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In this study, ensilaging of herring (Clupea harengus) filleting co-products was taken from lab-scale to pilot scale (1500 L) while monitoring the protein degree of hydrolysis (DH) and lipid oxidation. Subsequently, the possibility of recovering fish oil and protein hydrolysates using batch centrifugation at different g-forces/times was investigated. Around 38% DH was recorded after 2-day pilot-scale ensilaging of herring co-products at ambient temperature (i.e., ~ 22 °C), which was similar to the DH found in lab-scale (40% after 2 days; 22 °C). The lipid oxidation marker 2-thiobarbituric acid reactive substances (TBARS) reached 20 µmole TBARS/kg silage after 2-day ensilaging. Centrifugation of the silage at 3000–8500 × g for 2–20 min revealed successful separation into fish oil and protein hydrolysates. Heat-treating the silage (85 °C; 30 min) prior to centrifugation resulted in significantly higher oil and hydrolysates recoveries; the same being true for increased g-force. At 8500 × g, the recovery of oil and hydrolysates were 9.7 and 53.0% w/w, respectively, from heat-treated silage, while recoveries were 4.1 and 48.1% w/w, respectively, from non-heat treated silage. At 4500 × g, being a more scalable approach, corresponding numbers were 8.2 and 47.1% (w/w) as well as 2.0 and 40.2% (w/w). The recovered fish oil contained 8% EPA and 11% DHA of total fatty acids. Free fatty acids (FFA), peroxide value (PV), p-anisidine value (p-AV), and total oxidation (TOTOX) values of oils were in the range of 4–7% (FFA), 3.6–3.7 meq/kg oil (PV), 2.5–4.0 (p-AV), and 9.9–11.1 (TOTOX), respectively, which were within the acceptable limits for human consumption specified by the GOED voluntary monograph. The recovered protein hydrolysates contained peptides in the molecular weight range 0.3–6 kDa (~ 37%) and 11–34 kDa (~ 63%). Also, the remaining solids contained 15–17% (w/w) protein, having 44–45% essential amino acids. Overall, the results suggest that herring co-product silage is a valuable source of fish oil and protein hydrolysates, paving the way for ensilaging based-biorefining of herring co-products into multiple products.

A novel integrated process for recovery of protein-enriched biomasses from 5% presalting brines and spice brines of herring (Clupea harengus) was investigated by combining carrageenan- and/or acid-driven flocculation (F) plus dissolved air flotation (DAF). The F-DAF technique with carrageenan resulted in protein and lipid recoveries from 5% presalting brine of 78 and 38%, respectively. Without flocculation or with only acidification, protein and lipid recoveries in DAF were only 13 and 10%, respectively. Low protein and lipid recoveries, 8-12 and 1.8-8.2%, respectively, were also obtained when spice brine was subjected to only acidification and DAF. The protein content in dry biomasses from 5% presalting brine and spice brine was 36-43 and 13-16%, respectively. The corresponding lipid levels were 23-31 and 9-18%, respectively, with ash levels of 11-20 and 38-45%, respectively. Biomass proteins contained ≤45% essential amino acids, and the lipids had ≤16% long-chain n-3 polyunsaturated fatty acids. Freeze-dried spice brine biomasses were characterized by anchovy- and spice-related sensory attributes. 5% presalting brine biomasses were connected to fish and seafood attributes and showed gel forming capacity. The outlined F-DAF recovery system can thus recover both nutrients and interesting flavors from the herring process waters, which are currently lost from the food chain.

The seaweed Saccharina latissima is often blanched to lower iodine levels, however, it is not known how blanching affects protein extraction. We assessed the effect of blanching or soaking (80/45/12 °C, 2 min) on protein yield and protein extract characteristics after pH-shift processing of S. latissima. Average protein yields and extract amino acid levels ranked treatments as follows: blanching-45 °C ∼ control soaking ∼ blanching-80 °C. Although blanching-45 °C decreased protein solubilization yield at pH 12, it increased isoelectric protein precipitation yield at pH 2 (p 0.05). The former could be explained by a higher ratio of large peptides/proteins in the blanched biomass as shown by HP-SEC, whereas the latter by blanching-induced lowering of ionic strength, as verified by a dialysis model. Moreover, blanching-45 °C yielded a protein extract with 49 % less iodine compared with the control extract. We recommend blanching-45 °C since it is effective at removing iodine and does not compromise total protein extraction yield.

Cross-processing herring co-products with antioxidant-rich helpers including lingonberry-press-cake, shrimp-shells and seaweed was reported to mitigate lipid oxidation but reduce protein yield. Here, four strategies were used to counteract such yield-reduction; optimizing solubilization/precipitation pH, increasing raw-material-to-water-ratio, replacing single-stage-toothed- by radial-discharge- high-shear-mechanical-homogenization (RD-HSMH) and ultrasonication (US). The effects of RD-HSMH and US on lipid oxidation, protein structural and functional properties were studied. Combining four strategies improved total protein yield by 5-12 %, depending on helper type. More than the confirmed antioxidant effects, cross-processing also improved protein water solubility and emulsification activity but reduced gelation properties. RD-HSMH generally improved protein emulsifying and gelation properties but reduced protein water solubility. US reduced protein water solubility and gelation properties. Altogether, it was recommended for all helpers to increase solubilization pH to 12 and raw-material-to-water-ratio to 1:6 followed by RD-HSMH at 8000 rpm for 90 s, aiming for maximum protein yield and emulsifying and gelation properties.

