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- Preliminary investigation on antioxidant, antibacterial and antiproliferative activities on Pleurotus eryngii var. thapsiae, as potential source of bioactive compoundsPublication . Cirlincione, Fortunato; Cardoso, Rossana V.C.; Mandim, Filipa; Molina, Adriana K.; Pires, Tânia C.S.P.; Venturella, Giuseppe; Barros, LillianSince ancient times mushrooms have been appreciated as food, and in more recent years their medicinal properties have been increasingly exploited. Apart from their use as simple foods, mushrooms are being explored as functional ingredients in the development of novel foods, mushroom-based products, and food supplements. Several studies on medicinal mushrooms have highlighted their wide pharmacological activities, including antioxidant, antimicrobial, anti-inflammatory, and antitumor properties. These pharmacological effects can be attributed to the rich bioactive metabolites composition found in fungi, primarily in the mycelium and sporophores (1). The most studied molecules are phenolic compounds, proteins, fatty acids, terpenes, terpenoids, steroids, and vitamins (2). Several reports have also been conducted on polysaccharides, which are the structural components of the mushroom cell wall and exhibited a strong biological activity (3). Heteroglycans, peptidoglycans, and polysaccharide-protein complexes are polysaccharides that contribute to functional properties that mushroom are known to have, but the most abundant and well-known compounds are α- and β-glucans. Pleurotus (Fr.) P. Kumm. is one of the most widely taxonomic group and include different cultivated and economically important mushrooms. These mushrooms are widely used in human nutrition all over the world and are also recognized as medicinal mushrooms. The genus includes facultative saprotrophs or biotrophs which grow in association with plants of the Apiaceae family, the P. eryngii species complex is undoubtedly noteworthy (4). In this study, we focused on the use of P. eryngii var. thapsiae Venturella, Zervakis & Saitta, whose basidiomes grow in association with Thapsia garganica L. and have been currently reported so far only in Sicily (Italy). The basidiomes growing on two different substrates were used, the first based on wheat straw and the second based on a mixture of wheat straw and Aegilops spp., a common wheat weed. Aqueous crude extracts produced by two different techniques, conventional low-temperature, and ultrasound-assisted extraction, were used. These extracts were tested by radical scavenging activity (DPPH), thiobarbituric acid reactive substances (TBARS) formation inhibition, oxidative haemolysis inhibition assay (OxHLIA), antimicrobial (food and clinical bacteria and fungi) and antiproliferative assays on different human tumor cell lines. Preliminary results showed interesting antimicrobial activities against several Gram-positive and Gram-negative bacteria, including multi-resistant strains, as well as antifungal activities against Aspergillus brasiliensis. Regarding antiproliferative potential, very interesting results were observed against gastric (AGS), colorectal (CaCo-2), breast (MCF7), and liver (NCI-H460) cancer cell lines. Further evaluation of the biological activities and chemical characterization of polysaccharides within the extracts is in progress.
- Chemical and Bioactive Properties of Red Rice with Potential Pharmaceutical UsePublication . Baptista, Eugénia; Liberal, Ângela; Cardoso, Rossana V.C.; Fernandes, Ângela; Dias, Maria Inês; Pires, Tânia C.S.P.; Calhelha, Ricardo C.; García, Pablo A.; Ferreira, Isabel C.F.R.; Barreira, João C.M.Red rice has been proposed as a super-food. Accordingly, the nutritional properties (AOAC), as well as its chemical composition, including sugars (HPLC-RI), organic acids (UFLC-PDA), tocopherols (HPLD-FD), and phenolic compounds (LC-DAD-ESI/MSn), together with the main bioactive properties (antioxidant, cytotoxic, antiproliferative, and antibacterial activities), were evaluated to access its nutritional benefits and health improvement potential. The most abundant macronutrients found were carbohydrates (87.2 g/100 g dw), proceeded by proteins (9.1 g/100 g dw), fat (2.6 g/100 g dw), and ash (1.1 g/100 g dw). Sucrose and raffinose were the only detected sugars, with sucrose presenting the maximum concentration (0.74 g/100 g dw). MUFAs and PUFAs were the predominant fatty acids (40.7% and 31%, respectively). Among the two detected tocopherol isoforms, γ-tocopherol (0.67 mg/100 g dw) predominated over α-tocopherol. The phenolic compounds profile, majorly composed of flavan-3-ols, should be associated with the detected bioactivities, which may provide biological benefits to human health beyond the primary nutritional effect. Overall, the bioactive potential of red rice was comprehensively accessed.
