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Our work aimed to examine nanoparticle levels in 69 distinct pooled mussel samples along the Norwegian coastline, considering samples from different environmental contexts, including natural locations, potentially polluted hotspots, and mussel farms. Single-particle ICP-MS was utilized to determine particle mass and number concentrations at environmentally relevant levels in addition to the total content of 11 elements: aluminum, barium, cerium, copper, iron, manganese, lead, silicon, silver, titanium, and zirconium. Results showed nanoparticle mass concentrations of few ng/g up to tens of μg/g and number concentrations of 10 6 to 10 9 particles/g (wet weight). Certain urban and industrially impacted locations were linked to increased levels of, e.g., silver, lead, cerium, zirconium, and titanium NPs. Farmed mussels exhibited lower concentrations. However, natural variations were considerable, and impacted locations mostly did not differ from the highest levels in pristine areas. The study presents the first extensive survey of NPs of 11 different elements in marine biota and provides evidence of increased levels of NPs in areas with anthropogenic activities.

Demand for n-3 polyunsaturated fatty acids (n-3 PUFAs) exceeds supply. Large-scale studies on effects of season and geography of n-3 PUFAs in marine fish from the Northeast Atlantic Ocean (NEAO) may be used to optimize utilization and improve nutrition security. Using a sinusoid model, seasonal cycles of n-3 PUFAs were determined and found to be species-specific and clearly pronounced for the pelagic zooplankton feeding species. The Greenland halibut showed very little seasonal variation. The n-3 PUFA content in North Sea autumn-spawning (NSAS) herring peaked in summer, while Norwegian spring-spawning (NSS) herring and mackerel had their peak in autumn. A time shift of peaks in n-3 PUFAs between the two herring stocks was detected, likely due to different spawning strategies in addition to a delay of n-3 PUFAs flux in the northern regions of the NEAO. This study demonstrates that consideration of nutrient contents, such as n-3 PUFAs, when organizing and structuring fishery approaches may improve overall nutritional yield. Based on total annual Norwegian fish landings and seasonal variation in n-3 PUFA contents, n-3 PUFAs yield could theoretically be increased from 13.79 kilo ton per year from the current fishing tactics, to 15.54 if the pelagic species were only caught during the time of their seasonal n-3 PUFA peaks. Pelagic fish is a good source for dietary n-3 PUFAs, but harvest timing will also influence n-3 PUFAs intake by human consumers. One portion of fatty fish harvested during winter/spring may not meet the weekly intake reference nutritional guidelines for n-3 PUFAs. Marine n-3 PUFAs yields also varied geographically and decreased southwards, with the lowest values in Skagerrak. This study can serve as a model to understand patterns of reproductive cycles and geographical distribution of n-3 PUFAs in fatty fish from the NEAO and the novel approach may be useful to support sustainable, seasonal fishing programmes for optimization of n-3 PUFAs yields., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Marine fish from the North East Atlantic Ocean (NEAO) are nutrient rich and considered a valuable economic resource. However, marine fish are also a major dietary source of several contaminants, including persistent organic pollutants (POPs) and heavy metals. Using one of the world's largest seafood datasets (n > 25,000 individuals), comprising 12 commercially important fish species collected during 2006-2019 in the NEAO, we assessed the co-occurrence of elements and POPs, and evaluated potential risks to human consumers. Several positive correlations between concentrations of mercury (Hg), dioxins, polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) were observed. Concentrations of Hg, dioxins, PCBs and PBDEs increased from North to South and associations between marine sediment contamination, sea temperature, and fish Hg and POPs concentrations were identified using multi-linear regression (MLR) models. In general, Hg concentrations in fillet and liver of fish were positively associated with increases in both sediment contamination and sea temperature. POPs concentrations in both fillet and liver were positively associated with increases in sediment contamination, and only POPs concentrations in the liver of benthopelagic and demersal species were found to be positively correlated with sea temperature. Using a probabilistic approach to estimate human contaminant exposure from seafood, we showed that intake of pelagic species posed the highest risk of dioxins and dioxin-like PCBs (DL-PCBs) exposure, while intake of benthopelagic and demersal species posed the highest risk of Hg exposure. This study can serve as a model to further understand the distribution, co-occurrence, and trends of contaminants in seafood harvested from the NEAO and their potential risks to human consumers., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

