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- Sargassum muticum and Osmundea pinnatifida enzymatic extracts: chemical, structural, and cytotoxic characterizationPublication . Rodrigues, Dina; Costa-Pinto, Ana R.; Sousa, Sérgio; Vasconcelos, Marta W.; Pintado, Manuela M.; Pereira, Leonel; Rocha-Santos, Teresa A. P.; Costa, João P. da; Silva, Artur M. S.; Duarte, Armando C.; Gomes, Ana M. P.; Freitas, Ana C.Seaweeds, which have been widely used for human consumption, are considered a potential source of biological compounds, where enzyme-assisted extraction can be an efficient method to obtain multifunctional extracts. Chemical characterization of Sargassum muticum and Osmundea pinnatifida extracts obtained by Alcalase and Viscozyme assisted extraction, respectively, showed an increment of macro/micro elements in comparison to the corresponding dry seaweeds, while the ratio of Na/K decreased in both extracts. Galactose, mannose, xylose, fucose, and glucuronic acid were the main monosaccharides (3.2–27.3 mg/glyophilized extract) present in variable molar ratios, whereas low free amino acids content and diversity (1.4–2.7 g/100gprotein) characterized both extracts. FTIR-ATR and 1H NMR spectra confirmed the presence of important polysaccharide structures in the extracts, namely fucoidans from S. muticum or agarans as sulfated polysaccharides from O. pinnatifida. No cytotoxicity against normal mammalian cells was observed from 0 to 4 mglyophilized extract/mL for both extracts. The comprehensive characterization of the composition and safety of these two extracts fulfils an important step towards their authorized application for nutritional and/or nutraceutical purposes.
- Chemical composition of red, brown and green macroalgae from Buarcos bay in Central West Coast of PortugalPublication . Rodrigues, Dina; Freitas, Ana C.; Pereira, Leonel; Rocha-Santos, Teresa A. P.; Vasconcelos, Marta W.; Roriz, Mariana; Rodríguez-Alcalá, Luís M.; Gomes, Ana M. P.; Duarte, Armando C.Six representative edible seaweeds from the Central West Portuguese Coast, including the less studied Osmundea pinnatifida, were harvested from Buarcos bay, Portugal and their chemical characterization determined. Protein content, total sugar and fat contents ranged between 14.4% and 23.8%, 32.4% and 49.3% and 0.6–3.6%. Highest total phenolic content was observed in Codium tomentosum followed by Sargassum muticum and O. pinnatifida. Fatty acid (FA) composition covered the branched chain C13ai to C22:5 n3 with variable content in n6 and n3 FA; low n6:n3 ratios were observed in O. pinnatifida, Grateloupia turuturu and C. tomentosum. Some seaweed species may be seen as good sources of Ca, K, Mg and Fe, corroborating their good nutritional value. According to FTIR-ATR spectra, G. turuturu was associated with carrageenan seaweed producers whereas Gracilaria gracilis and O. pinnatifida were mostly agar producers. In the brown algae, S. muticum and Saccorhiza polyschides, alginates and fucoidans were the main polysaccharides found.
- Impact of enzyme- and ultrasound-assisted extraction methods on biological properties of red, brown, and green seaweeds from the Central West Coast of PortugalPublication . Rodrigues, Dina; Sousa, Sérgio; Silva, Aline; Amorim, Manuela; Pereira, Leonel; Rocha-Santos, Teresa A.P.; Gomes, Ana M. P.; Duarte, Armando C.; Freitas, Ana CristinaSeaweeds are an excellent source of bioactive compounds, and therefore the use of sustainable and food compatible extraction methods such as enzyme-assisted (EAE) and ultrasound-assisted extraction were applied on Sargassum muticum, Osmundea pinnatifida, and Codium tomentosum. Extracts were evaluated for proximate characterization and biological properties. Higher extraction yields were observed for C. tomentosum EAE (48-62%; p < 0.05 for Cellulase and Viscozyme), followed by O. pinnatifida (49-55%; p < 0.05 except Alcalase) and S. muticum (26-31%; p < 0.05). S. muticum extracts presented the highest nitrogen (25 +/- 2 mg/g(lyoph) (extract)) and total phenolics (261 +/- 37 mu g(catechol) (equiv)/gl(yoph) (extract)) contents, whereas higher sugars (78 +/- 14 mg(glucose) (equiv)/g(lyoph) (extract)) including sulfated polysaccharide (44 +/- 8 mg(Na2SO4) (acid)/g(lyoph) (extract)) contents characterized O. pinnatifida extracts. A higher effect on hydroxyl-radical scavenging activity (35-50%) was observed for all extracts, whereas S. muticum Alcalase and C. tomentosum Cellulase extracts exhibited higher prebiotic activity than fructooligosaccharides. O. pinnatifida and C. tomentosum EAE showed inhibitory potential against alpha-glucosidase (38-49%).