- Irradiação UV-C de cogumelos para obtenção de vitamina D2Publication . Cardoso, Rossana V.C.; Fernandes, Ângela; Barreira, João C.M.; González-Paramás, Ana María; Barros, Lillian; Ferreira, Isabel C.F.R.Inúmeras investigações descrevem o carater altamente nutritivo dos cogumelos, aliado às suas propriedades medicinais (e.g., antitumorais, antimicrobianas, hipocolesterolémicas, antioxidantes); propriedades estas atribuídas à presença de compostos bioativos nomeadamente o ergosterol (precursor da vitamina D2) e outros esteroides, os compostos fenólicos e as vitaminas. A vitamina D2, em particular, desempenha um papel importante na prevenção de diversas patologias e processos fisiopatológicos. Para além da suplementação e da dieta, as leveduras e os cogumelos são as únicas fontes de vitamina D2 (formada durante a exposição aos raios ultravioleta- UV) [1]. Uma vez que as fontes alimentares são escassas, a deficiência de vitamina D pode causar sérios problemas de saúde, tais como, raquitismo, osteoporose, esclerose múltipla, doenças cardiovasculares, entre outros [2]. Neste trabalho, avaliou-se o potencial da utilização de radiação UV-C no aumento do teor de vitamina D2 em amostras laminadas de Agaricus bisporus (J.E. Lange) Imbach e Agaricus bisporus Portobello. A irradiação foi realizada numa câmara UV com a intensidade de 0 (controlo), 200, 800 e 3200 mJ/cm2 e tempo de exposição 0, 2, 6 e 10 min. A vitamina D2 foi determinada por cromatografia líquida de alta eficiência acoplada a um detetor de UV [2,3]. Verificou-se uma interação significativa (p <0,050) entre os fatores (tempo de exposição x intensidade UV-C). A aplicação da radiação UV-C promoveu um aumento na quantidade de vitamina D2, provavelmente devido à conversão de parte do conteúdo de ergosterol naturalmente presente nos cogumelos. No que diz respeito ao tempo de exposição, o período de 6 min resultou como sendo a escolha mais adequada (125 µg/g DW de vitamina D2), uma vez que não foram alcançados aumentos significativos com o tempo máximo testado (10 min). Por outro lado, a dose recomendada de radiação UV-C seria a de 200 mJ/cm, considerando que essa opção poderia representar uma vantagem económica, sem comprometer o aumento da vitamina D2.
- Chemical features and bioactivities of lactuca canadensis L., an unconventional food plant from Brazilian cerradoPublication . Liberal, Ângela; Coelho, Carla T.P.; Fernandes, Ângela; Cardoso, Rossana V.C.; Dias, Maria Inês; Pinela, José; Alves, Maria José; Severino, Vanessa G.P.; Ferreira, Isabel C.F.R.; Barros, LillianThroughout the world, people’s diet is generally quite restricted regarding the variety of plants used in their daily regime. The Unconventional Food Plant (UFP) Lactuca canadensis L. is an edible species of wild lettuce sparsely described in literature and considered to be native from the eastern and central parts of North America. To valorize this species as potential alternative food, an analysis of its nutritional, chemical, and bioactive properties was performed. The results specify the occurrence of organic acids, mainly quinic acid (127.9 ± 0.6 g/kg dry weight (dw)), polyunsaturated fatty acids (65.3%), among which are linolenic acid (44.4 ± 0.4 %), and tocopherols, mostly α-tocopherol (61.2 ± 0.7 mg/kg dw). Additionally, eight phenolic compounds were also identified, among which luteolin-O-glucuronide was found in larger amounts in both infusion and hydroethanolic extracts (5.46 ± 0.09 and 4.6 ± 0.1 mg/g dw, respectively). Carbohydrates and proteins were the main macronutrients (603 ± 1 and 177.5 ± 0.3 g/kg dw, respectively), followed by ashes (166.5 ± 0.9), indicative of a great amount of minerals. Additionally, good antioxidant and antibacterial activities were detected in the analyzed extracts. In general, our results contribute to extend the range of different, unexploited, and nutritionally balanced plant foods, such as Lactuca canadensis, that can and should be included in the daily diet.
