Development of nutraceutical formulations based on mycelium of Pleurotus ostreatus and Agaricus bisporus

The importance of improving people's quality of life has aroused the interest of the food industry to develop new products with functional characteristics that may be associated with possible beneficial effects on human health, either preventive or therapeutic aspects. Functional foods and nutraceuticals present in its composition bioactive substances that provide medical benefits and, therefore, have aroused the interest of many consumers around the world. Mushrooms are a source of proteins, vitamins, minerals, and especially bioactive compounds. These compounds, including ergosterol, phenolic compounds, tocopherols, ascorbic acid and carotenoids, have been associated with antioxidant, anti-inflammatory, antimicrobial and cytotoxic properties, among others attributed to mushrooms and associated to its promoting health effects. Following this approach, the present work aimed to develope nutraceutical formulations based on mycelium of Agaricus bisporus L. and Pleurotus ostreatus (Jacq. Ex Fr.) P. Kumm. The present study highlights the potential of in vitro culture as a tool to improve production of bioactive compounds by two different mushroom species. Accordingly, A. bisporus and P. ostreatus were studied for their composition in phenolic acids and sterols, antioxidant activity (DPPH radicals scavenging, reducing power, β-carotene bleaching inhibition and TBARS formation inhibition), anti-inflammatory effect (by down-regulating LPS-stimulated NO in RAW264.7 cells) and cytotoxic activity (using MCF-7, NCI-H460, HeLa, HepG2 and PLP2 cell lines). Therefore, the mycelia of A. bisporus and P. ostreatus was cultured in different solid and liquid media, and further submitted to solid-liquid extraction processes; these assayed species showed differences in the growth rate and yielded biomass. Overall, P. ostreatus mycelia showed higher contents of ergosterol and phenolic compounds (but the mycelia of A. bisporus produced in PDA presented a higher amount of p-hydroxybenzoic acid) and stronger antioxidant activity than the corresponding fruting body. On the other hand, P. ostreatus and A. bisporus did not show anti-inflammatory activity. However, P. ostreatus showed cytotoxicity in human tumor cell lines in opposition to A. bisporus, that didn't present cytotoxic activity. In conclusion, the results show that these mushrooms are a good source of compounds with antioxidant and cytotoxic capacity, with variations among species.A importância de melhorar a qualidade de vida das pessoas tem despertado o interesse da indústria alimentar pelo desenvolvimento de novos produtos com características funcionais que possam estar associados a possíveis efeitos benéficos na saúde humana, quer a nível preventivo, quer terapêutico. Os alimentos funcionais e nutracêuticos apresentam na sua composição substâncias bioativas com efeitos benéficos e, portanto, tornam-se apelativos para muitos consumidores em todo o mundo. Os cogumelos são uma fonte de proteínas, vitaminas, minerais e, muitos outros compostos bioativos. Estes compostos, incluindo ergosterol, compostos fenólicos, tocoferóis, ácido ascórbico e carotenóides, têm sido associados a propriedades antioxidantes, anti-inflamatórias, antimicrobianas e citotóxicas, entre outras, relacionadas com os seus efeitos na promoção da saúde. Assim, o presente trabalho teve por objetivo desenvolver formulações nutracêuticas baseadas em micélio de Agaricus bisporus e Pleurotus ostreatus. O estudo destaca ainda o potencial da cultura in vitro como ferramenta para melhorar a produção de compostos bioativos em duas espécies de cogumelos diferentes. Por conseguinte, A. bisporus e P. ostreatus foram estudados quanto à sua composição em ácidos fenólicos e esteróis, atividade antioxidante (captação de radicais livres DPPH, poder redutor, inibição da descoloração do β-caroteno e inibição da formação de TBARS), efeito anti-inflamatório (inibição da produção de NO em células RAW264.7 estimuladas por LPS) e actividade citotóxica (usando Linhas celulares MCF-7, NCI-H460, HeLa, HepG2 e PLP2). Os micélios de A. bisporus e P. ostreatus foram cultivados em diferentes meios de cultura, sólidos e líquidos, e posteriormente submetidos a processos de extracção sólido-líquido; estas espécies analisadas apresentaram diferenças na taxa de crescimento e rendimento de biomassa. Em geral, o micélio de P. ostreatus apresentou teores mais elevados de compostos fenólicos e ergosterol (mas o micélio de A. bisporus produzido em PDA apresentou uma maior quantidade de ácido p-hidroxibenzóico) e maior atividade antioxidante do que o corpo de frutificação correspondente. Por outro lado, A. bisporus e P. ostreatus não mostraram nenhuma atividade anti-inflamatória. No entanto, P. ostreatus mostrou citotoxicidade em linhas celulares tumorais humanas, ao contrário de A. bisporus que não apresentou atividade citotóxica. Em conclusão, os resultados mostram que estes cogumelos são uma boa fonte de compostos com capacidade antioxidante e citotóxica, com variações entre espécies

