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Comparative monosaccharide profiling for taxon differentiation: An example of Icelandic edible seaweeds

Tengiliður

Guðjón Þorkelsson

Stefnumótandi sérfræðingur

gudjon.thorkelsson@matis.is

Edible seaweeds are usually sold as flakes or even powers, and morphological identification of seaweed taxa is difficult. Water-soluble polysaccharides (WSPs) in edible seaweeds are not only interesting functional food ingredients (e.g. gel-forming property, health benefits), but also useful for seaweed taxon differentiation. The current study aims to explore the utility of monosaccharide profiling of WSPs in seaweed differentiation. We developed a high-performance liquid chromatography-photodiode array detection for monosaccharide determination, and characterized monosaccharide profiles of WSPs from five edible seaweeds sold in Iceland (i.e. kombu Iceland – Laminaria hyperborea, sugar kelp – Saccharina latissima, dulse – Palmaria palmata and two types of nori labelled as Porphyra sp. and Pyropia sp.). Monosaccharide profiling data reflected both the complexity and quantitative differences of WSPs. The seaweed dulse showed the highest concentrations of total sugar (ca. 16 mM), followed by kombu (ca. 12 mM). From monosaccharide profiling of red algae, it was found that galactose (ca. 81–87% of total sugars) and xylose (ca. 65–77% of total sugars) are the dominant sugars in nori and dulse, respectively, which reflected the presence of galactans and xylans. In brown algae (i.e. kombu and sugar kelp), glucose (ca. 64–83%) and fucose (13–20%) are the main sugars, and they represent laminarins and fucoidans. The principal component analysis of sugar profiles showed the useful patterns ‘for seaweed taxon differentiation. The dendrogramresulting from hierarchical cluster analysis is congruent with phylogenetic tree, implying the chemotaxonomic value of seaweed monosaccharide profiles. This tool will be more useful in authentication of seaweed taxa when morphological and genetic identifications are not available.

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Environmental Impacts of Large-Scale Spirulina (Arthrospira platensis) Production in Hellisheidi Geothermal Park Iceland: Life Cycle Assessment

Tengiliður

Margrét Geirsdóttir

Verkefnastjóri

mg@matis.is

Spirulina algae (Spirulina platensis) cultivated in geothermally powered photobioreactors is here proposed as a potentially resource efficient, zero-carbon, and nutritious alternative to conventional beef meat. Employing a standard life cycle assessment, environmental impacts of large-scale Spirulina production in this facility are calculated. The production facility is sited in Orka náttúrunnar (ON Power) Geothermal Park, Iceland, and benefits from resource streams accessible through Hellisheiði (Hellisheidi) power station, including renewable electricity for illumination and power usage, hot and cold water streams for thermal management, freshwater for cultivation, and CO2 for biofixation. During cultivation, GHG-intensive ammonia-based fertilizers are replaced with macronutrients sourced from natural open mines. LCA results show that production of 1 kg of wet edible biomass in this facility requires 0.0378 m2 non-arable land, 8.36 m3 fresh water and is carbon neutral with − 0.008 CO2-eq GHG emissions (net zero). Compared with conventionally produced meat from beef cattle, Spirulina algae cultured in the ON Power Geothermal Park, referred to in this study as GeoSpirulina, requires less than 1% land and water and emits less than 1% GHGs. Considering food and nutritional security concerns, cultivation in a controlled environment agriculture system assures consistent nutritional profile year-round. Moreover, GeoSpirulina biomass assessed in this study contains all essential amino acids as well as essential vitamins and minerals. While keeping a balanced nutrition, for every kg beef meat replaced with one kg GeoSpirulina, the average consumer can save ~ 100 kg CO2-eq GHGs. It is concluded that environmental impacts of GeoSpirulina production in the Hellisheidi facility are considerably lower than those of conventionally produced ruminants.