Provided high product quality, ensilaging can be used to valorize fish filleting co-products into a silage suitable for food applications. However, a documented challenge for products from hemoglobin-rich fish raw materials is the high susceptibility to lipid oxidation, calling for stabilization by antioxidants. In a comparison among different rosemary-containing antioxidants and isoascorbic acid, we here found that the commercial mixture Duralox MANC-213 (MANC) provided the best protection against peroxide value and 2-thiobarbituric acid reactive substances (TBARS) development during ensilaging of herring filleting co-products (0–7 days, 22 °C), and also during subsequent heat-treatment (30 min, 85 °C). Increasing MANC concentration from 0.25 and 0.75 to 1.25% lowered TBARS values from 43.53 and 25.12 to 18.04 µmole TBARS/Kg silage, respectively, after 7 days of ensilaging. During storage at 4 °C/22 °C in presence of MANC, 1.25% provided the highest protection with 87–90% and 66–73% lower TBARS, at 4 °C and 22 °C, respectively, at 6 months compared to the controls. At this time point, heat-treated silages had lower protein degree of hydrolysis and free amino acids values than the non-heat-treated one. Regardless of antioxidant addition, total volatile basic nitrogen (TVB-N) formation still increased during the storage, but, overall, TVB-N values in silages were below the acceptable limit of 30 mg TVB-N/100 g fish for human consumption. Together with lipid oxidation data, this suggest that herring silage produced in presence of antioxidants can be used both for high quality feed and food applications.

Processes currently used for collagen extraction are complicated requiring a great deal of time and chemicals. Here, high shear mechanical homogenization (HSMH) and ultrasound (US) were integrated in the pretreatment step of collagen extraction from common starfish to reduce chemical use and time consumption. Effects of the assistant technologies on yield, structural integrity and functionality of collagen were also investigated. HSMH reduced the deproteinization time from 6 h to 5 min and its required amount of alkali 4 times, compared with classic methods. HSMH + US reduced the demineralization time from 24 h to 12 h and improved its efficiency in extraction of minerals. Collagen extraction with HSMH and HSMH + US resulted in similar yield as the classic method and did not affect triple helical structural integrity, polypeptide pattern, thermal stability or fibril-formation capacity of the collagens. Altogether, HSMH and US can effectively improve resource efficiency during collagen extraction without imposing negative effect on collagen quality.

This study aimed at limiting hemolysis of fish red blood cells (RBCs) as a strategy to limit hemoglobin (Hb)-induced lipid oxidation during post-mortem handling and processing. Effects of varying temperature, salinity, and mechanical impact were studied using washed resuspended RBCs (wr-RBCs) and whole blood (WB) from rainbow trout (Oncorhynchus mykiss) and herring (Clupea harengus). The wr-RBCs were most stable avoiding mechanical stress, keeping isotonic conditions (0.9–1.3% NaCl) and low temperature 0–6 °C, with predicted minimum at 2.5 °C. When compared at the same salinity, it was found that hemolysis was more pronounced in herring than trout wr-RBCs. Furthermore, WB was more stable than wr-RBCs, showing protecting the effects of blood plasma. Studying individual plasma components, stabilizing effects were found from glucose, proteins, and ascorbic acid. This study indicates that small adjustments in the early handling and processing of fish such as changing salinity of storage and rinsing solutions could minimize Hb contamination of the fish muscle and thereby improve quality.

A novel integrated process for the recovery of protein-enriched biomasses from shrimp boiling water (SBW) and shrimp peeling water (SPW) was investigated by combining flocculation (F) and dissolved air flotation (DAF) into an F-DAF process. Alginate and carrageenan were used as flocculants. It was found that the protein yield from SPW and SBW in the F-DAF process was 68-97% and 26-45%, respectively. This led to a reduction in protein content of the influent SBW and SPW (12.4 and 1.4 g/L, respectively) by up to 76% and 85%, resulting in outlets with 2.9 and 0.2 g/L protein, respectively. Further, the F-DAF recovery process concentrated the proteins of SBW and SPW up to 7 and 29 times, respectively, thus, generating a protein-enriched biomass. After spray drying, biomass from SBW had up to 61% proteins and out of the total amino acids, up to 47% were essential ones. Further, the spray dried powder contained up to 23% total lipids, 2.7% long chained n-3 polyunsaturated fatty acids and 49.7 mg/g total astaxanthin. The studied F-DAF recovery system can thus be successfully used for recovering nutrients from process waters generated during the production of peeled shrimps.

Ulva fenestrata is an economically and ecologically important green algal species with a large potential in seaweed aquaculture due to its high productivity, wide environmental tolerance, as well as interesting functional and nutritional properties. Here, we performed a series of manipulative cultivation experiments in order to investigate the effects of irradiance (50, 100, and 160 μmol photons m−2 s−1), temperature (13 and 18 °C), nitrate (pCO2 (200, 400, and 2500 ppm) on the relative growth rate and biochemical composition (fatty acid, protein, phenolic, ash, and biochar content) in indoor tank cultivation of Swedish U. fenestrata. High irradiance and low temperature were optimal for the growth of this northern hemisphere U. fenestrata strain, but addition of nutrients or changes in pCO2 levels were not necessary to increase growth. Low irradiance resulted in the highest fatty acid, protein, and phenolic content, while low temperature had a negative effect on the fatty acid content but a positive effect on the protein content. Addition of nutrients (especially nitrate) increased the fatty acid, protein, and phenolic content. High nitrate levels decreased the total ash content of the seaweeds. The char content of the seaweeds did not change in response to any of the manipulated factors, and the only significant effect of changes in pCO2 was a negative relationship with phenolic content. We conclude that the optimal cultivation conditions for Swedish U. fenestrata are dependent on the desired biomass traits (biomass yield or biochemical composition).

