Comparing in planta accumulation with microbial routes to set targets for a cost-competitive bioeconomy.

Plants and microbes share common metabolic pathways for producing a range of bioproducts that are potentially foundational to the future bioeconomy. However, in planta accumulation and microbial production of bioproducts have never been systematically compared on an economic basis to identify optimal routes of production. A detailed technoeconomic analysis of four exemplar compounds (4-hydroxybenzoic acid [4-HBA], catechol, muconic acid, and 2-pyrone-4,6-dicarboxylic acid [PDC]) is conducted with the highest reported yields and accumulation rates to identify economically advantaged platforms and breakeven targets for plants and microbes. The results indicate that in planta mass accumulation ranging from 0.1 to 0.3 dry weight % (dwt%) can achieve costs comparable to microbial routes operating at 40 to 55% of maximum theoretical yields. These yields and accumulation rates are sufficient to be cost competitive if the products are sold at market prices consistent with specialty chemicals ($20 to $50/kg). Prices consistent with commodity chemicals will require an order-of-magnitude-greater accumulation rate for plants and/or yields nearing theoretical maxima for microbial production platforms. This comparative analysis revealed that the demonstrated accumulation rates of 4-HBA (3.2 dwt%) and PDC (3.0 dwt%) in engineered plants vastly outperform microbial routes, even if microbial platforms were to reach theoretical maximum yields. Their recovery and sale as part of a lignocellulosic biorefinery could enable biofuel prices to be competitive with petroleum. Muconic acid and catechol, in contrast, are currently more attractive when produced microbially using a sugar feedstock. Ultimately, both platforms can play an important role in replacing fossil-derived products.

Extremophiles in a changing world.

Extremophiles and their products have been a major focus of research interest for over 40 years. Through this period, studies of these organisms have contributed hugely to many aspects of the fundamental and applied sciences, and to wider and more philosophical issues such as the origins of life and astrobiology. Our understanding of the cellular adaptations to extreme conditions (such as acid, temperature, pressure and more), of the mechanisms underpinning the stability of macromolecules, and of the subtleties, complexities and limits of fundamental biochemical processes has been informed by research on extremophiles. Extremophiles have also contributed numerous products and processes to the many fields of biotechnology, from diagnostics to bioremediation. Yet, after 40 years of dedicated research, there remains much to be discovered in this field. Fortunately, extremophiles remain an active and vibrant area of research. In the third decade of the twenty-first century, with decreasing global resources and a steadily increasing human population, the world's attention has turned with increasing urgency to issues of sustainability. These global concerns were encapsulated and formalized by the United Nations with the adoption of the 2030 Agenda for Sustainable Development and the presentation of the seventeen Sustainable Development Goals (SDGs) in 2015. In the run-up to 2030, we consider the contributions that extremophiles have made, and will in the future make, to the SDGs.

Valorisation of cotton post-industrial textile waste into lactic acid: chemo-mechanical pretreatment, separate hydrolysis and fermentation using engineered yeast.

BACKGROUND: The textile industry has several negative impacts, mainly because it is based on a linear business model that depletes natural resources and produces excessive amounts of waste. Globally, about 75% of textile waste is disposed of in landfills and only 25% is reused or recycled, while less than 1% is recycled back into new garments. In this study, we explored the valorisation of cotton fabric waste from an apparel textile manufacturing company as valuable biomass to produce lactic acid, a versatile chemical building block. RESULTS: Post-industrial cotton patches were pre-treated with the aim of developing a methodology applicable to the industrial site involved. First, a mechanical shredding machine reduced the fabric into individual fibres of maximum 35 mm in length. Afterwards, an alkaline treatment was performed, using NaOH at different concentrations, including a 16% (w/v) NaOH enriched waste stream from the mercerisation of cotton fabrics. The combination of chemo-mechanical pre-treatment and enzymatic hydrolysis led to the maximum recovery yield of 90.46 ± 3.46%, corresponding to 74.96 ± 2.76 g/L of glucose released, which represents a novel valorisation of two different side products (NaOH enriched wastewater and cotton textile waste) of the textile industry. The Saccharomyces cerevisiae strain CEN.PK m850, engineered for redirecting the natural alcoholic fermentation towards a homolactic fermentation, was then used to valorise the glucose-enriched hydrolysate into lactic acid. Overall, the process produced 53.04 g/L ± 0.34 of L-lactic acid, with a yield of 82.7%, being the first example of second-generation biomass valorised with this yeast strain, to the best of our knowledge. Remarkably, the fermentation performances were comparable with the ones obtained in the control medium. CONCLUSION: This study validates the exploitation of cotton post-industrial waste as a possible feedstock for the production of commodity chemicals in microbial cell-based biorefineries. The presented strategy demonstrates the possibility of implementing a circular bioeconomy approach in manufacturing textile industries.

