Lufenuron treatment temporarily represses gene expression and affects the SUMO pathway in liver of Atlantic salmon.

Lufenuron is a benzoylurea insecticide currently in use to combat sea lice infestation in salmon aquaculture in Chile. With pending approval in Norway, the aim of this work was to study the uptake and toxicity of lufenuron in liver tissue of Atlantic salmon. Juvenile salmon weighing 40 g were given a standard 7-day oral dose, and bioaccumulation and transcriptional responses in the liver were examined 1 day after the end-of-treatment (day 8) and after 1 week of elimination (day 14). Bioaccumulation levels of lufenuron were 29 ± 3 mg/kg at day 8 and 14 ± 1 mg/kg at day 14, indicating relatively rapid clearance. However, residues of lufenuron were still present in the liver after 513 days of depuration. The exposure gave a transient inhibition of transcription in the liver at day 8 (2437 significant DEGs, p-adj < .05), followed by a weaker compensatory response at day 14 (169 significant DEGs). Pathways associated with RNA metabolism such as the sumoylation pathway were most strongly affected at day 8, while the apelin pathway was most profoundly affected at day 14. In conclusion, this study shows that lufenuron easily bioaccumulates and that a standard 7-day oral dose induces a transient inhibition of transcription in liver of salmon.

Whole-animal accumulation, oxidative stress, transcriptomic and metabolomic responses in the pink shrimp (Pandalus montagui) exposed to teflubenzuron.

The benzoylurea chitin synthesis inhibitor teflubenzuron, widely used against sea lice in North Atlantic aquaculture, may pose an environmental threat to non-targeted crustaceans. In this experiment, laboratory acclimated pink shrimp (Pandalus montagui), a species found in fjords with Atlantic salmon farming, were exposed to dietary teflubenzuron for 46 days (control; low dose: 0.01 µg/g; high dose: 0.1 µg/g). The exposure doses represent 0.1% and 1% of a standard treatment dose for Atlantic salmon. Mortality and prevalence of deformities, pharmacokinetics, oxidative stress and transcriptomic and metabolomic profiling were used to assess the response to teflubenzuron exposure. Mortality in the high-dose group was 25% (five of 20 individuals). No control or low-dose group shrimps died. Phenotypic responses,i.e., leg deformities (0 control, 6 low, 8 high) and cloudy eyes (0 control, 3 low, 7 high), were observed in some surviving shrimps (control n = 15, low n = 17, high n = 15). Accumulated levels of teflubenzuron in shrimps from the high-dose group ranged from 4.7 to 369 ng/g wet weight. Transcriptomic profiling showed very few significantly altered genes in the exposed shrimps. Teflubenzuron-induced changes to the metabolome pointed to well-known effects of benzoylurea agents, with reduced levels of N-acetylglucosamine indicating an effect on chitin synthesis. The metabolomic profiling showed that teflubenzuron exposure was associated with reduced energy metabolism. Some metabolites pointed to increased necrosis and/or bacterial overgrowth in the teflubenzuron-exposed shrimps. In conclusion, this study shows that teflubenzuron causes phenotypic effects in P. montagui exposed to 0.1% of the treatment dose given to Atlantic salmon.

Lower levels of Persistent Organic Pollutants, metals and the marine omega 3-fatty acid DHA in farmed compared to wild Atlantic salmon (Salmo salar).

Contaminants and fatty acid levels in farmed- versus wild Atlantic salmon have been a hot topic of debate in terms of food safety. The present study determined dioxins (polychlorinated dibenzo-p-dioxin and dibenzofuran), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), organochlorine pesticides (OCPs), metals and fatty acids in wild and farmed Atlantic salmon. Contaminant levels of dioxins, PCBs, OCPs (DDT, dieldrin, lindane, chlordane, Mirex, and toxaphene), and mercury were higher in wild salmon than in farmed salmon, as were the concentrations of the essential elements selenium, copper, zinc and iron, and the marine omega-3 fatty acid docosahexaenoic acid (DHA). PBDE, endosulfan, pentachlorobenzene, hexachlorobenzene, cadmium and lead levels were low and comparable in both wild and farmed fish, and there was no significant difference in the marine omega-3 fatty acid eicosapentaenoic acid (EPA) concentration. The total fat content was significantly higher in farmed than wild salmon due to a higher content of both saturated and monounsaturated fatty acids, as well as a higher content of omega-6 fatty acids. The omega-3 to omega-6 fatty acid ratio was considerably lower in farmed than wild salmon due to the high level of omega-6 fatty acids. Contaminant concentrations in Atlantic salmon were well below maximum levels applicable in the European Union. Atlantic salmon, both farmed and wild, is a good source of EPA and DHA with a 200g portion per week contributing 3.2g or 2.8g respectively, being almost twice the intake considered adequate for adults by the European Food Safety Authority (i.e. 250mg/day or 1.75g/week).