In recent years, the interest in using seaweed for the sustainable production of commodities has been increasing as seaweeds contain many potentially worthwhile compounds. Thus, the extraction and refining processes of interesting compounds from seaweeds is a hot research topic but has been found to have problems with profitability for novel applications. To increase the economic potential of refining seaweed biomass, the content of the compounds of interest should be maximized, which can potentially be achieved through optimization of cultivation conditions. In this study, we studied how the monosaccharide composition of the green seaweed species Ulva fenestrata is influenced by the abiotic factors; irradiance, temperature, nitrate, phosphate, and pCO2. It was evident that lower nitrate concentration and cultivation at elevated temperature increased monosaccharide contents. A 70% increase in iduronic acid and a 26% increase in rhamnose content were seen under elevated irradiance and temperature conditions, though the absolute differences in monosaccharide concentration were small. Irradiance and nitrate impacted the ratio between iduronic and rhamnose, which is an indicator of the ulvan structure. These results could potentially be utilized to coax the ulvan towards specific bioactivities, and thus have a considerable impact on a potential biorefinery centered around Ulva.

Whole body saline-perfused rainbow trout (Oncorhynchus mykiss) was ice-stored for 4 weeks and compared with unwashed/washed minces from unbled and bled trout in terms of rancid odour, peroxide value (PV), thiobarbituric acid reactive substances (TBARS) and redness loss. Muscle from saline-perfused fish, which had 72% less total haem, was deficient in rancid odour during the whole storage, while bled (54% less haem) and unbled samples developed rancid odour already after ~4 and 2 days; higher intensity without bleeding. PV/TBARS also developed in the order unbled > bled > perfused samples; however, PV/TBARS were not as completely prevented as rancid odour after perfusion. Saline washing (3 × 3 volumes) of unbled mince removed 84% haem and yielded the second most stable sample while saline washing (1 × 1 volumes) destabilised unbled mince, despite 64% haem removal. Concurrent antioxidant removal during washing of minces obviously counteracted the effect of blood removal and washing fish mince with small volumes of solution should be used with great care.

The degradation of trout and bovine hemoglobin (Hb) and their pro-oxidant activities in washed cod muscle mince (WCM) were studied using simple pH-shifts to simulate gastrointestinal (GI) conditions (pH 7 → 6 → 3 → 7), as well as full static in vitro GI digestion. Following gastric acidification to pH 6, metHb formation increased, especially for trout Hb. Subsequent acidification to pH 3 promoted Hb unfolding and partial or complete heme group-loss. During full GI digestion, polypeptide/peptide analyses revealed more extensive Hb-degradation in the gastric than duodenal phase, without any species-differences. When digesting WCM +/-Hb, both Hbs strongly promoted malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE), and 4-hydroxy-2-nonenal (HNE) formation, peaking at the end of the gastric phase. Trout-Hb stimulated MDA and HHE more than bovine Hb in the first gastric phase. Altogether, partially degraded Hb, and/or free hemin -both mammal and fish-derived- stimulated oxidation of PUFA-rich lipids under GI-conditions, especially gastric ones.

Weight distribution, proximate composition, fatty acids, amino acids, minerals and vitamins were investigated in five sorted cuts (head, backbone, viscera + belly flap, tail, fillet) emerging during filleting of spring and fall herring (Clupea harengus). The herring co-product cuts constituted similar to 60 % of the whole herring weight, with backbone and head dominating. Substantial amounts of lipids (5.8-17.6 % wet weight, ww) and proteins (12.8-19.2 % ww) were identified in the co-products, the former being higher in fall than in spring samples. Co-product cuts contained up to 43.1 % long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) of total FA, absolute levels peaking in viscera + belly flap. All cuts contained high levels of essential amino acids (up to 43.3 %), nutritional minerals (e.g., iodine, selenium, calcium, and iron/heme-iron), and vitamins E, D, and B12. Co-products were, in many cases, more nutrient-rich than the fillet and could be excellent sources for both (functional) food and nutraceuticals.

Structural, functional and nutritional properties of protein recovered from brown seaweed, S. latissima with alkaline solubilization/isoelectric precipitation as a function of different post-harvest stabilization methods were studied. The latter included freezing at −20 °C/-80 °C, oven-drying, sun-drying, freeze-drying and ensilaging. Also, the efficacy of freeze/thaw-aided precipitation (F/T) in improving protein recovery of the process was evaluated. The freeze-dried, oven-dried, and −20 °C frozen seaweeds resulted in significantly higher protein yield than the −80°C-frozen, sun-dried and ensiled biomasses. F/T increased protein precipitation and doubled total protein yield. Sun-drying and −20°C-freezing caused extensive protein degradation as revealed by SDS-PAGE and HP-SEC, while oven-drying altered the seaweed protein structure with less α-helices. Functional properties of the seaweed proteins were remarkably affected by stabilization condition and F/T, but nutritional value of the proteins was only dependent on stabilization method. Thus, to efficiently recover seaweed proteins, its post-harvest stabilization condition must be carefully chosen based on the final application of the proteins.

The aims of this study were to investigate the role of hemoglobin (Hb) in lipid oxidation development during ensilaging of herring filleting co-products, and, to inhibit this reaction by pre-incubating the co-products in water or physiological salt, with/without different antioxidants. Results showed that both peroxide value (PV) and 2-thiobarbituric acid reactive substances (TBARS) gradually increased during 7 days of ensilaging at 22 °C in absence of antioxidants. The increase in TBARS was proportional to the Hb levels present, while PV was less affected. A Hb-fortified Tris-buffer model system adjusted to pH 3.50 confirmed that Hb changed immediately from its native oxyHb to the metHb state, which facilitated heme group release and thus probably explains the increased PV and TBARS during ensilaging. Pre-incubating the co-products for 30 s in a solution containing 0.5% rosemary extract was the most promising strategy to inhibit lipid oxidation both in the co-products during pre-processing storage and during the actual ensilaging. The solution could be re-used up to ten times without losing its activity, illustrating that this methodology can be a scalable and cost-effective strategy to extend the oxidative stability of herring co-products allowing for further value adding e.g., into a high-quality silage.

