 kilometres wide at the mouth, and is well sheltered from the wind in the most common directions. In Cuba as elsewhere, access to marine ingredients such as fat and protein for feed production is an important limiting factor for sustainable growth in fish farming. Cuba does not have large marine stocks that are INTERNATIONAL COOPERATION

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Aim to ensure iodine intake in Europe Cooperation: The project is led by the University Medicine Greifswald, Germany. Twenty-two of the member states of the EU participate in the project together with Iceland, Israel, Macedonia, Norway and Switzerland. Funding: The EU

suitable as feed ingredients, and the use of alternative animal or vegetable ingredients is a condition for future production growth. For example, the rapeseed plant can be used as a source of fat and the soya plant as a source of protein in the feed. In this context, Norway passes on the important information that farmed fish do not need feed ingredients, but nutrients. Cuba has good access to arable land. About 30 per cent of the country's land areas can be used for agricultural purposes, while, by comparison, it is estimated that only three per cent of Norway's land can be cultivated. It is too early to know how Cobia farming in Cuba will develop in the time ahead. However, it is already possible to say that this international cooperation has helped Cuba to succeed in producing its first Cobia. Initially, the produced fish will replace imported fish in the tourist segment, but, in the long term, marine farming can contribute to increased seafood consumption in Cuba. Globally speaking, seafood is an important part of a balanced diet, and NIFES is now starting work on analysing Cobia's content of important nutrients such as marine omega-3 fatty acids, proteins and minerals. This can help ensure good nutritional status, which can have a positive effect on public health in Cuba. 110

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Studies show that an increasing proportion of the population of Europe has an iodine deficiency. In the research project 'EUthyroid', the partners will work together to solve this problem. We know that a serious iodine deficiency can affect brain development, but new research indicates that even a mild iodine deficiency during pregnancy can affect the mental development of the foetus, and thereby negatively impact on its intelligence level. Iodine deficiency is the most important risk factor in developing thyroid disease among children and adults. It is recommended that pregnant and breastfeeding women get enough iodine to ensure optimal conditions for the development of their babies. The EU-funded project 'EUthyroid' started on 1 June 2015. It is the first joint European initiative to coordinate the information available on iodine intake in Europe, and thus provide expertise about the problem.

The three-year project will form the basis for developing expedient measures to improve iodine intake among Europe's population, in a collaboration between national iodine contacts. NIFES will provide data for one of the project's seven work packages. The 'EUthyroid' project consists of 30 partners from 27 countries, and thus gathers the expertise of a number of recognised experts in epidemiology, endocrinology, nutrition physiology and health economics. The global organisation Global Network (IGN) is also involved in the project. With its organisational partners and around 100 regional and national contacts, IGN is resolved to combat iodine deficiency. This network will help to ensure that the methods used and results obtained in the 'EUthyroid' project will be communicated and implemented by national health authorities in the individual countries.

SEMINARS Nordic iodine meeting Funding:

The Ministry of Trade, Industry and Fisheries

Nordic scientists are meeting for the third time to discuss iodine. In September 2015, 20 Nordic scientists gathered at NIFES to discuss and exchange expertise about the trace element iodine. The goal of the meeting was to update and motivate each other to continue work on ensuring the population of the Nordic countries get enough iodine. The meeting started with a brief update about the iodine research conducted in each country since the meeting last year. One of the main points of the meeting was the importance of studies on pregnant women and other groups that are vulnerable to iodine deficiency. Scientists discussed the types of data available, the data still needed, and agreed to continue work on updating and increasing the health authorities' awareness of the iodine status of the Nordic population. Another important point on the list was to make sure a Nordic iodine presentation is drawn up for the next Nordic Nutrition Conference in June 2016, in Gothenburg.

