Seafood has an important role to play to achieve a sustainable food system that provides healthy food to a growing world population. Future seafood production will be increasingly reliant on aquaculture where feed innovation is essential to reduce environmental impacts and minimize feed and food competition. This study aimed to investigate whether a novel single cell protein feed ingredient based on Paecilomyces variotii grown on a side stream from the forest industry could improve environmental sustainability of farmed rainbow trout (Oncorhynchus mykiss) by replacing the soy protein concentrate used today. A Life Cycle Assessment including commonly addressed impacts but also the rarely assessed biodiversity impacts was performed. Furthermore, feeding trials were included for potential effects on fish growth, i.e., an assessment of the environmental impacts for the functional unit ‘kg feed required to produce 1 kg live-weight rainbow trout’. Results showed that the best experimental diet containing P. variotii performed 16–73 % better than the control diet containing soy protein concentrate in all impact categories except for energy demand (21 % higher impact). The largest environmental benefits from replacing soy protein with P. variotii in rainbow trout diets was a 73 % reduction of impact on biodiversity and halved greenhouse gas emissions. The findings have high relevance for the aquaculture industry as the production scale and feed composition was comparable to commercial operations and because the effect on fish growth from inclusion of the novel ingredient in a complete diet was evaluated. The results on biodiversity loss from land use change and exploitation through fishing suggest that fishery can dominate impacts and exclusion thereof can greatly underestimate biodiversity impact. Finally, a novel feed ingredient grown on side streams from the forest industry has potential to add to food security through decreasing the dependence on increasingly scarce agricultural land resources. 

By-products from hydrothermal pretreatment of lignocellulosic biomass inhibit enzymatic saccharification and microbial fermentation. Three long-chain organic extractants (Alamine 336, Aliquat 336 and Cyanex 921) were compared to two conventional organic solvents (ethyl acetate and xylene) with regard to conditioning of birch wood pretreatment liquid (BWPL) for improved fermentation and saccharification. In the fermentation experiments, extraction with Cyanex 921 resulted in the best ethanol yield, 0.34 ± 0.02 g g−1 on initial fermentable sugars. Extraction with xylene also resulted in a relatively high yield, 0.29 ± 0.02 g g−1, while cultures consisting of untreated BWPL and BWPL treated with the other extractants exhibited no ethanol formation. Aliquat 336 was most efficient with regard to removing by-products, but the residual Aliquat after the extraction was toxic to yeast cells. Enzymatic digestibility increased by 19-33% after extraction with the long-chain organic extractants. The investigation demonstrates that conditioning with long-chain organic extractants has the potential to relieve inhibition of both enzymes and microbes. 

Trees constitute promising renewable feedstocks for biorefinery using biochemical conversion, but their recalcitrance restricts their attractiveness for the industry. To obtain trees with reduced recalcitrance, large-scale genetic engineering experiments were performed in hybrid aspen blindly targeting genes expressed during wood formation and 32 lines representing seven constructs were selected for characterization in the field. Here we report phenotypes of five-year old trees considering 49 traits related to growth and wood properties. The best performing construct considering growth and glucose yield in saccharification with acid pretreatment had suppressed expression of the gene encoding an uncharacterized 2-oxoglutarate-dependent dioxygenase (2OGD). It showed minor changes in wood chemistry but increased nanoporosity and glucose conversion. Suppressed levels of SUCROSE SYNTHASE, (SuSy), CINNAMATE 4-HYDROXYLASE (C4H) and increased levels of GTPase activating protein for ADP-ribosylation factor ZAC led to significant growth reductions and anatomical abnormalities. However, C4H and SuSy constructs greatly improved glucose yields in saccharification without and with pretreatment, respectively. Traits associated with high glucose yields were different for saccharification with and without pretreatment. While carbohydrates, phenolics and tension wood contents positively impacted the yields without pretreatment and growth, lignin content and S/G ratio were negative factors, the yields with pretreatment positively correlated with S lignin and negatively with carbohydrate contents. The genotypes with high glucose yields had increased nanoporosity and mGlcA/Xyl ratio, and some had shorter polymers extractable with subcritical water compared to wild-type. The pilot-scale industrial-like pretreatment of best-performing 2OGD construct confirmed its superior sugar yields, supporting our strategy. © 2023 The Authors. 