A genetic toolbox to empower Paracoccus pantotrophus DSM 2944 as a metabolically versatile SynBio chassis.

BACKGROUND: To contribute to the discovery of new microbial strains with metabolic and physiological robustness and develop them into successful chasses, Paracoccus pantotrophus DSM 2944, a Gram-negative bacterium from the phylum Alphaproteobacteria and the family Rhodobacteraceae, was chosen. The strain possesses an innate ability to tolerate high salt concentrations. It utilizes diverse substrates, including cheap and renewable feedstocks, such as C1 and C2 compounds. Also, it can consume short-chain alkanes, predominately found in hydrocarbon-rich environments, making it a potential bioremediation agent. The demonstrated metabolic versatility, coupled with the synthesis of the biodegradable polymer polyhydroxyalkanoate, positions this microbial strain as a noteworthy candidate for advancing the principles of a circular bioeconomy. RESULTS: The study aims to follow the chassis roadmap, as depicted by Calero and Nikel, and de Lorenzo, to transform wild-type P. pantotrophus DSM 2944 into a proficient SynBio (Synthetic Biology) chassis. The initial findings highlight the antibiotic resistance profile of this prospective SynBio chassis. Subsequently, the best origin of replication (ori) was identified as RK2. In contrast, the non-replicative ori R6K was selected for the development of a suicide plasmid necessary for genome integration or gene deletion. Moreover, when assessing the most effective method for gene transfer, it was observed that conjugation had superior efficiency compared to electroporation, while transformation by heat shock was ineffective. Robust host fitness was demonstrated by stable plasmid maintenance, while standardized gene expression using an array of synthetic promoters could be shown. pEMG-based scarless gene deletion was successfully adapted, allowing gene deletion and integration. The successful integration of a gene cassette for terephthalic acid degradation is showcased. The resulting strain can grow on both monomers of polyethylene terephthalate (PET), with an increased growth rate achieved through adaptive laboratory evolution. CONCLUSION: The chassis roadmap for the development of P. pantotrophus DSM 2944 into a proficient SynBio chassis was implemented. The presented genetic toolkit allows genome editing and therewith the possibility to exploit Paracoccus for a myriad of applications.

A Holistic Approach to Circular Bioeconomy Through the Sustainable Utilization of Microalgal Biomass for Biofuel and Other Value-Added Products.

Emissions from transportation and industry primarily cause global warming, leading to floods, glacier melt, and rising seas. Widespread greenhouse gas emissions and resulting global warming pose significant risks to the environment, economy, and society. The need for alternative fuels drives the development of third-generation feedstocks: microalgae, seaweed, and cyanobacteria. These microalgae offer traits like rapid growth, high lipid content, non-competition with human food, and growth on non-arable land using brackish or waste water, making them promising for biofuel. These unique phototrophic organisms use sunlight, water, and carbon dioxide (CO2) to produce biofuels, biochemicals, and more. This review delves into the realm of microalgal biofuels, exploring contemporary methodologies employed for lipid extraction, significant value-added products, and the challenges inherent in their commercial-scale production. While the cost of microalgae bioproducts remains high, utilizing wastewater nutrients for cultivation could substantially cut production costs. Furthermore, this review summarizes the significance of biocircular economy approaches, which encompass the utilization of microalgal biomass as a feed supplement and biofertilizer, and biosorption of heavy metals and dyes. Besides, the discussion extends to the in-depth analysis and future prospects on the commercial potential of biofuel within the context of sustainable development. An economically efficient microalgae biorefinery should prioritize affordable nutrient inputs, efficient harvesting techniques, and the generation of valuable by-products.