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Fish nutrition seminar Funding:

The Research Council of Norway

NIFES organised the Seminar on Fish Nutrition in Bergen After a 15-year break, a seminar on fish nutrition was again held in Bergen, on 5 and 6 November 2015. The aim of the seminar on fish nutrition is to give scientists and students who work in the field the opportunity to get together for an informal seminar every other year to discuss and share research results with colleagues in

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Norway. The seminar has traditionally been in the Norwegian language. The last seminar took place in Norheimsund in May 2000, but after a 15-year break, NIFES picked up the thread again and organised a new seminar. The two-day seminar took place at Clarion Admiral Hotel in Bergen, on 5 and 6 November 2015, with dinner at the historical Schøtstuene at Bryggen in the evening.

Representatives from research institutions and the industry in Norway participated in the seminar. There were 52 registered participants. The main theme on the first day of the seminar was 'The feed of the future for safe and healthy fish’ and on day two, 'Does the composition of feed affect salmon's health?’ There were 21 lectures in total. The seminar was a success and Nofima has been assigned the task of organising the next seminar on fish nutrition.

AQUACULTURE NUTRITION NIFES is chief editor of international aquaculture nutrition journal Cooperation: Wiley-Blackwell, Oxford, UK Funding: The Ministry of Trade, Industry and Fisheries, Wiley-Blackwell, Oxford, UK

Aquaculture Nutrition considered 295 submitted manuscripts in 2015. Just under half made it through. The journal Aquaculture Nutrition's second fully electronic edition, without an accompanying printed edition, was published in 2015. The journal also had a productive year and considered 295 submitted manuscripts from all over the world. 139 manuscripts were accepted during the same period. The journal had an impact factor at 1.4. This index indicates the amount of citations compared with other publications and is used to measure the quality of a journal. Aquaculture Nutrition is an internationally recognised journal in the field of one of NIFES' two research programmes: fish nutrition. The journal was published for the first time in 1995 and has since been the natural choice for scientists who wish to publish their findings in general fish nutrition and related fields.

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COLLABORATORS AND FUNDING SOURCES COLLABORATORS • Aarhus University, Denmark • AKVAgroup • Akvaplan NIVA • Alevines y Doradas, S.A ADSA, Spain • Alfred Wegener Institute Helmholtz Centre for Polar and Marine Research, Germany

• Instituto Español de Oceanografía (IEO), Spain

• Department of Biology, University of Bergen

• Instituto Português do Mar e da Atmosfera, Portugal

• Dor Dgey Yam Ltd (Dor), Israel • DTU Food (National Food Institute) • EURL • EWOS Innovation

• Aquaculture Forkys Ae, (FORKYS), Greece

• Facultes Universitaires Notre-Dame De La Paix De Namur (FUNDP), France

• Aquaculture management & Consulting

• Aranykarasz Mezogazdasagi Halaszaties Szaktanacsadoi Szolgaltato Bt (KARAS), Poland • Asialor Sarl (Asialor), France

• Institute of Physiology Academy of Sciences of the Czech Republic; Czech Republic

• Culmarex S.A.U., Spain

• Aqua Gen AS

• AquaTT, Ireland

• Flemish Institute for Technolology, Belgium • Frost Innovation • Fundacion Canaria Parque Cientifico Tecnologico De La Universidad De Las Palamas De Gran Canaria (FCPCT), Spain

• International Research Institute of Stavanger (IRIS), Norway • Irida Ae-Products for Animal Production-Services (IRIDA), Hellas • Israel Oceanographic and Limnological Research Limited (IOLR), Israel • IVL Swedish Environmental Research Institute, Sweden • Kentro Meleton Agoras Kai Koinis Gnomis Anonimi Emporiki Etairia (HRH), Greece • King's College London, UK • Laval University, Canada

• Asociación Empresarial De Productores De Cultivos Marinos (APROMAR), Spain

• Fundacion Centro Tecnologico Acuicultura De Andalucia (CTAQUA), Spain

• Lerøy Seafood Group

• Asociacion Nacional De Fabricantes De Conservas De Pescados Y Mariscos-Centro Tecnico Nacional De Conservacion De Productos De La Pesca, European Food Information Council Aisbl (EUFIC), Belgium

