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Edible filamentous fungi, a source of mycoprotein, are one of the sustainable alternative protein. This study compares protein digestibility (DH%) and amino acid and mineral accessibility in Rhizopus oligosporus cultivated in oat flour (OatRO) or glucose media (GluRO) by using the INFOGEST in vitro digestion protocol. Fungal total amino acids was higher in GluRO (39.0 ± 1.1 % dw) than OatRO (21.8 ± 1.3 % dw) which was also the case for calcium and magnesium content. After completed gastrointestinal digestion, there were no significant differences between GluRO and OatRO regarding DH% (27.21 ± 10.4 % and 29.4 ± 0.5 %), however, GluRO provided significantly higher amino acid accessibility compared to OatRO (64.3 ± 1.6 % and 55.1 ± 3.1 %). Mineral accessibility of GluRO was for Ca: 37.9 ± 1.8 %, Zn: 9.3 ± 0.4 %, Fe: 38.2 ± 1.9 %, Mg: 66.5 ± 1.4 % and Cu: 24.7 ± 1.3 % and for OatRO; Ca: −40.2 ± 2.4 %, Zn: −4.13 ± 0.15 %, Fe:14.6 ± 1.6 %, Mg: 74.5 ± 3.1 %, and Cu: 55.95 ± 0.8 %. Despite the low phytic acid content, OatRO thus showed antinutrient properties with respect to calcium, and zinc, suggesting that oat-derived fungi had antinutrients other than phytic acid. This study hereby revealed that the cultivation substrate affect amino acid and mineral accessibility of filamentous fungi and calls for deeper evaluations of antinutrients in oat-derived fungi.

A headspace solid–phase microextraction (HS-SPME) method coupled with gas chromatography/mass spectrometry with quadrupole time-of-flight (GC/MS-QTOF) was developed for analysis of volatile off-odor compounds, i.e., earthy/musty (2-methylisoborneol, 2-MIB) and rancid (aldehydes and alcohols), from farmed hybrid catfish (Clarias macrocephalus × Clarias gariepinus). The most efficient extraction of targeted volatiles was provided by 50 min at 70 °C with a CWR-PDMS fiber and 3 g of fish diluted to 5 mL with 1.5 g NaCl (30 % saturated NaCl). The maximum time-delay before extraction was 8 h to avoid spoilage and lipid oxidation during analysis. The final method showed good linearity, intraday repeatability of 5–9 %, interday reproducibility of 5–12 % and recoveries of 94–112 %. The implementation part proved that the developed method gave accurate quantitative results for oxidation-derived volatiles, several with high correlation to thiobarbituric acid reactive substances (TBARS). Altogether, our study provided an effective SPME-GC–MS method for the extraction and analysis of important off-odor compounds in catfish mince.

The green seaweed Ulva fenestrata is a future food candidate, however, both compositional, e.g. protein content, and sensory qualities depend on cultivation and harvest conditions. The aim was to explore sensory qualities of, and consumer attitudes to, U. fenestrata derived from three different cultivation conditions and three harvest times. U. fenestrata was cultivated at sea and in two different land-based tank settings, one mimicking seawater and one with added process water from fishing industry to increase protein content. The seaweed was subjected to sensory analysis performed by an analytical sensory panel and a focus group consisting of consumers. The former assessed the seaweed as dried whole blades (9 samples) and as emulsions (9 samples). Consumers assessed whole blades and a vegan spread enriched with powdered U. fenstrata. All seaweed samples were intensely green; had tastes of umami, salt, bitter, and sour; odours and flavours of grass and fresh seaweed; and had a crispy texture. Cultivation in tanks resulted in a crispier texture than sea cultivation (p

Sea lettuce (Ulva) species have been identified as a future protein source, and post-harvest techniques, including the sustainable integration of liquid food side streams, have been developed to further increase their protein content and commercial market value. This study explores the post-harvest treatment of commercially produced seaweed biomass from large-scale, sea-based cultivations of U. fenestrata with residual water streams emerging from industrial storage of herring – so called herring production tub water (TUB). Growth rates of U. fenestrata were significantly higher in TUB treated seaweeds compared to controls. Further, the crude protein content was 71.26 % higher when cultivated in TUB, compared to controls, reaching a crude protein content of 37.37 ± 1.83 % dry weight. Notably, there were no limiting amino acids, nor fish-related allergenic activity in the seaweed biomass. Our study demonstrates a new nutrient loop turning food waste into protein-rich biomass by applying sustainable seaweed cultivation.