- Lipolysis in probiotic and synbiotic cheese: The influence of probiotic bacteria, prebiotic compounds and ripening time on free fatty acid profilesPublication . Rodrigues, Dina; Rocha-Santos, Teresa A. P.; Gomes, Ana M.; Goodfellow, Brian J.; Freitas, Ana C.The influence of probiotic bacteria (Lactobacillus casei-01, Bifidobacterium lactis B94), prebiotic compounds (FOS and inulin) and ripening time (0–60 days) on the free fatty acid (FFA) profile of cheese, with special emphasis on the conjugated linoleic acid (CLA) content, was investigated. After 60 days of ripening, 109–1010 cfu g 1 cheese were recorded in both probiotic and synbiotic cheeses, despite harsh conditions of low pH values (4.1–5.1) and low moisture content (<30%, w/w). Increases in total FFA and CLA were observed throughout the ripening period, especially in synbiotic cheeses containing FOS and inulin (50:50) inoculated with B. lactis B94. The addition of FOS alone or combined with inulin did not significantly affect probiotic strain growth and viability during the ripening period; however, the advantage of the addition of prebiotic compounds in probiotic cheese manufacture is that it may allow the production of cheeses with improved performance as far as functional CLA compounds are concerned, as well as an improved nutritional quality reflected in a lower atherogenicity index.
- Chemical and structural characterization of Pholiota nameko extracts with biological propertiesPublication . Rodrigues, Dina; Freitas, Ana C.; Sousa, Sérgio; Amorim, Manuela; Vasconcelos, Marta W.; Costa, João P. da; Silva, Artur M. S.; Rocha-Santos, Teresa A. P.; Duarte, Armando C.; Gomes, Ana M. P.Edible mushrooms including Pholiota nameko are excellent sources of extractable bioactive compounds with much to explore. Enzymatic extractions with Cellulase and Viscozyme were responsible for highest extraction yields (67-77%). No strong antioxidant activity was observed although extracts were able to scavenge ABTS(+) and OH(+). Potential prebiotic activity was observed in all extracts, some increasing 1.4-2 Log cycles of Lactobacillus acidophilus La-5 and Bifidobacterium animalis BB12. 30-50% α-glucosidase inhibition was observed in ultrasound, Flavourzyme and Cellulase extracts. Flavourzyme and Cellulase extracts are richer in macro (Mg, K and P) and micro elements (Zn, Mn and Fe) in comparison to mushroom. Monosaccharides content and profile varied slightly among both extracts with predominance of glucose, galactose and mannose with no uronic acids detection; Flavourzyme extract reported higher free amino acids content. Presence of α and β-glycosidic structures such as glucans and glucan-protein complexes are among the polysaccharides found in both extracts.
- Development of a rice-based fermented probiotic beverage with lactobacillus spp. and streptococcus thermophilusPublication . Gomes, Ana Maria; Ramalhosa, Francisco; Sousa, Sérgio; Rodrigues, Dina; Freitas, Ana Cristina
- Development of probiotic tablets using microparticles: viability studies and stability studiesPublication . Sousa e Silva, J. P.; Sousa, Sérgio C.; Costa, Paulo; Cerdeira, Emília; Amaral, Maria H.; Lobo, José Sousa; Gomes, Ana M. P.; Pintado, Maria M.; Rodrigues, Dina; Rocha-Santos, Teresa; Freitas, A. C.Alternative vectors to deliver viable cells of probiotics, to those conferring limited resistance to gastrointestinal conditions, still need to be sought. Therefore the main goal of the study was to develop tablets able to protect entrapped probiotic bacteria from gastric acidity, thus providing an easily manufacturing scale-up dosage form to deliver probiotics to the vicinity of the human colon. Whey protein concentrate microparticles with Lactobacillus paracasei L26 were produced by spray-drying and incorporated in tablets with cellulose acetate phthalate and sodium croscarmellose. The viability of L. paracasei L. 26 throughout tableting as well as its gastric resistance and release from the tablets were evaluated. Storage stability of L. paracasei L26 tablets was also performed by evaluation of viable cells throughout 60 days at 23 degrees C and 33% relative humidity. A decrease of approximately one logarithmic cycle was observed after the acid stage and the release of L. paracasei L26 from the tablets occurred only after 4 h in the conditions tested. Microencapsulated L. paracasei L26 in tablets revealed some susceptibility to the storage conditions tested since the number of viable cells decreased 2 log cycles after 60 days of storage. However, the viability of L. paracasei L26 after 45 days of storage did not reveal significant susceptibility upon exposure to simulated gastrointestinal conditions. The developed probiotic tablets revealed to be potential vectors for delivering viable cells of L. paracasei L26 and probably other probiotics to persons/patients who might benefit from probiotic therapy.