Various extracts of the brown seaweed Cystoseira barbata with different compositions exert biostimulant effects on seedling growth of wheat.

Worldwide, where the demand for novel and greener solutions for sustainable agricultural production is increasing, the use of eco-friendly products such as seaweed-derived biostimulants as pre-sowing treatment represent a promising and important approach for the future. Cystoseira barbata, a brown seaweed species abundant in the Mediterranean Region, was collected from the Marmara Sea and subjected to water, alkali, and acidic extractions, and the biostimulant activity of these extracts was tested on wheat (Triticum durum cv. Saricanak-98) using different rates through application to the seeds or germination medium (substrate) applications. The different extracts were characterized by mineral, total phenolic, free amino acid, mannitol, polysaccharide, antioxidant concentrations and hormone-like activity. The effects of the extracts on growth parameters, root morphology, esterase activity, and mineral nutrient concentrations of wheat seedlings were investigated. Our results suggest that the substrate application was more effective in enhancing the seedling performance compared to the seed treatment. High rates of seaweed extracts applied to substrates increased the shoot length and fresh weight of wheat seedlings by up to 20 and 25%, respectively. The substrate applications enhanced the root fresh weights of wheat seedlings by up to 25% when compared to control plants. Among the biostimulant extract applications, the water extract at the highest rate yielded the most promising results in terms of the measured parameters. Cystoseira barbata extracts with different compositions can be used as effective biostimulants to boost seedling growth. The local seaweed biomass affected by mucilage problems, has great potential as a bioeconomy resource and can contribute to sustainable practices for agriculture.

Intensive Microalgal Cultivation and Tertiary Phosphorus Recovery from Wastewaters via the EcoRecover Process.

Mixed community microalgal wastewater treatment technologies have the potential to advance the limits of technology for biological nutrient recovery while producing a renewable carbon feedstock, but a deeper understanding of their performance is required for system optimization and control. In this study, we characterized the performance of a 568 m3·day-1 Clearas EcoRecover system for tertiary phosphorus removal (and recovery as biomass) at an operating water resource recovery facility (WRRF). The process consists of a (dark) mix tank, photobioreactors (PBRs), and a membrane tank with ultrafiltration membranes for the separation of hydraulic and solids residence times. Through continuous online monitoring, long-term on-site monitoring, and on-site batch experiments, we demonstrate (i) the importance of carbohydrate storage in PBRs to support phosphorus uptake under dark conditions in the mix tank and (ii) the potential for polyphosphate accumulation in the mixed algal communities. Over a 3-month winter period with limited outside influences (e.g., no major upstream process changes), the effluent total phosphorus (TP) concentration was 0.03 ± 0.03 mg-P·L-1 (0.01 ± 0.02 mg-P·L-1 orthophosphate). Core microbial community taxa included Chlorella spp., Scenedesmus spp., and Monoraphidium spp., and key indicators of stable performance included near-neutral pH, sufficient alkalinity, and a diel rhythm in dissolved oxygen.

Characterizing the Opportunity Space for Sustainable Hydrothermal Valorization of Wet Organic Wastes.

Resource recovery from wet organic wastes can support circular economies by creating financial incentives to produce renewable energy and return nutrients to agriculture. In this study, we characterize the potential for hydrothermal liquefaction (HTL)-based resource recovery systems to advance the economic and environmental sustainability of wastewater sludge, FOG (fats, oils, and grease), food waste, green waste, and animal manure management through the production of liquid biofuels (naphtha, diesel), fertilizers (struvite, ammonium sulfate), and power (heat, electricity). From the waste management perspective, median costs range from -193 $·tonne-1 (FOG) to 251 $·tonne-1 (green waste), and median carbon intensities range from 367 kg CO2 eq·tonne-1 (wastewater sludge) to 769 kg CO2 eq·tonne-1 (green waste). From the fuel production perspective, the minimum selling price of renewable diesel blendstocks are within the commercial diesel price range (2.37 to 5.81 $·gal-1) and have a lower carbon intensity than petroleum diesel (101 kg CO2 eq·MMBTU-1). Finally, through uncertainty analysis and Monte Carlo filtering, we set specific targets (i.e., achieve wastewater sludge-to-biocrude yield >0.440) for the future development of hydrothermal waste management system components. Overall, our work demonstrates the potential of HTL-based resource recovery systems to reduce the costs and carbon intensity of resource-rich organic wastes.

