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- Industry-friendly hydroethanolic extraction protocols for Grateloupia turuturu UV-shielding and antioxidant compoundsPublication . Félix, Rafael; Carmona, Ana M.; Félix, Carina; Novais, Sara C.; Lemos, Marco F.L.Grateloupia turuturu is an invasive macroalga on the Iberian coast, known to produce bioactive compounds with di erent cosmeceutical bioactivities, namelyUVshielding and antioxidants. The goal of this study was to optimize the extraction procedure of main bioactivities of this species with cosmetic potential, using Response Surface Methodology. Two Box–Behnken designs were used to evaluate the e ect of ethanol concentration (0–50%), liquid-solid ratio, time, pH, and temperature on yield, UV absorbance, and antioxidant activity. Both optimizations showed a similar trend: aqueous extracts have higher yields and extracts performed with ethanol as part of the solvent have higher activities concerning UV absorbance and antioxidant activity. For all the extracts an absorption peak between 320 and 340 nm was observed. This data now allows further studies by narrowing the extracts worthful of characterization. The development of industry-friendly extraction methods allows the valorization of this invasive species, contributing for the potential creation of natural and eco-friendly products by the cosmetic industry while contributing to the restoration of affected environments.
- Cosmeceutical potential of grateloupia turuturu: using low-cost extraction methodologies to obtain added-value extractsPublication . Félix, Carina; Félix, Rafael; Carmona, Ana M.; Januário, Adriana P.; Dias, Pedro D.M.; Vicente, Tânia F. L.; Silva, Joana; Alves, Celso; Pedrosa, Rui; Novais, Sara C.; Lemos, Marco F.L.The invasive macroalga Grateloupia turuturu is known to contain a diversity of bioactive compounds with different potentialities. Among them are compounds with relevant bioactivities for cosmetics. Considering this, this study aimed to screen bioactivities with cosmeceutical potential, namely, antioxidant, UV absorbance, anti-enzymatic, antimicrobial, and anti-inflammatory activities, as well as photoprotection potential. Extractions with higher concentrations of ethanol resulted in extracts with higher antioxidant activities, while for the anti-enzymatic activity, high inhibition percentages were obtained for elastase and hyaluronidase with almost all extracts. Regarding the antimicrobial activity, all extracts showed to be active against E. coli, S. aureus, and C. albicans. Extracts produced with higher percentages of ethanol were more effective against E. coli and with lower percentages against the other two microorganisms. Several concentrations of each extract were found to be safe for fibroblasts, but no photoprotection capacity was observed. However, one of the aqueous extracts was responsible for reducing around 40% of the nitric oxide production on macrophages, showing its anti-inflammatory potential. This work highlights G. turuturu’s potential in the cosmeceutical field, contributing to the further development of natural formulations for skin protection.
- Biotechnological potential of Grateloupia turuturu for the cosmeceutical industry – photoprotection screening of optimized extractsPublication . Carmona, Ana Marta Ferreira; Lemos, Marco Filipe Loureiro; Novais, Sara Calçada; Pedrosa, Rui Filipe PintoAs invasões biológicas são reconhecidas como uma alteração global de elevada importância e uma das maiores ameaças à biodiversidade aquática e integridade dos ecossistemas. Grateloupia turuturu é uma das 11 macroalgas invasoras documentadas em Portugal e na Galiza e a sua distribuição geográfica aumenta de ano para ano. Está também documentada a sua riqueza em polissacáridos, incluindo sulfatados, que representam uma vasta oportunidade para aplicações biotecnológicas. No âmbito do projeto AMALIA – Algae-to-Market Lab IdeAs –, financiado pela União Europeia, e numa abordagem focada para a indústria, extrações desta alga foram levadas a cabo, com recurso a baixos volumes de água e etanol, combinados com tempos de extração curtos, diferentes temperaturas, pH e rácios sólido-líquido. Os extratos mais promissores para a potencial incorporação em formulações cosméticas foram escolhidos após duas otimizações baseadas em Metodologia de Superfície de Resposta e com avaliação do rendimento de extração, da capacidade antioxidante in vitro e da absorvância UV. Os extratos selecionados foram então testados num simulador solar, com recurso à linha celular 3T3 para a determinação da sua potencial atividade fotoprotetora. No total, foram testados 30 extratos para otimizar as condições de extração com atividades mais promissoras. Destes, 6 condições foram testadas: 3 solventes diferentes (etanol 50%, etanol 25% e água), 3 temperaturas diferentes (20, 30 e 100°C), 3 pH diferentes (4, 7 e 9), 3 tempos de extração diferentes (20, 60 e 100 minutos) e 2 rácios sólido-líquido (1:10 e 1:40). O rendimento é superior nas extrações aquosas (43,370% e 50,841%), a absorvância UV é superior em extratos 50% etanólicos (5,823 e 4,060), bem como a atividade antioxidante testada pelo método de ORAC (153,088 e 102,279 μmol TE/g ext). No que toca aos ensaios celulares, a viabilidade é superior em extratos feitos a temperaturas baixas do que nos seus pares extraídos a 100°C e mantêm uma tendência decrescente à medida que a percentagem de etanol no solvente de extração diminui. No que toca à fotoproteção, os extratos mais promissores são dois extraídos com etanol 25% e um aquoso, com 38,7%, 33,2% e 32,4% de viabilidade celular (respetivamente) vs 6.0% de viabilidade celular do controlo negativo. O extrato 4 (etanol 25%, 20°C, pH 4, durante 100 minutos e rácio sólido-líquido de 1:10) aparenta ser o extrato mais promissor para posterior aplicação na indústria da fotoproteção, com 33,2% de viabilidade celular no ensaio de fotoproteção e 78,5% no ensaio de citotoxicidade. Contudo, o extrato 3 é relevante para a fotoproteção, pois mantém a morfologia celular o mais próximo da normalidade e mantém 38,7% de sobrevivência celular neste ensaio. São, no entanto, necessários testes adicionais para determinar os mecanismos de morte celular em citotoxicidade e de proteção na fotoproteção. Desta forma, este trabalho contribui para a transformação de uma ameaça numa oportunidade, utilizando um organismo invasor da costa noroeste portuguesa para o desenvolvimento de novos produtos, que pode ser recolhido diretamente da natureza, com processos direcionados para a indústria, diminuindo custos e ajudando na restauração do ecossistema.
