Comparing Different Packaging Conditions on Quality Stability of High-Pressure Treated Serra da Estrela Cheeses during Cold Storage.

Serra da Estrela cheese with a Protected Designation of Origin (PDO) is a traditional cheese that is wrapped in paper without vacuum. High-pressure processing (HPP), which requires vacuum packaging of the cheese, has been used for its cold pasteurization to overcome safety issues. In this study, two packaging systems were studied: non-vacuum greaseproof paper wrapping package and vacuum packaging in plastic film. Lactococci, lactobacilli, enterococci, and total mesophiles reached ca. 8 log cfu g-1 and 4-6 log cfu g-1 in control (unpasteurized) and HPP-treated cheeses, respectively, with no significant differences between packaging systems. Spoilage microorganisms' viable cell numbers were reduced to <3 log cfu g-1 (quantification limit) in HPP-treated cheeses, independently of the packaging system. Yeasts and molds reached >5 log cfu g-1 in non-vacuum paper-wrapped cheeses. A vacuum-packaging system enabled better control of cheese proteolysis, which was revealed to be closer to that of the original control cheese values at the end of the 10-month storage period. In addition, cheese stored under vacuum film packaging became harder than non-vacuum paper-wrapped cheeses at each time point. Overall, conventional non-vacuum paper wrapping is adequate for short storage periods (<3 months), but for long periods vacuum packaging in plastic film is preferable.

Optimization of Raw Ewes' Milk High-Pressure Pre-Treatment for Improved Production of Raw Milk Cheese.

Serra da Estrela protected designation of origin (PDO) cheese is manufactured with raw milk from Bordaleira and/or Churra Mondegueira da Serra da Estrela sheep breeds. Several socio-environmental shortcomings have reduced production capacity; hence, treatments that may contribute to its efficient transformation into cheese are welcome. High-pressure processing (HPP) milk pre-treatment may contribute to a cheese yield increment, yet optimization of processing conditions is warranted. An initial wide-scope screening experiment allowed for pinpointing pressure intensity, holding time under pressure and time after HPP as the most important factors influencing curd yield. Based on this, a more targeted screening experiment allowed for selecting the range of experimental conditions to be used for an experimental design study that revealed an HPP treatment at 121 MPa for 30 min as the optimum for milk processing to improve curd yield (>9%) and effectively maintain the beneficial cheese microbiota; the optimum was validated in a final experimental framework.

The Combined Effect of Pressure and Temperature on Kefir Production-A Case Study of Food Fermentation in Unconventional Conditions.

Food fermentation under pressure has been studied in recent years as a way to produce foods with novel properties. The purpose of this work was to study kefir production under pressure (7-50 MPa) at different temperatures (17-32 °C), as a case study of unconventional food fermentation. The fermentation time to produce kefir was similar at all temperatures (17, 25, and 32 °C) up to 15 MPa, compared to atmospheric pressure. At 50 MPa, the fermentation rate was slower, but the difference was reduced as temperature increased. During fermentation, lactic and acetic acid concentration increased while citric acid decreased. The positive activation volumes (Va) obtained indicate that pressure decreased the fermentation rate, while the temperature rise led to the attenuation of the pressure effect (lower Va). On the other hand, higher activation energies (Ea) were observed with pressure increase, indicating that fermentation became more sensitive to temperature. The condition that resulted in a faster fermentation, higher titratable acidity, and higher concentration of lactic acid was 15 MPa/32 °C. As the authors are aware, this is the second work in the literature to study the combined effect of pressure and temperature on a fermentative process.

Efficiency of purification methods on the recovery of exopolysaccharides from fermentation media.

De-Man Rogosa and Sharpe (MRS) is a complex medium commonly used to obtain exopolysaccharides (EPS) from lactic acid bacteria. However, the various nutrients (carbon and nitrogen sources) of media and supplements added to enhance the bacterial growth and EPS production, may interfere with the purification of the extracts resulting in an over-estimation of the EPS and erroneous structural assignments. The efficiency of trichloroacetic acid (TCA)/pronase and 5-sulfosalicylic acid - SSA methods was evaluated. In addition, the interference of the major MRS broth components as well as lactose was evaluated using xanthan gum as model control EPS. It was concluded that MRS medium is a major source of interfering compounds in the quantification of EPS, mainly glucose-rich material and to a lesser extent mannoproteins from yeast extract. This effect was found to be potentiated by the presence of lactose. TCA/pronase method was shown to be more efficient in eliminating interferents.

Sargassum muticum and Osmundea pinnatifida Enzymatic Extracts: Chemical, Structural, and Cytotoxic Characterization.

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 and structural characterization of Pholiota nameko extracts with biological properties.

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 ß-glycosidic structures such as glucans and glucan-protein complexes are among the polysaccharides found in both extracts.

