Evaluation of the maximum limits for selenium in Atlantic salmon feeds

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Quality of fish sludge as fertiliser to spring cereals: Nitrogen effects and environmental pollutants

The aim of this study was to contribute to development of organic fertiliser products based on fish sludge (i.e. feed residues and faeces) from farmed smolt. Four dried fish sludge products, one liquid digestate after anaerobic digestion and one dried digestate were collected at Norwegian smolt hatcheries in 2019 and 2020. Their quality as fertilisers was studied by chemical analyses, two 2-year field experiments with spring cereals and soil incubation combined with a first-order kinetics N release model. Cadmium (Cd) and zinc (Zn) concentrations were below European Union maximum limits for organic fertilisers in all products except one (liquid digestate). Relevant organic pollutants (PCB7, PBDE7, PCDD/F + DL-PCB) were analysed for the first time and detected in all fish sludge products. Nutrient composition was unbalanced, with low nitrogen/phosphorus (N/P) ratio and low potassium (K) content relative to crop requirements. Nitrogen concentration in the dried fish sludge products varied (27–70 g N kg-1 dry matter), even when treated by the same technology but sampled at different locations and/or times. In the dried fish sludge products, N was mainly present as recalcitrant organic N, resulting in lower grain yield than with mineral N fertiliser. Digestate showed equally good N fertilisation effect as mineral N fertiliser, but drying reduced N quality. Soil incubation in combination with modelling is a relatively cheap tool that can give a good indication of N quality in fish sludge products with unknown fertilisation effects. Carbon/N ratio in dried fish sludge can also be used as an indicator of N quality.publishedVersio

Selenium and selenium species in feeds and muscle tissue of Atlantic salmon

Selenium (Se) is an essential element for animals, including fish. Due to changes in feed composition for Atlantic salmon (Salmo salar), it may be necessary to supplement feeds with Se. In the present work, the transfer of Se and Se species from feed to muscle of Atlantic salmon fed Se supplemented diets was studied. Salmon were fed basal fish feed (0.35 mg Se/kg and 0.89 mg Se/kg feed), or feed supplemented either with selenised yeast or sodium selenite, at low (1–2 mg Se/kg feed) and high (15 mg Se/kg feed) levels, for 12 weeks. For the extraction of Se species from fish muscle, enzymatic cleavage with protease type XIV was applied. The extraction methods for Se species from fish feed were optimised, and two separate extraction procedures were applied, 1) enzymatic cleavage for organic Se supplemented feeds and 2) weak alkaline solvent for inorganic Se supplemented feeds, respectively. For selenium speciation analysis in feed and muscle tissue anion-exchange HPLC-ICP-MS for analysis of inorganic Se species and cation-exchange HPLC-ICP-MS for analysis of organic Se species, were applied. In addition, reversed phase HPLC-ICP-MS was applied for analysis of selenocysteine (SeCys) in selected muscle samples. The results demonstrated that supplemented Se (organic and inorganic) accumulated in muscle of Atlantic salmon, and a higher retention of Se was seen in the muscle of salmon fed organic Se diets. Selenomethionine (SeMet) was the major Se species in salmon fed basal diets and diets supplemented with organic Se, accounting for 91–118% of the total Se. In contrast, for muscle of salmon fed high inorganic Se diet, SeMet accounted for 30% of the total Se peaks detected. Several unidentified Se peaks were detected, in the fish fed high inorganic diet, and analysis showed indicated SeCys is a minor Se species present in this fish muscle tissue.publishedVersio

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.publishedVersio

Use of (Q)SAR genotoxicity predictions and fuzzy multicriteria decision-making for priority ranking of ethoxyquin transformation products

Ethoxyquin (EQ; 6-ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline) has been used as an antioxidant in feed for pets and food-producing animals, including farmed fish such as Atlantic salmon. In Europe, the authorization for use of EQ as a feed additive was suspended, due to knowledge gaps concerning the presence and toxicity of EQ transformation products (TPs). Recent analytical studies focusing on the detection of EQ TPs in farmed Atlantic salmon feed and fillets reported the detection of a total of 27 EQ TPs, comprising both known and previously not described EQ TPs. We devised and applied an in silico workflow to rank these EQ TPs according to their genotoxic potential and their occurrence data in Atlantic salmon feed and fillet. Ames genotoxicity predictions were obtained applying a suite of five (quantitative) structure–activity relationship ((Q)SAR) tools, namely VEGA, TEST, LAZAR, Derek Nexus and Sarah Nexus. (Q)SAR Ames genotoxicity predictions were aggregated using fuzzy analytic hierarchy process (fAHP) multicriteria decision-making (MCDM). A priority ranking of EQ TPs was performed based on combining both fAHP ranked (Q)SAR predictions and analytical occurrence data. The applied workflow prioritized four newly identified EQ TPs for further investigation of genotoxicity. The fAHP-based prioritization strategy described here, can easily be applied to other toxicity endpoints and groups of chemicals for priority ranking of compounds of most concern for subsequent experimental and mechanistic toxicology analyses.publishedVersio