Effects of aquaculture effluents on the slender sea pen Virgularia mirabilis.

This study aims to assess in situ the impact of effluents originating from an Atlantic salmon (Salmo salar) farm on a nearby slender sea pen (Virgularia mirabilis) field. We evidenced (1) the presence and persistence of emamectin residues (i.e. a common chemotherapeutants used for treating ectoparasites in salmons) in V. mirabilis tissue 56 days after treatment and (2) lethal and sublethal responses of V. mirabilis to effluents discharged by the salmon farm. Particularly, sea pens near the fish farm exhibited significant overproduction of mucus, contraction of polyps' tentacles, and disappearance of associated fauna. Furthermore, sea pens located directly underneath the farm showed substantial tissue necrosis and, in the most severe case, complete tissue loss and mortality. Our results suggest that lethal damages on sea pens occur directly below the farm, and that sublethal effects are visible up to 500 m from the farm. However, the presence of V. mirabilis below the studied farm, which has been active for more than twenty years, suggests that V. mirabilis population possesses the capacity to recover from the impacts of the farm, thereby preventing the complete disappearance from the area. In this context, it would be particularly interesting to run a temporal survey following the health state of V. mirabilis during an entire production cycle to have a more precise overview of fish farm impacts on this species, including during and after the post-production fallowing period.

Effects of the sea lice chemotherapeutant, emamectin benzoate, on metabolism and behaviour of the sea-pen Pennatula phosphorea.

Chemotherapeutants used to control infestations by sea lice can be released into the marine environment surrounding aquaculture farms. Among these therapeutic agents, emamectin benzoate is extensively utilized even though its impact on non-target taxa has not been thoroughly examined. In this context, we explored the effects of emamectin benzoate on a common Norwegian habitat-forming species: the phosphorescent sea-pen Pennatula phosphorea. Specifically, we examined P. phosphorea metabolic and responses before, during and after exposure to emamectin benzoate. Results indicate that an 8-day emamectin benzoate exposure (0.8 mg/L) did not induce P. phosphorea mortality or significant behavioural or metabolic modifications. However, we highlighted the presence and persistence of emamectin benzoate in exposed P. phosphorea tissue. These results indicate that emamectin benzoate is unlikely to adversely impact P. phosphorea populations in the environment. However, persistence of emamectin benzoate in tissue constitutes a potential for bioaccumulation with repeated treatments and should be examined in further studies.

Sampling of Atlantic salmon using the Norwegian Quality cut (NQC) vs. Whole fillet; differences in contaminant and nutrient contents.

Risk- benefit assessments of seafood require high-quality food composition data. In accordance with EU regulations, Atlantic salmon (Salmo salar) has conventionally been sampled using the Norwegian Quality Cut (NQC), a sub-cut of the middle section of the fish, in Norwegian surveillance programs. By comparing the contents of nutrients and contaminants in 34 samples of farmed Atlantic salmon, we aimed to evaluate the representativeness of the NQC compared with the whole fillet. Of the 129 analytes evaluated, eight single analytes, in addition to 25 different fatty acids, showed significant differences between the cuts. Significant differences were evident for total fat, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and sum PCB-6, but not for the sum of dioxins and dioxin-like PCBs. We further suggest that the NQC may still be used in large-scale sampling of Atlantic salmon, and that the whole fillet would be preferable when analysing the content of nutrients.

Developing a framework for open and FAIR data management practices for next generation risk- and benefit assessment of fish and seafood.

Risk and risk-benefit assessments of food are complex exercises, in which access to and use of several disconnected individual stand-alone databases is required to obtain hazard and exposure information. Data obtained from such databases ideally should be in line with the FAIR principles, i.e. the data must be Findable, Accessible, Interoperable and Reusable. However, often cases are encountered when one or more of these principles are not followed. In this project, we set out to assess if existing commonly used databases in risk assessment are in line with the FAIR principles. We also investigated how access, interoperability and reusability of data could be improved. We used the OpenFoodTox and the Seafood database as examples and showed how commonly used freely available open-source tools and repositories can be implemented in the data extraction process of risk assessments to increase data reusability and crosstalk across different databases.