Seaweed is a promising sustainable source of vegan protein as its farming does not require arable land, pesticides/insecticides, nor freshwater supply. However, to be explored as a novel protein source the content and nutritional quality of protein in seaweed need to be improved. We assessed the influence of pH-shift processing on protein degree of hydrolysis (%DH), protein/peptide size distribution, accessibility, and cell bioavailability of Ulva fenestrata proteins after in vitro gastrointestinal digestion. pH-shift processing of Ulva, which concentrated its proteins 3.5-times, significantly improved the %DH from 27.7±2.6% to 35.7±2.1% and the amino acid accessibility from 56.9±4.1% to 72.7±0.6%. Due to the higher amino acid accessibility, the amount of most amino acids transported across the cell monolayers was higher in the protein extracts. Regarding bioavailability, both Ulva and protein extracts were as bioavailable as casein. The protein/peptide molecular size distribution after digestion did not disclose a clear association with bioavailability.

Lipid oxidation in ice-stored sorted herring fractions (head, backbone, viscera+belly flap, tail, fillet) from spring and fall, and its association with endogenous prooxidants, antioxidants and lipid substrates were investigated. Peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) had increased significantly in all fractions after 1 day, but for both seasons, the most rapid PV and TBARS development occurred in head, which also had highest hemoglobin (Hb) levels and lipoxygenases (LOX) activity. Viscera+belly flap was overall the most stable part, and also had the highest α-tocopherol content. Pearson correlation analyses across all five fractions confirmed a significant impact of Hb, LOX and α-tocopherol on the lipid oxidation susceptibility, while content of total iron, copper, lipids or polyunsaturated fatty acids provided no significant correlation. Overall, the study showed which pro-oxidants that should be inhibited or removed to succeed with value adding of herring filleting co-products -and the fillet itself.

Herring and salmon filleting co-products were pH-shift processed together with seven antioxidant-containing raw materials ("helpers") including lingonberry-, apple-, oat-, barley- and shrimp-co-products, and two seaweeds (Saccharina latissima, Ulva fenestrata) to produce protein isolates stable towards lipid oxidation. Malondialdehyde (MDA) and 4-hydroxy-(E)-2-hexenal (HHE) levels revealed that all helpers, except shrimp shells, to different extents retarded lipid oxidation both during pH-shift-processing and ice storage. The three helpers performing best were: lingonberry press-cake apple pomace ∼ Ulva. Color of protein isolates was affected by helper-derived pigments (e.g., anthocyanins, carotenoids, chlorophyll) and lipid oxidation-induced changes (e.g., metHb-formation, pigment-bleaching). In conclusion, combining fish co-products with other food side-streams or seaweeds during pH-shift processing appears a promising new tool to minimize lipid oxidation of protein isolates, both during their production and subsequent storage. Lingonberry press-cake was the most efficient helper but provided dark color which may narrow product development possibilities, something which requires further attention.

Globally, there is a need for novel vegetarian protein sources. We recently showed that the pH-shift process, using alkaline protein solubilization followed by isoelectric precipitation, is an efficient way to produce extracts with high protein concentrations from Ulva lactuca (>50% on a dry matter basis). However, the total protein yield was low, and to improve this, the effects of adding ulvan lyase, preincubating the seaweed homogenate at pH 8.5 and using different protein extraction temperatures (8 °C, RT and 40 °C), were evaluated in this study. Addition of ulvan lyase reduced protein solubility but increased the precipitation. Incubation at pH 8.5, without ulvan lyase added, significantly increased both protein solubility and precipitation at 8 °C and RT. Temperature per se had no effect on protein solubility, while protein precipitation increased with decreasing temperature. Highest protein yield (29%) was achieved when keeping the process at 8 °C with a preincubation step at pH 8.5 for 1 h. By these process modifications, the yield was 3.2 times higher than achieved by the control process (9.2%).

The impacts of variation in fish filleting by-products origin including white muscle (cod), dark muscle (herring) and farmed fish (salmon) on physicochemical and gel-forming properties of protein recovered using the pH-shift method were studied. The effects of different solubilization pHs (acid or alkaline) on protein yield/composition, and its properties were also studied. Alkaline version (pH 11.5–12.5) resulted in maximum protein yield for the three resources which ranked them as: salmon > herring > cod. Increasing solubilization pH from 11.5 to 12.5 increased protein yield in salmon and herring, while maximum protein yield of cod was obtained at pH 12. However, increasing solubilization pH from 11.5 to 12.5 required a two-fold higher amount of alkali compared to the adjustment to 11.5. All recovered proteins had gel-forming capacity; however, cod gels showed higher WHC and breaking force compared with salmon and herring protein gels. Increasing solubilization pH from 11.5 to 12.5 negatively affected the breaking force and color of the salmon and cod protein gels, but improved heme pigment removal, breaking force and whiteness of herring protein gels. The pH-shift process thus showed good potential for recovering high quality protein from the by-products, but protein solubilization pH should be carefully selected based on the target species.