POLY: and perfluorinated alkyl substances (PFASs) are ubiquitously detected all around the world. Herein, for the first time, concentrations of 16 selected legacy and emerging PFASs are reported for sediment and edible fish collected from the Saudi Arabian Red Sea. Mean concentrations varied from 0.57 to 2.6 μg kg -1 dry weight (dw) in sediment, 3.89-7.63 μg kg -1 dw in fish muscle, and 17.9-58.5 μg kg -1 dw in fish liver. Wastewater treatment plant effluents represented the main source of these compounds and contributed to the exposure of PFAS to biota. Perfluorooctane sulfonate (PFOS) was the most abundant compound in sediment and fish tissues analysed, comprising between 42 and 99% of the ∑ 16 PFAS. The short chain perfluorobutanoate (PFBA) was the second most dominant compound in sediment and was detected at a maximum concentration of 0.64 μg kg -1 dw. PFAS levels and patterns differed between tissues of investigated fish species. Across all fish species, ∑ 16 PFAS concentrations in liver were significantly higher than in muscle by a factor ranging from 3 to 7 depending on fish species and size. The PFOS replacements fluorotelomer sulfonate (6:2 FTS) and perfluorobutane sulfonate (PFBS) exhibited a bioaccumulation potential in several fish species and 6:2 FTS, was detected at a maximum concentration of 7.1 ± 3.3 μg kg -1 dw in a doublespotted queenfish (Scomberoides lysan) liver. PFBS was detected at a maximum concentration of 2.65 μg kg -1 dw in strong spine silver-biddy (Gerres longirostris) liver. The calculated dietary intake of PFOS, perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorohexane sulfonic acid (PFHxS) exceeded the safety threshold established by the European Food Safety Authority (EFSA) in 2020 in doublespotted queenfish muscle, indicating a potential health risk to humans consuming this fish in Jeddah, Saudi Arabia., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Plastic pollution has become one of the most pressing environmental challenges and has received commensurate widespread attention. Although it is a top priority for policymakers and scientists alike, the knowledge required to guide decisions, implement mitigation actions, and assess their outcomes remains inadequate. We argue that an integrated, global monitoring system for plastic pollution is needed to provide comprehensive, harmonized data for environmental, societal, and economic assessments. The initial focus on marine ecosystems has been expanded here to include atmospheric transport and terrestrial and freshwater ecosystems. An earth-system-level plastic observation system is proposed as a hub for collecting and assessing the scale and impacts of plastic pollution across a wide array of particle sizes and ecosystems including air, land, water, and biota and to monitor progress toward ameliorating this problem. The proposed observation system strives to integrate new information and to identify pollution hotspots (i.e., production facilities, cities, roads, ports, etc.) and expands monitoring from marine environments to encompass all ecosystem types. Eventually, such a system will deliver knowledge to support public policy and corporate contributions to the relevant United Nations (UN) Sustainable Development Goals (SDGs).

We report the draft genome sequence of multidrug-resistant Pseudomonas protegens strain 11HC2, isolated from polypropylene collected from the water column near a beach in Øygarden, Norway. The genome sequence is 6,861,219 bp long, with a G+C content of 63.4%. Strain 11HC2 is resistant to cefotaxime, ampicillin, trimethoprim, and chloramphenicol., (Copyright © 2021 Radisic et al.)

The increase in the global population demands more biomass from the ocean as future food and feed, and the mesopelagic species might contribute significantly. In the present study, we evaluated the food and feed safety of six of the most abundant mesopelagic species in Norwegian fjords. Trace elements (i.e., arsenic, cadmium, mercury, and lead), organic pollutants (i.e., dioxins, furans, dioxin-like polychlorinated biphenyls, and polybrominated flame-retardants), and potentially problematic lipid compounds (i.e., wax esters and erucic acid) were analyzed and compared to existing food and feed maximum levels and intake recommendations. Furthermore, contaminant loads in processed mesopelagic biomass (protein, oil, and fish meal) was estimated using worst-case scenarios to identify possible food and feed safety issues. While most undesirables were low considering European food legislation, we identified a few potential food safety issues regarding high levels of fluoride in Northern krill, wax esters in glacier lanternfish, and long-chain monounsaturated fatty acids in silvery lightfish. Our estimates in processed biomass indicated high levels of undesirable trace elements in the protein fraction, frequently exceeding the maximum levels for feed ingredients. However, in fish meal, almost no exceedances were seen. In the oil fraction, dioxins and furans were above the maximum levels, given for food and feed ingredients. The present study is crucial to enable an evaluation of the value of these species; however, more data is needed before proceeding with large-scale harvesting of mesopelagic biomass.