Flour fortification for nutritional and health improvement: A review

Deficiencies of micronutrients, essentially vitamins and minerals, have mainly cause several illnesses, especially in children and women worldwide. Governments and world organizations have focused great efforts to address these shortcomings and improve the health of the populations. This malnutrition can be combated by fortifying basic foods that are consumed daily. Thus, flours, especially wheat, maize and rice, are a popular and common food in the world's populations diet and are gaining considerable attention as a suitable vehicle for micronutrient fortification. There are some studies that show the effectiveness of flour fortification in combating micronutrient deficiencies and several diseases and the flour fortification strategies are directed to meet nutritional and health needs of the populations. The main points discussed in this review are food fortification, with great focus in flour fortification, health benefits, and legislative issues. This review also highlights multifaceted issues related to flour fortification to meet nutritional needs and to improve the health of vulnerable populations.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), R.V.C. Cardoso grant (SFRH/BD/137436/2018). L. Barros and A. Fernandes also thank the national funding by FCT – Foundation for Science and Technology, P.I., through the institutional scientific employment program-contract for their contracts. This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®; and to FEEI through the Rural Development Program (PDR2020).info:eu-repo/semantics/publishedVersio

Development of nutraceutical formulations based on the mycelium of Pleurotus ostreatus and Agaricus bisporus

The present work is aimed at developing nutraceutical formulations based on the mycelium of Agaricus bisporus and Pleurotus ostreatus, highlighting the potential of in vitro culture as a tool to improve the production of bioactive compounds, namely phenolic acids and ergosterol. The mycelia of both species were cultured in different solid and liquid media in order to compare the growth rate and yielded biomass. Fruiting bodies, mycelia and culture media were compared regarding the antioxidant activity, anti-inflammatory effects in RAW264.7 cells and cytotoxicity in human tumor cell lines and non-tumor porcine liver cells. P. ostreatus mycelia showed higher contents of ergosterol and phenolic compounds, and stronger antioxidant activity than the corresponding fruiting body. P. ostreatus and A. bisporus did not show anti-inflammatory activity, and P. ostreatus was the only one showing cytotoxicity in tumor cell lines. The results show that these mushrooms provide compounds with antioxidant and cytotoxic capacities, with variations among species.The authors are grateful to the Fundação para a Ciência e a Tecnologia (FCT, Portugal) for financial support to CIMO (strategic project PEst-OE/AGR/UI0690/2013) and for L. Barros and R. C. Calhelha contracts.info:eu-repo/semantics/publishedVersio

A case study on surplus mushrooms production: extraction and recovery of vitamin D

The presented case study illustrates the possibility of adding value to the biological surplus remaining from the mushroom cultivation industry. In essence, the unused mushroom parts were submitted to UV-C irradiation, with the purpose of increasing the vitamin D2 content and validat-ing its extraction. Vitamin D2 concentration in three different mushroom species (Agaricus bisporus, A. bisporus Portobello, and Pleurotus ostreatus) was obtained by high-performance liquid chromatogra-phy (HPLC), by means of an ultraviolet (UV) detector. The method was validated using an A. bisporus Portobello sample, and its reproducibility and accuracy were confirmed. Independently of the UV-C irradiation dose, the effect on vitamin D2 concentration was significant, allowing it to increase from less than 4 µg/g dry weight (dw) to more than 100 µg/g dw in all mushroom species. These results are good indicators of the feasibility of industrial surplus mushrooms as sustainable vitamin D2 food sources, besides contributing to strengthen the circularity principals associated to the mushroom production chain.The authors are grateful to the Foundation for Science and Technology (Fundação para a Ciência e Tecnologia, Lisboa, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020), and R.V.C.C.’s PhD grant (SFRH/BD/137436/2018). The authors are also grateful to “Ponto Agricola” for providing the mushroom materials. A.F., J.C.M.B. and L.B. also thank the national funding by FCT, P.I. through the institutional scientific employment program contract. This work is funded by the ERDF through the Regional Operational Program North 2020, within the scope of Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®.info:eu-repo/semantics/publishedVersio