Temporal variations in the nutrient content of Norwegian farmed Atlantic salmon (Salmo salar), 2005-2020.

The changes in the feed of farmed Atlantic salmon (Salmo salar) towards a more plant-based diet affect the nutritional value of the fillets. By compiling the contents of a range of nutrients in 1108 samples of Norwegian farmed Atlantic salmon collected between 2005 and 2020, we found that the median contents of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) have decreased by > 60%. However, farmed Atlantic salmon remains a considerable source of EPA and DHA, with one and two portions being sufficient to meet the weekly adequate intake of EPA and DHA for adults (175 g) and two-year-olds (80 g), respectively. Farmed Atlantic salmon also remains a considerable source of protein, selenium, vitamin B12, and vitamin D3. Together, we demonstrate that farmed Atlantic salmon can contribute substantially to the nutrient intake of the consumers. These data are important for the Norwegian food composition table and future risk-benefit assessments on fatty fish consumption.

Co-occurrence of contaminants in marine fish from the North East Atlantic Ocean: Implications for human risk assessment.

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.

Analytical recovery of folate and its degradation products in human serum stored at -25 degrees C for up to 29 years.

Epidemiological studies on folate and chronic diseases often involve the use of frozen serum stored in biorepositories for decades. Folate instability may attenuate associations between folate status and study endpoints. In this cross-sectional study, we retrieved serum samples stored at -25 degrees C for 0, 4, 6, 17, or 29 y in the Janus biobank. Samples were obtained from a total of 650 men aged 40-49 y at the time of blood collection and were evenly distributed according to storage time. Folate was determined by a liquid chromatography tandem MS (LC-MS/MS) assay that measures 5-methyltetrahydrofolate (5mTHF), its oxidation product 4-alpha-hydroxy-5-methyltetrahydrofolate (hmTHF), and other folate species; by a Lactobacillus rhamnosus microbiological assay; and by LC-MS/MS as p-aminobenzoylglutamate (pABG) equivalents after oxidation and mild acid hydrolysis of the folate species. Concentrations of 5mTHF and microbiologically active folate were lower in samples that had been subjected to long-term storage and the data were consistent with a decrease of 3.2 and 2.8%/y, respectively. hmTHF was detected in all specimens but did not accumulate upon long-term storage (>4 y). Folate measured as pABG declined at a slow rate of 0.98%/y and approximately 80% of the folate was recovered after 29 y of storage. B-vitamin status did not differ between individuals delivering samples at different time points, as assessed by measuring total homocysteine, methylmalonic acid, and serum vitamin B-12. In conclusion, folate is substantially degraded in serum frozen for decades but can to a large extent be recovered as pABG equivalents. The pABG assay appears to be the method of choice for the determination of folate status in archival serum samples.

Liquid chromatography-tandem mass spectrometry analysis of folate and folate catabolites in human serum.

BACKGROUND: Folate status is associated with several chronic diseases; thus accurate assessment of folate status has become important in the clinical setting and in epidemiological studies. The diversity of folate forms complicates the task of assaying endogenous folate. We developed and validated an assay that measures various forms of folate in addition to folate catabolites in human serum. METHODS: We added ascorbic acid to serum samples from 168 healthy blood donors and 39 patients with renal failure, and precipitated the proteins with acetonitrile containing (13)C-labeled folate forms as internal standards. The supernatant was evaporated and the analytes redissolved in water. We then used liquid chromatography-tandem mass spectrometry to quantify 5-methyltetrahydrofolate (5mTHF), 4-alpha-hydroxy-5-methyltetrahydrofolate (hmTHF), folic acid (FA), 5-formyltrahydrofolate (5fTHF), p-aminobenzoylglutamate (pABG), and p-acetamidobenzoylglutamate (apABG). RESULTS: Detection limits were 0.07-0.52 nmol/L, and the assay was linear to 140 nmol/L for all analytes. The mean serum folate concentration from 168 blood donors was 22.7 nmol/L, of which 85.8% was 5mTHF, 12.1% hmTHF, 2.1% FA, and 0.0% 5fTHF. In the same individuals, the mean concentrations of pABG and apABG were 0.07 nmol/L and 0.47 nmol/L, respectively. The concentrations of folate catabolites were 22-30 times higher in 39 patients with renal failure. This folate assay correlated well with the microbiologic assay (r(2) = 0.92) and with measurement of serum folate as pABG equivalents (r(2) = 0.93). CONCLUSIONS: This method based on liquid chromatography-tandem mass spectrometry measures the most abundant folate species and 2 folate catabolites in human serum.