- Tailoring shrimp aquafeed to tackle Acute Hepatopancreatic Necrosis Disease by inclusion of industry-friendly seaweed extractsPublication . Félix, Rafael; Félix, Carina; Januário, Adriana P.; Carmona, Ana M.; Baptista, Teresa; Gonçalves, Rui A.; Sendão, João; Novais, Sara C.; Lemos, Marco F.L.Shrimp farming has been severely affected by Vibrio infections in southeastern Asia, with severe economic impacts. Invasive seaweeds are an increasing threat that can be strategically valorized by biotechnology, promoting economic development while contributing to ecologic remediation. Asparagopsis armata and Sargassum muticum are two such species that have been frequently reported as excellent bioactive compounds producers, namely anti-bacterial and antioxidant. Thus, A. armata and S. muticum were extracted in an industry-focused approach, using low amounts of ethanol and short extraction times at room temperature. Yield, in vitro antioxidant capacity by the 2,2-diphenyl−1-picrylhydrazyl radical scavenging assay, and in vitro antibacterial activity against Vibrio parahaemolyticus (microdilution) were optimized using Response Surface Methodology (RSM). Optimal extracts (one from each seaweed species) were incorporated in feed, and an in vivo trial using the whiteleg shrimp Penaeus vannamei was performed in industrial facilities using the supplemented feed (1.5, 3.5 and 7.5 g extract per kg feed). Feed quality (along an 8-week period under regular storage conditions) and shrimp performance (40-day trial) were studied, as well as post-challenge (infection by virulent Vibrio parahaemolyticus) mortality rate and hepatopancreas condition. The results from the RSM showed that lower extraction times and ethanol volumes maximized extracts' bioactive properties (A. armata – 20 min, 10 mL solvent per gram seaweed; S. muticum – 100 min, 10 mL.g−1). The seaweed extracts did not impact shrimp performance (no statistically significant differences in weight gain, feed conversion rate or survival). Asparagopsis armata selected extract at 7.5 g per kg feed was able to decrease feed contamination by fungi along time and reduce shrimp mortality by up to 50% upon challenge with V. parahaemolyticus. Despite lacking statistical significance, a difference in hepatopancreas condition after challenge could be identified in shrimp fed the supplemented feed.
- Cosmeceutical potential of crude extracts from Grateloupia turuturuPublication . Carmona, Ana M.; Félix, Rafael; Silva, Joana; Alves, Celso M.; Novais, Sara C.; Pedrosa, Rui P.; Lemos, Marco F.Biological invasions are recognised as a major global change and one of the most important threats to aquatic biodiversity and ecosystem integrity. Grateloupia turuturu is one of the 11 invasive macroalgae documented in Portugal and Galicia and its geographical span is growing every year. It has also been documented that this seaweed is a great source of polysaccharides, sulphated or not, presenting vast biotechnological opportunities. Under the scope of EU-funded project AMALIA – Algae-to-Market-Lab-IdeAs, and in an industry-focused approach, extractions of this alga were performed using low volumes of water and ethanol combined with short extraction times and different temperatures. The most promising extracts to be fitted into a cosmetic formulation with photoprotection activities, and collagenase and elastase inhibitory effects, were chosen after a Response Surface Methodology-based optimization of yield, in vitro antioxidant capacity, and UV-absorbance. The chosen extracts were then tested with a solar simulatorand using a 3T3 cell line for the assessment of their potential photoprotection activity. The same extracts were tested for the inhibitoryactivity of the enzymes collagenase and elastase, responsible for UV radiation-induced skin aging, showing promising activities in all the tests performed.