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Tomato by-products from processing industries have a higher potential to be reused as a source of bioactive compounds. Reliable national data on tomato by-products and physicochemical characterisation that will inform and find effective planning on tomato waste management in Portugal is absent. To help obtain this knowledge, selected Portugal companies were recruited to obtain representative samples of by-products generation, and physicochemical composition was evaluated. Furthermore, an environmental-friendly method (the ohmic heating (OH) method, which allows the recovery of bioactive compounds in absence of hazardous reagents) was also used and compared with conventional methods to explore new safe value-added ingredients. Total antioxidant capacity and total and individual phenolic compounds were also evaluated by spectrophotometric and high-performance liquid chromatography (HPLC), respectively. Tomato processing by-products have revealed a higher potential since both collected samples from companies were rich in protein (between 16.3 to 19.4 g/100 g DW, with fibre content ranging from 57.8 to 59.0 g/100 g DW). In addition, these samples contain 17.0 g/100 g of fatty acids (mainly polyunsaturated, monounsaturated and saturated, such as linoleic, oleic, and palmitic acid, respectively). Also, they present mainly chlorogenic acid and rutin as phenolic compounds. After understanding its composition, the OH was applied to determine added-value solutions to tomato by-products. With extractions, two types of fractions were obtained, namely liquid fraction rich in phenols, free sugars, and carotenoids and a solid fraction rich in fibre bound to phenols and carotenoids. This treatment has been shown to have the ability to preserve carotenoids, such as lycopene relative to conventional methods. Nevertheless, new molecules were identified by LC-ESI-UHR-OqTOF-MS analysis, such as phene-di-hexane and N-acethyl-D-tryptophan. According to the results, the OH boosts the potential of tomato by-products and can be directly introduced into the process, contributing to the circular economy and zero by-products. [ABSTRACT FROM AUTHOR]

The by-products of the winemaking process can represent chances for the development of new products. This study focused on the "zero waste" strategy development for by-products generated within winemaking from white and red grape varieties cultivated in the north of Portugal. The phytochemical properties of by-products were identified and characterized. Ohmic heating (OH) as a green extraction method was also applied to grape pomace due to their unknown effects on centesimal and phytochemical compositions. Both protein and carbohydrates were shown to be higher in grape bagasse than in stems. Additionally, red bagasse is richer in bioactive compounds (BC) than white bagasse. The sugar content was 21.91 and 11.01 g/100 g of DW in red and white grape bagasse, respectively. The amount of protein was 12.46 g/100 g of DW for red grape bagasse and 13.18 g/100 g of DW for white. Regarding the extraction methods, two fractions were obtained, a liquid fraction and solid (the remainder after the methodology application). OH presented a higher antioxidant capacity than a conventional (CONV) method. In addition, both extracts presented similar contents of anthocyanins, e.g., delphinidin-3-O-glucoside, petunidin-3-O-glucoside, and peonidin-3-O-glucoside. The solid fraction presented higher amounts of protein and phenols bound to fiber than CONV, which allows its use as a functional ingredient. In conclusion, OH can be an alternative extraction method compared with CONV methods, avoiding non-food grade solvents, thus contributing to circular economy implementation. [ABSTRACT FROM AUTHOR]

Carob fruit is native to the Mediterranean region and produced mainly in Portugal, Italy, Morocco and Turkey. The production of the carob fruit in Portugal is highly extensive and sustainable. Currently, carob flour (CF) production is mainly achieved after pulp separation, despite it having been demonstrated that the seeds improve the extraction efficiency of bioactive compounds such as polyphenols, promoting human health. This study aimed to produce an integral CF through an innovative process and assess its physicochemical and bioactive properties at different particle sizes throughout simulated gastrointestinal tract (GIT) digestion. The sugar content profile obtained throughout GIT digestion indicated that sucrose, the sugar present at the highest concentration in undigested CF, was digested and broken down into simple sugars, namely glucose and fructose. The total phenolic content (TPC) and antioxidant activity obtained for the ≤100 µm fraction were in accordance and gastric digestion promoted an increase in the TPC value compared to the undigested sample. The >100 µm fractions displayed a distinct profile from the ≤100 µm fraction. This study showed that the particle size affects the sugar, antioxidant and total phenolic content of CFs and also their gastrointestinal tract digestion. The ≤100 µm fraction demonstrated the most suitable profile as a functional food ingredient. [ABSTRACT FROM AUTHOR]