The microbial safety of seaweed as a feed component for black soldier fly (Hermetia illucens) larvae

Farmed insects can offer an environmentally sustainable aquafeed or livestock feed ingredient. The value of black soldier fly (Hermetia illucens) (BSF) larvae however, could be improved by enrichment in omega-3 through the dietary inclusion of seaweed. However, the industry practice of drying seaweed at low temperatures to retain nutritional properties may benefit the survival of human pathogenic bacteria, particularly if the seaweed has been harvested from contaminated water. Here we have demonstrated that E. coli and E. coli O157:H7 died-off in seaweed dried at 50 °C, although both were detected in the dried powder following 72 h storage. V. parahaemolyticus fell below the level of detection in stored seaweed after drying at ≥ 50 °C, but L. monocytogenes remained detectable, and continued to grow in seaweed dried at ≤60 °C. Therefore, drying seaweed at low temperatures risks pathogen carry-over into insects destined for animal feed. BSF larvae reared on an artificially contaminated seaweed-supplemented diet also became contaminated by all four bacteria present in the supplement. Water quality at seaweed harvesting sites, seaweed desiccation, and insect rearing practices, represent critical points where development of regulatory standards could achieve targeted control of pathogenic hazards

Program for overvåking av fôr til landdyr - Årsrapport for prøver analysert av Havforskningsinstituttet i 2021

Overvåknings- og kartleggingsprogrammet for landdyr fôr utføres på vegne av Mattilsynet som en del av Norges oppfølging av nasjonalt og Europeisk regelverk på dyrefôr. Havforskningsinstituttet utførte i 2021 et utvalg av analyseporteføljen i dette overvåkingsprogrammet. I 2021 ble det mottatt 41 prøver fôr til drøvtyggere. Prøvene ble analysert for prosesserte animalske proteiner (PAP) og Salmonella ved Havforskningsinstituttet. Resultatene viste ingen funn av PAP eller bakterien Salmonella i fôrene som ble undersøkt i 2021.publishedVersio

Zinc uptake in fish intestinal epithelial model RTgutGC: Impact of media ion composition and methionine chelation

Apical uptake of zinc as ionic Zn(II) or as Zn-methionine (Zn-Met) was studied in RTgutGC cell line in vitro under media compositions mirroring the gut luminal ionic concentration of freshwater (FW) and seawater (SW) acclimated salmonids. Viability of the RTgutGC cells exposed to experimental media preparations showed a time-dependent decrease in SW treated cells, with the effect being significant at 48 h (P < 0.01), but not at 12 h or 24 h. Half effective concentration of Zn exposure over 12 h (EC50, in μM) was not differentially affected by media composition (FW, 59.7 ± 12.1 or SW, 83.2 ± 7.2; mean ± SE, P = 0.43). Zinc (65Zn) influx in RTgutGC was not different between FW or SW treated cells, but increased significantly in the presence of methionine (2 mM, L-Met or DL-Met). An interaction effect was observed between Zn concentration and media ionic composition on the impact of Met on apical Zn uptake (L-met, P < 0.001; DL-met, P = 0.02). In the presence of Met, apical Zn uptake in SW medium was significantly lower compared to FW, but only at higher Zn concentrations (12 and 25 μM, P < 0.01). Further, Met facilitated Zn uptake was reduced in cells treated with an amino acid transport system blocker with the effect being more significant and stereospecific in SW ionic conditions. The findings of this study showed that (i) Zn speciation in the presence of Met improved apical Zn uptake in RTgutGC cells and Zn-Met species were possibly taken up through Met uptake system. (ii) The effect was differentially affected by the ionic composition of the medium. Implications and limitations of the observations towards practical Zn nutrition of salmonids are discussed.publishedVersio