Inhibitors formed during pretreatment impair lignocellulose bioconversion by making enzymatic saccharification and microbial fermentation less efficient, but conditioning of slurries and hydrolysates can improve fermentability and sometimes also enzymatic digestibility. Conditioning of pretreated softwood using four industrial reducing agents (sodium sulfite, sodium dithionite, sodium borohydride, and hydrogen) was compared with standard methods, such as overliming and treatment with activated charcoal. A dosage of approx. 1 mM sulfur oxyanion (sulfite or dithionite) per percent water-insoluble solids (WIS) in the slurry was found to result in good fermentability. Treatment of 10–20% WIS slurries with 15 mM sulfur oxyanion under mild reaction conditions (23°C, pH 5.5) resulted in sulfonation of the solid phase and saccharification improvements of 18–24% for dithionite and 13–16% for sulfite. Among the different conditioning methods studied, treatment of slurries with sodium sulfite was superior with respect to cost-efficient improvement of fermentability. Treatments of slurry or pretreatment liquid with 15 mM sulfite or dithionite resulted in 58–76% reduction of the content of formaldehyde. The comparison indicates that conditioning of pretreated biomass using sulfur oxyanions warrants further attention. 

Conditioning of lignocellulosic hydrolysates with sulfur oxyanions, such as dithionite, is one of the most potent methods to improve the fermentability by counteracting effects of inhibitory by-products generated during hydrothermal pretreatment under acidic conditions. The effects of pH, treatment temperature, and dithionite dosage were explored in experiments with softwood hydrolysates, sodium dithionite, and Saccharomyces cerevisiae yeast. Treatments with dithionite at pH 5.5 or 8.5 gave similar results with regard to ethanol productivity and yield on initial glucose, and both were always at least ~20% higher than for treatment at pH 2.5. Experiments in the dithionite concentration range 5.0–12.5 mM and the temperature range 23–110◦ C indicated that treatment at around 75◦ C and using intermediate dithionite dosage was the best option (p ≤ 0.05). The investigation indicates that selection of the optimal temperature and dithionite dosage offers great benefits for the efficient fermentation of hydrolysates from lignin-rich biomass, such as softwood residues. © 2021 by the authors. 

Background: Aquaculture represents a solution to the future world demand for healthy protein while challenges that require urgent solutions are emerging in feed production, such as the rising costs of feed protein and massive imports. From a European perspective, a large proportion of the protein demand is met with imported protein. This article will focus on the development of protein-rich microorganisms (i.e. Single cell protein) as a novel raw material in fish feed which can be produced as an important co-product in wood-based biorefineries, increasing sustainability and the utilization of organic waste material. Scope and Approach: Developing a safe and sustainable protein resource from local organic waste-material represents an opportunity for Europe to decrease its reliance on nutritional imports, and address mounting food sector sustainability concerns and a growing protein deficit. At the same time, the nutrient recycling industry represents a growing industry, addressing waste valorization and protein feed production concerns at once. Key Findings and Conclusion: An industry and research collaboration has focused on selecting which microorganisms and residual streams from a wood-biorefinery site that would be best suited for production of SCP. The study showed that 38-68% of the fishmeal in a Nile tilapia (Oreochromis niloticus) feed could be replaced with SCP while maintaining a similar or slightly improved fish growth. As reported by FAO, aquaculture production of Nile tilapia in 2014 was 3.7 million tonnes, making it one of the most produced fish species in the world.