The cleaning or removal of manganese stains on Cultural Heritage has not been much tested or successful so far. The aim of this article was to assess a new green cleaning gel for Mn-rich black-blue stains on different substrates. The black-blue stains were characterized at optical and chemical level through colour-related data, optical microscope, FTIR, XRF and XPS. Mn-stained granite found on historical churches at Vila Real (North of Portugal) and glass jars of Leclanché cells, belonging to the ISEP's Museum (Portugal) collection, were the ideal case studies to test the efficiency of chitosan: thiourea dioxide (TD) cleaning gel. TD proved to be the best candidate to reduce insoluble manganese oxides, over Hydroxylamine Hydrochloride and Hydroxymethanesulfinic Acid. Cleaning assays performed on stained granite samples collected at a historical quarry and in situ application on stained granite churches allowed removal of the stains to a satisfactory level. Similar results were obtained on stained glass jars. [ABSTRACT FROM AUTHOR]

The 'Bravo de Esmolfe' apple (BE) is rich in antioxidants and represents one of the most important traditional cultivars in Portugal. A large amount of non-compliant BE, with no market value, it is generated every year. Thus, an effort to find value-added solutions is of utmost importance. For the first time, green extraction (microwave-assisted-extraction (MAE) and ultrasound-assisted-extraction (UAE)) were compared against conventional extraction (with (CET) and without temperature (CE)) to select the most efficient process to produce a natural antioxidant extract from non-compliant BE. The results showed that MAE and CET are good methodologies for developing a BE antioxidant extract. However, MAE is more sustainable than CET, requiring less time to reach high temperatures. The MAE-extract showed great antioxidant activity (14.80 ± 0.70 mg TE/g DE by ORAC, 3.92 ± 0.25 mg AAE/g DE by ABTS, and 5.11 ± 0.13 mg TE/g DE by DPPH). This extract revealed high amount of chlorogenic acid (0.48 ± 0.07 mg/g DE), (−)-epicatechin (0.30 ± 0.02 mg/g DE) and phloridzin (0.13 ± 0.01 mg/g DE). This study shows that non-compliant BE is a useful source of antioxidants, being a sustainable way for the recovery of value-added compounds from the rejected fruit in line with sustainable and circular bioeconomy principles. [ABSTRACT FROM AUTHOR]

Wine and beer consumption are an integral part of European culture: Southern Europe is associated with wine and Northern Europe is associated with beer. When consumed in moderation, these alcoholic beverages can be part of a balanced and healthy diet. In the 1990s, non-alcoholic beer (NAB), which has no cultural roots, became available in the market. This review identifies determinants for consumption of wine, beer, and NAB, using data on consumption patterns from Portugal and the Netherlands. Since the 1960s the image of Portugal as a wine country declined, whereas the image of the Netherlands as a beer country remained stable. In each country beer is now the most consumed alcoholic beverage and is mainly a men's beverage, whereas wine is the second most consumed and is consumed by both genders. Cultural differences define Portuguese as "outdoors, everyday drinkers", within a meal context, and Dutch as "at home, weekend drinkers." Wine is perceived as the healthiest beverage, followed by NAB, and regular beer. Motivation for consumption is related to context: wine for special occasions, beer for informal occasions, and NAB for occasions when alcohol is not convenient. Moderate wine and beer consumption seems to be surrounded by positive emotions. This review is relevant for public health, for industry market strategies, and identifies opportunities of future research on drinking behaviour. [ABSTRACT FROM AUTHOR]

