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- Evaluation of the Biological Potential of Himanthalia elongata (L.) S.F.Gray and Eisenia bicyclis (Kjellman) Setchell Subcritical Water ExtractsPublication . Gomes, Inês; Rodrigues, Helena; Rodrigues, Carla; Oliveira, Marta Marques; Paíga, Paula; Paiva, Alexandre; Simões, Pedro; Fernandes, Virgínia Cruz; Vieira, Mónica; Delerue-Matos, Cristina; Soares, Cristina; Grosso, ClaraNeuroprotection is a need that remains unmet in treating chronic neurodegenerative disorders, despite decades of extensive research. To find new neuroprotective compounds, extracts of Himanthalia elongata (L.) S.F.Gray and of Eisenia bicyclis (Kjellman) Setchell were obtained through subcritical water extraction applying a four-step temperature gradient. The fractions obtained were screened against brain enzymes involved in neurodegenerative etiology, namely in Alzheimer’s and Parkinson’s diseases, and against reactive oxygen and nitrogen species, all contributing factors to the progression of neurodegeneration. Results showed no significant enzyme inhibition but strong radical scavenging activities, particularly in the fourth fraction, extracted at the highest temperature (250 °C), highlighting their ability to retard oxidative and nitrosative stresses. At higher temperatures, fractions were composed of phenolic compounds and Maillard reaction products, a combination that contributed to their antioxidant activity and, consequently, their neuroprotective properties. All fractions were evaluated for the presence of iodine, 14 organochlorine and 7 organophosphorus pesticides, and pharmaceuticals used in Alzheimer’s and Parkinson’s diseases (14), psychiatric drugs (8), and metabolites (8). The fractions studied did not present any of the screened contaminants, and only fraction 1 of E. bicyclis should be used with caution due to iodine content.
- Bioactive Lipids of Seaweeds from the Portuguese North Coast: Health Benefits versus Potential ContaminationPublication . Soares, Cristina; Sousa, Sara; Machado, Susana; Vieira, Elsa; Carvalho, Ana P.; Ramalhosa, Maria João; Morais, Simone; Correia, Manuela; Oliva-Teles, MT; Domingues, Valentina; Delerue-Matos, CristinaThe total lipid content and lipidic profile of seaweeds harvested in the North Coast and purchased in Portugal were determined in this paper. The amount of total lipids in the different species of seaweeds varied between 0.7 ± 0.1% (Chondrus crispus) and 3.8 ± 0.6% (Ulva spp.). Regarding the fatty acid content, polyunsaturated fatty acids (PUFA) ranged between 0–35%, with Ulva spp. presenting the highest amount; monounsaturated fatty acids (MUFA) varied between 19 and 67%; and saturated fatty acids (SFA) were predominant in C. crispus (45–78%) and Gracilaria spp. (36–79%). Concerning the nutritional indices, the atherogenicity index (AI) was between 0.4–3.2, the thrombogenicity index (TI) ranged from 0.04 to 1.95, except for Gracilaria spp., which had a TI of 7.6, and the hypocholesterolemic/hypercholesterolemic ratio (HH) values ranged between 0.88–4.21, except for Gracilaria spp., which exhibited values between 0.22–9.26. The n6/n3 ratio was below 1 for most of the species evaluated, except for Ascophyllum nodosum, which presented a higher value, although below 2. Considering the PUFA/SFA ratio, seaweeds presented values between 0.11–1.02. The polycyclic aromatic hydrocarbons (PAHs) and aliphatic hydrocarbons (AHCs) contamination of seaweeds under study was also quantified, the values found being much lower than the maximum levels recommended for foodstuff.
- The influence of the extraction temperature on polyphenolic profiles and bioactivity of chamomile (Matricaria chamomilla L.) subcritical water extractsPublication . Cvetanović, Aleksandra; Švarc-Gajić, Jaroslava; Zeković, Zoran; Jerković, Jelena; Zengin, Gokhan; Gašić, Uroš; Tešić, Živoslav; Mašković, Pavle; Soares, Cristina; Barroso, M. Fátima; Delerue-Matos, Cristina; Đurović, SašaThe main goal of this research was to determine the relationship among chemical structure, bioactivity and temperature of chamomile during subcritical water extraction in isobaric conditions (45 bar) at seven different temperatures (65-210 °C). The influence of temperature on phenolic profiles was defined by UHPLC-HESI-MS/MS. The overall results indicate that the presence of conjugated double bonds, side chains, glucose moiety or ether moiety in molecules influence the efficiency of polyphenols' extraction in subcritical water. In terms of antioxidant activity, the extracts were the most active towards ABTS radicals (IC50 = 7.3-16.8 µg/mL), whereby temperature of 150 °C was optimal. On the other hand, the extracts obtained at 115 °C showed highest cytotoxicity. Inhibition of α-amylase and α-glucosidase was the highest at 65 and 85 °C, i.e. 0.51 and 4.13 mmolAE/g, respectively. Activity against tyrosinase was the highest at 210 °C (17.92 mgKAE/g). The data showed that different non-phenolic compounds may also participate in bio-activities of the extracts.