Analytical recovery of folate degradation products formed in human serum and plasma at room temperature.

Folate is not stable in serum and plasma. This may impair laboratory diagnostics and distort the outcome of epidemiological studies on folate and chronic diseases. The present study was designed to determine the kinetics of folate loss in human serum and plasma (collected into tubes containing EDTA, heparin, or citrate) at room temperature and the recovery of folate as 4-alpha-hydroxy-5-methyltetrahydrofolate (hmTHF) or p-aminobenzoylglutamate (pABG) equivalents. Different folate species and pABG were determined by liquid chromatography-tandem MS and microbiologically active folate was measured by a Lactobacillus rhamnosus assay. Concentrations of 5mTHF and microbiologically active folate had a parallel and rapid decrease in EDTA plasma to approximately 60% of the initial concentration after 24 h. In serum, heparin plasma, and citrate plasma, folate decreased more slowly to approximately 50% after 192 h. The loss of 5mTHF that occurred within 48 h was totally recovered as hmTHF. Folate measured as pABG equivalents decreased slowly to approximately 80% in 192 h and the decline was essentially matrix independent. In conclusion, the degradation of 5mTHF and microbiologically active folate in serum and plasma at room temperature can largely be corrected for by determining hmTHF or measuring folate as pABG equivalents. Moreover, results obtained using conventional folate assays may be biased by improper sample handling or if samples contained high concentrations of hmTHF.

Identification of ethoxyquin and its transformation products in salmon after controlled dietary exposure via fish feed.

Ethoxyquin (EQ) is an additive present in fish feed and its fate in fish should be carefully characterized due to food safety concerns regarding this compound. Therefore, the objective of this work was to identify the transformation products (TPs) of EQ in Atlantic salmon. Salmon in independent tanks were given feed containing ethoxyquin concentrations of 0.5 mg/kg, 119 mg/kg or 1173 mg/kg for 90 days. After the feeding trial, salmon fillets were extracted in acetonitrile and analyzed by liquid chromatography with traveling-wave ion mobility spectrometry coupled to high resolution mass spectrometry (UHPLC-TWIMS-QTOFMS). EQ was transferred from the feed to salmon fillets and 23 TPs were characterized, resulting from dimerization, oxygenation, cleavage, cleavage combined with oxygenation, cleavage combined with conjugation, and other uncategorized alterations. Moreover, EQ and some TPs were also detected in commercial salmon randomly sampled from different Norwegian fish farms. This study confirmed that the dimer 1,8'-EQDM was the main TP of EQ and, together with previous research, brought the overall number of characterized TPs to a total of 47.

Natural and anthropogenic radionuclides in Norwegian farmed Atlantic salmon (Salmo salar).

Norway is one of the main producers of farmed fish and the world's second-largest exporter of seafood. Farmed Atlantic salmon (Salmo salar) represents the most exported species. This is the first comprehensive survey of anthropogenic (137Cs, 90Sr, 238Pu, 239,240Pu and 241Am) and natural (40K, 226Ra, 228Ra, 210Pb, 210Po) radionuclides in farmed salmon and manufactured fish feed from Norway. The only anthropogenic radionuclide detected in salmon and fish feed was 137Cs. The levels were low with arithmetic means in salmon and feed of 0.13 and 0.30 Bq/kg fresh weight (fw), respectively. The natural radionuclide 40K exhibited the highest levels with arithmetic means in salmon and feed of 115 and 239 Bq/kg fw, respectively. The arithmetic means of 210Po and 210Pb in salmon were 0.013 and 0.044 Bq/kg fw, respectively, with a mean 210Po:210Pb activity ratio of 0.32. For fish feed, the situation was reversed: the arithmetic means of 210Po and 210Pb were 3.8 and 0.67 Bq/kg fw, respectively, with a mean 210Po:210Pb activity ratio of 5.7. The radionuclide levels found in farmed salmon in the present study are comparable to or lower than the levels found in other fish species in the North Atlantic Ocean. A highly conservative dose estimate for consumption showed that doses were no higher than 1.2 µSv/year for toddlers and 4.0 µSv/year for adults. This suggests that the risk associated with radioactivity in farmed salmon is very low even when considering individuals with high consumption and the highest radionuclide levels found in this study.

