Desenvolvimento e aplicação de um protótipo para o tratamento de resíduos de cortantes e perfurantes

Tese de mestrado. Inovação e Empreendedorismo Tecnológico. Faculdade de Engenharia. Universidade do Porto. 201

Solid-state fermentation of winery and olive mill wastes for lignocellulolytic enzymes and antioxidant compounds production

Solid-state fermentation (SSF) is a process that can use agroin-dustry solid residues as substrate for microbial growth. During SSF, value-added products and fermented biomass are generated con-tributing for the reutilization of otherwise pollutant agroindustry wastes. By SSF, fungi can hydrolyze anti-nutritional factors of plant-based residues, as the lignocellulosic matrix, and produce enzymes, phenolic compounds and other bioactive compounds, making this a high interest bioprocess for the aquafeed industry. The present study optimized (simplex centroid design) the mixture of winery and olive mill wastes to produce lignocellulolytic enzymes and to extract phenolic compounds through SSF by Aspergillus niger. The substrate mixture that maximized the production of lignocellu-lolytic enzymes, antioxidant and phenolic compounds was 0.44 g exhausted olive pomace/gds; 0.3 g exhausted grape marc/gds; 0.15 g vine-shoot trimmings and 0.1 g crude olive pomace/gds. The crude extract will be applied as additive in meagre (Argyrosomus regius) diets, to improve the feed utilization efciency of low-sh meal-based diets, contributing to the sustainability of aquafeeds.Funded by Programa Operacional Mar2020, Portugal 2020 under the project InovFeed (ref.MAR-02.01.01-FEAMP-0111) and BioTec Norte operation (NORTE-01-0145-FEDER-000004) funded by ERDF under the scope of Norte 2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Improved lignocellulolytic enzyme production and antioxidant extraction using solid-state fermentation of olive pomace mixed with winery waste

Olive pomace is characterized by its low nutritional value and high phenolic content, which hinders its direct use as animal feed, fertilizer, or as a substrate in bioprocesses such as solidstate fermentation (SSF). A possible strategy for bioprocessing olive pomace by SSF is the mixture of olive mill wastes with other wastes produced in the same region, such as winery wastes. This may improve the production of bioactive compounds like enzymes and antioxidant phenolics. A simplexcentroid design was used to evaluate the use of olive mill and winery wastes alone or in combination as a substrate for SSF with Aspergillus niger and Aspergillus ibericus. Synergistic effects of combinations of crude olive pomace (COP), exhausted olive pomace (EOP), vine trimming shoots (VTS), and exhausted grape marc (EGM) were observed in the production of xylanases, cellulases, glucosidases, and in the variation in total phenolics and antioxidant activity of SFF extracts. A multiple response optimization was carried out, leading to the following optimal mixture of substrates: for A. niger, 23% (w/w) COP, 30% EGM, 33% VTS, 14% EOP; for A. ibericus, 30% EGM, 36% VTS, 34% EOP. The scaleup to tray bioreactor with optimal substrate made it possible to achieve the maximum xylanase, cellulase, and glucosidase production of 189.1 ± 26.7, 56.3 ± 2.1 and 10.9 ± 0.8 U/g, respectively. The antioxidant activity of fermented wastes was also improved 2.2fold as compared with unfermented wastes. Thus, a combination of olive mill and winery wastes in SSF is a potential strategy to increase their value and to develop a circular strategy in these industries. © 2019 Society of Chemical Industry and John Wiley & Sons, Ltd.José Manuel Salgado was supported by grant CEB/N2020 – INV/01/2016 from Project ‘BIOTECNORTE – Underpinning Biotechnology to foster the north of Portugal bioeconomy’ (NORTE-01-0145-FEDER-000004). This study was supported by SPO3 project, reference POCI-010145-FEDER-030377, funded by European Regional Development Fund (ERDF) and Portuguese Foundation for Science and Technology (FCT), by the InovFeed project, reference MAR-02.01.01- FEAMP-0111. This study was supported by the FCT under the scope of the strategic funding of UID/BIO/04469/2019 unit and by the BioTecNorte operation (NORTE-01-0145- FEDER-000004) funded by the ERDF under the scope of North 2020 – Northern Regional Operational Program. Helena Fernandes and Carolina Castro were supported by PhD grant SFRH/BD/131219/2017 and post-doctoral grant SFRH/BDP/114942/2016, funded by the Portuguese Foundation for Science and Technology (FCT), respectivelyinfo:eu-repo/semantics/publishedVersio

Solid-state fermentation of distiller’s dried grains with solubles improves digestibility for european seabass (dicentrarchus labrax) juveniles

