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- Assessment of groundwater contamination in an agricultural peri-urban area (NW Portugal): an integrated approachPublication . Barroso, M. Fátima; Ramalhosa, Maria João; Olhero, A.; Antão, M. C.; Pina, M. F.; Guimarães, L.; Teixeira, J.; Afonso, M. J.; Delerue-Matos, Cristina; Chaminé, Helder I.The excessive use of pesticides and fertilisers in agriculture has generated a decrease in groundwater and surface water quality in many regions of the EU, constituting a hazard for human health and the environment. Besides, on-site sewage disposal is an important source of groundwater contamination in urban and peri-urban areas. The assessment of groundwater vulnerability to contamination is an important tool to fulfil the demands of EU Directives. The purpose of this study is to assess the groundwater vulnerability to contamination related mainly to agricultural activities in a peri-urban area (Vila do Conde, NW Portugal). The hydrogeological framework is characterised mainly by fissured granitic basement and sedimentary cover. Water samples were collected and analysed for temperature, pH, electrical conductivity, chloride, phosphate, nitrate and nitrite. An evaluation of groundwater vulnerability to contamination was applied (GOD-S, Pesticide DRASTIC-Fm, SINTACS and SI) and the potential nitrate contamination risk was assessed, both on a hydrogeological GIS-based mapping. A principal component analysis was performed to characterised patterns of relationship among groundwater contamination, vulnerability, and the hydrogeological setting assessed. Levels of nitrate above legislation limits were detected in 75 % of the samples analysed. Alluvia units showed the highest nitrate concentrations and also the highest vulnerability and risk. Nitrate contamination is a serious problem affecting groundwater, particularly shallow aquifers, especially due to agriculture activities, livestock and cesspools. GIS-based cartography provided an accurate way to improve knowledge on water circulation models and global functioning of local aquifer systems. Finally, this study highlights the adequacy of an integrated approach, combining hydrogeochemical data, vulnerability assessments and multivariate analysis, to understand groundwater processes in peri-urban areas.
- Polycyclic aromatic hydrocarbons bioaccessibility in seafood: Culinary practices effects on dietary exposurePublication . Fogaça, Fabíola Helena dos Santos; Soares, Cristina; Oliveira, Marta; Alves, Ricardo N.; Maulvault, Ana L.; Barbosa, Vera L.; Anacleto, Patrícia; Magalhães, João Avelar; Bandarra, Narcisa M.; Ramalhosa, Maria João; Morais, Simone; Marques, AntónioThis work aimed to determine the effect of culinary practices on the contamination level and bioaccessibility of polycyclic aromatic hydrocarbons (PAHs) in seafood. The selected farmed seafood species (marine shrimp, clams and seaweed) were commercially available in Portugal. The mean concentrations of PAHs varied between 0.23 and 51.8 µg kg-1, with the lowest value being observed in raw shrimp and the highest in dried seaweed. The number of compounds detected in seaweed and clams (naphthalene, acenaphthene, fluorene, phenanthrene, benzo(b)fluoranthene and benzo(j)fluoranthene) were higher than in shrimp (fluorene and pyrene). Among the PAHs measured, fluorene was the predominant one. There was a significant interaction effect between species and culinary treatment (p < 0.05), thus boiled and dried seaweed samples presented the lowest and the highest levels of fluorene (0.13 and 1.8 µg kg-1), respectively. The daily intake of PAHs decreased with bioaccessibility, varying from 22% for benzo(k)fluoranthene (in raw clam) to 84% for phenanthrene (in steamed clam). According to the potency equivalent concentrations, screening values and bioaccessibility of PAHs, the consumption of marine shrimp, clam and seaweed is considered as safe for consumers.
- Total antioxidant capacity of plant infusions: Assessment using electrochemical DNA-based biosensor and spectrophotometric methodsPublication . Barroso, M. Fátima; Ramalhosa, Maria João; Alves, Rita C.; Dias, Alexandra; Soares, Cristina M. D.; Oliva-Teles, M. Teresa; Delerue-Matos, CristinaNine plant infusions were assessed in terms of their polyphenolic compounds and antioxidant capacity properties by performing a “battery” of assays. Bioactive compounds (total phenolic and flavonoid content) were measured using spectrophotometric procedures and the total antioxidant capacity was determined by DPPHradical dot-RSA, FRAP and DNA-based biosensor against the reactive oxygen species: HOradical dot and H2O2. Although the optical methods are based on the donation of hydrogen or electrons between antioxidants and radicals, the DNA-based biosensor intends to simulate an in vivo assay by exposing DNA layer to biological ROS and antioxidants. In general, green and black tea showed higher polyphenolic content and antioxidant capacity evaluated by the described methods. All the analysed plant beverages showed to be a good source of bioactive compounds, particularly the green and black tea, peppermint and senna.