The sensory quality of seaweed, whether as a whole biomass or as a protein ingredient, plays a crucial role in its successful commercialization. This study explored the effect of different Ulva species, biomass washing, and pH-shift-based protein extraction on the sensory quality of emulsions with 0, 5, and 10 % oil. A trained panel assessed the sensory profile, complemented by analyses of volatile compounds, total ash, and amino acids. Saltiness emerged as the primary distinction between emulsions with unwashed U. linza and U. fenestrata, due to higher ash in the former. Washing U. fenestrata retained sensory qualities despite reduced ash and increased content of the lipid oxidation-marker pentanal. Protein extraction up-concentrated total amino acids 2.9-fold, and yielded emulsions with reduced particle sensation and grassy flavor, while bitterness, sourness, dark color, pentanal, hexanal, and 2-ethylfuran increased. Increased oil content of emulsions lowered their grassy odor which correlated with reduced hexanal content. Overall, these findings can contribute to the development of food products containing seaweed or protein ingredients thereof that match consumer preferences.

The effects of cross-processing lingonberry press cake (LPC) (2.5–30 %, dw/dw) with herring co-products on protein yield, oxidative stability and color of pH-shift-produced protein isolates were investigated. Even at 2.5 % LPC, the formation of volatile oxidation-derived aldehydes, including hexanal, (E)-2-hexenal, heptanal, octanal, and 2,4-heptadienal, were prevented during the actual protein isolate production. Adding 10 % LPC successfully prevented formation of all these aldehydes also during eight days ice storage which was explained by the partitioning of phenolics, especially ideain (1.09 mg/g dw) and procyanidin A1 (65.5 mg/g dw), into isolates. Although higher amounts of LPC (20–30 %) further prolonged the oxidation lag phase, it reduced total protein yield, increased the consumption of acid and base, and darkened protein isolates. Therefore, it is recommended to use 10 % LPC when pH-shift-processing sensitive fish raw materials as a route to mitigate lipid oxidation and at the same time promote industrial symbiosis and more circular food production.

Abstract The relationship between hemolysis and lipid oxidation was explored in red blood cell (RBCs)-spiked washed cod mince (WCM). At pH 6.8 and 3 ± 1 °C, intact RBCs (71 µM Hb) delayed lipid oxidation by 1 day compared to WCM with partly or fully lysed RBCs which oxidized immediately. Intact RBCs also lowered peak peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) with up to 59.5% and 48.1%, respectively. Adding 3% (v/w) blood plasma to RBC-spiked WCM delayed the lipid oxidation onset from 1 to 3–4 days without delaying hemolysis. At pH 6.4 the oxidation onset in RBC-WCM was the same as for pH 6.8 while at pH 7.2–7.6 lipid oxidation was suppressed for 7 days. Micrographs revealed RBC-lysis from day 2 at pH 6.4 but at pH 7.6, RBC stayed intact for ≥ 7 days. Thus, assuring presence of plasma-derived antioxidants and/or elevating muscle pH to avoid hemolysis can aid valorization of blood rich underutilized fish raw materials.