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A Comparison of Fresh and Frozen Lamb Meat-Differences in Technological Meat Quality and Sensory Attributes

Tengiliður

Guðjón Þorkelsson

Stefnumótandi sérfræðingur

gudjon.thorkelsson@matis.is

Technological meat quality and sensory attributes of fresh and frozen lamb meat were compared. Samples were collected from two abattoirs (one small-scale, one large-scale) that use different slaughter methods in terms of chilling regime and electrical stimulation. The fresh and frozen meat samples included products from both slaughter systems. Ten twin pairs of ram lambs were used in the study, with one of each twin slaughtered at each abattoir. Fresh meat was analysed after chilling and frozen meat was stored frozen for three months and analysed after thawing. The Musculus longissimus thoracis et lumborum was analysed for colour, cooking loss, sensory attributes, Warner-Bratzler shear force (WBSF) and distribution of water and lipid within each meat sample. Meat samples analysed after frozen storage were darker, less red and more yellow than the fresh meat. Freezing and frozen storage increased fluid loss and WBSF compared with the fresh meat, due to protein denaturation. Frozen storage affected sensory attributes by increasing fatty odour, frying flavour, sour flavour, fatty flavour and liver flavour, and by reducing juicy texture and mushy texture.

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Microbial Metabolism of Amino Acids – Biologically Induced Removal of Glycine and the Resulting Fingerprint as a Potential Biosignature

Tengiliður

Viggó Marteinsson

Fagstjóri

viggo@matis.is

The identification of reliable biomarkers, such as amino acids, is key for the search of extraterrestrial life. A large number of microorganisms metabolize, synthesize, take up and excrete amino acids as part of the amino acid metabolism during aerobic and/or anaerobic respiration or in fermentation. In this work, we investigated whether the anaerobic microbial metabolism of amino acids could leave a secondary biosignature indicating biological activity in the environment around the cells. The observed fingerprints would reflect the physiological capabilities of the specific microbial community under investigation. The metabolic processing of an amino acid mixture by two distinct anaerobic microbial communities collected from Islinger Mühlbach (ISM) and Sippenauer Moor (SM), Germany was examined. The amino acid mixture contained L-alanine, β-alanine, L-aspartic acid, DL-proline, L-leucine, L-valine, glycine, L-phenylalanine and L-isoleucine. In parallel, an amino acid spiked medium without microorganisms was used as a control to determine abiotic changes over time. Liquid chromatography mass spectrometry (LC-MS) was used to track amino acid changes over time. When comparing to the control samples that did not show significant changes of amino acids concentrations over time, we found that glycine was almost completely depleted from both microbial samples to less than 3% after the first two weeks- This results indicates a preferential use of this simple amino acid by these microbial communities. Although glycine degradation can be caused by abiotic processes, these results show that its preferential depletion in an environment would be consistent with the presence of life. We found changes in most other amino acids that varied between amino acids and communities, suggesting complex dynamics with no clear universal pattern that might be used as a signature of life. However, marked increases in amino acids, caused by cellular synthesis and release into the extracellular environment (e.g., alanine), were observed and could be considered a signature of metabolic activity. We conclude, that substantial anomalous enhancements of some amino acids against the expected abiotic background concentration may be an agnostic signature of the presence of biological processes.

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Production of acetone, butanol, and ethanol by fermentation of Saccharina latissima: Cultivation, enzymatic hydrolysis, inhibitor removal, and fermentation

Tengiliður

Guðmundur Óli Hreggviðsson

Stefnumótandi sérfræðingur

gudmundo@matis.is

Seaweed (or macroalgae) produced sustainably at large scale opens opportunities as source of fuels, chemicals and food. The production does not directly compete with terrestrial food production and may make use of anthropogenic sources of carbon dioxide and nitrogen. Seaweed biomass can be transformed into a suitable substrate for fermentation using a biorefinery approach. In this study the entire process of biofuel production from seaweed is described: starting with cultivation and harvest, the seaweed is dried and cut, enzymatically hydrolysed, demineralized, detoxified, and finally fermented into acetonebutanol, and ethanol (ABE). Juvenile Saccharina latissimawas directly seeded on AlgaeTex® nets and cultivated in the North East Atlantic off the west coast of Scotland for 6 months. Sun dried seaweed was hydrolysed with different enzymes, looking for optimal glucose release, solid/liquid ratio, and enzyme load. Using Cellic® CTec2 in combination with alginate lyases, approximately 80% of available glucose was released. The hydrolysis was scaled up to 100 L, using only Cellic® CTec2. Part of the hydrolysate was demineralized using ion-exclusion chromatography, removing over 90% of minerals while recovering 92% of glucose and mannitol. A fraction of the demineralized hydrolysate was additionally detoxified using a hydrophobic resin to remove hydrophobic components to a concentration below detection limit. The three hydrolysates (untreated, demineralized, and demineralized followed by detoxification) were used as substrate for ABE production by a newly developed strain of Clostridium acetobutylicum adapted to grow on Slatissima hydrolysate. Demineralization reduced the lag phase of fermentation from 72 h (untreated) to 24–48 h. Further detoxification of the hydrolysate led to immediate fermentation, resulting in a yield of 0.23 ± 0.02 gABE/gsugarsimilar to control fermentation in control medium (0.19 gABE/gsugar).