- Potential of Portuguese vine shoot wastes as natural resources of bioactive compoundsPublication . Moreira, Manuela M.; Barroso, M. Fátima; Porto, João Vasconcellos; Ramalhosa, Maria João; Švarc-Gajić, Jaroslava; Estevinho, Letícia; Morais, Simone; Delerue-Matos, CristinaSince annually a high amount of wastes is produced in vine pruning, the aim of this study was to evaluate the potential of vine shoots from two Portuguese grape varieties (Touriga Nacional - TN and Tinta Roriz - TR) to be used as a natural source of phenolic compounds. To reach this goal, three techniques were explored, namely microwave-assisted extraction (MAE), subcritical water extraction (SWE) and conventional extraction (CE). The phenolic composition of the extracts, antioxidant and biological activities were evaluated by spectrophotometry and chromatography. MAE and SWE produced the highest concentrated extracts. TR vine shoot variety had the highest antioxidant activity and total phenolic (32.1±0.9mggallicacidequivalents/g dry sample), as well as flavonoid content (18.7±1.2mgepicatechinequivalents/g dry sample). For the first time, the biological activity of the vine shoot extracts was tested. Results demonstrated that all of them had antimicrobial potential against different bacteria and yeasts, and the ability of inhibiting α-amylase and acetylcholinesterase enzymes, with MAE TR extracts being the most efficient. HPLC analysis enabled the identification of different phenolic compounds, with gallic acid, catechin, myricetin and kaempferol-3-O-rutinoside being the main contributors to the phenolic composition. Portuguese vine shoot wastes could serve as easily accessible source of natural antioxidants for the food or pharmaceutical industries.
- Honey as an Adjuvant in the Treatment of COVID-19 Infection: A ReviewPublication . Soares, Sónia; Bornet, Mélina; Grosso, Clara; Ramalhosa, Maria João; Gouvinhas, Irene; Garcia, Juliana; Rodrigues, Francisca; Delerue-Matos, CristinaSince ancestor times, honey has been used to promote human health due to its medicinal, and nutritious properties, mainly due to bioactive compounds present, such as phenolic compounds. The emergence of COVID-19, caused by the SARS-CoV-2 virus, led to the pursuit of solutions for the treatment of symptoms and/or disease. Honey has proven to be effective against viral infections, principally due to its potential antioxidant and anti-inflammatory activities that attenuate oxidative damage induced by pathogens, and by improving the immune system. Therefore, the aim of this review is to overview the abilities of honey to attenuate different COVID-19 symptoms, highlighting the mechanisms associated with these actions and relating the with the different bioactive compounds present. A brief, detailed approach to SARS-CoV-2 mechanism of action is first overviewed to allow readers a deep understanding. Additionally, the compounds and beneficial properties of honey, and its previously application in other similar diseases, are detailed in depth. Despite the already reported efficacy of honey against different viruses and their complications, further studies are urgently needed to explain the molecular mechanisms of activity against COVID-19 and, most importantly, clinical trials enrolling COVID-19 patients.
- Chemical Characterization and In Vitro Bioactivity of Apple Bark Extracts Obtained by Subcritical WaterPublication . Švarc-Gajić, Jaroslava; Cerdà, Víctor; Delerue-Matos, Cristina; Mašković, Pavle; Clavijo, Sabrina; Suarez, Ruth; Cvetanović, Aleksandra; Ramalhosa, Maria João; Barroso, M. Fátima; Moreira, Manuela; Morais, Simone; Withouck, Hannes; Boeykens, AnnickRecently, the value of wood residues has received increasing attention due to the presence of pharmacologically active compounds. In the present study, apple bark was treated with subcritical water (SWE) with the aim of bioactivity determination and chemical analysis by GC–MS applying two different sample preparation protocols and by HPLC. Extracts were screened for their total antioxidant capacity by means of total phenolic (TPC) and flavonoid contents (TFC), ferric reducing antioxidant power (FRAP) and DPPH-radical scavenging activity (DPPH-RSA). Anti-microbial and anti-proliferative activities of the apple bark subcritical water extracts were also examined. Different lignin and cellulose degradation products were identified by GC–MS, as well as naturally present compounds. Identified compounds encompassed organic and fatty acids, alcohols, sugars, phenol, benzaldehyde, cinnamic acid, cinnamaldehyde and azulene derivatives, as well as other organic compounds. Phenolic profiles of apple bark extracts defined by HPLC–PDA analysis revealed that the main contributors were gallic acid (9.4 ± 0.5 mg/g DW) and catechin (8.3 ± 0.4 mg/g DW). The diversity of the extracts composition supports the fact that this medium is able to extract compounds of different polarities, simultaneously acting as catalyst and reactant. The characterized apple bark extracts demonstrated good antioxidant (TPC = 31.47 ± 1.86 mg GAE/g; TFC = 17.40 ± 1.89 mg EE/g; FRAP = 22.45 ± 2.22 mg AAE/g; DPPH-RSA = 22.57 ± 2.24 mg Trolox E/g DW), antimicrobial (MIC = 19.53–156.25 μg/mL) and anti-proliferative activity (IC50 = 19.88–47.44 μg/mL). The findings of this study showed that SWE can be used as an environmentally friendly and safe technique for the valorization of apple tree wood residues, applicable in food and pharmaceutical industries.