ABSTRACTNutrient and toxicant levels as well as their bioavailability in S. latissima and Ulva species (fenestrata, intestinalis, rigida) were reviewed. Nutritional quality was assessed by nutrient contribution to daily reference intake (DRI) per portion (5 g dry weight), nutrient density score NRF21.3, and comparisons to reference foods. Toxicological assessments comprised tolerable daily intake (TDI)-levels. Based on mean %DRI per portion, S. latissima and Ulva species were good sources (%DRI >15) of calcium, magnesium, iron, selenium, and vitamin B12. Mean %DRI was

The possibility of reducing the amount of fresh water used during alkaline the pH-shift processing of salmon head (SH) and herring frame (HF) was evaluated with ultrasound (US) as a tool to mitigate its negative effects on protein yield. The role of water ratio and US for homogenate viscosity, mass yield, crude composition, functional properties and lipid oxidation of the SH and HF protein isolates were also investigated. Applying US during the solubilization step of the pH-shift process completely compensated for the reduced protein yield coming from using 3 rather than 6 volumes of water for HF, but not for SH. Using US had no negative effect on the composition and protein functionality of the HF protein isolate. However, it slightly increased its level of secondary lipid oxidation products. Altogether, applying US during the pH-shift processing at low water ratios can be a promising solution for more resource-smart valorization of herring side streams.

Fermentation can be a powerful tool for developing new sustainable foods with increased nutritional value and fermented microbial biomass derived from filamentous fungi is a promising example. This study investigates the nutritional profile of edible Aspergillus oryzae biomass produced under submerged fermentation (SmF) using oat flour as a substrate. The fermentation occurred in a 1m3 airlift bioreactor during 48 h at 35 °C and the nutritional profile of the produced fungal biomass in terms of amino acids, fatty acids, minerals (Fe, Zn, Cu, Mn), vitamins (E, D2), and dietary fiber was compared to oat flour as well as pure fungal biomass grown on semi-synthetic medium. The total amount of amino acids increased from 11% per dry weight (dw) in oat flour to 23.5% dw in oat fungal biomass with an improved relative ratio of essential amino acids (0.37 to 0.42). An increase in dietary fibers, minerals (Fe, Zn, Cu), vitamin E, as well as vitamin D2 were also obtained in the oat fungal biomass compared to oat flour. Moreover, the short chain omega-3 α-linolenic acid (ALA) and omega-6 linoleic acid (LA) values increased from 0.6 to 8.4 and 21.7 to 68.4 (mg/g dry weight sample), respectively, in oat fungal biomass. The results indicate that fungal biomass grown on oat flour could have a potential application in the food industry as a nutritious source for a wide variety of products.

Abstract Hemoglobin (Hb) is a powerful promoter of lipid oxidation, particularly in muscle of small pelagic fish species and fish by-products, both having high Hb-levels and highly unsaturated lipids. As Hb is located within the red blood cells (RBCs) it is here hypothesized that the perishable polyunsaturated fatty acids (PUFAs) can be protected from oxidation by limiting hemolysis during early fish processing. Using a model system consisting of washed-resuspended trout (Oncorhynchus mykiss) RBCs (wr-RBCs), the aim of this study was to evaluate how RBC lysis under cold storage was affected by selected parameters linked to blood or muscle: bacterial growth, energy status, pH, RBC membrane lipid oxidation and colloidal osmotic pressure (COP). The results indicated that bacterial growth had a modest effect on hemolysis while pH-values typical for post mortem fish muscle (6.4–6.8), and absence of glucose or albumin stimulated hemolysis. The rapid hemolysis observed at pH 6.4–6.8 correlated with lipid oxidation of the RBC membrane, while the lower hemolysis at pH 7.2–8.0 occurred with low, or without any RBC membrane lipid oxidation. When hemin was added to the RBCs at pH 6.8 hemolysis was induced without parallel RBC membrane oxidation, pointing at Hb-autoxidation and hemin-release per se as important events triggering lysis in fish muscle. Altogether, the study provided valuable findings which ultimately can aid development of new tools to combat lipid oxidation in post mortem fish muscle by limiting hemolysis.