There is a strong need in animal production for sustainable protein sources used in feed. The aim of the present study was to evaluate the potential of the pH-shift process as a method to concentrate proteins from complex bone-rich herring (Clupea harengus) by-products. By-product parts (frame, head, guts), protein solubilization pH (pH 2.8 or 11.3) and separation g-force (800 or 8000 g) were varied. The findings revealed that protein recovery yields of up to 60% were obtained and that higher protein yield was obtained from the alkaline than the acid process. By-product part and g-force had no significant impact on protein yield. Protein levels reached in the concentrates were 75–80 % (dw basis). Protein concentrates contained a similar amino acid profile as have been reported for fishmeal, with ~40% essential amino acids and up to 7.8% and 2.8% of lysine and methionine. Residual levels of LC n-3 PUFA as well as calcium, magnesium, potassium and phosphorous were also found in the produced protein concentrates which could be an added value. Acid produced concentrates were particularly high in calcium potassium and phosphorous. Provided a careful evaluation, the pH-shift process could become a successful alternative to classic fish meal production for increasing the use of fish by-products for production of protein ingredients for feed.

Meat analogs based on plant protein extracts are rising in popularity as meat consumption declines. A dietary shift away from meat, which has a high iron bioavailability, may have a negative effect on the amount of iron absorbed from the diet. Iron absorption from legumes cultivated in regions not suitable for soy production, such as fava bean, has not yet been explored. The aim of this study was to evaluate non-heme iron absorption from a meal with texturized fava bean protein compared to beef and cod protein meals. The study included two single-blinded iron isotope trials in healthy Swedish women of the ages 18–45 years, each of whom served as their own control. The participants were served matched test meals containing beef and fava bean protein (Study 1) or cod and fava bean protein (Study 2) with radiolabeled non-heme iron 55Fe and 59Fe. The absorption of non-heme iron from test meals was measured by whole-body counting and erythrocyte incorporation. The absorption of non-heme iron, measured as erythrocyte incorporation ratio, from beef protein meal was 4.2 times higher compared to texturized fava bean meal, and absorption from cod protein meal was 2.7 times higher compared to the fava bean meal. The adjusted non-heme iron absorption, normalized to a 40% reference dose uptake, was 9.2% for cod protein meal, 21.7% for beef protein meal, and 4.2% for texturized fava bean meal. A fava bean protein meal has markedly lower iron bioavailability in healthy females compared with a meal of beef or cod protein. Therefore, a dietary shift from meat and fish protein to fava bean protein may increase the risk of iron deficiency.

In this study, we investigated biological effects associated with ingestion of polystyrene (PS) microplastic (MPs) in fish. We examined whether ingestion of contaminated PS MPs (100-400 μm) results in chemical stress in rainbow trout (Oncorhynchus mykiss) liver and we explored whether this exposure can affect the oxidative stability of the fillet during ice storage. Juvenile rainbow trout were fed for 4 weeks with four different experimental diets: control (1) and feeds containing virgin PS MPs (2) or PS MPs exposed to sewage (3) or harbor (4) effluent. A suite of ecotoxicological biomarkers for oxidative stress and xenobiotic-related pathways was investigated in the hepatic tissue, and included gene expression analyses and enzymatic measurements. The potential impact of MPs exposure on fillet quality was investigated in a storage trial where lipid hydroperoxides, loss of redness and development of rancid odor were assessed as indications of lipid peroxidation. Although, chemical analysis of PS MPs revealed that particles sorb environmental contaminants (e.g., PAHs, nonylphenol and alcohol ethoxylates and others), the ingestion of relatively high doses of these PS MPs did not induce adverse hepatic stress in fish liver. Apart from small effect on redness loss in fillets of fish exposed to PS MPs, the ingestion of these particles did not affect lipid peroxidation or rancid odor development, thus did not affect fillet's quality.

The effect of microbial transglutaminase (M‐TGase) (0–0.6 units g−1 sample) and setting condition (25 °C/180 min, 30 °C/120 min, 35 °C/60 min and 40 °C/30 min) on gel properties of blend protein isolate of gutted kilka and silver carp was studied. The protein isolate provided a good substrate for M‐TGase activity so that a low amount of M‐TGase (0.2 unit g−1 sample) substantially improved textural properties and water holding capacity (WHC) of the gels. Breaking force of the gels was positively affected by M‐TGase up to 0.6 unit g−1 sample, but it negatively affected their WHC. Prior setting at 25–35 °C increased the breaking force of proteins compared to directly heated gel, resulting in maximum breaking force at 35 °C/60 min. However, the setting at 40 °C/30 min caused proteolysis, which was reflected in higher amounts of TCA‐soluble peptides and gel weakening. Denser microstructure and higher myosin heavy chain polymerisation observed in the gels which experienced the setting was well correlated with improvement in textural properties.

Earthy off-odour in farm-raised freshwater fish is considered a quality defect. This study aimed to investigate the potential of pH-shift processing to remove off-odours from farm-raised hybrid catfish while at the same time documenting de-novo formation of other volatile compounds. In comparison with crude mince and conventional surimi, the alkali pH-shift process gave larger reductions in geosmin, 2-methylisoborneol, undesirable volatile compounds (e.g. hexanal, (E)-2-nonenal, (E)-2-heptenal, 2-butanone, and hexadecane), lipids, myoglobin, total volatile basic nitrogen, and TCA-soluble peptides (p 0.05). The acid-produced protein isolate showed the highest TBARS and processing-induced evolution of the following volatiles: octanal, nonanal, decanal, 2-butyl-2-octenal, pentadecanal, 1-hexanol, 1-octanol, 1-octen-3-ol, and 2,3-octanediol (p 0.05). Alkali-aided process provided better overall gelling characteristics (i.e. breaking force, deformation, and texture profile) and gave lower fishy, earthy, and rancid off-odour scores (p 0.05). Thus, alkali pH-shift process can be used to isolate gel-forming proteins from hybrid catfish while minimizing the accumulation of undesirable volatile compounds.