Aquaculture requires new, economical, and eco-friendly protein sources to replace traditional fisheries and plant ingredients. Using agriculture by-products as protein sources would reduce land-based feed production pressure and waste production, promoting a circular economy and sustainable aquaculture. Distiller’s dried grains with solubles (DDGS) is the main by-product of bioethanol production. Corn DDGS has a high protein level, but its high fiber content limits its use as a feed ingredient, particularly for carnivorous fish. Solid-state fermentation (SSF) uses lignocellulosic-rich substrates, such as DDGS, for microbial growth in the near absence of water, promoting enzyme production that degrades the lignocellulosic matrix, increasing free reducing sugars, protein, and antioxidant levels of the substrate. In the present work, the SSF of corn DDGS with Aspergillus carbonarius, A. ibericus, and A. uvarum was tested. Then, the digestibility of the most promising fermented DDGS (in terms of upgraded nutritional composition) was tested by including it in a reference diet (70% of a reference diet; 48% crude protein; 15% crude lipids) for European seabass (Dicentrarchus labrax) juveniles (171 g averaged weight; trial duration of 52 days). Among the fungi tested, Aspergillus ibericus led a generally higher upgrading of the DDGS nutritional composition, leading to a high amount of protein (from 42.7 to 49.7 g N/kg DM), phenolic compounds (1.49 to 4.86 mg/g caffeic acid equivalents), free sugars (9.5 to 31.9 mg/g), and enzyme production (45 U/g and 68 U/g of cellulase and xylanase, respectively), and a high reduction in acid detergent fiber and neutral detergent fiber content (up to 29 and 43%, respectively). Compared to the unfermented DDGS, fermented DDGS presented increased protein, lipids, starch, and energy digestibility, while phosphorous digestibility was similar. Compared to the reference diet, dietary inclusion of unfermented or fermented DDGS increased trypsin and chymotrypsin activities. The activity of digestive enzymes was not affected by the inclusion of fermented DDGS, except for amylase activity, which was lower with the fermented DDGS than with the unfermented DDGS diet. In conclusion, SSF of DDGS enhanced its nutritional value, increasing DDGS digestibility when included in diets for European seabass juveniles.This research was funded by the R&D&I project MB4Aqua, reference FCT.2022.06587.PTDC from Fundação para a Ciência e Tecnologia (FCT) and by the structural projects CEBpest (UIDB/04469/2020) and LABBELS (LA/P/0029/2020). Diogo Filipe was supported by a grant reference 2020.05525info:eu-repo/semantics/publishedVersio

Individual and mixture toxicity evaluation of three pharmaceuticals to the germination and growth of Lactuca sativa seeds

This work aims to assess, individually and in mixtures, possible phytotoxic effects of three pharmaceuticals (paracetamol, ibuprofen and amoxicillin) on germination and early growth of Lactuca sativa seeds. Pharmaceuticals are an important group of emerging contaminants, whose presence has been described in several environmental compartments, including soils. However, knowledge on their possible impact in terrestrial organisms is still sparse and even more when mixtures are considered. Germination tests are important to evaluate the quality of soil and the toxic effects that contaminants can pose to plants. The acute effects of individual pharmaceuticals as well as binary and ternary mixtures were assessed using different endpoints, namely: percentage of seed germination, root elongation, shoot and leaf length, after an exposure time of five days. Overall, in the exposure of L. sativa seeds to individual pharmaceuticals there are indications of acute toxicity in the early plant growth. However, this inhibitory effect tends to be cancelled in the acute exposure to mixtures. This study shows the importance of evaluating the toxicity of mixtures of pharmaceuticals, since they might have distinct toxic effects when compared to the single compounds, and also because, probably, it is the closest scenario to the reality that can be found in the environment.This work received financial support from the European Regional Development Fund (ERDF) through COMPETE – Operational Competitiveness Programme and national funds provided by FCT – Foundation for Science and Technology under the projects UID/QUI/50006/2013.info:eu-repo/semantics/publishedVersio

Evaluation of the seaweeds Chondrus crispus and Ulva lactuca as functional ingredients in gilthead seabream (Sparus aurata)

