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Cardoso Freitas Lopes de Freitas, Ana Cristina
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- Biodegration of microplastics by marine fungiPublication . Silva, A.A.; Bastos, A.S.V.; Paço, A.; Costa, J.P.; Santos, P.; Duarte, K.; Rocha-Santos, T.; Freitas, A. C.
- 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.
- Effects of different ripening procedures on the final characteristicsPublication . Freitas, A. C.; Malcata, F. X.Picante da Beira Baixa (or Picante) cheese is a hard, piquant, salted traditional cheese manufactured in Portugal from raw sheep’s and goat’s milks. The purpose of this work was to quantitatively assess the influence of various ripening procedures on the final characteristics of Picante cheese. Two alternative ripening protocols were considered, the traditional one and another with controlled environmental conditions via use of maturation chambers set at different preselected temperatures. The experimental cheeses were characterised in terms of microbiological, physicochemical, biochemical, sensorial and textural properties. Ripening time and temperature were statistically significant parameters for all microflora. The two ripening methods led to statistically significant differences in all physicochemical and biochemical parameters, especially the moisture content and the soluble nitrogen fractions (i.e. water loss was slower and proteolysis was faster in cheeses ripened via the traditional method). Differences in microbiological, physicochemical and biochemical properties were probables implicated in differences in textural and sensorial properties, especially cheese hardness and flavour. It was concluded that the standard ripening method was closest to the traditional one in terms of final cheese characteristics when the ripening temperature was above 11.5 7C.
- Formulation strategies for enhancing growth of akkermansia muciniphila and its survival through lyophilisation and storage at air ambientPublication . Freitas, A.; Almeida, D.; Machado, D.; Mendo, S.; Gomes, A.Gut bacterium Akkermansia muciniphila has been recognized as a new potential probiotic due to promising outcomes in the prevention and treatment of several human diseases. However, despite its multiple beneficial effects, its viability is compromised by the presence of oxygen. This has so far precluded its application namely in the development of nutraceutical/therapeutic formulations. In order to enhance growth of A. muciniphila and its survival through lyophilisation and storage at air ambient, formulation strategies are presented based on use of prebiotic compounds, antioxidants and cryoprotectants. According to our best knowledge, there is no information regarding lyophilisation procedures involving A. muciniphila as well as its subsequent storage at ambient air.
- 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.
- Biodegradation of polyethylene microplastics by the marine fungus zalerion maritimumPublication . Paço, Ana; Duarte, Kátia; Costa, João P. da; Santos, Patrícia S.M.; Pereira, R.; Pereira, M. E.; Freitas, Ana C.; Duarte, Armando C.; Rocha-Santos, Teresa A. P.Plastic yearly production has surpassed the 300milliontons mark and recycling has all but failed in constituting a viable solution for the disposal of plastic waste. As these materials continue to accumulate in the environment, namely, in rivers and oceans, in the form of macro-, meso-, micro- and nanoplastics, it becomes of the utmost urgency to find new ways to curtail this environmental threat. Multiple efforts have been made to identify and isolate microorganisms capable of utilizing synthetic polymers and recent results point towards the viability of a solution for this problem based on the biodegradation of plastics resorting to selected microbial strains. Herein, the response of the fungus Zalerion maritimum to different times of exposition to polyethylene (PE) pellets, in a minimum growth medium, was evaluated, based on the quantified mass differences in both the fungus and the microplastic pellets used. Additionally, molecular changes were assessed through attenuated total reflectance Fourier transform Infrared Spectroscopy (FTIR-ATR) and Nuclear Magnetic Resonance (NMR). Results showed that, under the tested conditions, Z. maritimum is capable of utilizing PE, resulting in the decrease, in both mass and size, of the pellets. These results indicate that this naturally occurring fungus may actively contribute to the biodegradation of microplastics, requiring minimum nutrients.
- Role of dominant microflora of Picante cheese on proteolysis and lipolysisPublication . Freitas, A. Cristina; Pintado, Ana I. E.; Pintado, Manuela; Malcata, F. XavierFour species of bacteria (Enterococcus faecium and E. faecalis, Lactobacillus plantarum and Lb. paracasei) and three species of yeasts (Debaryomyces hansenii, Yarrowia lipolytica and Cryptococcus laurentii) isolated from Picante cheese were assayed for proteolytic and lipolytic activities. The milk type (caprine or ovine), the ripening time (0–65 d) and the concentration of NaCl (0–14% (w/v)) have been studied in terms of their effects upon in vitro curdled milk. Proteolytic and peptidolytic activities were demonstrated to be high for Y. lipolytica, and at much lower levels for the other strains. Milk type, ripening time and content of NaCl appeared to be statistically significant processing factors in terms of proteolysis. Clear lipolytic activity was detected for Y. lipolytica, but release of free fatty acids to lesser extents was observed for the other strains under study. Ripening time was statistically significant for lipolysis but milk type was not. Lipolytic activities were strongly affected by NaCl content and the extent of fat hydrolysis was affected by the increase of NaCl from 0 to 7% (w/v) more than by change from 7 to 14% (w/v). In view of the experimental results, a mixed-strain starter for Picante cheese including Lb. plantarum, E. faecium (or E. faecalis) and D. hansenii (and/or Y. lipolytica) is of potential interest.
- 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%).
- Development of a non-dairy probiotic fermented rice food with bifidobacterium animalis and streptococcus thermophilusPublication . Gomes, A; Rodrigues, D.; Baltazar, B.; Sousa, S.; Vasconcelos, Marta; Pimenta, Ana; Freitas, A.
- Incorporation and survival of probiotic bacteria in whey cheese matricesPublication . Madureira, Ana R.; Gião, Maria S.; Pintado, Manuela E.; Gomes, Ana M. P.; Freitas, A. Cristina; Malcata, F. XavierThe viabilities of probiotic strains of Lactobacillus acidophilus, Lactobacillus paracasei, Bifidobacterium animalis, and Lactobacillus brevis were studied following incorporation in a whey cheese matrix. Experimental production of plain, as well as sugar-added or salt-added whey cheeses, was based on the traditional manufacture protocol of Requeijäo, a Portuguese whey cheese that essentially results from protein denaturation via heating of whey at about 85°C. After inoculation, the experimental whey cheeses were incubated at 7°C for 28 d. Our results have shown that all strains considered were able to maintain (or even increase) their initial viable numbers; L. paracasei ssp. paracasei strain LCS-1 and L. acidophilus strain Ki exhibited the highest cell viability in plain Requeijão by the end of the storage period—an increase of ca. 2 log cycles in their viable numbers was actually recorded. Among the parameters studied, bacterial species and matrix nature had the most important effect upon viable counts, whereas time of storage was the least important.