Seaweed is often recognized for its potential health benefits, attributed to its abundance of dietary fibers, protein, and polyphenols. While human observational studies have shown promise, the collective evidence from human intervention trials remains limited. This narrative review aims to comprehensively analyze the effects of seaweed intake on humans, while critically assessing the methodology, including Cochrane risk-of-bias assessment. A search was conducted in online databases, including PubMed, Scopus, and Google Scholar, covering the period from 2000 to May 2023. The focus was on randomized controlled clinical trials (RCTs) evaluating the impact of whole seaweed, either consumed as capsules, integrated into food products or as part of meals. Various health outcomes were examined, including appetite, anthropometric measures, cardiometabolic risk factors, thyroid function, markers of oxidative stress, and blood mineral concentrations. Out of the 25 RCTs reviewed, the findings revealed limited yet encouraging evidence for effects of seaweed on blood glucose metabolism, blood pressure, anthropometric measures, and, to a lesser extent, blood lipids. Notably, these favorable effects were predominantly observed in populations with type-2 diabetes and hypertension. Despite most trials selecting a seaweed dose aligning with estimated consumption levels in Japan, considerable variability was observed in the pretreatment and delivery methods of seaweed across studies. Moreover, most studies exhibited a moderate-to-high risk of bias, posing challenges in drawing definitive conclusions. Overall, this review highlights the necessity for well-designed RCTs with transparent reporting of methods and results. Furthermore, there is a need for RCTs to explore seaweed species cultivated outside of Asia, with a specific emphasis on green and red species. Such studies will provide robust evidence-based support for the growing utilization of seaweed as a dietary component in regions with negligible seaweed consumption, e.g., Europe.

The overall goal of this study was to provide solutions to innovative microalgae-based technology for wastewater remediation in a cold-water recirculating marine aquaculture system (RAS). This is based on the novel concept of integrated aquaculture systems in which fish nutrient-rich rearing water will be used for microalgae cultivation. The produced biomass can be used as fish feed, while the cleaned water can be reused, to create a highly eco-sustainable circular economy. Here, we tested three microalgae species Nannochloropis granulata (Ng), Phaeodactylum tricornutum (Pt), and Chlorella sp (Csp) for their ability to remove nitrogen and phosphate from the RAS wastewater and simultaneously produce high-value biomass, i.e., containing amino acids (AA), carotenoids, and polyunsaturated fatty acids (PUFAs). A high yield and value of biomass were achieved for all species in a two-phase cultivation strategy: i) a first phase using a medium optimized for best growth (f/2 14x, control); ii) a second “stress” phase using the RAS wastewater to enhance the production of high-value metabolites. Ng and Pt performed best in terms of biomass yield (i.e., 5-6 g of dry weight, DW.L-1) and efficient cleaning of the RAS wastewater from nitrite, nitrate, and phosphate (i.e., 100% removal). Csp produced about 3 g L-1 of DW and reduced efficiently only nitrate, and phosphate (i.e., about 76% and 100% removal, respectively). The biomass of all strains was rich in protein (30-40 % of DW) containing all the essential AA except Methionine. The biomass of all three species was also rich in PUFAs. Finally, all tested species are excellent sources of antioxidant carotenoids, including fucoxanthin (Pt), lutein (Ng and Csp) and β-carotene (Csp). All tested species in our novel two-phase cultivation strategy thus showed great potential to treat marine RAS wastewater and provide sustainable alternatives to animal and plant proteins with extra added values.

The possibility of replacing the very time and resource demanding salting out (SO) method with isoelectric precipitation (IP) during collagen extraction from common starfish and lumpfish was investigated. The effect of IP on yield, structural and functional properties of the collagens was therefore compared with SO. Application of IP resulted in a higher or similar collagen mass yield compared with SO from starfish and lumpfish, respectively. However, the purity of collagens recovered with IP was lower than those recovered with SO. Replacing SO with IP did not affect polypeptide pattern and tropohelical structural integrity of collagen from the two resources as revealed with SDS-PAGE and FTIR analysis. Thermal stability and fibril formation capacity of collagens recovered with IP were also well preserved. Overall, the results showed that the IP can be a promising resource smart alternative for the classic SO precipitation during collagen extraction from marine resources.