- Biofortified Diets Containing Algae and Selenised Yeast: Effects on Growth Performance, Nutrient Utilization, and Tissue Composition of Gilthead Seabream (Sparus aurata)Publication . Ferreira, Mariana; Ribeiro, Pedro C.; Ribeiro, Laura; Barata, Marisa; Domingues, Valentina F.; Sousa, Sara; Soares, Cristina; Marques, Alexandra; Pousão-Ferreira, Pedro; Dias, Jorge; Castro, L. Filipe C.; Marques, António; Nunes, Maria L.; Valente, Luisa M. P.Efforts have been made to find natural, highly nutritious alternatives to replace fish meal (FM) and fish oil (FO), which can simultaneously promote fish health and improve the nutritional quality of filets for human consumption. This study evaluated the impact of biofortified diets containing microalgae (as replacement for FM and FO), macroalgae (as natural source of iodine) and selenised yeast (organic source of selenium) on gilthead seabream growth, nutrient utilization, tissue composition and gene expression. A control diet (CTRL) with 15% FM and 5.5% FO was compared with three experimental diets (AD1, AD2, and AD3), where a microalgae blend (Chlorella sp., Tetraselmis sp., and DHA-rich Schizochytrium sp.) replaced 33% of FM. Diet AD1 contained 20% less FO. Diets were supplemented with Laminaria digitata (0.4% AD1 and AD2; 0.8% AD3) and selenised yeast (0.02% AD1 and AD2; 0.04% AD3). After feeding the experimental diets for 12 weeks, growth was similar in fish fed AD1, AD2, and CTRL, indicating that microalgae meal can partially replace both FM and FO in diets for seabream. But AD3 suppressed fish growth, suggesting that L. digitata and selenised yeast supplementation should be kept under 0.8 and 0.04%, respectively. Despite lower lipid intake and decreased PUFAs bioavailability in fish fed AD3, compared to CTRL, hepatic elovl5 was upregulated resulting in a significant increase of muscle EPA + DHA. Indeed, filets of fish fed AD2 and AD3 provided the highest EPA + DHA contents (0.7 g 100 g–1), that are well above the minimum recommended values for human consumption. Fish consuming the AD diets had a higher retention and gain of selenium, while iodine gain remained similar among diets. Upregulation of selenoproteins (gpx1, selk, and dio2) was observed in liver of fish fed AD1, but diets had limited impact on fish antioxidant status. Overall, results indicate that the tested microalgae are good sources of protein and lipids, with their LC-PUFAs being effectively accumulated in seabream muscle. Selenised yeast is a good fortification vehicle to increase selenium levels in fish, but efforts should be placed to find new strategies to fortify fish in iodine
- Removal of diclofenac and sulfamethoxazole from aqueous solutions and wastewaters using a three-dimensional electrochemical processPublication . Soares, Cristina; Correia-Sá, Luísa; Paíga, Paula; Barbosa, Carlos; Remor, Paula Verônica; Freitas, Olga; Moreira, Manuela M.; Nouws, Henri; Correia, Manuela; Ghanbari, Amir; Rodrigues, António J.; Oliveira, Carlos M.; Figueiredo, Sónia; Delerue-Matos, CristinaThe three-dimensional (3D) electrochemical treatment process was studied for the removal of two pharmaceuticals, diclofenac (anti-inflammatory) and sulfamethoxazole (antibiotic), in mono and bi-component systems. Adsorption and conventional two-dimensional electrochemical processes were initially studied and then combined to develop the 3D process. The influence of different operating parameters on the removal efficiency was studied: the distance between the cathode and the anode, the pharmaceutical and electrolyte (NaCl) concentrations, the pH, and the (carbon-based) adsorbent used as particulate electrode (biochar and commercial activated carbon, granulometry, and amount). The energy consumption and the electric energy per order were evaluated. The results demonstrate the efficiency of the 3D process for the removal of diclofenac and sulfamethoxazole from aqueous solutions, both for mono- and bi-component systems, achieving their complete removal respectively in 10 and 30 min, using a Mixed Metal Oxide anode (titanium-coated with RuO2-IrO2-TiO2), a stainless steel cathode, a biochar particulate electrode (1–2 mm), an initial pharmaceutical concentration of 10 mg/L, an inter-electrode distance of 7.5 cm, a pH value of 7 and a current density of 7 mA/cm2. The optimised 3D process was also successfully applied to a wastewater treatment plant effluent, but lower removal efficiencies were observed (after 30 min) for bi-component fortified samples; 49% for DCF and 86% for SMX, with energy consumptions of 1224 and 613 Wh/g and an electric energy per order of 19.1 and 8.77 kWh/m3 respectively. On the other hand, the pharmaceuticals were completely removed from the effluent when real concentrations (i.e. without their addition) were used