Effectiveness of gamma and electron beam irradiation as preserving technologies of fresh Agaricus bisporus Portobello: a comparative study

The main objectives of the present work were as follows: (a) to develop an analytical procedure to quantify free sugars on chestnuts based on green-chemistry principles and (b) to evaluate the effect of hot air convective drying (50 °C along 10 hr) on the carbohydrate profile of sliced chestnuts (Longal and Judia varieties). The analytical method allowed the quantification of several sugars in chestnut extracts, with low detection limits and good precision. In fresh, Judia variety had higher sucrose, fructose, and glucose contents than Longal, together with amylose. Nevertheless, when applying hot air convective drying to chestnut slices, a significant increase in fructose contents was observed with time, particularly after 10 hr drying in Longal variety. Nonetheless, a significant increase in glucose content was observed on both varieties, probably related to starch thermal hydrolysis. Practical applications: An environment-friendly methodology for free sugars quantification, based on ethanol/water extraction (green solvents), ion-exchange high-performance liquid chromatography separation with water as mobile phase, and evaporative light scattering detection, was developed. The method allowed the quantification of raffinose, sucrose, glucose, and fructose in chestnut extracts, with detection limits of 6 μg for sucrose and 0.3 μg for other sugars. Good precision was achieved with intra-day and inter-day coefficients of variation below 5%. Regarding chestnuts' drying, hot air convective drying might be applied to chestnut slices but it may induce modifications on the carbohydrate composition of the samples, particularly when long drying periods are applied, requiring some attention because it might induce both visual and sensory changes.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), C2TN (UID/Multi/ 04349/2013), L. Barros and J. Barreira contracts, and A. Fernandes post-doc grant (SFRH/BPD/114753/2016). This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®; and to FEEI through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). This work was also developed within the Coordinated Research Project D61024 “Development of New Applications of Machine Generated Food Irradiation Technologies” financed by the International Atomic Energy Agency (IAEA).info:eu-repo/semantics/publishedVersio

Influence of calcium silicate on the chemical properties of pleurotus ostreatus var. Florida (Jacq.) P. Kumm

Supplementation of mushroom substrates has been linked to a higher resistance against insect pests, although few studies show the impact of this supplementation on the different agronomical parameters of mushrooms or even their chemical composition. In this work, the variation in the biological and chemical composition of oyster mushroom (Pleurotus ostreatus var. florida (Jacq.) P. Kumm) was analysed after varying the substrate supplementation of calcium silicate (0, 0.5, 1, 2, and 4% (w/w%)) during two harvest flushes. Overall, supplementation did not change the weight, the number of fruiting bodies, biological efficiency, yield ratio, and productivity rate of the mushrooms, although the harvest flushes did show significant differences. Furthermore, slight changes were found in the chemical composition with an increase in vitamin D2 and tocopherols for the mushrooms with higher amounts of calcium silicate. Overall, the substrate supplementation did not seem to induce expressive changes or decrease production yields, and can, therefore, continue to be researched as a potential application to fight agronomical pests.This work is funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®; and to European Agricultural Fund for Rural Development (EAFRD), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101-031472). Acknowledgments: The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020), and R.V.C. Cardoso’s PhD grant (SFRH/BD/137436/2018). L. Barros and A. Fernandes also thank the national funding by FCT, P.I. through the institutional scientific employment program-contract, and M. Carocho thanks FCT, P.I. through the individual scientific employment program-contract.info:eu-repo/semantics/publishedVersio