Unlocking the potential of biosurfactants: Production, applications, market challenges, and opportunities for agro-industrial waste valorization.

Biosurfactants are eco-friendly compounds with unique properties and promising potential as sustainable alternatives to chemical surfactants. The current review explores the multifaceted nature of biosurfactant production and applications, highlighting key fermentative parameters and microorganisms able to convert carbon-containing sources into biosurfactants. A spotlight is given on biosurfactants' obstacles in the global market, focusing on production costs and the challenges of large-scale synthesis. Innovative approaches to valorizing agro-industrial waste were discussed, documenting the utilization of lignocellulosic waste, food waste, oily waste, and agro-industrial wastewater in the segment. This strategy strongly contributes to large-scale, cost-effective, and environmentally friendly biosurfactant production, while the recent advances in waste valorization pave the way for a sustainable society.

Biobased heterogeneous renewable catalysts: Production technologies, innovations, biodiesel applications and circular bioeconomy.

The growing population and waste biomass accumulation are leading to increased environmental pollution and climate change. Waste biomass comprising of nutrient rich components has promising potential to produce value-added products for sustainable environmental solutions. This review explores the critical role of bio-based heterogeneous catalysts in enabling sustainable waste biomass utilization. In industrial chemical transformations, over 95% involve catalysts, with more than 90% being heterogeneous systems, prized for their robustness, ease of product separation, and reusability. Bio-based heterogeneous catalysts address the pressing need for sustainable waste biomass management, allowing the conversion of diverse waste biomasses into biodiesel as valuable products. Research on these catalysts, particularly for biodiesel production, has shown yields exceeding 90% with enhanced catalyst reusability. This surge in research is evident from the increasing number of published articles, notably in 2022 and 2023, highlighting growing interest and importance in the scientific community. The synthesis of these catalysts is examined, including novel approaches and techniques to enhance their efficiency, selectivity, and stability. The challenges with their feasible solutions of heterogeneous catalysts in catalyst-based processes are addressed. Altogether, this review underscores the immense potential of bio-based heterogeneous catalysts in sustainable waste biomass utilization, aligning with resource efficiency and environmental conservation goals while offering distinct insights and perspectives on the latest innovations in the field.

Evaluating the feasibility of medium-chain oleochemical synthesis using microbial chain elongation.

Chain elongating bacteria are a unique guild of strictly anaerobic bacteria that have garnered interest for sustainable chemical manufacturing from carbon-rich wet and gaseous waste streams. They produce C6-C8 medium-chain fatty acids, which are valuable platform chemicals that can be used directly, or derivatized to service a wide range of chemical industries. However, the application of chain elongating bacteria for synthesizing products beyond C6-C8 medium-chain fatty acids has not been evaluated. In this study, we assess the feasibility of expanding the product spectrum of chain elongating bacteria to C9-C12 fatty acids, along with the synthesis of C6 fatty alcohols, dicarboxylic acids, diols, and methyl ketones. We propose several metabolic engineering strategies to accomplish these conversions in chain elongating bacteria and utilize constraint-based metabolic modelling to predict pathway stoichiometries, assess thermodynamic feasibility, and estimate ATP and product yields. We also evaluate how producing alternative products impacts the growth rate of chain elongating bacteria via resource allocation modelling, revealing a trade-off between product chain length and class versus cell growth rate. Together, these results highlight the potential for using chain elongating bacteria as a platform for diverse oleochemical biomanufacturing and offer a starting point for guiding future metabolic engineering efforts aimed at expanding their product range. ONE-SENTENCE SUMMARY: In this work, the authors use constraint-based metabolic modelling and enzyme cost minimization to assess the feasibility of using metabolic engineering to expand the product spectrum of anaerobic chain elongating bacteria.

Mixotrophic Cultivation of Arthrospira platensis (Spirulina) under Salt Stress: Effect on Biomass Composition, FAME Profile and Phycocyanin Content.