BACKGROUND: More extensive utilization of bacterial nanocellulose (BNC) is severely restricted by the low efficiency and small scale of the traditional static cultivation. Submerged fermentation in stirred-tank reactors (STRs) is potentially favourable for large-scale production of BNC, but scale-up of cultivation remains challenging. Even though the STR is most commonly used for submerged cultivation in the fermentation industry, there are few previous attempts to scale-up production of BNC to pilot scale using an STR. Furthermore, the question of how scale-up of submerged cultivation affects the properties of the BNC has received very little attention. RESULTS: Four strains were compared in 250-mL shake flasks. Strain DHU-ATCC-1 displayed the highest volumetric productivity, 0.56 g L−1 d−1, and was then cultivated in a 400-mL STR, showing a similar productivity of 0.55 g L−1 d−1. Scale-up using a 75-L STR pilot bioreactor resulted in enhancement of the BNC production rate from 0.056 g d−1 in the shake flasks to 17.3 g d−1 in the 75-L STR, although the productivity decreased to 0.43 g L−1 d−1. During scale-up from shake flasks to 400-mL STR and further on to 75-L STR, the BNC fibers formed more bundles, whereas the fiber diameter decreased from 25.6 to 21.7 nm. The BNC from the 75-L STR exhibited a higher degree of polymerization, specifically 3230, higher degree of crystallinity, specifically 83%, larger crystallites, and improved strength including higher tensile energy absorption index and superior stretch at break. CONCLUSION: It is possible to enhance BNC production, and maintain or improve its properties when scaling up submerged cultivation in STRs.

Utilization of bacterial nanocellulose (BNC) for large-scale applications is restricted by low productivity in static cultures and by the high cost of the medium. Fiber sludge, a waste stream from pulp and paper mills, was enzymatically hydrolyzed to sugar, which was used for the production of BNC by the submerged cultivation of Komagataeibacter xylinus. Compared with a synthetic glucose-based medium, the productivity of purified BNC from the fiber sludge hydrolysate using shake-flasks was enhanced from 0.11 to 0.17 g/(L × d), although the average viscometric degree of polymerization (DPv) decreased from 6760 to 6050. The cultivation conditions used in stirred-tank reactors (STRs), including the stirring speed, the airflow, and the pH, were also investigated. Using STRs, the BNC productivity in fiber-sludge medium was increased to 0.32 g/(L × d) and the DPv was increased to 6650. BNC produced from the fiber sludge hydrolysate was used as an additive in papermaking based on the chemithermomechanical pulp (CTMP) of birch. The introduction of BNC resulted in a significant enhancement of the mechanical strength of the paper sheets. With 10% (w/w) BNC in the CTMP/BNC mixture, the tear resistance was enhanced by 140%. SEM images showed that the BNC cross-linked and covered the surface of the CTMP fibers, resulting in enhanced mechanical strength.

Increased demand of fish in combination with overexploitation of the fish stocks of the oceans has led to an increased production of fish through aquaculture. Today, fishmeal is the main protein source in fish feed for most aquaculture species. However, fishmeal is soon expected to fall short of demand as its production is associated with environmental problems. This shortage must therefore be met by sustainable alternative protein sources. Protein-rich microorganisms (i.e. Single cell protein) is an interesting option as a fishmeal substitute in fish feed which, in addition, can be produced as an important co-product in wood-based biorefineries. In the current study, four different microorganisms were cultivated on five different residual streams from Swedish wood-based biorefineries. Screening experiments were carried out in shake flasks, optimization experiments in benchtop bioreactors, and scale-up experiments were performed in a 50-litre pilot bioreactor. In addition, a demo-scale experiment was carried out in the Swedish Biorefinery Demo Plant. Microbial biomass from the scale-up experiments was collected and used for production of different fish feed formulations which, in turn, were used in feeding trials of the freshwater fish Tilapia. Fishes fed with feed, in which part of the fishmeal had been substituted with Single cell protein, showed similar or better growth than fishes fed with a fishmeal-based control feed.