Plastic pollution has become a major environmental concern due to its omnipresence and degradation to smaller particles. The potential toxicological effects of micro- and nanoplastic on biota have been investigated in a growing number of exposure studies. We have performed a comprehensive review of the main determining factors for plastic particle toxicity in the relevant exposure systems, from publications until including the year 2018. For a focused scope, effects of additives or other pollutants accumulated by the plastic particles are not included. In summary, current literature suggests that plastic particle toxicity depends on concentration, particle size, exposure time, particle condition, shape and polymer type. Furthermore, contaminant background, food availability, species, developmental stage and sex have major influence on the outcome of plastic particles exposures. Frequently reported effects were on body and population growth, energy metabolism, feeding, movement activity, physiological stress, oxidative stress, inflammation, the immune system, hormonal regulation, aberrant development, cell death, general toxicity and altered lipid metabolism. Several times reported were increased growth and food consumption, neuro-, liver- or kidney pathology and intestinal damage. Photosynthesis disruption was reported in studies investigating effects on phytoplankton. For the currently unquantified plastic particles below 10 μm, more toxic effects were reported in all aquatic life, as compared to plastic particles of larger size., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

The projected increase in global population will demand a major increase in global food production. There is a need for more biomass from the ocean as future food and feed, preferentially from lower trophic levels. In this study, we estimated the mesopelagic biomass in three Norwegian fjords. We analyzed the nutrient composition in six of the most abundant mesopelagic species and evaluated their potential contribution to food and feed security. The six species make up a large part of the mesopelagic biomass in deep Norwegian fjords. Several of the analyzed mesopelagic species, especially the fish species Benthosema glaciale and Maurolicus muelleri , were nutrient dense, containing a high level of vitamin A1, calcium, selenium, iodine, eicopentaenoic acid (EPA), docosahexaenoic acid (DHA) and cetoleic acid. We were able to show that mesopelagic species, whose genus or family are found to be widespread and numerous around the globe, are nutrient dense sources of micronutrients and marine-based ingredients and may contribute significantly to global food and feed security.

Limited availability of fish oils (FO), rich in n-3 long-chain (≥C20) PUFA, is a major constraint for further growth of the aquaculture industry. Long-chain n-3 rich oils from crops GM with algal genes are promising new sources for the industry. This project studied the use of a newly developed n-3 canola oil (DHA-CA) in diets of Atlantic salmon fingerlings in freshwater. The DHA-CA oil has high proportions of the n-3 fatty acids (FA) 18 : 3n-3 and DHA and lower proportions of n-6 FA than conventional plant oils. Levels of phytosterols, vitamin E and minerals in the DHA-CA were within the natural variation of commercial canola oils. Pesticides, mycotoxins, polyaromatic hydrocarbons and heavy metals were below lowest qualifiable concentration. Two feeding trials were conducted to evaluate effects of two dietary levels of DHA-CA compared with two dietary levels of FO at two water temperatures. Fish increased their weight approximately 20-fold at 16°C and 12-fold at 12°C during the experimental periods, with equal growth in salmon fed the FO diets compared with DHA-CA diets. Salmon fed DHA-CA diets had approximately the same EPA+DHA content in whole body as salmon fed FO diets. Gene expression, lipid composition and oxidative stress-related enzyme activities showed only minor differences between the dietary groups, and the effects were mostly a result of dietary oil level, rather than the oil source. The results demonstrated that DHA-CA is a safe and effective replacement for FO in diets of Atlantic salmon during the sensitive fingerling life-stage.

The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently, concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat, and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time.