- Chemical composition and nutritive value of Pleurotus citrinopileatus var cornucopiae, P. eryngii, P. salmoneo stramineus, Pholiota nameko and Hericium erinaceusPublication . Rodrigues, Dina M. F.; Freitas, Ana C.; Rocha-Santos, Teresa A. P.; Vasconcelos, Marta W.; Roriz, Mariana; Rodríguez-Alcalá, Luís M.; Gomes, Ana M. P.; Duarte, Armando C.The chemical composition and nutritive value of five mushrooms species, some less studied such as Pleurotus citrinopileatus var. cornucopiae, P. salmoneo stramineus or Pholiota nameko, were determined. Protein, sugar and fat contents ranged between 16.2 to 26.6, 52.7 to 64.9 and 2.3 to 3.5 g/100g(dry mushroom), respectively. Highest total phenolic content was observed for P. citrinopileatus var. cornucopiae with 1140 mu g cathecol equiv./g(dry mushroom). Higher content in mono and polyunsaturated fatty acids (FA) than saturated FA characterized mushrooms FA profile with high linoleic acid concentration (> 30 gFA/100g(fat)). In addition, these mushrooms may be considered good sources of K, Mg, P, highlighting K (2627-3736 mg/Kg(dry mushroom)) as the most predominant, and of Zn, Cu and Fe; some contributing over 15 % of their recommended daily intakes. beta-glucans, alpha-glucans and evidence of glucan-protein complexes were identified by FTIR-ATR. The reported values emphasize the nutritional potential of the five species to be consumed in a healthy diet.
- Encapsulation of probiotic strains in plain or cysteine-supplemented alginate improves viability at storage below freezing temperaturesPublication . Sousa, Sérgio; Gomes, Ana M.; Pintado, Maria M.; Malcata, Francisco X.; Silva, José P.; Sousa, José M.; Costa, Paulo; Amaral, Maria H.; Rodrigues, Dina; Rocha-Santos, Teresa A. P.; Freitas, Ana C.Four probiotic bacteria (Lactobacillus paracasei L26, L. casei-01, L. acidophilus Ki, and Bifidobacterium animalis BB-12 R ) were encapsulated in plain alginate or alginate supplemented with L-cysteine·HCl, and resulting microcapsules were stored at different temperatures, namely 21, 4, −20, or −80◦C for a period of up to6months. The results showed that the encapsulation in calcium alginate microcapsules was only effective in promoting protection at freezing temperatures, independently of the sensitivity of the strain. Storage of calcium alginate microcapsules at −80◦C indicated a protective effect upon viability of all four probiotic strains and the presence of L-cysteine·HCl in the alginate matrix improved protection upon cell viability of B. animalis BB-12 R . An increase in storage temperature of encapsulated bacteria caused an increase in rate of loss in their viability that was strain dependent. This study suggests that microencapsulation of probiotic cells in calcium alginate can be suitable for sustaining the viability of probiotics in food products that require storage below freezing temperatures, even in the absence of cryoprotectors, contributing to an increased shelf life.
- Analytical strategies for characterization and validation of functional dairy foodsPublication . Rodrigues, Dina; Rocha-Santos, Teresa A.P.; Freitas, Ana C.; Gomes, Ana M.P.; Duarte, Armando C.Functional foods (FFs) are food products to be consumed as part of a balanced diet. They provide physiological benefits or reduce the risk of chronic disease beyond basic nutritional functions. Functional foods containing probiotics and/or prebiotics have gained much interest in recent years due to their health-promoting capacity. The main objective of this review is to discuss the analytical strategies that have been used to validate FFs associated with dairy products containing probiotics and/or prebiotics. In these products, the biochemical events, carried out by enzymes of different sources (milk, bacteria, rennet) leading to the transformation of milk to diverse products (e.g., yoghurt and cheese), are glycolysis, proteolysis and lipolysis. We present the analytical methodologies used to study the microbial probiotic flora and to evaluate the biochemical transformations, the associated functionality in terms of intestinal microbiome and the safety of such FFs. We address the analytical figures of merit. We cover the advantages and the disadvantages of such analytical methodologies and comment on future applications and potential research interest within this field.