- Valorization of cereal by-products from the milling industry as a source of nutrients and bioactive compounds to boost resource-use efficiencyPublication . Cardoso, Rossana V.C.; Fernandes, Ângela; Pinela, José; Dias, Maria Inês; Pereira, Carla; Pires, Tânia C.S.; Carocho, Márcio; Fernández Vasallo, Esteban; Ferreira, Isabel C.F.R.; Barros, LillianCereal by-products (wheat germ, maize bran–germ mixture, rye bran, and wheat bran) from the flour milling industry were characterized for their nutritional value and chemical composition, as well as for antioxidant and antibacterial activities. Carbohydrates (including sucrose) were the major nutritional constituents (56.35–78.12 g/100 g dw), followed by proteins (11.2–30.0 g/100 g dw). The higher energy value (432.3 kcal/100 g dw) was presented by the wheat germ. This by-product also presented the highest citric acid content (0.857 g/100 g dw), the most abundant organic acid detected. Unsaturated fatty acids predominated in all samples given the high content of linoleic (53.9–57.1%) and oleic (13.4–29.0%) acids. Wheat germ had the highest levels of tocopherols (22.8 mg/100 g dw) and phenolic compounds (5.7 mg/g extract, with a high apigenin-C-pentoside-C-hexoside content). In turn, while the wheat bran extract was particularly effective in inhibiting the formation of thiobarbituric acid reactive substances (TBARS), the rye bran extract was the only sample capable of protecting erythrocytes from oxidative hemolysis. Regarding antibacterial properties, in general, the lowest minimum inhibitory concentrations were observed against methicillin-resistant Staphylococcus aureus. These results highlight the characterized by-products as sustainable ingredients for the development of novel bakery and functional food products and contribute to a better bioresource-use efficiency and circularity.
- Gamma irradiation preserves nutritional and chemical composition of Agaricus bisporus PortobelloPublication . Cardoso, Rossana V.C.; Fernandes, Ângela; Barreira, João C.M.; Cabo Verde, Sandra; Antonio, Amilcar L.; Santos, Pedro M.P.; González-Paramás, Ana María; Barros, Lillian; Ferreira, Isabel C.F.R.Mushrooms are being increasingly produced and consumed, but their high perishability still represents a major drawback for commercial purposes. Besides the main effect of increasing the product shelf-life, the conservation technologies should be the most innocuous possible for the consumers. Among the current approaches, irradiation technologies seem to represent interesting alternatives [1, 2]. The present work reports the effects of gamma radiation (1, 2 and 5 kGy) and storage time (0, 4 and 8 days at 5 °C) on the nutritional (moisture, fat, proteins, ash, carbohydrates and energy) and chemical composition (free sugars, ergosterol, tocopherols, organic acids and fatty acids) of Agaricus bisporus Portobello fresh samples, andthe results were compared with control samples (0 kGy, non-irradiated). The irradiation was performed in a Co-60 experimental four sources chamber. The proximate composition was evaluated by AOAC official procedures. Free sugars, ergosterol and tocopherols were determined using high performance liquid chromatography coupled to a refraction index, UV and fluorescence detector, respectively; organic acids were determined using ultrafast liquid chromatography coupled to a diode array detector, while fatty acids were determined using a gas chromatography coupled to a flame ionization detector [3]. Irradiation dose did not exert any remarkably negative effect over the nutritional parameters of stored (up to 8 days) Portobello samples, specifically the irradiated samples at 5 kGy indicated a preserved nutritional profile with the highest levels of protein. It was possible to observe the maintenance of fructose, mannitol and trehalose contents in the irradiated samples, along storage time. All the applied doses also preserved the total organic acids (oxalic, quinic and malic acids) in relation to the control sample. A higher ergosterol level was found in samples irradiated at 2 kGy, while the highest percentages of C16:0 (8.6%) were found in samples treated with 5 kGy. Gamma irradiation was effective in maintaining nutritional and chemical profiles throughout the assayed time intervals. Accordingly, this technology might represent an effective preservation approach for Portobello mushrooms.