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Adaptive Introgression Facilitates Adaptation to High Latitudes in European Aspen (Populus tremula L.)

Tengiliður

Sæmundur Sveinsson

Fagstjóri

saemundurs@matis.is

Understanding local adaptation has become a key research area given the ongoing climate challenge and the concomitant requirement to conserve genetic resources. Perennial plants, such as forest trees, are good models to study local adaptation given their wide geographic distribution, largely outcrossing mating systems, and demographic histories. We evaluated signatures of local adaptation in European aspen (Populus tremula) across Europe by means of whole-genome resequencing of a collection of 411 individual trees. We dissected admixture patterns between aspen lineages and observed a strong genomic mosaicism in Scandinavian trees, evidencing different colonization trajectories into the peninsula from Russia, Central and Western Europe. As a consequence of the secondary contacts between populations after the last glacial maximum, we detected an adaptive introgression event in a genome region of ∼500 kb in chromosome 10, harboring a large-effect locus that has previously been shown to contribute to adaptation to the short growing seasons characteristic of Northern Scandinavia. Demographic simulations and ancestry inference suggest an Eastern origin—probably Russian—of the adaptive Nordic allele which nowadays is present in a homozygous state at the north of Scandinavia. The strength of introgression and positive selection signatures in this region is a unique feature in the genome. Furthermore, we detected signals of balancing selection, shared across regional populations, that highlight the importance of standing variation as a primary source of alleles that facilitate local adaptation. Our results, therefore, emphasize the importance of migration–selection balance underlying the genetic architecture of key adaptive quantitative traits.

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Use of Probiotic Bacteria and Bacteriocins as an Alternative to Antibiotics in Aquaculture

Tengiliður

Viggó Marteinsson

Fagstjóri

viggo@matis.is

In addition to their use in human medicine, antimicrobials are also used in food animals and aquaculture, and their use can be categorized as therapeutic against bacterial infections. The use of antimicrobials in aquaculture may involve a broad environmental application that affects a wide variety of bacteria, promoting the spread of bacterial resistance genes. Probiotics and bacteriocins, antimicrobial peptides produced by some types of lactic acid bacteria (LAB), have been successfully tested in aquatic animals as alternatives to control bacterial infections. Supplementation might have beneficial impacts on the intestinal microbiota, immune response, development, and/or weight gain, without the issues associated with antibiotic use. Thus, probiotics and bacteriocins represent feasible alternatives to antibiotics. Here, we provide an update with respect to the relevance of aquaculture in the animal protein production sector, as well as the present and future challenges generated by outbreaks and antimicrobial resistance, while highlighting the potential role of probiotics and bacteriocins to address these challenges. In addition, we conducted data analysis using a simple linear regression model to determine whether a linear relationship exists between probiotic dose added to feed and three variables of interest selected, including specific growth rate, feed conversion ratio, and lysozyme activity.

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Protein Recovery of Tra Catfish (Pangasius hypophthalmus) Protein-Rich Side Streams by the pH-Shift Method