Screening has revealed that modern-day feeds used in Atlantic salmon aquaculture might contain trace amounts of agricultural pesticides. To reach slaughter size, salmon are produced in open net pens in the sea. Uneaten feed pellets and undigested feces deposited beneath the net pens represent a source of contamination for marine organisms. To examine the impacts of long-term and continuous dietary exposure to an organophosphorus pesticide found in Atlantic salmon feed, we fed juvenile Atlantic cod ( Gadus morhua ), an abundant species around North Atlantic fish farms, three concentrations (0.5, 4.2, and 23.2 mg/kg) of chlorpyrifos-methyl (CPM) for 30 days. Endpoints included liver and bile bioaccumulation, liver transcriptomics and metabolomics, as well as plasma cholinesterase activity, cortisol, liver 7-ethoxyresor-ufin-O-deethylase activity, and hypoxia tolerance. The results show that Atlantic cod can accumulate relatively high levels of CPM in liver after continuous exposure, which is then metabolized and excreted via the bile. All three exposure concentrations lead to significant inhibition of plasma cholinesterase activity, the primary target of CPM. Transcriptomics profiling pointed to effects on cholesterol and steroid biosynthesis. Metabolite profiling revealed that CPM induced responses reflecting detoxification by glutathione-S-transferase, inhibition of monoacylglycerol lipase, potential inhibition of carboxylesterase, and increased demand for ATP, followed by secondary inflammatory responses. A gradual hypoxia challenge test showed that all groups of exposed fish were less tolerant to low oxygen saturation than the controls. In conclusion, this study suggests that wild fish continuously feeding on leftover pellets near fish farms over time may be vulnerable to organophosphorus pesticides., (Copyright © 2019 Olsvik, Larsen, Berntssen, Goksøyr, Karlsen, Yadetie, Sanden and Kristensen.)

Liver and muscle tissue of tusks ( Brosme brosme) have been analyzed for their THg and MeHg concentrations and Hg isotopic signatures for tracing Hg pollution along the Norwegian coast. Clear differences between tissue types and locations were established. At five of the eight locations, the Hg concentration in muscle exceeded the maximum allowable level of 0.5 mg kg -1 wet weight. δ 202 Hg values in both tissue types indicated that Hg speciation affects the bulk Hg isotopic signature. Tusk liver seems to be more sensitive to immediate changes and to anthropogenic inorganic Hg, while the muscle rather reflects the Hg accumulated over a longer period of exposure. The δ 202 Hg values of liver and muscle also enabled different sources of Hg and exposure pathways to be distinguished. δ 202 Hg muscle -δ 202 Hg liver showed a clear correlation with the % MeHg in tusk liver for the coastal waters, but not for the fjords. The absence of significant differences in Δ 199 Hg values between both tissues of tusk from the same location suggests that in vivo metabolic processes are the underlying reason for the differences in Hg speciation and in δ 202 Hg values. This work highlights the importance of selecting different tissues of marine fish in future Hg monitoring programs.

The European Food Safety Authority (EFSA) published a risk assessment of erucic acid (22:1n-9) in 2016, establishing a Tolerable Daily Intake (TDI) for humans of 7 mg kg -1 body weight per day. This report largely excluded the contribution of erucic acid from fish and seafood, due to this fatty acid often not being reported separately in seafood. The Institute of Marine Research (IMR) in Norway analyzes erucic acid and has accumulated extensive data from analyses of fish feeds, farmed and wild fish, and seafood products. Our data show that rapeseed oil (low erucic acid varieties) and fish oil are the main sources of erucic acid in feed for farmed fish. Erucic acid content increases with total fat content, both in farmed and wild fish, and it is particularly high in fish liver, fish oil, and oily fish, such as mackerel. We show that the current TDI could be exceeded with a 200 g meal of mackerel, as at the maximum concentration analyzed, such a meal would contribute 143% to the TDI of a 60 kg person. These data cover a current knowledge gap in the scientific literature regarding the content of erucic acid in fish and seafood.