Pharmacokinetic Data Show That Oxolinic Acid and Flumequine Are Absorbed and Excreted Rapidly From Plasma and Tissues of Lumpfish.

This study examined the uptake, tissue distribution and elimination of the antibacterial agents oxolinic acid and flumequine in lumpfish (Cyclopterus lumpus L.) by use of LC-MS/MS following a single oral administration of 25 mg/kg fish given in feed. Lumpfish are increasingly used as cleaner fish for removal of sea lice on commercially farmed salmon. The production of lumpfish is successful, but there are challenges with bacterial infections and the number of antibacterial treatments has increased in recent years. As the lumpfish is a novel species to farming, there is a need for pharmacokinetic data and establishment of protocols for efficient antibacterial treatment. The current study describes the pharmacokinetic properties of oxolinic acid and flumequine in lumpfish. Absorption of oxolinic acid was moderate and was characterized by a calculated peak plasma concentration (Cmax) of 2.12 µg/ml after 10.3 h (Tmax) and an elimination half-life (t1/2ß) of 21 h. Area under curve (AUC) and AUC from 0 to 24 h (AUC0-24h) were calculated to be 60.9 and 34.0 h µg/ml, respectively. For flumequine, plasma Cmax was found to be 2.77 µg/ml after 7.7 h (Tmax) with t1/2ß of 22 h. The area under the curve (AUC) and AUC from 0 to 24 h (AUC0-24) were calculated as 104.3 and 50.3 h µg/ml, respectively. Corresponding Cmax values in muscle, liver, and head-kidney for oxolinic acid were 4.01, 3.04, and, 4.68 µg/g, respectively and Tmax of 11.1, 9.2, and 10.0 h, respectively. For flumequine, Cmax values of 4.16, 4.01, and 7.48 µg/g were obtained in muscle, liver, and head kidney, respectively, with corresponding Tmax values of 10.2, 10.3, and 6.0 h. Antimicrobial susceptibility values as determined by minimum inhibitory concentration (MIC) analyses against 28 isolates of Aeromonas salmonicida isolated from diseased lumpfish ranged from 0.06 to 15 µg/ml for oxolinic acid and 0.024 to 6.25 µg/ml for flumequine. Bimodal distributions in susceptibility to both oxolinic acid and flumequine were observed. The combination of pharmacokinetic properties and MIC data make possible calculation of efficient treatment doses, which are needed to improve the welfare of lumpfish and minimize development of antibiotic resistant bacteria.

Effects of exercise and dietary protein sources on adiposity and insulin sensitivity in obese mice.

Low-fat diets and exercise are generally assumed to ameliorate obesity-related metabolic dysfunctions, but the importance of exercise vs. dietary changes is debated. Male C57BL/6J mice were fed a high-fat/high-sucrose (HF/HS) diet to induce obesity and then either maintained on the HF/HS or shifted to low-fat (LF) diets containing either salmon or entrecote. For each diet, half of the animals exercised voluntarily for 8 weeks. We determined body composition, glucose tolerance, insulin sensitivity and hepatic triacylglycerol levels. The microbiota composition in cecal and fecal samples was analyzed using 16S ribosomal RNA gene amplicon sequencing. Voluntary exercise improved insulin sensitivity but did not improve glucose tolerance. Voluntary exercise did not reduce adiposity in mice maintained on an HF/HS diet but enhanced LF-induced reduction in adiposity. Hepatic triacylglycerol levels were reduced by voluntary exercise in LF- but not HF/HS-fed mice. Voluntary exercise induced shifts in the cecal and fecal microbiota composition and functional potential in mice fed LF or HF/HS diets. Whereas voluntary exercise improved insulin sensitivity, a switch to an LF diet was the most important factor related to body weight and fat mass reduction.

Modelling the long-term feed-to-fillet transfer of leuco crystal violet and leuco malachite green in Atlantic salmon (Salmo salar).