Evidences and perspectives in the utilization of CLNA isomers as bioactive compounds in foods.

Conjugated alpha linolenic acid (CLNA) isomers are promising lipids owing to their similarities with conjugated linoleic acid (CLA) but exerting their bioactivity at lower doses; some isomers also belong to omega 3 family. This review aims to summarize the state of the art about the utilization of CLNA as a functional ingredient. Indeed, in vitro and in vivo studies reported that CLNA exerted anticancer, anti-inflammatory, anti-obese, and antioxidant activities. However, CLNA has not been tested in humans. These compounds are naturally present in meat and milk fat from ruminants but the highest concentrations are found in vegetable oils. Their incorporation in foodstuffs is one of the most effective strategies to elaborate CLNA-enriched products together with the microbiological production. Lactobacilli, propionibacteria, and bifidobacteria strains have been assayed to produce CLNA isomers but at the current moment there are not high CLNA concentration products elaborated using these strains. Furthermore, it is known that CLNA isomers are highly prone to oxidation when compared with linoleic acid and CLA, but the possible effects of elaboration and storage on high CLNA productsare unknown.The utilization of CLNA as a functional compound still remains a challenge and requires more research to address all of its technological and bioactivity aspects.

Influence of the addition of Lactobacillus acidophilus La-05, Bifidobacterium animalis subsp. lactis Bb-12 and inulin on the technological, physicochemical, microbiological and sensory features of creamy goat cheese.

The effects of the addition of Lactobacillus acidophilus LA-05, Bifidobacterium animalis subsp. lactis BB-12 and inulin on the quality characteristics of creamy goat cheese during refrigerated storage were evaluated. The manufactured cheeses included the addition of starter culture (Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris - R-704) (CC); starter culture, L. acidophilus LA-05 and inulin (CLA); starter culture, B. lactis BB-12 and inulin (CBB); or starter culture, L. acidophilus LA-05, B. lactis BB-12 and inulin (CLB). In the synbiotic cheeses (CLA, CBB and CLB), the counts of L. acidophilus LA-05 and B. lactis BB-12 were greater than 6log CFU g-1, the amount of inulin was greater than 6 g per 100 g, and the firmness was reduced. The cheeses evaluated had high brightness values (L*), with a predominance of yellow (b*). CC had higher contents of proteins, lipids and minerals compared to the other cheeses. There was a decrease in the amount of short-chain fatty acids (SCFAs) and an increase of medium-chain (MCFAs) and long-chain fatty acids (LCFAs) in the synbiotic cheeses compared to CC. The amount of conjugated linoleic acid increased in CLA, CBB and CLB. The highest depth of proteolysis and the greatest changes in the release of free amino acids were found in CLB. The addition of inulin and probiotics, alone or in co-culture, did not affect the cheese acceptance. Inulin and probiotics can be used together for the production of creamy goat cheese without negatively affecting the general quality characteristics of the product, and to add value because of its synbiotic potential.

Chemical composition of red, brown and green macroalgae from Buarcos bay in Central West Coast of Portugal.

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 Portugal.

Seaweeds 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/glyoph extract) and total phenolics (261 ± 37 µgcatechol equiv/glyoph extract) contents, whereas higher sugars (78 ± 14 mgglucose equiv/glyoph extract) including sulfated polysaccharide (44 ± 8 mgNa2SO4 acid/glyoph 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 α-glucosidase (38-49%).

Strategies based on silica monoliths for removing pollutants from wastewater effluents: a review.

Silica monoliths have been used for more than half a century in a wide variety of applications, such as stationary phases for microextraction fibers, capillary columns for chromatography, in the encapsulation of biomolecules for affinity chromatography and for microfluidic or microarray chips and, more recently, and less well known for wastewater treatment. The main objective of this review article is to specifically overview the strategies that use silica monoliths for the removal of chemical pollutants from wastewater effluents or prepared solutions. The discussion of advantages and drawbacks of such strategies will be supported with the main studies carried out so far which have been performed in laboratory environment only. The application and potential research interest in several strategies using composites and biocomposites based silica monoliths as cleaning systems are also discussed.

Marine biotechnology advances towards applications in new functional foods.

The marine ecosystem is still an untapped reservoir of biologically active compounds, which have considerable potential to supply food ingredients towards development of new functional foods. With the goal of increasing the availability and chemical diversity of functional marine ingredients, much research has been developed using biotechnological tools to discover and produce new compounds. This review summarizes the advances in biotechnological tools for production of functional ingredients, including enzymes, for the food industry. Tools involving biotechnological processes (bioreactors, fermentations, bioprocessing) and those involving genetic research designated as molecular biotechnology are discussed highlighting how they can be used in the controlled manipulation and utilization of marine organisms as sources of food ingredients, as well as discussing the most relevant shortcomings towards applications in new functional foods.