Abstract: The main purpose of this study was to isolate and briefly characterize proteolytic bacteria from a poorly known habitat – raw wool. Fleece samples were accordingly collected from Merino raw wool – a Portuguese ewe breed, at three distinct areas of their body, from animals exhibiting no symptoms or signs of abnormalities; they were then subjected to enumeration and isolation of a total of 158 bacterial strains. Said isolates were screened for protease activity, using the spot technique, on Calcium Caseinate Agar containing 1% (w/v) skim milk. The 36 isolates displaying the highest protease activity underwent a more refined assessment of enzymatic performance – by examining their cell-free supernatant extracts, using casein as substrate. Two Bacillus isolates were eventually chosen owing to their highest proteolytic activities (24.6 and 15.9U/mL), and identified using molecular biology tools. [Copyright &y& Elsevier]

This article discloses a rare and outstanding type of Mn-rich black-blue patina found on mural painting and granite ashlars located in the church of Sta. Marinha, north of Portugal, and conjectures the phenomenon associated to the appearance of such patinas in different surface materials. This Mn-patina reported on mural painting and their origin is probably assigned to manganese leaching from building materials (i.e., granite and phyllites). Stained mural painting and granite examined by XPS and SEM have showed patinas enriched with manganese (IV) oxide, potentially catalysed by a microbiota, like fungi, observed in SEM micrographs. The pigments used to depict mural painting and groundwater were also analysed by micro-Raman and ICP-MS, respectively, indicating that they are unlikely manganese sources. Unstained building materials, such as granite ashlars, historic joints, mortars and phyllite rocks, were also analysed by ICP-MS showing that historic joints and mortars present significant concentrations of manganese, possibly associated to their absorbing feature. The main materials with potential to impart manganese to Mn-rich patinas are granite ashlars and phyllites. The aim of this investigation is to reveal and ascertain the hypothetical sources and the phenomenon responsible for the Mn-rich black-blue patina appearance, both on mural painting and granite ashlars. [ABSTRACT FROM AUTHOR]

In northern Portugal it was identified that some mural paintings from the 15th and 16th centuries disclose a chromatic alteration in the form of black stains possibly of microbiological origin. In the search for its cause, samples were collected and microorganisms were isolated from selected areas of the pictorial sets of three case studies that displayed dark pigmentation. Four representative fungal isolates were selected due to their pigmentation and growth characteristics – Alternaria alternata , Alternaria tenuissima , TZ 10.2.2 (possibly Pestaloptia sp.) and TZ 8.2.2 (possibly Penicillium sp.). An attempt was made to encounter an innovative environmentally innocuous and green safe alternative biocide to replace the chemical-based toxic compounds currently in use. Essential oils with confirmed antifungal potential and seldom employed in the conservation of cultural heritage and even less in mural painting (Rosmarinus officinalis , Foeniculum vulgare , Citrus lemmon , Ocimum basilicum and Salvia officinalis), were tested against commercial biocides (Benzalkonium chloride, Preventol R80, CaSoPaL® and Durolith®), to evaluate their in vitro efficacy and the determination of the minimum inhibitory concentration. Basil was able to inhibit growth on all tested strains at 1.25 μl ml−1. From the commercial biocides, Benzalkonium chloride and Preventol R80 were the ones with the best performance but needed higher concentrations than basil to achieve the same result enabling this essential oil a useful alternative to commercial biocides. The literature regarding the use of essential oils in mural paintings is, apart from two known authors, practically non-existent making it a novel approach. Image 1 • 15th-16th century fresco wall paintings are located in N. Portugal granite churches. • Black stains chromatic alterations possibly originated by microorganisms. • Four fungal isolates selected for in vitro assays with essential oils and biocides. • Basil was the only essential oil able to inhibit growth of all fungal isolates. • BC and Preventol R80 inhibit growth of all fungi at higher concentrations than basil. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was determine (i) the chemical composition (ii) the antimicrobial activity (antibacterial and antifungal) and (iii) the antioxidant activity by means of four different antioxidant tests (DDPH, FIC, FRAP and TBARS) of the EOs of three aromatic herbs, Coriander (Coriandrum sativum), celery (Apium graveolens) and bush-basil (Ocimum minimum) widely used in Portugal. There is a great variability of the compounds presented in the three tested essential oils. Bush-basil EO had the highest total phenolic content (794.9 mg GAE/L) while coriander EO had the lower total phenolic content (52.3 mg GAE/L). Since bush-basil had the highest TPC it was expected to present a very high antioxidant profile, which was verified in 3 of the 4 assays (DPPH inhibition of 95.9%; FRAP values of 2.7 mmol Trolox/L; TBARS inhibition of 87.2%); coriander, despite the low TPC showed the highest inhibition in the FIC assay (94.1%). The bush-basil EO showed the highest antimicrobial activity, with MIC ranging between 0.6 and 5 μL/mL against bacteria and 0.04–2.5 μL/mL against yeasts. Both celery and coriander EO had a very similar antimicrobial activity against all the tested strains. The antifungal activity was higher in the bush-basil EO against Mucor racemosus and Penicillium chrysogenum since it was the only EO that showed growth inhibition on all the tested concentrations. Alternaria alternata showed great resistance against all the tested essential oils. [Copyright &y& Elsevier]