Brominated flame-retardants (BFRs) such as polybrominated diphenyl ethers (PBDE) and hexabromocyclododecane (HBCD) are considered hazardous to human health. Due to their persistence, they are still present in the environment and in biota and seafood is major contributor of BFRs to human exposure. Here, we used data from >9700 samples of wild and farmed fish, fish feed and fish feed ingredients collected from the North Atlantic between 2006 and 2016 aiming to investigate factors influencing the risk assessments of BFRs. Due to most representative number of analyses, PBDEs were the main focus of investigation. Mean ∑PBDE in fillet samples ranged from below quantification in Atlantic cod fillet to 2.0 μg kg -1 in Atlantic halibut. The main congener contributing to the ∑PBDE in all species was BDE 47. Factors affecting the level of BFR in seafood were multifaceted, and the levels were within species mainly determined by fish age, geographical origin and time of sampling. BDE 47, 99, 153 and HBCD were selected for margin of exposure (MOE) evaluation. When other sources of BFR than seafood were excluded, our risk assessment showed low risk at the current dietary intake of seafood. However, the dietary intake of BDE 99 may be of concern for toddlers when all sources are considered. The choice of fish species, dietary studies, choice of statistics, as well as exposure from other sources than seafood, were all factors that influenced the final MOE of BFRs. We propose the use of regression on order statistics as a tool for risk assessment, to illustrate means and spreads in large surveillance datasets to avoid the issue of measurements below the limit of quantification. A harmonized, updated evaluation of the risk associated with exposure to BFRs from diet, air and dust is warranted, where the fish species most commonly consumed also is taken into consideration., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The salmon feed composition has changed the last decade with a replacement of traditionally use of fish oil and fishmeal diets with vegetable ingredients and the use decontaminated fish oils, causing reduced concentrations of dioxins and dioxin-like PCBs in farmed Norwegian Atlantic salmon. The development of novel salmon feeds has prompted the need for prediction on dioxins and dl-PCB concentrations in future farmed salmon. Prediction on fillet dioxins and dl-PCB concentrations from different feed composition scenarios are made using a simple one-compartmental transfer model based on earlier established dioxin and dl-PCB congener specific uptake and elimination kinetics rates. The model is validated with two independent feeding trials, with a significant linear correlation (r(2) = 0.96, y = 1.0x, p < 0.0001, n = 116) between observed and predicted values. Model fillet predictions are made for the following four scenarios; (1) general feed composition of 1999, (2) feed composition of 2013, (3) future feed composition with high fish oil and meal replacement, (4) future feed composition with high fish oil and meal replacement and decontaminated fish oil. Model predictions of fillet dioxin and dl-PCB concentrations from 1999 (1.05 ng WHO2005-TEQs kg(-1)ww) and 2013 (0.57 ng WHO2005-TEQs kg(-1)ww) are in line with the data observed in national surveillance programs of those years (1.1 and 0.52 ng WHO2005-TEQs kg(-1)ww, respectively). Future use of high replacement and decontaminated oils feeds gave predicted fillet concentrations of 0.27 ng WHO2005-TEQs kg(-1)ww, which is near the limit of quantification., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The present study was conducted to evaluate the effects on Atlantic salmon hepatic lipid metabolism when fed diets with increasing substitution of fish oil (FO) with a vegetable oil (VO) blend. Four diets with VOs replacing 100, 90, 79 and 65 % of the FO were fed for 5 months. The levels of eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) in the experimental diets ranged from 1.3 to 7.4 % of fatty acids (FAs), while cholesterol levels ranged from 0.6 to 1.2 g kg(-1). In hepatocytes added [1-(14)C] α-linolenic acid (ALA, 18:3n-3), more ALA was desaturated and elongated to EPA and DHA in cells from fish fed 100 % VO, while in fish fed 65 % VO, ALA was elongated to eicosatrienoic acid (ETE; 20:3n-3), indicating reduced Δ6 desaturation activity. Despite increased desaturation activity and activation of the transcription factor Sp1 in fish fed 100 % VO, liver phospholipids contained less EPA and DHA compared with the 65 % VO group. The cholesterol levels in the liver of the 100 % VO group exceeded the levels in fish fed the 65 % VO diet, showing an inverse relationship between cholesterol intake and liver cholesterol content. For the phytosterols, levels in liver were generally low. The area as a proxy of volume of lipid droplets was significantly higher in salmon fed 100 % VO compared with salmon fed 65 % VO. In conclusion, the current study suggests that suboptimal dietary levels of cholesterol in combination with low levels of EPA and DHA (1.3 % of FAs) can result in minor metabolic perturbations in the liver of Atlantic salmon.