• Galway-Mayo Institute of Technology, Ireland

• Ludvig-Maximillians University (LMU), Tyskland

• GEOMAR Helmholtz-Zentrum für Ozeanforschung, Germany

• Lund University, Sweden

• Ayuntamiento De A Coruna (Mc2), Spain

• Geophysics, Trieste, Italy

• Azienda Agricola Ittica Caldoli (ITTICAL), Italy

• Gerdts, Gunnar; Alfred-Wegener Institut, Germany

• Beijing Genome Institute (BGI), China

• Ghent University, Belgium

• Benchmark holding plc

• Gildeskål forskningsstasjon (GIFAS)

• Bergen University College

• Havella, Geir Agnar Landøy

• Marine Harvest • Marine Systems Institute at Tallinn University of Technology, Estonia • Matís, Iceland • Medical University of Vienna, Austria • Ministerio des Pescas (Department of fisheries), Angola

• BIOMAR, Denmark

• Heinrich Heine University, Germany

• National Food Institute (DTU), Denmark

• Bundesverband Der Deutschen Fischindustrie Und Des Fischgrosshandels E.V. (BVFI), Germany

• Hellenic Center for Marine Research (HCMR), Greece

• Canarias Explotaciones Marinas Sl (CANEXMAR), Spain

• Hofgaard, Sven Marius; Fisher in the Oslofjord

• National Institute of Oceanography and Experimental East China Normal University (ECNU), China

• Center for Development Cooperation in Fisheries • Centre for Ecological and Evolutionary Synthesis (CEES), Department of Biosciences, University of Oslo • Centre Investigaciones Pesqueras, Cuba • Centre of Aquaculture Competence (CAC) • Centre of Marine Sciences CCMA, Portugal

• Hokkaido University, Japan • Hordafôr AS • Hungarian Aquaculture Association (MASZ), Hungary • Ichthyokalliergeies Argosaronikou Anonymi Etairia (ARGO), Greece • Institut De Recerca I Tecnologia Agroalimentaries (IRTA), Spanin

• Centre wallon de Recherches agronomiques (CRAW), Belgium

• Institut Francais De Recherche Pour L'exploitation De La Mer (IFREMER), France

• CNR-IAMC, Italy

• Institut National de la Recherche Agronomique (INRA), France

• CNRS-LOV, France • Consejo Superior de Investigaciones Científicas (CSIC), Spain

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• Conselleria Do Mar - Xunta De Galicia (CMRM), Spain

COLLABORATORS AND FUNDING SOURCES

• National Oceanic and Atmospheric Administration (NOAA), USA • National Research Institute of Fisheries Science, Japan • National University of Ireland Galway (NUI), Ireland • Nikøy Eigedom AS • NMBU Veterinary Science • Nofima, Norway • Nord University • NOREL S.A., Spain • NorInsect AS

• Institut Universitari de Plaguicides i Aigües. Universitat Jaume I Avda (IUPA), Spania,

• Norwegian Environment Agency

• Institute of Marine Research (IMR)

• Norwegian Food Safety Authority

• Norwegian Institute for Air Research (NILU) • Norwegian Institute for Water Research (NIVA)

• Syndesmos Ellhnikon Thalassokalliergeion Somateo (FGM), Greece

• Völzke, Henry; University Medicine Greifswald, Tyskland

• Norwegian Institute of Bioeconomy Research (NIBIO)

• Technische Universiteit Eindhoven, Netherlands

• Wageningen University WU, Nederland

• The Arctic University of Norway (UiT)

• Wiley-Blackwell

• Norwegian Institute of Public Health

• The Norwegian Coastal Administration

• Norwegian Seafood Council.

