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  • Manufacturing of fermented goat milk with a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus in a controlled bioreactor
    Publication . Kongo, J. M.; Gomes, Ana M. P.; Malcata, F. Xavier
    Aims: This work was undertaken to study the feasibility and the characteristics of a fermented product made of goat milk, using a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus under controlled conditions, and to determine their survival in the fermented milk during refrigerated storage. Methods and Results: Goat milk was inoculated with Lact. acidophilus and Bif. animalis mixed starter, fermented in a glass bioreactor with controlled temperature (37 C) and anaerobiosis, and monitored for growth and acidification. The fermented milk was then stored for 10 days under refrigeration, and monitored daily for starter microflora survival and pH changes. Lact. acidophilus viable counts reached a maximum of 7Æ1 · 108 colony-forming units (CFU) ml)1, and Bif. animalis a maximum of 6Æ3 · 107 CFU ml)1 by 20 h of fermentation. During refrigerated storage, both strains exhibited a good survival, with viable numbers remaining essentially constant throughout the experiment, whereas the pH of the fermented milk dropped slightly. Conclusions: Mixed cultures of Bif. animalis and Lact. acidophilus may be used to produce fermented goat milk with high counts of both probiotic strains. Significance and Impact of the Study: Goat milk fermented with Bif. animalis and Lact. acidophilus can be manufactured as an alternative probiotic dairy product.
  • Sargassum muticum and Osmundea pinnatifida enzymatic extracts: chemical, structural, and cytotoxic characterization
    Publication . 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.
  • Antimicrobial Susceptibility of Enterococci Isolated from Terrincho Cheese
    Publication . Pimentel, L. L.; Novais, C.; Pintado, M. E.; Gomes, Ana M. P.; Coque, T.; Sousa, J. C; Malcata, F. Xavier; Peixe, L.
  • Survival of probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing in the gastrointestinal tract
    Publication . Madureira, A. R.; Pereira, C. I.; Truszkowska, K.; Gomes, Ana M. P.; Pintado, M. E.; Malcata, F. X.
    Various foods may be used to deliver probiotic bacteria into the gastrointestinal tract; one such example is Requeijão, a Portuguese whey cheese. Survival and stability of Bifidobacterium animalis strains BLC-1, Bb-12, and Bo, Lactobacillus acidophilus strains LAC-1 and Ki, L. paracasei ssp. paracasei strain LCS-1 and L. brevis strain LMG 6906 inoculated into Requeijão, when exposed to simulated gastrointestinal tract conditions, were assessed. Homogenates of inoculated whey cheese in 0.85% (w/v) sterile saline water were exposed to a solution of hydrochloric acid (pH 2.5–3.0) and pepsin (1000 unitsmL–1) at 371C, and then to 0.3% (w/v) bile salts after 60 or 120 min of acid exposure. All bacterial strains retained their initial viable cell numbers. Bifidobacterium animalis strains Bb-12 and Bo, and L. brevis strain LMG 6906 exhibited the highest viable cell numbers when exposed to bile salts, whereas the other strains had variable death rates.
  • Viability of bifidobacterium la ctis and lactobacill us acidophilus in milk: sodium chloride concentration and storage temperature
    Publication . Gomes, Ana M. P.; Teixeira, M. G. M.; Malcata, F. Xavier
    The growth and viability of Bifidobacterium lactis and Lactobacillus acidophilus per se and for both strains as a coculture were studied in reconstituted skim milk so as to assess their dependence upon temperature (5, I0 and 15C), NaCl concentration (0, 0.51 and 1.03 mol L-'), and storage time (0–8 weeks). Cultures of B. lads exhibited no signijkant loss of viability either with increasing NaCl concentration or increasing storage temperature within the ranges studied On the other hand, viability of L. acidophilus decreased with increasing temperature and increasing NaCl concentration under similar conditions. When cocultured with L. acidophilus, B. lactis was signiJicantly less tolerant to higher NaCl levels and higher temperatures than when in pure culture, although rial numbers were still above the threshold required for commercial application. Coculturing with B. lactis had no detrimental effect on viability of L. acidophilus. A mechanistic model, which considers the behavior of the pure and mixed microbial populations is described by specijic death rates which vary with temperature following Arrhenius relationships) and NaCl levels following simple inhibition kinetics) in the milk medium. Activation energies for the death rates of B. lactis and L. acidophilus were 14 kcal mot' and 9–15 kcal mol-I, respectively.
  • Effects of added Lactobacillus acidophilus and Bifidobacterium lactis probiotics on the quality characteristics of goat ricotta and their survival under simulated gastrointestinal conditions
    Publication . Meira, Quênia Gramile Silva; Magnan, Marciane; Medeiros Júnior, Francisco Cesino de; Queiroga, Rita de Cássia Ramos do Egito; Madruga, Marta Suely; Gullón, Beatriz; Gomes, Ana Maria; Pintado, Maria Manuela Estevez; Souza, Evandro Leite de
    This study evaluated the effects of incorporating the probiotics Bifidobacterium animalis subsp. lactis Bb-12 (B. lactis) or Lactobacillus acidophilus La-05 (L. acidophilus) into goat ricotta on the technological, physicochemical, physical and sensory parameters of this product during refrigerated storage, as well as the protective effects of the goat ricotta on the survival of the tested probiotics during exposure to simulated gastrointestinal conditions. Incorporating the tested probiotics did not affect the yield or syneresis of the obtained goat ricotta. The counts of L. acidophilus and B. lactis during the chosen storage period were approximately 6 log CFU/g. The ricotta samples containing a probiotic strain presented smaller and greater amounts of lactose and lactic acid, respectively, and exhibited greater hardness and lower brightness after storage compared with the samples lacking a probiotic. No differences were observed in the fatty acid profiles of the goat ricotta containing or not containing a probiotic. All of the ricotta samples were described as a soft cheese with a homogeneous texture; however, the goat ricotta cheeses containing L acidophilus or B. lactis were described as having a more acidic flavor. At the end of a challenge using experimental human digestive conditions, the counts of each of the tested probiotic strains were approximately 6 log CFU/g if it had been incorporated into goat ricotta. These results demonstrated the feasibility of incorporating L. acidophilus or B. lactis into goat ricotta because these probiotics did not negatively affect the quality characteristics of this product and suggested that goat ricotta is an efficacious food matrix for maintaining the viability of these probiotics during storage and under the stressful conditions imposed by the human gastrointestinal tract. (C) 2015 Elsevier Ltd. All rights reserved.