The effect of a dietary incorporation of the seaweedsUlva lactucaandChondrus crispusas functional ingredients was evaluatedin gilthead seabream (Sparus aurata) juvenile growth, feed efficiency, muscle lipid composition, intermediary metabolism,oxidative status and gut histomorphology. Fish (15 g initial body weight) were fed with isoproteic (46%) and isolipidic (18%)diets with fish meal (FM) and plant feedstuffs (PF) (27:73 protein from FM:PF) as main protein sources (control diet). Three otherdiets were formulated similar to the control but including 5%U. lactuca,5%C. crispusor 2.5% of both algae (dietsUlva,Chondrusand Mix, respectively). Dietary incorporation ofUlvadid not affect growth performance (P> 0.05) while it leads tohigher muscleΣSFA (saturated fatty acids); and liver lipid peroxidation (LPO). Moreover, lower muscleΣPUFA (polyunsatu-rated fatty acids) and plasma glucose was observed. Dietary incorporation ofChondrusleads to lower growth; whole-body drymatter and lipid content; muscleΣPUFA andω3 fatty acids; and plasma glucose. In addition, higher muscleΣMUFA (mono-unsaturated fatty acids); and liver LPO was observed. Dietary incorporation of both algae (Mix) led to lower growth; whole-bodylipid content; muscleΣPUFA andω3 fatty acids; plasma glucose; and hepatic PK (pyruvate kinase) and HOAD (3-hydroxyacyl-CoA dehydrogenase). Furthermore, it leads to higher muscleΣSFA andΣMUFA; and liver LPO. Overall, no beneficial effects ofincluding the tested seaweeds in the diets was observed, while oxidative status was negatively affected in all dietary treatments.This work was supported by the StructuredR&D&I Project INNOVMAR (ref. NORTE-01-0145-FEDER-000035)within the research line INSEAFOOD, founded by the NorthernRegional Operational Programme (NORTE2020) through the EuropeanRegional Development Fund (ERDF). The Fundação para a Ciência e aTecnologia (FCT) and the FEDER, under programme PT2020 (ProjectUID/QUI/50006/2013) are also acknowledged for the financial funding.I.G., R.M. and A.C. were supported by FCT grants (SFRH/BPD/114959/2016, SFRH/BD/115870/2016 and SFRH/BPD/101354/2014, respec-tively). F.C. was a recipient of a grant within the Project ZEBRALGRE(PTDC/CVT-WEL/5207/2014).info:eu-repo/semantics/publishedVersio

Production of enzymatic extracts for aquafeeds by solid-state fermentation with Aspergillus ibericus of winery and olive mill wastes

[Excerpt] Introduction The replacement of fishmeal by plant ingredients in aquafeeds imposed new formulation strategies to overcome some nutritional restrictions associated with these alternative ingredients. Thus, supplementation of plant-based diets with feed additives, as exogenous enzymes and antioxidants compounds, has recently attracted increasing interest to improve feed utilization and to promote animal health. Solid state fermentation (SSF) of agro-industrial wastes has a high potential for the production of these additives, being a practical, economical, and environmentally-friendly process. Olive mill and winery wastes have valuable compounds that may be valorized thought SSF and that may be used as additives for aquafeeds. This study was conducted to optimize the production of non-starch carbohydrases through the SSF of the olive mill and winery wastes and to test its efficacy to improve the release of pentoses during digestion of a plant-based diet in European seabass.[...]Work supported by FCT PhD grant SFRH/BD/131219/2017, SFRH/BDP/114942/2016, IJFCT-POCI 01-0145-FEDER-030377 and MAR-02.01.01-FEAMP-0111info:eu-repo/semantics/publishedVersio

Novel fish feed supplements: giving value to agroindustrial wastes

Sustainable aquaculture requires the reduction of fish meal use in diets, by replacing it with ecofriendly ingredients, as that vegetal feedstuff not desirable for human consumption. However, these pose challenges due to the presence of antinutritional factors. The supplementation with some supplements, like exoenzymes and antioxidants, has shown potential to reduce their nutritional drawbacks. The solid-state fermentation (SSF) of agro-industrial wastes may be used to obtain such compounds. The goal of this work was to test the applicability of an SSF-extract, obtained through the SSF of an optimized mixture of three agro-industrial wastes with Aspergilus ibericus (30% EGM, 36% VTS, 34% EOP), determined previously with a simplex-centroid mixture design. The activity of this extract was measured to be xylanase: 1866.9 U/g (lyophilized extract); cellulose: 1563.6 U/g; β-glucosidase: 736.455 U/g, and total antioxidants: 439.4 μmol Trolox equivalents/g lyophilized extract; total phenols: 28.1 mg caffeic acid equivalents/g lyophilized extract. The efficiency of this extract as a feed additive in plant-based diets evaluated by an in vitro digestibility trial, analyzing its efficiency to improve the release of pentoses and antioxidants during digestion. The extract was used to supplement diets for Dicentrarchus labrax, with increasing levels of extract: 0, 4 and 8 U cellulase/g diet. Dietary supplementation with the extract led to the increase of pentose release, during alkaline digestion. The effect of the supplementation with the SSF extract on the release of antioxidants and phenols compounds are being evaluated and will be presented.Project SPO3 (ref. POCI-01-0145-FEDER-030377; FEDER-Operational Programme Competitiveness and Internationalization and FCT); project InovFeed (ref. MAR-02.01.01-FEAMP0111; Programa Operacional Mar2020); BioTecNorte (NORTE-01-0145-FEDER-000004; ERDF under the scope of Norte2020-Programa Operacional Regional do Norte)info:eu-repo/semantics/publishedVersio