- Lipid content of frozen fish: comparison of different extraction methods and variability during freezing storagePublication . Ramalhosa, Maria João; Paíga, Paula; Morais, Simone; Alves, M. R.; Delerue-Matos, Cristina; Oliveira, M. Beatriz P. P.In this work, a microwave-assisted extraction (MAE) methodology was compared with several conventional extraction methods (Soxhlet, Bligh & Dyer, modified Bligh & Dyer, Folch, modified Folch, Hara & Radin, Roese-Gottlieb) for quantification of total lipid content of three fish species: horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), and sardine (Sardina pilchardus). The influence of species, extraction method and frozen storage time (varying from fresh to 9 months of freezing) on total lipid content was analysed in detail. The efficiencies of methods MAE, Bligh & Dyer, Folch, modified Folch and Hara & Radin were the highest and although they were not statistically different, differences existed in terms of variability, with MAE showing the highest repeatability (CV = 0.034). Roese-Gottlieb, Soxhlet, and modified Bligh & Dyer methods were very poor in terms of efficiency as well as repeatability (CV between 0.13 and 0.18).
- Seasonal and Spatial Comparison of Polycyclic Aromatic Hydrocarbons Among Decapod Shrimp from Coastal PortugalPublication . Maia, Maria Luz; Paíga, Paula; Ramalhosa, Maria João; Delerue-Matos, Cristina; Calhau, Conceição; Domingues, ValentinaPolycyclic aromatic hydrocarbons (PAHs) are ubiquitous global pollutants that are lipophilic, bioaccumulative and can be both endocrine disruptors and carcinogens. In this study, we measured 18 PAHs among decapod shrimp samples (Palaemon spp.) collected during the spring and autumn from multiple locations along the Portuguese coastal, including local markets and aquaculture farms. ΣPAH concentrations in samples ranged from 3.14 to 320.48 ng/g (ww) with the highest value in this range detected in a sample provided from an aquaculture facility. Fifteen of the 18 scanned PAHs were present in shrimp samples. However, detectable concentrations (0.07 ng/g ww) of the carcinogenic benzo-a-pyrene occurred for a single sample only and approached method detection limits. No significant differences in shrimp ΣPAH concentrations were evident between the spring and autumn seasons. Target hazard quotient estimation suggested low probability of adverse health effects to consumers through consumption of shrimp from the study collection locations.
- Development of a modified acetonitrile-based extraction procedure followed by ultra-high performance liquid chromatography–tandem mass spectrometry for the analysis of psychiatric drugs in sedimentsPublication . Santos, Lúcia H. M. L. M.; Ramalhosa, Maria João; Ferreira, Marta; Delerue-Matos, CristinaAn analytical method based on a modified Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) extraction procedure followed by ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) was developed for the analysis of psychiatric drugs in sediments. An optimized approach was applied in sample preparation by using basic acetonitrile as extraction solvent. Extraction was followed by a clean-up using dispersive solid phase extraction (dSPE) to remove matrix interfering compounds. The analytical method was validated in terms of sensitivity, linearity, recovery, intra- and inter-day precisions and method detection and quantification limits. Under optimized conditions, limits of detection ranged from 0.01 ng g−1 to 2.08 ng g−1; and recoveries between 47 and 110% with relative standard deviation (RSD) below 5%. The developed methodology was applied to sediments of two Portuguese rivers (Douro and Lima rivers) and nine out of eleven psychiatric drugs were detected in sediments at concentrations up to 26.4 ng g−1 (dry weight). To the best of our knowledge, it was the first time that the human metabolites norfluoxetine and norsertraline were detected in river sediments at levels of few nanograms per gram.
- Polycyclic aromatic hydrocarbons in commercial squids from different geographical origins: levels and risks for human consumptionPublication . Gomes, Filipa; Oliveira, Marta; Ramalhosa, Maria João; Delerue-Matos, Cristina; Morais, SimoneThe concentrations of 18 polycyclic aromatic hydrocarbons (PAHs) were determined in five commercially valuable squid species from different geographical origins (Atlantic, Indic and Pacific Oceans). Out of the 18 quantified PAHs (the 16 PAHs considered by US EPA as priority pollutants, dibenzo(a,l)pyrene and benzo(j)fluoranthene) only dibenz(a,h)anthracene was not detected. The total concentrations of PAHs varied by a factor of more than 100-fold, from 0.22 (Loligo gahi) to 60.9 lg/kg ww (Loligo reynaudii). Intraand inter-specific variability of PAH levels was statistically assessed. Nine carcinogenic (probable/possible) PAHs accounted for 1% (L. reynaudii) to 26% (Loligo opalescens) of the total PAHs content being the main contributors naphthalene (in Loligo duvaucelii, L. reynaudii and Loligo vulgaris species), chrysene (in L. opalescens) and indeno(1,2,3-cd)pyrene (in L. gahi). PAHs source analysis indicated that four of the five zones of capture of the different squid species are significantly affected by both petrogenic and pyrolytic sources. Assessment of the target carcinogenic risks, established by the US EPA, suggested that L. gahi (Atlantic Ocean) and L. opalescens (from Pacific Ocean) may pose additional risks for consumers, if not eaten in moderation, derived from benzo(a)pyrene ingestion.
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