  • Antimicrobial activity and lipid profile of seaweed extracts from the North Portuguese coast
    Publication . Mendes, M.; Pereira, R.; Pinto, I. Sousa; Carvalho, A. P.; Gomes, A. M.
    Seaweeds or marine macroalgae are a potential renewable resource in the marine environment and can represent a source of new natural compounds for human nutrition. This experimental work focused on the selection of a suitable extraction method, using Gracilaria vermiculophylla as a model, followed by the characterization of the antimicrobial activity of different solvent extracts of Gracilaria vermiculophylla, Porphyra dioica and Chondrus crispus, both from wild and from an integrated multi-trophic aquaculture system (IMTA), collected in the north of Portugal. Additionally, the fatty acid profile of the seaweed extracts was also investigated. Extraction tests included the definition of the physical state of the seaweed, temperature and type of solvents to be used during the process. Results revealed that test organisms (Gram negative and Gram positive bacteria as well as one yeast species) were more sensitive to extracts obtained with dried algae, processed continuously at higher temperatures. Results from antimicrobial activity of wild and IMTA seaweed extracts showed stronger antimicrobial activity in extracts of ethyl acetate when compared with those from methanol and diethyl ether; furthermore, among the type of microorganisms tested, there was tendency for inhibition of the Gram positive ones. In general, there appears to be a higher antimicrobial activity for the microorganisms under study in extracts obtained from aquaculture species, when compared with the wild ones. The fatty acid profile of ethyl acetate extracts revealed a predominance of saturated fatty acids (SFA), especially palmitic acid (16:0), followed by polyunsaturated fatty acids (PUFA) and monounsaturated fatty acids (MUFA). A higher content of fatty acids in aquaculture extracts of Gracilaria vermiculophylla and Porphyra dioica was also observed.
  • Antioxidant properties of sterilized yacon (Smallanthus sonchifolius) tuber flour
    Publication . Sousa, Sérgio; Pinto, Jorge; Rodrigues, César; Gião, Maria; Pereira, Cláudia; Tavaria, Freni; Malcata, F. Xavier; Gomes, Ana; Pacheco, M. T. Bertoldo; Pintado, Manuela
    The objective of this research work was to investigate the antioxidant properties of sterilized yacon tuber flour. The results revealed for the first time the high antioxidant activity of sterilized yacon flour. The best extract obtained by boiling 8.9% (w/v) of yacon flour in deionised water for 10 min exhibited a total antioxidant capacity of 222 ± 2 mg (ascorbic acid equivalent)/100 g DW and a total polyphenol content of 275 ± 3 mg (gallic acid equivalent)/100 g DW associated to the presence of four main phenolic compounds: chlorogenic acid, caffeic acid, coumaric acid and protocatechuic acid, as well as the amino acid tryptophan. The most abundant was chlorogenic acid, followed by caffeic acid. Biological assays revealed that the extract had indeed antioxidant protection, and no pro-oxidant activity. In conclusion, sterilized yacon tuber flour has the potential to be used in the food industry as a food ingredient to produce functional food products.
  • Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines
    Publication . Pinho, Olivia; Pintado, Ana I. E.; Gomes, Ana M. P.; Pintado, M. E.; Malcata, F. Xavier; Ferreira, Isabel M. P. L. V. O.
    Changes in the microbiological, physicochemical, and biochemical characteristics of Terrincho cheese as represented by native microflora, pH, water activity, soluble nitrogen fractions, free amino acids, and biogenic amines (e.g., ethylamine, dimethylamine, tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, cystamine, and spermine) during ripening were monitored. Terrincho is a traditional Portuguese cheese manufactured from raw ewe's milk. The main groups of microorganisms (lactococci, lactobacilli, enterococci, pseudomonads, staphylococci, coliforms, yeasts, and molds) were determined following conventional microbiological procedures. Free amino acids and biogenic amines were determined by reverse-phase high-performance liquid chromatography, following extraction from the cheese matrix and derivatization with dabsyl chloride. The total content of free amino acids ranged from 1,730 mg/kg of dry matter at the beginning of the ripening stage to 5,180 mg/kg of dry matter by day 60 of ripening; such an increase was highly correlated with the increase of watersoluble nitrogen in total nitrogen, 12% trichloroacetic acid–soluble nitrogen in total nitrogen, and 5% phosphotungstic acid– soluble nitrogen in total nitrogen throughout ripening. Histamine was consistently present at very low levels, whereas putrescine, cadaverine, and tryptamine were the dominant biogenic amines and increased in concentration during ripening. Ethylamine, tryptamine, phenylethylamine, and cystamine reached maxima by 30 days of ripening and decreased thereafter. Significant correlations between amino acid precursors and corresponding biogenic amines, as well as between biogenic amines and microbial viable numbers, were observed.