Solid-state fermentation as green technology to improve the use of plant feedstuffs as ingredients in diets for european sea bass (Dicentrarchus labrax) juveniles

This study aimed to evaluate the utilization by juvenile European sea bass of a SSFed PF mixture with Aspergillus niger CECT 2088. A 22-day digestibility and a 50-day growth trial were performed testing four diets, including 20 or 40% of an unfermented or SSFed PF mixture (rapeseed, soybean, rice bran, and sunflower seed meals, 25% each). SSF of the PF added cellulase and β-glucosidase activity to the diets. Mycotoxin contamination was not detected in any of the experimental diets except for residual levels of zearalenone and deoxynivalenol (100 and 600 times lower than that established by the European Commission Recommendation-2006/576/EC). In diets including 20% PF, SSF did not affect growth but increased apparent digestibility coefficients of protein and energy, feed efficiency, and protein efficiency ratio. On the contrary, in diets including 40% PF, SSF decreased growth performance, feed intake, feed and protein efficiency, and diet digestibility. SSF decreased the intestinal amylase activity in the 40% SSFed diet, while total alkaline proteases decreased in the 20% and 40% SSFed diets. Hepatic amino acid catabolic enzyme activity was not modulated by SSF, and plasma total protein, cholesterol, and triglyceride levels were similar among dietary treatments. In conclusion, dietary inclusion of moderate levels of the SSFed PF, up to 20%, improves the overall feed utilization efficiency without negatively impacting European sea bass growth performance. The replacement of PF with the SSFed PF mixture may contribute to reducing the environmental footprint of aquaculture production.Simple Summary: The rapid growth of the world’s population has increased the demand for seafood, leading to the expansion of aquaculture to fulfill these needs and reduce the pressure on wild fish stocks. Plant feedstuffs (PFs) are often used as the main protein source in aquafeeds due to their wide availability and low cost. However, PFs usually contain high levels of non-starch polysaccharides that limit their utilization in aquafeeds, mainly for carnivorous fish. Solid-state fermentation (SSF) is a cost-effective technological process that may reduce anti-nutritional factor levels while improving nutrient digestibility and the production of several bioactive compounds, enhancing feedstuffs’ nutritional value in aquafeeds. Hence, this study evaluated the effects of using a PF mixture (rapeseed, soybean, rice bran, and sunflower seed meals, 25% each) solid-state fermented (SSFed) with Aspergillus niger CECT 2088 at two inclusion levels (20% and 40%) on European sea bass juveniles’ growth performance, feed digestibility, digestive and catabolic enzyme activity, and plasma metabolites. Overall, the SSFed PF mixture improved the overall feed digestibility, and utilization efficiency, when included at balanced level (20%) without negatively impacting fish growth performance, but not at the higher level (40%).Fundação para a Ciência e a Tecnologia | Ref. UIDB/04423/2020Fundação para a Ciência e a Tecnologia | Ref. UIDP/04423/2020Fundação para a Ciência e a Tecnologia | Ref. 2020.05525.BDFundação para a Ciência e a Tecnologia | Ref. SFRH/BD/115870/2016Fundação para a Ciência e a Tecnologia | Ref. SFRH/BD/137919/2018Fundação para a Ciência e a Tecnologia | Ref. SFRH/BD/143614/201

Assessing antimicrobial potential of agroindustrial byproducts' extracts produced through solid state fermentation in marine fish

[Excerpt] Sustainability is definitely the 21st century word. The future global food provision is dependent on fish and on sustainable expansion of aquaculture production. For that, alternative nutritional strategies for aquaculture disease management are required, reducing the use of synthetic drugs. Agroindustrial byproducts(brewery, winery and olive oil industry) constitute a circular and ecofriendly potential source of add-value bioactive compounds. Solid state fermentation(SSF) is a biotechnological low cost process that may be applied to these byproducts to effectively increase bioavailability of their bioactive compounds and so their antimicrobial potential against fish pathogens. The aim of this study was to evaluate antibacterial properties of agroindustry(beer, wine and oil processing industries) byproducts' extracts produced through SSF with Aspergillus ibericus. [...]SPO3(ref.POCI-01-0145-FEDER-030377;FCT) & InovFeed(ref. MAR-02.01.01-FEAMP0111;Mar2020)info:eu-repo/semantics/publishedVersio