Arthrospira platensis holds promise for biotechnological applications due to its rapid growth and ability to produce valuable bioactive compounds like phycocyanin (PC). This study explores the impact of salinity and brewery wastewater (BWW) on the mixotrophic cultivation of A. platensis. Utilizing BWW as an organic carbon source and seawater (SW) for salt stress, we aim to optimize PC production and biomass composition. Under mixotrophic conditions with 2% BWW and SW, A. platensis showed enhanced biomass productivity, reaching a maximum of 3.70 g L-1 and significant increases in PC concentration. This study also observed changes in biochemical composition, with elevated protein and carbohydrate levels under salt stress that mimics the use of seawater. Mixotrophic cultivation with BWW and SW also influenced the FAME profile, enhancing the content of C16:0 and C18:1 FAMES. The purity (EP of 1.15) and yield (100 mg g-1) of PC were notably higher in mixotrophic cultures, indicating the potential for commercial applications in food, cosmetics, and pharmaceuticals. This research underscores the benefits of integrating the use of saline water with waste valorization in microalgae cultivation, promoting sustainability and economic efficiency in biotechnological processes.

Valorization of waste engine oil to mono- and di-rhamnolipid in a sustainable approach to circular bioeconomy.

This study aims to valorize waste engine oil (WEO) for synthesizing economically viable biosurfactants (rhamnolipids) to strengthen the circular bioeconomy concept. It specifically focuses on investigating the influence of key bioprocess parameters, viz. agitation and aeration rates, on enhancing rhamnolipid yield in a fed-batch fermentation mode. The methodology involves conducting experiments in a stirred tank bioreactor (3 L) using Pseudomonas aeruginosa gi |KP 163922| as the test organism. Central composite design and response surface methodology (CCD-RSM) are employed to design the experiments and analyze the effects of agitation and aeration rates on various parameters, including dry cell biomass (DCBM), surface tension, tensoactivity, and rhamnolipid yield. It is also essential to determine the mechanistic pathway of biosurfactant production followed by the strain using complex hydrophobic substrates such as WEO. The study reveals that optimal agitation and aeration rates of 200 rpm and 1 Lpm result in the highest biosurfactant yield of 29.76 g/L with minimal surface tension (28 mN/m). Biosurfactant characterization using FTIR, 1H NMR, and UPLC-MS/MS confirm the presence of dominant molecular ion peaks m/z 543.9 and 675.1. This suggests that the biosurfactant is a mixture of mono- and di-rhamnolipids (RhaC10C10, RhaRhaC10C12:1, RhaRhaC12:1C10). The findings present a sustainable approach for biosurfactant production in a fed-batch bioreactor. This research opens the possibility of exploring the use of pilot or large-scale bioreactors for biosurfactant production in future investigations.

Phytomanagement of trace element polluted fields with aromatic plants: supporting circular bio-economies.

Trace elements pollution of soils became a global concern because of their persistence in the environment which can lead to accumulation in food chains up to toxic levels. At the same time, there is a shortage of arable land for growing food, fodder and industrial crops, which highlights the need for remediation/use of polluted land. Restoration of degraded lands has been included as a vital component of UN Sustainable Development Goals (SDGs). We summarize various sources of entry of important trace elements in the environment, available biological reclamation and management strategies and their limitations. Recent advances in phytomanagement approaches using aromatic crops to obtain economically valuable products such as essential oils and revalorize such polluted areas are reviewed. The worldwide application of this strategy in the last 10 years is illustrated through a choropleth map. Finally, the emerging concept of phytomanagement as a restorative and regenerative circular bio-economy is also discussed.

Phytomanagement studies have remarkably increased during the last 4 years. However, there were limited studies on field application of this strategy. France is the leading country in phytomanaged polluted fields using aromatic plants. Applying aromatic plants based degraded land management models are a sustainable approach toward circular economy and to achieve the objectives of sustainable development goals (SDGs).

A Drug Discovery Approach to a Reveal Novel Antioxidant Natural Source: The Case of Chestnut Burr Biomass.