Abstract: In Portugal Atlantic codfish (Gadus morhua) is dry-salted with food-grade marine salt for 6days. During this process, codfish incorporates salt and drains away water up to 22%(w/w) of its weight, which contains important compounds, such as free amino acids, peptides and proteins. Hence, the chemical profile of such water, composed of various soluble nitrogen fractions, was thoroughly determined. Along the salting process, the ratio of water released to trichloroacetic acid-soluble nitrogen remained constant, but the proportion between water released and phosphotungstic acid-soluble nitrogen decreased because of proteolysis. The concentration of free amino acids increased from 2.68g/l to 5.41g/l within 6days, but myofibrillar proteins were found to be approximately constant, ca. 3.31g/l. Creatine, aspartic and glutamic acids, glycine, taurine and tryptophan were the dominant free amino acids, the release of which was successfully modelled. Total biogenic amines concentration in wastewater at the end of the salting process was ca. 100mg/kg. [ABSTRACT FROM AUTHOR]

Abstract: The bacteriophage P22/Salmonella Typhimurium system, as well as human erythrocytes have been used to assay for protection, against forced oxidation caused by hydrogen peroxide, brought about by several aqueous extracts of selected adventitious plants grown in Portugal. This study proved, for the first time, that the aforementioned bacteriophage-based system is a suitable method to assess the antioxidant activity of plant extracts; among the 12 plants tested, raspberry (Rubus idaeus), sage (Salvia sp.), savory (Satureja montana) and yarrow (Achillea millefolium) were found to effectively protect against oxidative damage caused by H2O2. Haemolysis was inhibited via pre-treatment with every plant extract tested, except heath at 0.1%(w/v). The two analytical methods produced different results – and for some plants, there was a dependence (either direct or inverse) of the quantitative protection effect on extract concentration, whereas for others no significant dependence was found at all. Savory yielded the most promising results, using either method. Therefore, the P22/Salmonella system can be used as a suitable in vivo assay, and human erythrocytes as a suitable in vitro assay to confirm (or not) the antioxidant capacity of plant extracts in biological matrices. [Copyright &y& Elsevier]