Currently, side-streams (e.g., head, backbone, tail, and intestines) generated in the fish processing industry often end up as low-value products for feed applications or even as waste. In order to upcycle such side-streams, they need to be preserved to avoid oxidative degradation of the lipids between the generation point and the valorization plant. In the cod filleting industry, three main solid side-streams: viscera, heads, and backbones, are obtained. Hence, this study aimed to identify the most efficient antioxidant for preserving the cod side-streams using a dipping-based strategy prior to pre-valorization storage at low temperatures (ice and frozen storage). The dipping solutions evaluated contained: (i) a lipophilic rosemary extract (0.05% and 0.2% in 0.9% NaCl), (ii) Duralox MANC (a mixture of rosemary extract, ascorbic acid, tocopherols, and citric acid; 2% in 0.9% NaCl), and (iii) NaCl (0.9%) w/w solution. One group was not dipped. No dipping and dipping in NaCl were included as controls. The results showed a positive effect of dipping with solutions containing antioxidants as measured by peroxide value (PV), TBA-reactive substances (TBARS), and sensory profiling, e.g., rancid odor. Moreover, the oxidative stability increased with decreased storage temperature. The cod side-streams were in general most efficiently preserved by Duralox MANC, followed by the lipophilic rosemary extract (0.2%), compared to no dipping and dipping in NaCl solution and the lower concentration of the lipophilic rosemary extract (0.05%). The efficiency of the antioxidant treatments was independent of the side-stream fraction and storage temperature. Thus, using antioxidant dipping combined with low temperature storage is an efficient preservation method for maintaining the quality of the lipids in cod solid side-streams during their pre-valorization storage.

A massive amount of crustaceans and bivalves are consumed each year, leading to millions of tons of processing side streams from the seafood industry. Considering the current trend of (bio)circular and zero-waste food production, crustacean and bivalve processing side streams (CBPS) seem a promising and emerging resource for producing high-value-added products. This paper highlights the general composition of CBPS with high commercial values, namely, protein, lipids, carotenoids, minerals and chitins. The extraction strategies of these fractions, including conventional chemical and environmentally friendly methods, are also discussed. This review presents and summarises CBPS as raw materials for developing fast time-to-market products complying with specific EU regulations, including animal feeds, bio-pesticide/stimulants, and cosmetic ingredients. This paper also provides insights into challenges of applying CBPS as raw materials to generate products for human consumption.

High acid-consumption and lipid oxidation are challenges when recovering functional proteins from herring co-products via pH-shift-processing. Here, lingonberry press-cake (LP), which is abundant in organic acids and phenolics, was added to alkali-solubilized herring-co-product-proteins (2.5–30 % LP per dry weight) aiming to aid protein precipitation, save hydrochloric acid (HCl) and provide oxidative stability. The results revealed 5–30 % LP addition reduced HCl-consumption by 13–61 % and 19–79 % when precipitating proteins at pH 5.5 and 6.5, respectively. Higher LP% decreased protein content and lightness of protein isolates but raised the lipid content. Precipitation at pH 6.5 used less acid, reduced total protein yield and raised moisture content and darkness of isolates. Contrary to controls, lipid oxidation-derived volatiles did not develop in protein isolates precipitated with 10 % and 30 % LP, neither during the process itself nor during 21 days on ice. Altogether, LP was identified as a promising all-natural processing-aid to use during herring protein isolation.

Abstract 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.

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 ∼ 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.

One approach to improve the sustainability of food processing is the recovery of valuable compounds from food industry’s side streams. In this study a life cycle assessment (LCA) was used to quantify the potential environmental impacts of cross-processing herring side streams with different antioxidant-rich biomasses, so-called helpers, for the extraction of a protein ingredient that is stable against lipid oxidation. New primary experimental data was combined with literature values to model cross-processing of herring with different helpers, namely, lingonberry pomace, apple pomace, and brown and green seaweed. Different addition ratios and delayed addition of the pomace were also assessed for cross-processing herring with lingonberry pomace. The environmental performance of the resulting protein ingredients were assessed on a mass and delivered protein basis. Potential environmental impacts for climate change, energy consumption, land occupation, and depletion of marine resources were addressed. No ingredient performed better in all environmental impact categories, but delaying the helper addition had the most significant influence in reducing the product’s environmental impacts. This study’s outcomes enable analysts to direct research towards the most relevant parameters for producing a protein ingredient with lower environmental impact.