Currently, many environmental and energy-related problems are threatening the future of our planet. In October 2022, the Worldmeter recorded the world population as 7.9 billion people, estimating that there will be an increase of 2 billion by 2057. The rapid growth of the population and the continuous increase in needs are causing worrying conditions, such as pollution, climate change, global warming, waste disposal, and natural resource reduction. Looking for novel and innovative methods to overcome these global troubles is a must for our common welfare. The circular bioeconomy represents a promising strategy to alleviate the current conditions using biomass-like natural wastes to replace commercial products that have a negative effect on our ecological footprint. Applying the circular bioeconomy concept, we propose an integrated in silico and in vitro approach to identify antioxidant bioactive compounds extracted from chestnut burrs (an agroforest waste) and their potential biological targets. Our study provides a novel and robust strategy developed within the circular bioeconomy concept aimed at target and drug discovery for a wide range of diseases. Our study could open new frontiers in the circular bioeconomy related to target and drug discovery, offering new ideas for sustainable scientific research aimed at identifying novel therapeutical strategies.

Review of methodological decisions in life cycle assessment (LCA) of biorefinery systems across feedstock categories.

The application of life cycle assessment (LCA) to biorefineries is a necessary step to estimate their environmental sustainability. This review explores contemporary LCA biorefinery studies, across different feedstock categories, to understand approaches in dealing with key methodological decisions which arise, including system boundaries, consequential or attributional approach, allocation, inventory data, land use changes, product end-of-life (EOL), biogenic carbon storage, impact assessment and use of uncertainty analysis. From an initial collection of 81 studies, 59 were included within the final analysis, comprising 22 studies which involved dedicated feedstocks, 34 which involved residue feedstocks (including by-products and wastes), and a further 3 studies which involved multiple feedstocks derived from both dedicated and secondary sources. Many studies do not provide a comprehensive LCA assessment, often lacking detail on decisions taken, omitting key parts of the value chain, using generic data without uncertainty analyses, or omitting important impact categories. Only 28% of studies included some level of primary data, while 39% of studies did not undertake an uncertainty or sensitivity analysis. Just 8% of studies included data related to dLUC with a further 8% including iLUC, and only 14% of studies considering product end of life within their scope. The authors recommend more transparency in biorefinery LCA, with justification of key methodological decisions. A full value-chain approach should be adopted, to fully assess burdens and opportunities for biogenic carbon storage. We also propose a more prospective approach, taking into account future use of renewable energy sources, and opportunities for increasing circularity within bio-based value chains.

A decision support procedure for the bioeconomy transition: A Colombian case study.

An increasing number of countries and regions consider the bioeconomy transition a strategic policy priority. When approached through the lens of a circular economy perspective, investments in bioeconomy have the potential to enhance resource utilisation efficiency, preserve biodiversity and ecosystems, and foster sustainable development with low emissions. At the same time, if requirements and contextual factors of bioeconomy strategies are not formally analysed, bioeconomic investments might lead to unintended negative consequences. This paper proposes a decision support procedure to design, assess, prioritise, and monitor bioeconomy investments and policies. The flexibility and scalability of our decision support procedure is tested in Colombia to foster a regional and local transition to bioeconomy initiatives that consider the local capital assets and the stakeholders' views. The heterogeneous character of the Colombian environment, economy, society and culture represents an ideal condition to test the strength of the decision support procedure to promote bioeconomy in low and middle-income countries. Our empirical results highlight the benefit of adopting a formal assessment framework that includes strategic national indicators, regional features and stakeholders' views. In terms of the Colombian regional bioeconomy ambitions, we highlight the need for expanding knowledge hubs and participatory stakeholder networks and buttressing appropriate financial mechanisms.

Comprehensive assessment of cow manure hydrothermal treatment products for land application and energy recovery.