Leuco crystal violet (LCV) and leuco malachite green (LMG) are the main metabolites of two dyes that are forbidden for use in food production, but can be present at low background concentration in novel Atlantic salmon feed ingredients such as processed animal proteins (animal by-product [ABP]). In this study, the potential transfer of dietary LCV or LMG to the fillet of farmed Atlantic salmon was investigated. The uptake and elimination rate kinetics were determined in seawater-adapted Atlantic salmon (initial weight 587 ± 148 g) fed two levels of either LCV- or LMG-enriched diets (~500 and 4000 µg kg-1, respectively) for 40 days, followed by a 90-day depuration period with feeding on control diets (<0.15 µg kg-1 LCV and LMG). A three-compartmental model was developed, based on a fillet fat, fillet muscle and a central body compartment comprising all other tissues. Model calibrations showed a good fit with measured values during overall uptake and elimination period; however, the model poorly predicted the short-term (days) peak measured values at the end of the exposure period. The model was used to simulate the long-term (>16 months) LCV and LMG feed-to-fillet transfer in Atlantic salmon under realistic farming conditions such as the seasonal fluctuations in feed intake, growth and fillet fat deposition. The model predictions gave highest expected LCV and LMG fillet concentrations of approximately 0.12 and 0.45 µg kg-1, depending on the dietary levels of ABP and background level of LCV and LMG contamination. These levels are under the reference point for action of 2 µg kg-1 for the sum of MG and LMG that EFSA assessed as adequate to protect public health. However, for LCV, the predicted highest levels exceed the analytical decision limit (CCα) of 0.15 µg kg-1 for the method used in this paper.

Factors influencing risk assessments of brominated flame-retardants; evidence based on seafood from the North East Atlantic Ocean.

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.

Investigation of pharmaceuticals in processed animal by-products by liquid chromatography coupled to high-resolution mass spectrometry.

There is an on-going trend for developing more sustainable salmon feed in which traditionally applied marine feed ingredients are replaced with alternatives. Processed animal products (PAPs) have been re-authorized as novel high quality protein ingredients in 2013. These PAPs may harbor undesirable substances such as pharmaceuticals and metabolites which are not previously associated with salmon farming, but might cause a potential risk for feed and food safety. To control these contaminants, an analytical strategy based on a generic extraction followed by ultra-high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS) using quadrupole time-of-flight mass analyzer (QTOF MS) was applied for wide scope screening. Quality control samples, consisting of PAP commodities spiked at 0.02, 0.1 and 0.2 mg/kg with 150 analytes, were injected in every sample batch to verify the overall method performance. The methodology was applied to 19 commercially available PAP samples from six different types of matrices from the EU animal rendering industry. This strategy allows assessing possible emergent risk exposition of the salmon farming industry to 1005 undesirables, including pharmaceuticals, several dyes and relevant metabolites.

Distribution and persistence of the anti sea-lice drug teflubenzuron in wild fauna and sediments around a salmon farm, following a standard treatment.

The salmon louse (Lepeoptheirus salmonis) is a challenge in the farming of Atlantic salmon (Salmo salar). To treat an infestation, different insecticides are used like the orally administered chitin synthetase inhibitor teflubenzuron. The concentrations and distribution of teflubenzuron were measured in water, organic particles, marine sediment and biota caught in the vicinity of a fish farm following a standard medication. Low concentrations were found in water samples whereas the organic waste from the farm, collected by sediment traps had concentrations higher than the medicated feed. Most of the organic waste was distributed to the bottom close to the farm but organic particles containing teflubenzuron were collected 1100 m from the farm. The sediment under the farm consisted of 5 to 10% organic material and therefore the concentration of teflubenzuron was much lower than in the organic waste. Teflubenzuron was persistent in the sediment with a stipulated halflife of 170 days. Sediment consuming polychaetes had high but decreasing concentrations of teflubenzuron throughout the experimental period, reflecting the decrease of teflubenzuron in the sediment. During medication most wild fauna contained teflubenzuron residues and where polychaetes and saith had highest concentrations. Eight months later only polychaetes and some crustaceans contained drug residues. What dosages that induce mortality in various crustaceans following short or long-term exposure is not known but the results indicate that the concentrations in defined individuals of king crab, shrimp, squat lobster and Norway lobster were high enough shortly after medication to induce mortality if moulting was imminent. Considering food safety, saith and the brown meat of crustaceans contained at first sampling concentrations of teflubenzuron higher than the MRL-value set for Atlantic salmon. The concentrations were, however, moderate and the amount of saith fillet or brown meat of crustaceans to be consumed in order to exceed ADI is relatively large.