Incorporation of probiotic bacteria in whey cheese: decreasing the risk of microbial contamination.

For dairy products that are consumed fresh, contamination by spoilage microorganisms and pathogens from the environment is a major concern. Contamination has been associated with a number of outbreaks of foodborne illnesses; however, consistent data pertaining to the microbial safety of whey cheeses specifically have not been reported. Hence, the goals of this research effort were (i) to manufacture a probiotic whey cheese with Bifidobacterium animalis and Lactobacillus casei and (ii) to assess the antimicrobial activity of these probiotics against a set of foodborne pathogens (Listeria innocua, Salmonella Enteritidis, and Staphylococcus aureus) and food spoilage microorganisms (Pseudomonas aeruginosa and Escherichia coli). Three ranges of these microbial contaminants were used for inoculation of cheeses: 10(3) to 10(4), 10(4) to 10(6), and 10(6) to 10(8) CFU/g. Inoculation in plain culture medium served as a control. The inhibition produced by the probiotics was calculated, and the major effect was found to be bacteriostatic. In specific cases, full inhibition was observed, i.e., by B. animalis against P. aeruginosa and by L. casei against Salmonella Enteritidis and L. innocua. Conversely, the least inhibition was detected for L. casei against P. aeruginosa. Our results suggest that use of these probiotic strains can extend the shelf life of whey cheeses and make them safer by delaying or preventing growth of common contaminant bacteria.

Influence of bacterial dynamics upon the final characteristics of model Portuguese traditional cheeses.

The microbiological profile in raw milk cheeses is typically characterized by a multitude of microbial groups, with interactions among them throughout ripening that are not fully understood to date. Incidence of undesired microorganisms in raw cheesemaking milk, as is the case of either spoilage or even pathogenic ones, is a common trait in Portuguese traditional cheeses. Hence, they will likely contribute to the physicochemical changes occurring therein and, consequently, to the characteristics of the final product. In order to gain insight into their role, model cheese systems, manufactured as far as possible according to artisanal practices (except that the initial microbial load and biodiversity were controlled), were experimentally tested. Single contaminants, or a consortium thereof, were inoculated at two levels in sterilized raw ewe's milk, and duly combined with inocula containing one or two lactic acid bacteria normally found in those traditional cheeses. The physicochemical composition, organic acid profile, and evolution of both protein breakdown and rheology were monitored throughout a 60 d-ripening period. Modifications brought about within the cheese matrix as a result of microbial metabolism, especially those arising from the interaction between lactic acid bacteria and unwanted microorganisms, included the enhanced release of peptides and free amino acids, which in turn led to higher viscoelastic moduli. The final model cheeses could be well discriminated, based on the impact of the various inocula considered upon the levels of organic acids. Conversely, proteolysis and viscoelastic properties appeared to be essentially independent of the initial microflora.

Bacterial dynamics in model cheese systems, aiming at safety and quality of Portuguese-style traditional ewe's cheeses.

An experiment using model ewe's milk cheeses was designed to characterize microbial interactions that arise in actual raw milk cheese environments. These model cheeses were manufactured according to Portuguese artisanal practices, except that the microbial load and biodiversity were fully controlled: single potential pathogens and spoilage bacteria, or a combination thereof, were combined at various initial inoculum levels in sterilized raw ewe's milk with several lactic acid bacteria (LAB) normally found in traditional cheeses. Viable microbial counts were monitored throughout a 60-day ripening period. Two alternative mathematical approaches were used to fit the experimental data generated in terms of population dynamics: percent of inhibition and D-values. These were able to explain the complex competitive interactions between the contaminant microorganisms and the LAB adventitious populations. In general, the tested LAB were less able to inhibit contaminants present in combination and in higher concentrations. Lactococcus lactis, with its strong acidifying potential, was the most effective factor in controlling the unwanted bacterial population, especially single Staphylococcus aureus. The two lactobacilli studied, especially Lactobacillus brevis, were shown to be less effective; Escherichia coli and Listeria innocua were the contaminants least inhibited by the LAB.

Proteolysis in model Portuguese cheeses: Effects of rennet and starter culture.

To shed further light onto the mechanisms of proteolysis that prevail throughout ripening of Portuguese cheeses, model cheeses were manufactured from bovine milk, following as much as possible traditional manufacture practices - using either animal or plant rennet. The individual role upon proteolysis of two (wild) strains of lactic acid bacteria - viz. Lactococcus lactis and Lactobacillus brevis, which are normally found to high viable numbers in said cheeses, was also considered, either as single or mixed cultures. Our experimental results confirmed the influence of rennet on the proteolysis extent, but not on proteolysis depth. On the other hand, the aforementioned strains clearly improved release of medium- and small-sized peptides, and contributed as well to the free amino acid pool in cheese.

Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines.

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 water-soluble 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.