Abstract 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 aquaculture can provide the growing human population with a sustainable source of proteins. Sea-based cultivation is an effective method for farming seaweeds on a large scale and can yield high biomass output. However, the quality and biochemical composition of the biomass is seasonally dependent, which limits the harvests to certain periods of the year. Here we show the possibility to extend the sea-based cultivation season of Ulva fenestrata when aiming for high protein levels, by post-harvest treatment in herring production process waters. We harvested U. fenestrata at an optimal period in terms of yield, but suboptimal in terms of protein content. We then cultivated the seaweed in onshore tank systems with the nutrient-rich process waters for 14 days. We monitored biomass yield, crude protein content, amino acid composition, and content of the health concerning metals arsenic, mercury, lead, and cadmium, as well as the sensory properties of the dried biomass. After cultivation in the process waters, biomass yields were 30 - 40% higher (210 – 230 g fresh weight) compared to in seawater (160 g fresh weight). Also, the crude protein and amino acid content increased three to five times in the process waters, reaching 12 - 17 and 15 – 21% dry weight, respectively. The protein enriched biomass followed food graded standards for heavy metal content, and consumption of the biomass does not exceed health based reference points. Additionally, no sensory attributes regarded as negative were found. This rapid, post-harvest treatment can help extend the cultivation season of sea-based seaweed farms, maximizing their output of sustainable proteins.

Ulva spp. (sea lettuce) has recently gained attention as a sustainable protein source due to its high productivity and many nutritional properties interesting for the food industry. In this study, we explored a possible industrial symbiosis between herring production processing industries and Ulva fenestrata cultivation. We show that U. fenestrata cultivated in herring production process waters had four to six times higher biomass yields (27.17 - 37.07 g fresh weight vs. 6.18 g fresh weight) and three times higher crude protein content (> 30% dry weight vs. 10% dry weight) compared to U. fenestrata cultivated in seawater. Along with the elevation of protein, the herring production process waters also significantly increased levels of all essential amino acids in the seaweed biomass. The content of some heavy metals (arsenic, mercury, lead, and cadmium) was well below the maximum allowed levels in foodstuff. Therefore, quantities of biomass around 100 g dry weight could be consumed daily following the US Environmental Protection Agency’s reference doses. Combined, the results show that cultivation of U. fenestrata in herring production process waters has great potential to produce sustainable proteins for the growing world population. At the same time, nutrients of currently discarded process waters are circulated back to the food chain.

Sea lettuce (Ulva) aquaculture has increased the last decade due to high productivity, wide environmental tolerance, and interesting functional and nutritional properties of the crop. Research focus has mainly been on adult biomass production, but knowledge of performance and biochemical content of early developmental stages – which are the basis to any large-scale production - is still limited. The life-history of Ulva alternates between a diploid sporophytic life-stage and a haplontic gametophytic life-stage. Whereas the sporophyte give raise to recombinant gametophytes through zoids, gametophytes can give raise to parthenogenetically developing, clonal gametes in absence of a mating partner. This study shows that recombinant gametophytes have a faster ontogenetic development, higher growth rate, as well as higher protein, fatty acid, and pigment contents compared to clonal gametophytes of the crop Ulva fenestrata. Nutrient addition is required for a normal development, but temperature and swarmer density have relatively small effects on the hatchery success, relative growth rate and biochemical profile of the juvenile biomass. Our study reveals that the selection of the life-history-phase in novel sea lettuce crop strains could largely contribute to the emerging seaweed aquaculture sector.