Applying value-adding techniques to fish filleting co-products is rendered difficult due to their high susceptibility to lipid oxidation, microbial spoilage, and amine formation. In this study, a recyclable dipping strategy was developed and investigated for its ability to stabilize herring (Clupea harengus) co-products (head, backbone, caudal fin, intestines, belly flap, skin, and in some cases roe) against oxidation and microbial spoilage. From initial screening of seven antioxidative components/formulas in minced herring co-products during ice storage, an oil-soluble rosemary extract (RE-B) and isoascorbic acid (IAA) were identified as most promising candidates. These compounds were then formulated to a recyclable solution to be used for dipping of the herring co-products. The commercial Duralox MANC antioxidant mixture was used as a positive control. Dipping in 0.2% RE-B solution ± 0.5% IAA or in 2% Duralox MANC solutions remarkably increased the oxidation lag phase from 12 days during subsequent storage on ice (0–1 °C) of minced or intact co-products, respectively, even when the antioxidant solutions were re-used up to 10 times. The dipping also reduced microbiological growth and total volatile basic nitrogen, but the effect became weaker with an increased number of re-using cycles. The presented dipping strategies could hereby facilitate more diversified end use of herring co-products from current fish meal to high-quality minces, protein isolates, or oils for the food industry.

This chapter focuses on the oxidative stability of n-3 polyunsaturated fatty acids (n-3 PUFAs) delivery systems and n-3 PUFAs fortified foods during digestion, with specific attention given to the long-chain n-3 PUFAs. The topic is of importance since the formation of reactive lipid oxidation products in the gastrointestinal tract (GIT) can lead to unwanted reactions e.g., with proteins, phospholipids, and DNA of the intestinal epithelial layer as well as decrease the amount of n-3 PUFAs being bioavailable in the GIT. Initially, the use of in vitro digestion models to study lipid oxidation is reviewed, after which available literature about the susceptibility of different n-3 PUFA delivery systems—neat oils, emulsions, capsules, and n-3 PUFA-rich foods—to oxidation in the GIT is summarized. In the next part of the chapter, the link between digestion-induced oxidation and initial oxidative status of the n-3 PUFA system, the degree of lipolysis as well as the presence of different pro- and antioxidants is discussed. Based on available findings, some future directions in the current research area are finally given.

Applying value-adding techniques to fish co-products is rendered difficult due to their high susceptibility to hemoglobin (Hb)-mediated lipid oxidation. In this study, we investigated a dipping technology with a solution containing Duralox MANC 213- a mixture of rosemary extract, ascorbic acid, tocopherols and citric acid – to control lipid oxidation during storage at 0 °C and 20 °C. The possibilities to re-use the antioxidant solution was also analyzed, along with studies on the link between Duralox MANC and Hb-form. Dipping in Duralox MANC largely increased the oxidation lag phase; from 3.5 d at 20 °C, and from 11 d at 0 °C. Even after re-use of the solution up to 10 times, lipid oxidation was completely inhibited at 0 °C. Duralox MANC could prevent auto-oxidation and hemin loss of herring Hb; which are suggested as the main mechanisms behind the observed stabilization of herring co-products against lipid oxidation.

The influence of recovery technique (pH-shift processing vs mechanical separation), antioxidant addition and endogenous factors on lipid oxidation in protein-enriched products from herring, salmon and cod backbones was investigated. Salmon-derived products were very stable during both ice and -20 °C storage. Contrary, peroxide value and TBA-reactive substances in herring- and cod-derived products increased rapidly during ice storage, with the pH-shift-produced protein isolates (PI) being most susceptible to oxidation in case of cod. Duralox MANC (0.5%) however largely increased the oxidation lag phase in both PI and mechanically separated meat (MSM); from1 day to15 days. At -20 °C, mainly the herring products oxidized, and particularly the MSM. Pearson correlation tests showed that endogenous levels of Hb, total Fe, ascorbic acid and α-tocopherol correlated significantly (p 0.05) with lipid oxidation development. Evaluating the role of pre-processing storage indicated that fish co-products should be processed immediately after the filleting process unless antioxidants are added.

The pH-shift process for isolation of gel-forming proteins from fish processing by-products was extended to allow parallel isolation of fish oil. Subjecting the floating emulsion layer formed during pH-shift processing of salmon by-products to pH-adjustment or freeze/thawing efficiently released the emulsified oil at 4 °C. However, for herring by-products higher temperature (10 °C) and a combination of the emulsion-breaking techniques was required for efficient oil release. Oil recovery yield using the adjusted pH-shift process was lower than with classic heat-induced oil isolation (90 °C/20 min), but pH-shift-produced oils had higher amounts of n-3 polyunsaturated fatty acids (n-3 PUFA). Also, alkaline pH-shift processing produced oils with remarkably less oxidation products and free fatty acids compared with acid pH-shift process or heat-induced isolation. Extending the pH-shift process with emulsion breaking techniques can thus be a promising biorefinery approach for parallel cold production of high-quality fish oil and gel-forming proteins from fish by-products.