Aiming to re-evaluate current recommendations for nutrient supplementations when Atlantic salmon are fed diets based on plant ingredients, two regression experiments, with parr and post-smolt, were conducted. A control diet was included to evaluate if ingredients supplied sufficient nutrients without any added nutrient package (NP). The nutrient package consisted of vitamins B, C, E, minerals, cholesterol, methionine, taurine and histidine. This paper focus on B-vitamins. In parr, growth, health and welfare parameters responded on NP additions, but this was not observed in the seawater stage. During three months of feeding, parr tripled their weight. Parr given diets added the NP above NRC (2011) showed improved protein retention, and reduced liver and viscera indices. Post-smolt fed the same diets during five months showed a doubling of weight, but did not respond to the variation in NP to the same extent as parr. Significant regressions were obtained in body compartments for several of the B-vitamins in the premix. Whole body biotin concentration was unaffected by micronutrient premix level, and mRNA expression of the enzymes dependent of biotin showed only weak increases with increased biotin. Muscle thiamine plateaued at a diet level similar to NRC (2011) recommendation in freshwater, and showed stable values independent on premix addition in seawater. The mRNA expression of the enzyme G6PDH (glucose-6-phosphate dehydrogenase) is sensitive to thiamine availability; results did not indicate any need to add thiamine above levels recommended for fish in general. Niacin showed a steady increase in whole body concentrations as feed niacin increased. Muscle riboflavin peaked at a diet level of 12.4 mg kg -1 . Sufficient riboflavin is important to avoid e.g., development of cataract. Cataract was not registered to be any problem, neither in fresh- nor in seawater. Cobalamin (B 12 ) in muscle and liver was saturated at 0.17 mg kg -1 diet. Muscle pyridoxine showed a dose-dependent level in muscle, and peaked around 10 mg kg -1 diet. White muscle ASAT (asparagine amino transferase) activity steadily increased, with indications of stable values when dietary pyridoxine was around 10-16 mg kg -1 diet. Pantothenic acid increased in gill tissue up to a level of 5.5 mg kg -1 soft gill tissue; at a dietary level of 22 mg kg -1 . Improved performance, and coverage of metabolic need for niacin was at a dietary level of 66 mg kg -1 , riboflavin 10-12 mg kg -1 , pyridoxine 10 mg kg -1 and panthotenic acid 22 mg kg -1 . Based on these results, recommended B-vitamin supplementation in plant based diets for Atlantic salmon should be adjusted., Competing Interests: Kristin Hamre is an Academic Editor for PeerJ, otherwise there are no competing interests. Joana Silva is an employee of BioMar, Norway, and has no competing interest or any conflict with this study and publication.

This study aimed to investigate whether dioxin (TCDD) and methylmercury (MeHg) pose a threat to offspring of fish exposed to elevated concentrations of these chemicals via epigenetic-based mechanisms. Adult female zebrafish were fed diets added either 20 μg/kg 2,3,7,8 TCDD or 10 mg/kg MeHg for 47 days, or 10 mg/kg 5-aza-2'-deoxycytidine (5-AZA), a hypomethylating agent, for 32 days, and bred with unexposed males in clean water to produce F1 and F2 offspring. Global DNA methylation, promoter CpG island methylation and target gene transcription in liver of adult females and in 3 days post fertilization (dpf) F1 and F2 embryos were determined with HPLC, a novel CpG island tiling array containing 54,933 different probes and RT-qPCR, respectively. The results showed that chemical treatment had no significant effect on global DNA methylation levels in F1 (MeHg and TCDD) and F2 (MeHg) embryos and only a limited number of genes were identified with altered methylation levels at their promoter regions. CYP1A1 transcription, an established marker of TCDD exposure, was elevated 27-fold in F1 embryos compared to the controls, matching the high levels of CYP1A1 expression observed in F0 TCDD-treated females. This suggests that maternal transfer of TCDD is a significant route of exposure for the F1 offspring. In conclusion, the selected doses of TCDD and MeHg, two chemicals often found in high concentrations in fish, appear to have only modest effects on DNA methylation in F1 (MeHg and TCDD) and F2 (MeHg) embryos of treated F0 females., (Copyright © 2014 Elsevier Inc. All rights reserved.)