- Biochemical approaches on commercial strains of Agaricus subrufescens growing under two environmental cultivation conditionsPublication . Almeida, Daiana; Cardoso, Rossana V.C.; Pereira, Carla; Alves, Maria José; Ferreira, Isabel C.F.R.; Zied, Diego Cunha; Vieira Junior, Wagner Gonçalves; Caitano, Cinthia E.C.; Fernandes, Ângela; Barros, LillianIn the present work, the effect of the cultivation process, in the field and under a con- trolled environment, on biochemical parameters by using commercial strains of A. subrufescens were evaluated. The results obtained revealed that the strains cultivated in the field presented higher levels for most of the parameters evaluated (organic acids (20.5–48.0 g/100 g dw), tocopherols (107.0–198.6 μg/100 g dw), and phenolic acids and related compounds (245.2–359.0 μg/100 g dw and 10.6–23.7 μg/100 g dw, respectively)), except for the carbohydrates (53.4–72.6 g/100 g dw), energetic value (373–380 Kcal/100 g dw), and total free sugars (28.8–43.1 g/100 g dw), parameters in which the strains grown in a controlled environment present better results. For both cultivation systems, similar results were obtained regarding saturated, monounsaturated, and polyunsaturated fatty acids, as well as antioxidant and antimicrobial activities. These data contribute to the knowledge and highlight the characterized strains and the cultivation process, which can be used to obtain ingredients with potential applicability as a source of functional compounds.
- Influência do silicato de cálcio nas propriedades químicas de Pleurotus ostreatus var. FlóridaPublication . Cardoso, Rossana V.C.; Carocho, Márcio; Fernandes, Ângela; Zied, Diego Cunha; Valuenzuela Cobos, Juan Diego; González-Paramás, Ana María; Barros, Lillian; Ferreira, Isabel C.F.R.Os cogumelos do género Pleurotus spp. são conhecidos e apreciados pelas suas aplicações na gastronomia, mas também pela adaptação a diferentes tipos de substratos. A suplementação de substratos (ex. silicato de cálcio) para a produção de cogumelos tem sido utilizada, uma vez que confere uma maior resistência a pragas. Poucos estudos evidenciam o impacto da suplementação do substrato no perfil químico dos cogumelos e neste sentido, foi avaliada a suplementação em amostras de Pleurotus ostreatus var. Flórida com silicato de cálcio a 0, 0.5, 1, 2 e 4% em duas colheitas seguidas do mesmo substrato. Os perfis em ergosterol e vitamina D2 foram determinados por cromatografia líquida de alta eficiência (HPLC) acoplada a um detetor UV, e os tocoferóis foram determinados por HPLC acoplada a um detetor de fluorescência. Foram encontradas pequenas alterações no perfil químico, verificando-se um aumento nos teores da vitamina D2 (779 μg/100 g ms) e tocoferóis totais (0,39 mg/100g ms) nas amostras com 2% de suplementação. Neste sentido, a suplementação com silicato de cálcio não parece induzir mudanças significativas nos perfis químicos destes cogumelos.