In this study, cow manure was hydrothermally treated in a 2-litre reactor for 1 h at temperatures between 100 °C and 260 °C. Both the raw manure and the solid and liquid products of the hydrothermal treatment were characterized to understand the fate of the inorganic elements and to assess the suitability of the products for land applications and energy recovery. Satisfactory elemental balances were obtained for the organic and most inorganic elements and indicated that most inorganic elements were incorporated into the solids with lower solubility, with the exception of potassium and sodium, which were mostly solubilized in the process water; calcium and chlorine were also solubilized to a lesser extent in the process water. Elemental composition and surface functional groups showed that hydrochar produced within the hydrothermal carbonization range (180-260 °C) seemed better suited for utilization as a soil amendment than raw cow manure. The potential for energy recovery lies in the anaerobic digestion of the process water, from which higher methane yields can be obtained than from raw cow manure. Lower temperatures in hydrothermal carbonization are considered a compromise for the safe land applications of cow manure, energy recovery from the process water, and enhanced dewaterability. These findings can help to eliminate bottlenecks in the upscaling of cow manure hydrothermal treatment and promote the circular bio-economy.

Unlocking pilot-scale green diesel production from microalgae.

Microalgae have emerged as a promising source of biomass to produce renewable biofuels due to their ability to synthesize high-energy density compounds of commercial interest. This study proposes an approach for pilot-scale oil extraction, purification by fractional distillation, hydrocarbon characterization by gas chromatography coupled to mass spectrometry (GC-MS), evaluation of physicochemical parameters of the produced hydrocarbons, preliminary cost analysis, and challenges and future opportunities for green diesel on a commercial scale. Here, the microalgae Tetradesmus obliquus was cultivated in 12 m³ photobioreactors using biodigested swine waste as a culture medium. The resulting biomass was subjected to drying and harvesting, followed by oil extraction using a hot solvent extraction method, followed by distillation to purify the compounds. Three different extraction and distillation experiments were conducted, each using different solvent combinations. The results obtained revealed that extraction with a solvent blend, composed of hexane and ethanol, provided more significant yields compared to extraction with pure hexane. GC-MS analysis showed the presence of alkanes and alkenes in the oil samples, and the proportion of solvent used in the extraction directly influenced the production of alkanes. Additionally, specific hydrocarbons such as 4-methyl-1-decene, 8-heptadecene, 1-pentadecene, 9-heneicosene, and 2-dodecene were identified. The evaluation of the physicochemical parameters demonstrated that the calorific value of the distilled oil samples is within the range of typical values for petroleum diesel. However, it was observed that the distilled oil samples had higher sulfur content compared to conventional diesel. Regarding the cost analysis, it was found that it varies depending on the experimental conditions. In particular, the process using a solvent mixture of 70% hexane and 30% ethanol proved to be more economical than the others, since it extracted a greater quantity of oil with a lower initial biomass requirement. In summary, this microalgae-derived hydrocarbon production process is promising and offers insights for compound purification and future biofuel applications.

Transfer and bioaccumulation of chemical and biological contaminants in the marine polychaete Hediste diversicolor (OF müller 1776) when reared on salmon aquaculture sludge.

Side streams from aquaculture production such as fish sludge poses ample opportunities for biological upcycling, as the sludge contains high amounts of nutrients, energy and valuable biochemicals, making it an ideal food for extractive species. Sludge has been proposed as a feed stock for polychaete production, which in turn can be utilized live in shrimp aquaculture or as an aquafeed ingredient. However, the biosafety of such value chains has not yet been addressed. We conducted an experiment exposing the polychaete Hediste diversicolor to aquaculture sludge spiked with four different fish pathogens (Mycobacterium salmoniphilum, Yersinia ruckeri, Infectious Pancreatic Necrosis (IPN) and Infectious Salmon Anaemia (ISA)) known to cause diseases in Atlantic salmon (Salmo salar L.). Moreover, we assessed whether heavy metals and other potentially hazardous elements present in fish sludge bioaccumulates in the polychaetes. Neither of the bacteria nor viruses could be detected in the polychaetes after 14 days of continuous exposure. Seven of the 15 elements we analysed showed bioaccumulation factors significantly below one, meaning biodilution, while the other eight did not differ from one, meaning no bioaccumulation. None of the elements showed a significant bioaccumulation. Further on, none of the heavy metals found in the polychaetes at the end of our experiment exceeded the EU regulatory maximum levels for fish feed ingredients. The current results suggest that a H. diversicolor can reared on aquaculture sludge, and aquaculture sludge may serve as feed stock for polychaete production without the product exceeding EU regulations for contaminants in animal feed.