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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.

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The Arctic as a food-producing region. Chapter in Renewable Economies in the arctic 

Tengiliður

Ólafur Reykdal

Verkefnastjóri

olafur.reykdal@matis.is

This chapter provides a summary of food production in three regions of the Arctic. These include: the entirety of Iceland; Norway’s three northernmost counties—Nordland, Troms, and Finnmark; and northern Canada, including Yukon, Northwest Territories, Nunavut, Nunavik, and Labrador. In 2016, the Sustainable Development Working Group endorsed The Arctic as a Food Producing Region research project. The export of fish and fish products are by far the most important export items from Iceland and contribute significantly to the Icelandic economy. From 2012 to 2016, Norway exported fish products worth 339,207,335,000 Norwegian krone. Marine products accounted for 89% of northern Canada’s total food export. There are over 3,000 sheep farms in Iceland. These farms tend to be small and family owned. The sheep farming is as old as the human settlement of Iceland. The largest agricultural production systems in Northern Norway are based on meat production. The agricultural production in northern Norway is the northernmost active agricultural system in the world.

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Medium development and production of carotenoids and exopolysaccharides by the extremophile Rhodothermus marinus DSM16675 in glucose-based defined media  

Tengiliður

Þórdís Kristjánsdóttir

Verkefnastjóri

thordis@matis.is

Background

The marine thermophilic bacterium Rhodothermus marinus can degrade many polysaccharides which makes it interesting as a future cell factory. Progress using this bacterium has, however, been hampered by limited knowledge on media and conditions for biomass production, often resulting in low cell yields and low productivity, highlighting the need to develop conditions that allow studies of the microbe on molecular level. This study presents development of defined conditions that support growth, combined with evaluation of production of carotenoids and exopolysaccharides (EPSs) by R. marinus strain DSM 16675.

Results

Two defined media were initially prepared: one including a low addition of yeast extract (modified Wolfe’s medium) and one based on specific components (defined medium base, DMB) to which two amino acids (N and Q), were added. Cultivation trials of R. marinus DSM 16675 in shake flasks, resulted in maximum cell densities (OD620 nm) of 2.36 ± 0.057, cell dry weight (CDW) 1.2 ± 0.14 mg/L, total carotenoids 0.59 × 10–3 mg/L, and EPSs 1.72 ± 0.03 mg/L using 2 g/L glucose in DMB. In Wolfe’s medium (supplemented by 0.05 g/L yeast extract and 2.5 g/L glucose), maximum OD620 nm was 2.07 ± 0.05, CDW 1.05 ± 0.07 mg/L, total carotenoids 0.39 × 10–3 mg/L, and EPSs 1.74 ± 0.2 mg/L. Growth trials at 5 g/L glucose in these media either failed or resulted in incomplete substrate utilization. To improve reproducibility and increase substrate utilization, a screening of macroelements (e.g. phosphate) in DMB, was combined with use of trace elements and vitamins of the modified Wolfe’s medium. The resulting defined minimal R. marinus medium, (DRM), allowed reproducible cultivations to a final OD620nm of 6.6 ± 0.05, CDW 2.85 ± 0.07 mg/L, a maximum specific growth rate (µmax) of 0.26 h−1, total carotenoids 0.77 × 10–3 mg/L and EPSs 3.4 ± 0.17 mg/L in cultivations supplemented with up to 5 g/L glucose.

Conclusion

A minimal defined medium (DRM) was designed that resulted in reproducible growth and an almost doubled formation of both total carotenoids and EPSs. Such defined conditions, are necessary for systematic studies of metabolic pathways, to determine the specific requirements for growth and fully characterize metabolite production.

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Torula yeast in the diet of Atlantic salmon Salmo salarand the impact on growth performance and gut microbiome

Tengiliður

Birgir Örn Smárason

Fagstjóri

birgir@matis.is

Atlantic salmon aquaculture is expanding, and with it, the need to find suitable replacements for conventional protein sources used in formulated feeds. Torula yeast (Cyberlindnera jadinii), has been identified as a promising alternative protein for feed and can be sustainably cultivated on lignocellulosic biomasses. The present study investigated the impact of torula yeast on the growth performance and gut microbiome of freshwater Atlantic salmon. A marine protein base diet and a mixed marine and plant protein base diet were tested, where conventional proteins were replaced with increasing inclusion levels of torula yeast, (0%, 10%, 20%). This study demonstrated that 20% torula yeast can replace fish meal without alteration to growth performance while leading to potential benefits for the gut microbiome by increasing the presence of bacteria positively associated with the host. However, when torula yeast replaced plant meal in a mixed protein diet, results suggested that 10% inclusion of yeast produced the best growth performance results but at the 20% inclusion level of yeast, potentially negative changes were observed in the gut microbial community, such as a decrease in lactic acid bacteria. This study supports the continued investigation of torula yeast for Atlantic salmon as a partial replacement for conventional proteins.

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Feeding black soldier fly larvae (Hermetia illucens) reared on organic rest streams alters gut characteristics of Atlantic salmon (Salmo salar)

Tengiliður

Birgir Örn Smárason

Fagstjóri

birgir@matis.is

The Atlantic salmon (Salmo salar) aquaculture industry is growing, and with it, the need to source and optimise sustainable ingredients for aquafeeds. Black soldier fly (BSF) larvae (Hermetia illucens) have received increasing research attention since they are a good source of protein that can efficiently convert a wide range of low-value organic material into valuable resources. This study investigated the impact of three differently processed BSF meals, an untreated BSF diet (BSFC+), a dechitinated BSF diet (BSFC-) and a fermented BSF diet (BSFC+P+) at a 10% inclusion level replacing fish meal in a fish meal control (FM) and a marine and soy protein concentrate based control diet (SPC). Growth performance, gut microbiome and gut histology of salmon fry was assessed. The inclusion and processing methods of BSF showed no adverse impacts on either growth performance or gut histology. However, the gut microbiome of fish was significantly altered by both the protein source and the processing method of the BSF larvae. Fish fed BSFC+, had an increased diversity and evenness of the community compared with conventional protein sources alone, and compared with the other BSF processing methods. However, control diets had a greater presence of lactic acid bacteria and genera associated with faster growing hosts. Fish fed BSF had a high relative abundance of the genus, Exiguobacterium, a chitin-degrading bacterium and in BSFC+P+ fed fish this bacterium completely dominated the community, indicating the presence of dysbiosis. Future studies should determine, why Exiguobacterium has dominated the community for the BSFC+P+ diet, and if it provides a digestive function to the host and identify bacteria that are indicators of optimal host performance and resilience. The results confirmed that BSF is a promising fish meal replacement for salmon, and it demonstrated that BSFC+ has a potential prebiotic impact on the gut microbiome of Atlantic salmon.

IS