In the present study, zebrafish (Danio rerio) were fed casein/gelatin-based diets containing either 19% Bt (Bacillus thuringiensis)-maize or its parental non-Bt (nBt)-maize control for two generations (F0: sixty fish; F1: forty-two to seventy fish per treatment). The study focused on growth and reproductive performance, liver CuZn superoxide dismutase (SOD) enzyme activity, gene transcript levels targeting important cellular pathways in the liver and mid-intestine, histomorphological evaluation of the intestine, differential leucocyte counts, offspring larva swimming activity and global DNA methylation in offspring embryos. No significant effects were observed in the parental generation. The offspring were either fed the same diets as those fed to their parents (Bt-Bt or nBt-nBt) or switched from the Bt diet to the nBt diet (Bt-nBt). The Bt-Bt offspring exhibited a significantly higher body mass increase, specific growth rate and feed utilisation than fish fed the nBt-nBt diet and/or fish fed the Bt-nBt diet. Liver and mid-intestinal gene transcript levels of CuZn SOD were significantly higher in fish fed the nBt-nBt diet than in those fed the Bt-Bt diet. Liver gene transcript levels of caspase 6 were significantly lower for the nBt-nBt group than for the Bt-Bt group. Overall, enhanced growth performance was observed in fish fed the Bt diet for two generations than in those fed the nBt diet for one and two generations. Effects observed on gene biomarkers for oxidative stress and the cell cycle (apoptosis) may be related to the contamination of nBt-maize with fumonisin B1 and aflatoxin B1. In conclusion, it is suggested that Bt-maize is as safe and nutritious as its nBt control when fed to zebrafish for two generations.

The effects of feeding six diets spiked with increasing levels of DON for 45 days to zebrafish (Danio rerio) on performance and liver gene biomarkers were investigated. In addition long term effects on fecundity, offspring larvae swimming activity and global DNA methylation in embryos were investigated. Zebrafish performance was not affected. Liver CYP1A mRNA levels were significantly higher in fish fed 2.0 ppm DON compared to the control group, 0.1, 0.5 and 1.5 ppm group. Gene transcripts of CuZn SOD and Cyclin G1 increased with increasing content of dietary DON. The percentage of 5-methylcytosine in embryos did not differ and was 7.0-7.1% across the groups. Fecundity showed a biphasic response pattern. Interestingly, fish fed 1.5 ppm DON had 22% higher fecundity compared to control. A trend towards increased larvae swimming activity was seen in the high DON group. Our data suggest that DON is detoxified in the liver through the phase 1 system resulting in a disturbance in the oxidative balance. We do not know if effects observed on fecundity and larvae swimming activity are attributed to a direct interaction of DON with the reproductive organ or secondary to the maternal/paternal liver oxidative imbalance., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The objective of the present study was to investigate the effects of replacing high levels of marine ingredients with vegetable raw materials and with emphasis on lipid metabolism and net production of long-chain polyunsaturated ω-3 fatty acids (EPA + DHA). Atlantic salmon were fed three different replacement vegetable diets and one control marine diet before sensory attributes, β-oxidation capacity, and fatty acid productive value (FAPV) of ingested fatty acids (FAs) were evaluated. Fish fed the high replacement diet had a net production of 0.8 g of DHA and a FAPV of 142%. Fish fed the marine diet had a net loss of DHA. The present work shows that Atlantic salmon can be a net producer of marine DHA when dietary fish oil is replaced by vegetable oil with minor effects on sensory attributes and lipid metabolism.

The present study was conducted to follow up on apparent differences in growth, relative organ sizes, cellular stress and immune function in Atlantic salmon fed feed containing GM Bacillus thuringiensis maize compared with feed containing the non-modified parental maize line. Gene expression profiling on the distal intestinal segment and liver was performed by microarray, and selected genes were followed up by quantitative PCR (qPCR). In the liver, qPCR revealed some differentially regulated genes, including up-regulation of gelsolin precursor, down-regulation of ferritin heavy subunit and a tendency towards down-regulation of metallothionein (MT)-B. This, combined with the up-regulation of anti-apoptotic protein NR13 and similar tendencies for ferritin heavy chain and MT-A and -B in the distal intestine, suggests changes in cellular stress/antioxidant status. This corresponds well with and strengthens previous findings in these fish. To exclude possible confounding factors, the maize ingredients were analysed for mycotoxins and metabolites. The GM maize contained 90 μg/kg of deoxynivalenol (DON), while the non-GM maize was below the detection limit. Differences were also observed in the metabolite profiles of the two maize varieties, some of which seemed connected to the mycotoxin level. The effects on salmon observed in the present and previous studies correspond relatively well with the effects of DON as reported in the literature for other production animals, but knowledge regarding effects and harmful dose levels in fish is scarce. Thus, it is difficult to conclude whether the observed effects are caused by the DON level or by some other aspect of the GM maize ingredient.