One Gram-stain-positive, non-motile, non-spore-forming, catalase-negative, and coccobacilli-shaped strain, designated c10Ua161M T , was isolated from a urine sample from a reproductive-age healthy woman. Comparative 16S rRNA gene sequence analysis indicated that strain c10Ua161M T belonged to the genus Lactobacillus . Phylogenetic analysis based on pheS and rpoA gene sequences strongly supported a clade encompassing strains c10Ua161M T and eight other strains from public databases, distinct from currently recognized species of the genus Lactobacillus. In silico Average Nucleotide Identity (ANI) and Genome-to-Genome Distance Calculator (GGDC), showed 87.9 and 34.3 % identity to the closest relative Lactobacillus jensenii , respectively. The major fatty acids of strain c10Ua161M T were C 18 : 1 ω 9c (65.0%), C 16 : 0 (17.8%), and summed feature 8 (10.2 %; comprising C 18 : 1 ω7 c , and/or C 18 : 1 ω6 c ). The DNA G+C content of the strains is 34.2 mol%. On the basis of data presented here, strain c10Ua161M T represents a novel species of the genus Lactobacillus , for which the name Lactobacillus mulieris sp. nov. is proposed. The type strain is c10Ua161M T (=CECT 9755 T =DSM 108704 T ).

Sea urchin population harvest in the North Atlantic coast of Portugal was characterized in terms of gonad yield, nutritional composition and important market-related traits, over one reproductive cycle (March 2016 to March 2017). Most of the quality attributes showed a seasonal variation strongly dependent on sea urchin sex. Maximum gonad yield (18%) was observed in March 2017. A single spawning event occurred between May and July. Gonads are rich sources of protein (12-18% WW) with low fat content (≤6% WW), that increase during the gametogenic stages of recovery and growing (November-December). Polyunsaturated fatty acids were the dominant class in both sexes (4.2-14.7 mg.g -1 WW), being preferentially accumulated in females. Total gonads carotenoid varied seasonally, with the highest level being observed in males during spawning season. Echinenone was the main pigment present in gonads, showing highest concentrations in males during spawning and gonad recovering. During the growing and early maturation period gonads were more reddish, yellowish and brighter, as well as more firm, irrespectively of the sex. Based on all seasonal changes affecting gonad yield and quality, the period between November and February seems the most suitable to harvest high quality gonads in the Atlantic coast of Portugal.

A Gram-positive, rod-shaped, endospore-forming Bacillus isolate, Bi.(FFUP1) (T), recovered in Portugal from a health product was subjected to a polyphasic study and compared with the type strains of Bacillus pumilus, Bacillus safensis, Bacillus altitudinis and Bacillus xiamenensis, the phenotypically and genotypically most closely related species. Acid production from cellobiose, D-glucose and D-mannose and absence of acid production from D-arabinose, erythritol, inositol, maltose, mannitol, raffinose, rhamnose, sorbitol, starch and L-tryptophan discriminated this new isolate from the type strains of the most closely related species. Additionally, a significant different protein and carbohydrate signature was evidenced by spectroscopic techniques, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and Fourier transform IR spectroscopy with attenuated total reflectance. Using a chemometric approach, the score plot generated by principal component analysis clearly delineated the isolate as a separate cluster. The quinone system for strain Bi.(FFUP1) (T) comprised predominantly menaquinone MK-7 and major polar lipids were diphosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. Strain Bi.(FFUP1) (T) showed ≥ 99% 16S rRNA gene sequence similarity to B. safensis FO-036b(T), B. pumilus (7061(T) and SAFR-032), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T). Differences in strain Bi.FFUP1 (T) gyrB and rpoB sequences in comparison with the most closely related species and DNA-DNA hybridization experiments with Bi.FFUP1 (T) and B. pumilus ATCC 7061(T), B. safensis FO-036b(T), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T) gave relatedness values of 39.6% (reciprocal 38.0%), 49.9% (reciprocal 42.9%), 61.9% (reciprocal 52.2%) and 61.7% (reciprocal 49.2%), respectively, supported the delineation of strain Bi.(FFUP1) (T) as a representative of a novel species of the genus Bacillus, for which the name Bacillus invictae sp. nov. is proposed, with strain Bi.(FFUP1) (T) ( =DSM 26896(T) =CCUG 64113(T)) as the type strain., (© 2014 IUMS.)