• The Norwegian Reference Fleet

• Norwegian University of Life Sciences (NMBU), Norway

• The Royal Norwegian Society for Development (NorgesVel)

• Norwegian University of Science and Technology (NTNU)

• The Shellfish Industry

FUNDING SOURCES • CAC (Centre of Aquaculture Competence) • CEN • EU

• NOVA.ID FCT, Caparica, Portugal

• The University Court Of The University Of Aberdeen (UNIABDN), Scotland

• Havella, Geir Agnar Landøy

• Nutricontrol, Netherlands

• Uni Research

• Jakob Hatteland Resources AS

• Ocean Forest

• Universidad De La Laguna (ULL), Spain

• Ministry of Trade, Industry and Fisheries

• Oslo Universitety Hospital

• Universidad de Las Palmas de Gran Canaria (ULPGC), Spain

• Nikøy Eigedom AS • NOFIMA AS

• Outi Seatala, Finnish Environment Institute, Finland

• Universidade da Coruña (UDC)-Instituto, Spain

• Plymouth University, Great Britain

• Universitá Degli Studi Di Bari Aldo Moro (UNIBA), Italy

• Norwegian Food Safety Authority

• Protix Biosystems BV, Netherlands

• Norad

• Radboud Universitet Nijmegen (RUN), Netherlands

• Università dell’Insubria (USI), Italy

• Norwegian Research Council

• Université De Lorraine (UL), France

• Ocean Forest, VestMarin

• Rap.ID Particle Systems GmbH, Germany

• University of Arkansas, USA

• Salmobreed AS, Norway

• Raunes fiskefarm

• University of Barcelona, Spain

• SINTEF Fisheries and Aquaculture

• Regional Center for Child and Adolescent Mental Health, Eastern and Southern Norway (RBUP)

• University of Bayreuth, Germany

• Skretting ARC, Norway

• University of Bergen (UiB), Norway

• The Ministry for Foreign Trade and Foreign Investment of Cuba (MINCEX).

• Regional Centre for Child and Youth Mental Health and Child Welfare will, Uni Research Helse • Research institute for fisheries, aquaculture and irrigation HAKI, Hungary • Rijks instituut voor volksgezondheid en milieu (RIVM), Netherlands

• University of California, USA • University of Copenhagen, Denmark • University of Copenhagen, Denmark • University of Gothenburg, Sweden • University of Liège, Belgium

• Røe Communications

• University of Maine, France

• SalmoBreed AS

• University of Nordland

• Seaweed AS

• University of Oldenburg, Germany

• SINTEF

• University of Oslo (UiO)

• SINTEF Fisheries and Aquaculture

• University of Stirling (UoS), Scotland

• Sir Alister Hardy Foundation for Ocean Science (SAHFOS), Great Britain

• University of Victoria, Canada

• The Norwegian Coastal Administration • The Norwegian Embassy of Angola. • The Norwegian Seafood Research Fund (FHF), Norway • The Royal Norwegian Society for Development (NorgesVel) • Værlandet Fishing equipment AS

• Værlandet Fiskeredskap AS

• Skretting Aquaculture Research Centre (Skretting ARC)

• Vas. Geitonas & Co Ltd Ee (Gei), Greece

• SPAROS Lda, Portugal

• Vekst i Stordal

• Sterling White Halibut

• Vienna University of Technology, Austria

• Stichting Dienst Landbouwkundig Onderzoek (DLO), Belgium

• Viviers de Sarrance VDS, France

COLLABORATORS AND FUNDING SOURCES

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Research News offers extensive information about the seafood of the long Norwegian Coast.

NIFES is a national reference laboratory, and provides advice to national authorities, industry and the public sector. Our research provides the basis for measures designed to ensure that seafood is safe and healthy food, and we have a comprehensive programme of publishing our results via both scientific and popular media. Research News informs you about our most recent and current research projects and on-going monitoring programmes.

ADDRESSES

Street address: Strandgaten 2295004 Bergen Phone: 55 90 51 00 Fax: 55 90 52 99 E-mail: postmottak@ nifes.no

Post address: PB 2029 Nordnes 5817 Bergen

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Invoicing address: Fakturamottak DFØ PB 4104 2307 Hamar Business Register Nr.: 979529783

www.nifes.no

NIFES 2016 • Photo: Eivind Senneset, Helge Skodvin, Emil Weatherhead Breistein, Geir Gundersen, UW Photo, Getty Images, Øyvind Lothe, SalmoBreed/Bolaks, Shutterstock, NIFES

IS IT SAFE TO EAT SEAFOOD ? WHY IS FISH A HEALTHY FOOD ?