This study aimed to evaluate the shelf-life of mechanically filleted well-fed Atlantic mackerel during frozen storage at −25 °C and effect of treatment with antioxidants (sodium erythorbate and a polyphosphate mixture) and different antioxidant application methods (dipping, spraying and glazing). Both physicochemical measurements and sensory analysis were applied. Antioxidant treatments prolonged shelf-life of mackerel. Sensory analysis indicated that untreated fillets had a shelf-life of less than 2.5 months, while all antioxidant treated fillets exceeded that. The most effective treatment, dipping fillets into a sodium erythorbate solution, yielding a shelf-life of 15 months. Physicochemical methods used to evaluate degradation of lipids in the fillets were free fatty acids (FFA), lipid hydroperoxides (PV) and thiobarbituric acid reactive substances (TBARS). They did not correlate with sensory results and might therefore be a questionable choice for evaluation of oxidation and development of rancid flavour and odour in complex matrixes such as Atlantic mackerel.

Publisher's version (útgefin grein), This study investigated the efficacy of three commercially relevant packaging methods (vacuum with water glazing VAC-G; vacuum with seawater VAC-S; shatter-layer packaging SL) to improve frozen storage stability of mechanically filleted Atlantic mackerel at − 25 °C, in comparison to water glazing alone (GL) and storage as whole unglazed, block frozen fish. Besides proximate composition and pH of raw material, quality changes were analysed by free fatty acid content (FFA), water holding capacity (WHC), cooking yield, lipid oxidation (lipid hydroperoxides, PV; non-protein bound thiobarbituric acid reactive substances, TBARS) and sensory profiles of cooked samples after 3.5, 8, 10 and 12 months of frozen storage. Vacuum-packaging was effective in mitigating the PV and TBARS as well as rancid odour and flavour. The inclusion of seawater in VAC-S altered the sensory textural attributes of the mackerel fillet to be more juicy, tender and soft and increased the attribute of salty flavour in the sample. SL delayed rancid odour and flavour by 2 months compared to GL. Processing of mackerel under industrial conditions, including filleting, handling, double-freezing and glazing accelerated the formation of FFA as well as losses of WHC and cooking yield in the fillet regardless the packaging methods., Open Access funding provided by Nofima the food research institute. The authors would like to gratefully acknowledge the financial support of Nordic Innovation (MAR14306). Prof. María Guðjónsdóttir at University of Iceland is acknowledged for her valuable feedback on the manuscript.

The potential of cross-processing herring or salmon by-products with brown seaweed, shrimp peeling by-products and lingonberry press cake (called “helpers”) to minimize lipid oxidation during acid/alkaline pH-shift protein isolation was evaluated. Lingonberry press cake remarkably reduced levels of lipid hydroperoxides, malondialdehyde (MDA) and 4-hydroxy-trans2-hexenal (HHE) in both herring and salmon protein isolates (PI) while seaweed and shrimp by-product mitigated generation of MDA and HHE in herring PI. The salmon PI, however, by itself had very low oxidation levels. Cross-processing reduced protein yield compared with the classical pH-shift process by diminishing protein solubility, but still, the alkaline process version was promising. The color of the PIs varied greatly between different process combinations, reflecting partitioning of natural pigments into isolates. Altogether, cross-processing seems a promising tool for inhibition of lipid oxidation during pH-shift processing of sensitive fish by-products, with the alkaline version and lingonberry press cake being closest at hand for further studies.

Nutritional, structural, functional, and sensorial properties of protein isolate developed from salmon (Salmo salar), cod (Gadus morhua), and herring (Clupea harengus) by-products using the pH-shift method was studied. Function of the proteins in an emulsion system in terms of viscoelastic properties was also evaluated. Regardless of origin, the proteins showed satisfying nutritional value as reflected in their high essential amino acid content. The proteins contained significantly (p

peer-reviewed The demand for vegetable proteins increases globally and seaweeds are considered novel and promising protein sources. However, the tough polysaccharide-rich cell walls and the abundance of polyphenols reduce the extractability and digestibility of seaweed proteins. Therefore, food grade, scalable, and environmentally friendly protein extraction techniques are required. To date, little work has been carried out on developing such methods taking into consideration the structural differences between seaweed species. In this work, three different protein extraction methods were applied to three Swedish seaweeds (Porphyra umbilicalis, Ulva lactuca, and Saccharina latissima). These methods included (I) a traditional method using sonication in water and subsequent ammonium sulfate-induced protein precipitation, (II) the pH-shift protein extraction method using alkaline protein solubilization followed by isoelectric precipitation, and (III) the accelerated solvent extraction (ASE®) method where proteins are extracted after pre-removal of lipids and phlorotannins. The highest protein yields were achieved using the pH-shift method applied to P. umbilicalis (22.6 ± 7.3%) and S. latissima (25.1 ± 0.9%). The traditional method resulted in the greatest protein yield when applied to U. lactuca (19.6 ± 0.8%). However, the protein concentration in the produced extracts was highest for all three species using the pH-shift method (71.0 ± 3.7%, 51.2 ± 2.1%, and 40.7 ± 0.5% for P. umbilicalis, U. lactuca, and S. latissima, respectively). In addition, the pH-shift method was found to concentrate the fatty acids in U. lactuca and S. latissima by 2.2 and 1.6 times, respectively. The pH-shift method can therefore be considered a promising strategy for producing seaweed protein ingredients for use in food and feed.

Here we evaluated the levels of lipid oxidation products, fatty acids, ascorbic acid and colour of Porphyra and Ulva after oven-drying at 40 °C, and during subsequent storage for ≥370 days under light, semi-light and dark conditions. Part of the seaweed was pre-soaked in freshwater or pre-coated with a whey protein mixture. Controls consisted of freeze-dried seaweeds. Throughout storage there was a moderate development of the lipid oxidation-derived aldehydes, malondialdehyde, 4-hydroxy-trans-2-hexenal and 4-hydroxy-trans-2-nonenal, while there was a great loss of unsaturated fatty acids and ascorbic acid. Light storage and freeze-drying stimulated the fatty acid loss as well as pigment bleaching, seen as increased a*-values. For Ulva, the coating reduced malondialdehyde, 4-hydroxy-trans-2-hexenal and 4-hydroxy-trans-2-nonenal formation during drying and slightly prevented loss of polyunsaturated fatty acids during light storage. Pre-soaking in freshwater had no effect on the seaweed stability, although it reduced the ash content and thereby increased the relative content of ascorbic acid and fatty acids of the biomasses.