The aim of this study was to investigate potential differences in liver protein expression of Atlantic salmon fed genetically modified (GM) Roundup Ready soy at a high inclusion level (25% inclusion, constituting 21% of crude protein in the diet) for 7 months or a compositionally similar non-GM diet. The liver was selected as the target organ due to its importance in the general metabolism, and 2D gel electrophoresis used as a screening tool. Samples from 12 individual fish from each diet group were evaluated. Of a total of 781 analysed protein spots, only 36 were significantly different by ANOVA (p < 0.05) in abundance between the diet groups. All these spots had low fold differences (1.2-1.6) and high false discovery rate (q = 0.44), indicating minor differences in liver protein synthesis between fish fed GM and non-GM soy. Additionally, low fold differences were observed. Four protein spots were analyzed by liquid chromatography tandem mass spectrometry and identified using a combination of online searches in NCBI and searches in an inhouse database containing salmonid expressed sequence tags and contigs. Follow-up on these proteins by real-time polymerase chain reaction did not identify differences at the transcriptional level.

Background: The aim of the study was to examine the intestinal cellular localization of proliferating cell nuclear antigen (PCNA) and cytochrome P450 A1 (CYP1A) expression in Atlantic salmon Salmo salar L. exposed to a model toxicant. The stress response was induced by intraperitoneal injection of four salmon with a single dose (50 mg/kg) of the CYP1A inducer beta-naphthoflavone (BNF) and intestinal tissue (mid and distal intestine; MI and DI) was sampled seven days later. Samples for histology and gene transcription analysis were collected from four exposed fish and four control fish. PCNA was assessed by immunohistochemistry, CYP1A mRNA was studied by in situ hybridization (ISH) and finally the transcription of five genes was quantified by real-time quantitative RT-PCR (real-time RT-PCR); two detoxifying genes (CYP1A and glutathione S-transferase; GST), a stress marker gene (heat shock protein 70; HSP70), PCNA and a gene marker of apoptosis (caspase 6A)., Results: PCNA protein and CYP1A mRNA were successfully localized in the intestinal cells (MI) of both experimental groups. At the cellular level, BNF significantly lowered intestinal cell proliferation and increased the CYP1A mRNA levels compared to the control group. The real-time RT-PCR data, which showed an increased mRNA expression both in the MI and DI of 139- and 62-fold, respectively, confirmed the increased cellular CYP1A mRNA levels detected using ISH. HSP70 expression was also up-regulated in the exposed fish. The other examined genes did not show any differential regulation in the experimental fish group., Conclusion: This study showed that CYP1A mRNA had a specific intestinal cellular transcription pattern in Atlantic salmon exposed to BNF. At the cellular level CYP1A mRNA expression was always observed at or around the cell nucleus close to the basolateral cell membrane and at the tissue level CYP1A mRNA expression was most frequently observed in the basal and apex area of the intestinal folds. Taken together, a link between the intestinal detoxification system (CYP1A) and cell renewal system (PCNA) is indicated with these two processes being inversely correlated in BNF exposed fish.

Background: The aim of this work was to study how evenly detoxifying genes are transcribed spatially in liver tissue of fish. Ten Atlantic salmon Salmo salar were intraperitoneally injected with 50 mg/kg of the strong CYP1A inducer beta-naphthoflavone and liver tissue harvested seven days later. The liver from 10 control and 10 exposed fish were split into eight sections, RNA extracted and three reference (beta-actin, elongation factor 1AB (EF1AB)) and two detoxifying genes (CYP1A and GST) quantified with real-time RT-PCR. The cellular localization of the EF1AB and CYP1A mRNA in the liver of control and beta-naphthoflavone treated fish was then determined by in situ hybridization (ISH) using EF1AB and CYP1A biotinylated oligonucleotide probes., Results: The study shows that genes encoding phase I and phase II conjugating enzymes are unevenly transcribed in different parts of the liver of Atlantic salmon seven days after a single-dose of beta-naphthoflavone exposure. Transcription of CYP1A and GST was higher in the middle section of the liver compared to the distal and proximal parts of the organ. The ISH data suggest that CYP1A transcription happens mainly in hepatocyte cells in the liver, and that hepatocytes in the vicinity of blood vessels respond stronger to beta-naphthoflavone than cells further away from the blood supply., Conclusion: Overall, the qRT-PCR and ISH results reported here suggest that gene expression analysis should be performed on as pure cell populations as possible. If bulk tissue samples are to be used, one should always check how evenly the target genes are expressed in tissue sections and organs in every study.