Valorization of cereal by-products from the milling industry as a source of nutrients and bioactive compounds to boost resource-use efficiency

Cereal 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.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020) and national funding by FCT, P.I., through the institutional scientific employment program-contract for L. Barros, A. Fernandes, M.I. Dias, and C. Pereira and through the individual scientific employment program-contract for J. Pinela (CEECIND/01011/2018) and M. Carocho (CEECIND/00831/2018). To FEDER-Interreg España-Portugal program for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_P.info:eu-repo/semantics/publishedVersio

Electron-beam irradiation preserves nutritional profile of Agaricus bisporus Portobello

All over the world, mushrooms are highly consumed due to their high contents of digestible proteins, carbohydrates, vitamins and fibers. Mushrooms contain about 90% of water, which leads to a faster deterioration due to senescence, browning, water loss and microbial attack. Despite of the immense popularity of this food worldwide, data regarding alternative technologies to increase mushrooms shelf-life are scarce. Ionizing radiation processing is recognized as a safe and effective method for conservation, being widely used to extend the shelf-life of raw foods. The present work reports the effects of storage time (0, 4 and 8 days) and electron-beam irradiation on the nutritional composition (moisture, fat, proteins, ash, carbohydrates and energy) of Agaricus bisporus Portobello samples. The irradiation was performed with a 10 MeV energy irradiator at the doses of 1, 2 and 5 kGy. The proximate composition was evaluated by AOAC official procedures. Storage time (ST) and electron-beam (EB) irradiation showed a significant interaction in all cases (p-value > 0.05), indicating that the effects potentially exerted by ST depended on EB dose and vice-versa. Considering the individual effect of each factor, EB induced more changes than ST, which in fact had a significant effect only for protein and carbohydrates. Among different EB doses, despite the significant differences, it is evident that the nutritional profiles were not greatly changed with any applied dose. Therefore, up to 5 kGy, EB seems to be a suitable conservation treatment for A. bisporus Portobello.Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), C2TN (UID/Multi/04349/2013), grant to A. Fernandes (SFRH/BPD/114753/2016) and L. Barros contract; European Structural and Investment Funds (ESIF) (Regional Operational Program Norte 2020, Project ValorNatural®); Rural Development Program (Project MicoCoating, PDR2020-101-031472); International Atomic Energy Agency (IAEA) Coordinated Research Project D61024 DEXAFI.info:eu-repo/semantics/publishedVersio

Chemical features and bioactivities of lactuca canadensis L., an unconventional food plant from Brazilian cerrado

Throughout 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.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020) and national funding by FCT, P.I., through the institutional scientific employment program-contract for L.B., A.F. and M.I.D., and through the individual scientific employment program-contract for J.P. (CEECIND/01011/2018). The authors are also grateful to the project TRANSCoLAB (0612_TRANS_CO_LAB_2_P), to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth, Norte-01-0145-FEDER-000042. The authors C.T.P.C. and V.G.P.S. are grateful to the international mobility departments of the Federal University of Goiás and the Polytechnic Institute of Bragança for the opportunity for student exchange of C.T.P.C. and developing of the research with L. canadensis species.info:eu-repo/semantics/publishedVersio

Chemical and Bioactive Properties of Red Rice with Potential Pharmaceutical Use

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.This work was supported by national funds through FCT/MCTES (PIDDAC): CIMO, UIDB/00690/2020 (DOI: 10.54499/UIDB/00690/2020), UIDP/00690/2020 (DOI: 10.54499/UIDP/ 00690/2020), and SusTEC, LA/P/0007/2020 (DOI: 10.54499/LA/P/0007/2020). A. Fernandes, M.I. Dias, R.C. Calhelha, and J.C.M. Barreira are also grateful for the national funding by FCT. P.I. through the institutional and individual program contract. The authors are grateful to FEDER Cooperación Interreg VI A Espanha—Portugal (POCTEP) 2021–2027 for its financial support provided through the project TRANSCoLAB PLUS 0112_TRANSCOLAB_PLUS_2_P.info:eu-repo/semantics/publishedVersio