Tengiliður

Sigurjón Arason

Yfirverkfræðingur

sigurjon.arason@matis.is

Increasing protein demand has led to growing attention being given to the full utilization of proteins from side streams in industrial fish processing. In this study, proteins were recovered from three protein-rich side streams during Tra catfish (Pangasius hypophthalamus) processing (dark muscle; head-backbone; and abdominal cut-offs) by an optimized pH-shift process. Physicochemical characteristics of the resulting fish protein isolates (FPIs) were compared to industrial surimi from the same raw material batch. The pH had a significant influence on protein extraction, while extraction time and the ratio of the extraction solution to raw material had little effect on the protein and dry matter recoveries. Optimal protein extraction conditions were obtained at pH 12, a solvent to raw material ratio of 8, and an extraction duration of 150 min. The resulting FPI contained <10% of the fat and <15% of the ash of the raw material, while the FPI protein recovery was 83.0−88.9%, including a good amino acid profile. All FPIs had significantly higher protein content and lower lipid content than the surimi, indicating the high efficiency of using the pH-shift method to recover proteins from industrial Tra catfish side streams. The FPI made from abdominal cut-offs had high whiteness, increasing its potential for the development of a high-value product.

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Protein Characteristics and Bioactivity of Fish Protein Hydrolysates from Tra Catfish (Pangasius hypophthalmus) Side Stream Isolates

Tengiliður

Sigurjón Arason

Yfirverkfræðingur

sigurjon.arason@matis.is

Enzymatic hydrolysis is a novel method to recover highly potent bioactive fish protein hydrolysates (FPHs) from fish processing side-streams. The common way of producing FPHs directly from fish side-streams may be inappropriate due to the excess of lipids and pro-oxidants, especially in lipid-rich streams, as obtained from Tra catfish. This study aimed to optimise the hydrolysis conditions for a commercial enzyme (Alcalase® 2.4 L) (enzyme concentrate, temperature, and time) in FPH production from the fish protein isolate obtained from Tra catfish dark muscle (DM-FPI) using the pH-shift method. The degree of hydrolysis (DH), protein recovery (PR), and antioxidant properties, including DPPH radical scavenging activity (DPPH-RSA) and total reducing power capacity (TRPC), were measured to evaluate the effects of the hydrolysis conditions on the FPHs. Optimal hydrolysis was obtained at an enzyme/substrate protein ratio of 3% (v/w) and a hydrolysis temperature of 50 °C for 3 h. The FPHs obtained from different substrates, including DM-FPI, abdominal cut-off (ACO) FPI, and head and backbone blend (HBB) FPI, had similar DHs under these optimum conditions, ranging from 22.5% to 24.0%. However, the FPH obtained from abdominal cut-off isolate (ACO-FPH) showed the highest PR of 81.5 ± 4.3% and the highest antioxidant properties, with a DPPH-RSA of 86.1 ± 1.6% and a TRPC of 6.4 ± 0.4 equivalent mg vitamin C/g protein. The resulting FPHs present a natural source of antioxidants with great potential for food applications, especially the ACO-FPH. In addition, all FPHs had excellent amino acid profiles, indicating strong potential for their use as supplements. Tra catfish protein-rich side-streams can thus be processed into high-value bioactive FPHs using Alcalase for human consumption.

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Changes in Protein and Non-Protein Nitrogen Compounds during Fishmeal Processing – Identification of Unoptimized Processing Steps

Tengiliður

Sigurjón Arason

Yfirverkfræðingur

sigurjon.arason@matis.is

Quality changes of protein and non-protein nitrogen compounds during industrial fishmeal processing of fatty pelagic species (mackerel/herring rest material blend, MHB) and lean fish (whole blue whiting, BW) were studied to identify processing steps that require optimization to allow production of products for human consumption. Samples from protein-rich processing streams throughout the fishmeal production were analyzed for proximate composition, salt soluble protein content (SSP), biogenic amines (BA), total volatile basic nitrogen (TVB-N), trimethylamine (TMA), and dimethylamine (DMA). Mass flows throughout processing were balanced based on the total mass and proximate composition data. The quality of the final fishmeal products was highly dependent on the fish species being processed, indicating that the processes require optimization towards each raw material. The chemical composition changed in each processing step, resulting in different properties in each stream. Most of the non-protein nitrogen compounds (including BA, TVB-N, TMA, and DMA) followed the liquid streams. However, the concentrate contributed less than 20% to the produced fishmeal quantity. Mixing of this stream into the fishmeal processing again, as currently carried out, should thus be avoided. Furthermore, the cooking, separating, and drying steps should be optimized to improve the water and lipid separation and avoid the formation of undesired nitrogen compounds to produce higher-value products intended for human consumption.

IS