Seaweed biomass is a renewable resource with multiple applications. Sea-based cultivation of seaweeds can provide high biomass yields, low construction, operation, and maintenance costs and could offer an environmentally and economically sustainable alternative to land-based cultivations. The biochemical profile of sea-grown biomass depends on seasonal variation in environmental factors, and the optimization of harvest time is important for the quality of the produced biomass. To identify optimal harvest times of Swedish sea-based cultivated sea lettuce (Ulva fenestrata), this study monitored biomass yield, morphology, chemical composition, fertility, and biofouling at five different harvesting times in April – June 2020. The highest biomass yields (approximately 1.2 kg fw [m rope]–1) were observed in late spring (May). The number and size of holes in the thalli and the amount of fertile and fouled tissue increased with prolonged growth season, which together led to a significant decline in both biomass yield and quality during summer (June). Early spring (April) conditions were optimal for obtaining high fatty acid, protein, biochar, phenolic, and pigment contents in the biomass, whereas carbohydrate and ash content, as well as essential and non-essential elements, increased later in the growth season. Our study results show that the optimal harvest time of sea-based cultivated U. fenestrata depends on the downstream application of the biomass and must be carefully selected to balance yield, quality, and desired biochemical contents to maximize the output of future sea-based algal cultivations in the European Northern Hemisphere.

In vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant pancreatic enzymes such as proteases, amylase and lipase. Rabbit gastric extracts (RGE) have been recently introduced to provide gastric lipase in addition to pepsin. Before preparing simulated gastric and pancreatic extracts with targeted enzyme activities as described in in vitro digestion protocols, it is important to determine the activities of enzyme preparations using validated methods. The purpose of this inter-laboratory study within the INFOGEST network was to test the repeatability and reproducibility of lipase assays using the pH-stat technique for measuring the activities of gastric and pancreatic lipases from various sources. Twenty-one laboratories having different pH-stat devices received the same protocol with identical batches of RGE and two pancreatin sources. Lipase assays were performed using tributyrin as a substrate and three different amounts (50, 100 and 200 µg) of each enzyme preparation. The repeatability results within individual laboratories were satisfactory with coefficients of variation (CVs) ranging from 4 to 8% regardless of the enzyme amount tested. However, the inter-laboratory variability was high (CV > 15%) compared to existing standards for bioanalytical assays. We identified and weighted the contributions to inter-laboratory variability of several parameters associated with the various pH-stat equipment used in this study (e.g. reaction vessel volume and shape, stirring mode and rate, burette volume for the automated delivery of sodium hydroxide). Based on this, we established recommendations for improving the reproducibility of lipase assays using the pH-stat technique. Defining accurate and complete recommendations on how to correctly quantify activity levels of enzyme preparations is a gateway to promising comparison of in vitro data obtained from different laboratories following the same in vitro digestion protocol.

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.

The growing world population demands an increase in sustainable resources for biorefining. The opening of new farm grounds and the cultivation of extractive species, such as marine seaweeds, increases worldwide, aiming to provide renewable biomass for food and non-food applications. The potential for European large-scale open ocean farming of the commercial green seaweed crop Ulva is not yet fully realized. Here we conducted manipulative cultivation experiments in order to investigate the effects of hatchery temperature (10 and 15 °C), nutrient addition (PES and 3xPES) and swarmer density (500 and 10,000 swarmers ml−1) on the biomass yield and biochemical composition (fatty acid, protein, carbohydrate, pigment and phenolic content) of off-shore cultivated Ulva fenestrata in a Swedish seafarm. High seedling densities were optimal for the growth of this northern hemisphere crop strain and significantly increased the mean biomass yield by ~84% compared to low seedling densities. Variations of nutrients or changes in temperature levels during the hatchery phase were not necessary to increase the subsequent growth in an open-water seafarm, however effects of the factors on the thallus habitus (thallus length/width) were observed. We found no significant effect of the environmental factors applied in the hatchery on the total fatty acid or crude protein content in the off-shore cultivated Ulva. However, low seedling density and low temperature increased the total carbohydrate content and furthermore, high temperature in combination with high nutrient levels decreased the pigment content (chlorophyll a, b, carotenoids). Low temperature in combination with high nutrient levels increased the phenolic content. Our study confirms the successful and sustainable potential for large-scale off-shore cultivation of the Scandinavian crop U. fenestrata. We conclude that high seedling density in the hatchery is most important for increasing the total biomass yield of sea-farmed U. fenestrata, and that changing temperature or addition of nutrients overall does not have a large effect on the biochemical composition. To summarize, our study contributes novel insights into the large-scale off-shore cultivation potential of northern hemisphere U. fenestrata and underpins suitable pre-treatments during the hatchery phase of seedlings to facilitate a successful and cost-efficient large-scale rope cultivation.