Saccharina latissima biomass cultivated along the Swedish west coast was subjected to four different scalable preservation methods after harvest; freezing, sun-drying, oven-drying and ensiling. Freeze-drying and freezing at −80 °C were also included to provide dry and wet references. The effects of the different preservation methods on the composition of Saccharina biomass (on dry weight, DW, basis), and the recovery as well as properties of high-quality protein, alginate and biogas were evaluated. Sun-drying significantly reduced protein, alginate and fatty acid content of the seaweeds and thereby concentrated ash in the biomass compared to the other methods. Protein/amino acids and fatty acids were significantly concentrated in ensiled biomass, while mannitol and laminarin were reduced compared to the other biomasses. Oven-drying and −20 °C freezing affected the composition the least, with lower ash content and alterations in some specific amino and fatty acids. Sun-drying and ensiling resulted in significantly lower protein solubility at high pH compared to the other biomasses which translated into the lowest total seaweed protein recovery using the pH-shift process. Highest protein yield was obtained with the freeze-dried reference. Ensiling lead to a significant decrease in the molecular weight of alginate, while sun-drying caused a negative effect on alginate by inducing a shift in the guluronic and mannuronic acids composition of alginate. Sun-drying gave the lowest methane yield in the anaerobic digestion experiments while freezing at −80 °C gave the highest yield, closely followed by freezing at −20 °C and ensiling. To conclude, preservation methods must be carefully chosen to protect the valuable component in Saccharina latissima, and to achieve an efficient downstream processing ultimately yielding high quality products as part of a seaweed biorefinery.

This study aimed to evaluate how blending pH-shift produced protein isolates from gutted kilka (Clupeonella cultriventris) and silver carp (Hypophthalmichthys molitrix) affected dynamic rheological and chemical properties of the proteins as well as microstructural and physico-mechanical properties of produced gels. Studied variables were protein solubilization pH (acid vs. alkaline) and blending step (before or after protein precipitation). Comparisons were made with conventionally washed minces from kilka and silver carp fillets; either alone or after blending. Rheological studies revealed that blending alkali-produced protein isolates before precipitation resulted in rapid increase of G' reflecting the formation of intermolecular protein-protein interactions with higher rate. Furthermore, blending of alkali-produced protein isolates and washed minces, respectively, of kilka and silver carp improved physico-mechanical properties of the resultant gels compared to pure kilka proteins. However, the pH-shift method showed higher efficacy in development of blend surimi at the same blending ratio compared to the conventional washing.

In this study, fish muscle press juice, PJ, was evaluated as an antioxidative liquid for glazing of fish fillets prior to frozen storage. PJ has in previous studies efficiently protected fish mince lipids against haemoglobin- mediated oxidation. Frozen herring fillet were glazed with water, PJ or 50 mM phosphate buffer of the same pH as herring PJ (6.5). Over 52 weeks, peroxide value (PV), TBA-reactive substances (TBARS), rancid odour and colour were monitored. From 26 to 52 weeks, water-glazed fillets were significantly (P < 0.05) more oxidised compared to fillets glazed with PJ or buffer, most clearly shown by PV and TBARS data. The protective effects of both PJ and buffer compared to water could indicate a role, for example, of the physiological ionic strength. The finding that PV became elevated in PJ which was frozen stored alone in a thin layer indicates that PJ stripped from, for example, trace lipids and haemoglobin would provide better protection.

Flocculation and sedimentation of a protein-rich biomass from shrimp boiling water (SBW) using food grade polysaccharides (carrageenan, alginate and carboxymethyl cellulose (CMC)) as flocculants was investigated at different pH-values. The effect of flocculant concentration on particle size and viscosity of SBW was also evaluated. Flocculation with carrageenan (0.45 g/L) at pH = 4 exhibited the most efficient protein sedimentation; protein concentration of the upper phase was here reduced by 77%, allowing 86% protein to be sedimented from SBW. Flocculation by alginate and CMC at pH = 4 showed 67% and 60% protein reduction of the upper phase at concentrations of 0.5 and 0.2 g/L, respectively. Contrary to alginate and CMC, carrageenan concentration affected the size distribution of flocs. Finally, carrageenan at 0.45 g/L and pH = 4 was successfully tested in a scaled up trial (5L) providing 78.5% protein recovery and a biomass with 75% protein on dry weight basis.

The formation of malondialdehyde (MDA), 4-hydroxy-2-hexenal (HHE), 4-hydroxy-2-nonenal (HNE), and 4-oxo-2-nonenal (ONE) in cod liver-, anchovy-, krill-, and algae oil during in vitro digestion with human gastrointestinal fluids was investigated. Adding rabbit gastric lipase, lipase inhibitor (orlistat) and tocopherols to cod liver oil, lipolysis and oxidation was also studied. Among the marine oils, the highest aldehyde levels (18 µM MDA, 3 µM HHE and 0.2 µM HNE) were detected after digestion of cod liver oil, while the lowest levels were detected in krill and algae oils. Addition of rabbit gastric lipase significantly increased the release of HNE during the digestion. Orlistat significantly reduced lipolysis and MDA formation. Formation of MDA and HHE was delayed by tocopherols, the tocopherol mix Covi-ox® T 70 EU being more effective than pure α-tocopherol.