- Efeito do silicato de cálcio na composição química de Pleurotus ostreatus var. FloridaPublication . Cardoso, Rossana V.C.; Fernandes, Ângela; Carocho, Márcio; Zied, Diego Cunha; Barreira, João C.M.; González-Paramás, Ana María; Barros, Lillian; Ferreira, Isabel C.F.R.Os cogumelos são um alimento muito apreciado desde tempos imemoriais pelo seu sabor e elevado valor nutritivo. Uma das espécies que tem suscitado maior interesse nos últimos anos é Pleurotus ostreatus, devido à sua facilidade de cultivo, potencial económico e qualidade nutricional, bem como pelas suas propriedades terapêuticas e biológicas [1]. O silício (Si) desempenha um papel importante na nutrição mineral dos cogumelos e plantas; é utilizado com o intuito de aumentar a produtividade uma vez que aumenta a disponibilidade de nutrientes, favorece o crescimento da biomassa, e confere resistência ao stresse biótico e abiótico [2]. São vários os estudos que descrevem a influência da suplementação de cálcio e silício na qualidade do substrato para o cultivo do P. ostreatus [3]. Neste estudo, amostras de Pleurotus ostreatus var. Florida foram suplementadas com silicato de cálcio (0.5, 1, 2 e 4%) com o intuito de verificar o efeito na composição química e nutricional. Os açúcares livres e tocoferóis foram analisados por cromatografia líquida de alta eficiência acoplada a um detetor de índice de refração e fluorescência, respetivamente; e os ácidos gordos foram determinados por cromatografia gasosa acoplada a um detetor de ionização em chama [4]. Foram identificados três açúcares livres, sendo o manitol o maioritário. Verificou-se um aumento nos teores de açúcares totais com a suplementação de 2% de silicato de cálcio. Nos tocoferóis foram identificadas 3 isoformas (α, β e γ-tocoferol), e verificou-se um aumento de quase 30x do teor de γ-tocoferol nas amostras suplementadas com 2% de silicato de cálcio (143 µg/100 g massa seca) comparativamente com o controlo (5 µg/100 g ms). Relativamente aos ácidos gordos, não se obtiveram diferenças acentuadas em comparação com o controlo. Verificou-se que o silicato de cálcio contribuiu para uma melhoria do perfil químico, nos parâmetros avaliados, de P. ostreatus. Desta forma, esta substância poderá ser utilizada para suplementar o substrato de crescimento dos cogumelos, obtendo compostos essenciais para a dieta.
- Optimization and recovery of Vitamin D2 from spurplus production of Agaricus bisporus PortobelloPublication . Cardoso, Rossana V.C.; Fernandes, Ângela; Barreira, João C.M.; Abreu, Rui M.V.; Mandim, Filipa; González-Paramás, Ana María; Ferreira, Isabel C.F.R.; Barros, LillianDuring mushroom production a percentage as high as 20% of surplus can be generated. These unused mushrooms have high nutritional value and valuable chemical compounds. In this sense, finding innovative alternatives to valorizing this surplus mushroom production needs to be explored [1,2]. Irradiation of mushrooms surplus to obtain vitamin D2 is a sustainable strategy to increment vitamin D availability. Under this perspective, the objective of this study was setting the UV-C irradiation and extraction conditions that maximize vitamin D2 contents in surplus mushrooms production (Agaricus bisporus Portobello). The bioactive effects and potential toxicity of vitamin D2-enriched extracts were also evaluated. The surplus production from A. bisporus Portobello were supplied by the Ponto Agrícola, Baião, North of Portugal. The irradiation was performed using an ultraviolet (UV-C at 200 mJ/cm2, 800 mJ/cm2 and 3200 mJ/cm2) radiation chamber with different exposure times: 0, 2, 6 and 10 min. Sensitivity and linearity of the high performance liquid chromatography (HPLC) coupled to ultraviolet detector (UV) were determined and the method was validated by the instrumental precision, repeatability and accuracy, and the extracts rich in vitamin D2 were also quantified by HPLC-UV. The cytotoxicity of the vitamin D2 extract were evaluated using three tumoral cell lines (MCF-7 - breast adenocarcinoma, NCI-H460 - non-small cell lung cancer and AGS - gastric cancer) and one non-tumoral cell line of bone origin (h-FOB 1.19 - human osteoblasts). The chromatographic method revealed a great reproducibility and accuracy, thus being the method validated. Independently of the UV-C irradiation dose, the effect in the conversion of vitamin D2 concentration was very high, allowing it to increase from ~3 μg/g dw to more than 100 μg/g dw in A. bisporus portobello. The extract enriched in vitamin D2 presented an effective activity in AGS (82 μg/mL) tumoral cell line and a moderate activity in NCI-H460 (293 μg/mL) and CaCo (377 μg/mL) tumoral cell lines. Furthermore, the extract did not show cytotoxicity against the non-tumor bone cell h-FOB 1.19 (GI50> 400 μg/mL). Accordingly, development food applications of mushroom extracts enriched in vitamin D2, from surplus mushroom production, can be considered and valorized, supporting and adding value to the agriculture sector or pharmaceutical industries.