Bioactivity of cod (Gadus morhua) and chicken (Gallus domesticus) protein hydrolysates before and after in vitro gastrointestinal (GI) digestion was investigated using yeast Saccharomyces cerevisiae as a model organism. Both hydrolysates were exposed to in vitro GI digestion prior to cellular exposure to simulate digestion conditions in the human body and therefore investigate the role of modulations in the GI tract on the cell response. The effect of digested and undigested hydrolysates on intracellular oxidation, cellular metabolic energy and proteome level was investigated. No difference in the effect on intracellular oxidation activity was obtained between cod and chicken hydrolysates, while higher affect on intracellular oxidation was provided by digested hydrolysates, with relative values of intracellular oxidation of cod of (70.2±0.8) and chicken of (74.5±1.4) % than by undigested ones, where values of cod and chicken were (95.5±1.2) and (90.5±0.7) %, respectively. Neither species nor digestion had any effect on cellular metabolic energy. At proteome level, digested hydrolysates gave again significantly stronger responses than undigested counterparts; cod peptides here also gave somewhat stronger response than chicken peptides. The knowledge of the action of food protein hydrolysates and their digests within live cells, also at proteome level, is important for further validation of their activity in higher eukaryotes to develop new functional food ingredients, such as in this case chicken and cod muscle-derived peptides.

Impacts of processing technology (mechanical separation and pH-shift processing) on protein recovery from salmon, herring and cod backbones and the content of macro- and micronutrients in the recovered protein enriched products were investigated. Mechanical separation led to higher protein recovery compared with the pH-shift process and using both techniques, recovery ranked the species as herring > salmon > cod. However, the pH-shift process up-concentrated protein from herring and salmon backbones more efficiently than mechanical separation by removing more fat and ash. This consequently reduced n-3 PUFA and vitamin D content in their protein isolates compared with the backbones and mechanically separated meat (MSM). Cod protein isolate, however, contained higher levels of these nutrients compared with MSM. Mechanical separation concentrated vitamins E and C in salmon MSM but not for cod and herring. Opposite, pH-shift processing reduced levels of these two vitamins for cod and herring backbones, while vitamins D and C were reduced for salmon. For minerals, selenium, calcium, magnesium, and potassium were lower in protein isolates than MSM, while copper, zinc, iron and manganese were similar or higher. Overall, there is a major potential for upcycling of fish backbones to food ingredients, but processing technology should be carefully balanced against the desired nutrient profile and final application area.

In vitro digestion of marine oils has been reported to promote lipid oxidation, including the formation of reactive aldehydes (e.g., malondialdehyde (MDA) and 4-hydroxy-2-hexenal (HHE)). We aimed to investigate if human in vitro digestion of supplemental levels of oils from algae, cod liver, and krill, in addition to pure MDA and HHE, affect intestinal Caco-2 cell survival and oxidative stress. Cell viability was not significantly affected by the digests of marine oils or by pure MDA and HHE (0–90 μM). Cellular levels of HSP-70, a chaperone involved in the prevention of stress-induced protein unfolding was significantly decreased (14%, 28%, and 14% of control for algae, cod and krill oil, respectively; p ≤ 0.05). The oxidoreductase thioredoxin-1 (Trx-1) involved in reducing oxidative stress was also lower after incubation with the digested oils (26%, 53%, and 22% of control for algae, cod, and krill oil, respectively; p ≤ 0.001). The aldehydes MDA and HHE did not affect HSP-70 or Trx-1 at low levels (8.3 and 1.4 μM, respectively), whilst a mixture of MDA and HHE lowered Trx-1 at high levels (45 μM), indicating less exposure to oxidative stress. We conclude that human digests of the investigated marine oils and their content of MDA and HHE did not cause a stress response in human intestinal Caco-2 cells.