- Subcritical water extraction as an environmentally-friendly technique to recover bioactive compounds from traditional Serbian medicinal plantsPublication . Nastić, Nataša; Švarc-Gajić, Jaroslava; Delerue-Matos, Cristina; Barroso, M. Fátima; Soares, Cristina; Moreira, Manuela M.; Morais, Simone; Mašković, Pavle; Srček, Višnja Gaurina; Slivac, Igor; Radošević, Kristina; Radojković, MarijaSubcritical water extraction (SWE) has become a popular green extraction technique for the isolation of different classes of compounds from natural matrices. Low price, safety and green character of water, good yields of target compounds and reduced energy consumption, make this technique favorable for potential industrial applications. The purpose of this study was to evaluate antioxidant, antimicrobial and cytotoxic activity of four medicinal plants traditionally used in folk medicine of Serbia. Black mulberry (Morus nigra L.), wall germander (Teucrium chamaedrys L.), wild geranium (Geranium macrorrhizum L.) and comfrey (Symphytum officinale L.) were extracted by subcritical water at different temperatures. Antioxidant activity of the extracts was defined by conventional spectrophotometric methods, such as the total phenolic content (TPC), DPPH-radical scavenging activity (DPPH-RSA), ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) assessed by a DNA-based sensor. Additionally, the main phenolic compounds contributing to the antioxidant activity of the produced extracts were also identified and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Antimicrobial properties of extracts were evaluated against eight microbial strains. Furthermore, the cytotoxic activity was observed for two human cancer cell lines and a cell line derived from murine fibroblast.
- Development of New Canned Chub Mackerel Products Incorporating Edible Seaweeds—Influence on the Minerals and Trace Elements CompositionPublication . Vieira, Elsa F.; Soares, Cristina; Machado, Susana; Oliva-Teles, MT; Correia, Manuela; Ramalhosa, Maria João; Carvalho, A.; Domingues, Valentina; Antunes, Filipa; Morais, Simone; Delerue-Matos, CristinaThis study aimed to develop new canned chub mackerel products incorporating edible seaweeds (Ascophyllum nodosum, Fucus spiralis, Saccorhiza polyschides, Chondrus crispus, Porphyra sp. and Ulva sp.) harvested in the Portuguese North-Central coast, with simultaneous sensory improvement and minerals enrichment. Two processes were compared, namely the addition of seaweeds in i) the canning step and ii) in the brining step (as the replacement for salt). The concentrations of four macrominerals (Na, K, Ca and Mg), chloride, and twelve trace elements (Co, Cu, Fe, I, Li, Mn, Mo, Rb, Se, Sr, V and Zn) were determined by high-resolution continuum source flame atomic absorption spectrometry (HR-CS-FAAS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Results showed that canned chub mackerel incorporating C. crispus and F. spiralis was found to be the preferred sensory option, also exhibiting contents enriched with Cl, Co, Cu, Fe, I, Li, Mg, Mn, Mo, Na, Rb, Se, and Sr. This effect was more pronounced when both seaweed species were added to replace the salt added in the brining step.
- Investigating the Antioxidant Capacity of Fruits and Fruit Byproducts through an Introductory Food Chemistry Experiment for High SchoolPublication . Soares, Cristina; Correia, Manuela; Delerue-Matos, Cristina; Barroso, M. FátimaThis paper reports a laboratorial internship included in the Portuguese Science and Technology promotion program “Internships for Young People in Laboratories (Ciência Viva no Laboratório)”, which provided high school students an opportunity to approach the reality of scientific and technological research in a higher education institution. During this internship, students acquired knowledge related to the assessment of the total antioxidant capacity (TAC) of fruits and their byproducts while learning techniques, such as molecular spectrophotometry, as an analytical methodology to measure TAC using the ferric reducing antioxidant power (FRAP) and the total phenolic content (TPC) assays. First, the students were introduced to solid–liquid extraction as a conventional technique to extract antioxidants from the selected matrices. In order to optimize the extraction yield, different solvents, temperatures, and extraction times were used. Then, the students developed skills on TPC and FRAP assays by performing calibration curves using standard antioxidants, namely, gallic acid and ascorbic acid, prior to the measurement of the TAC of fruits (apple and orange) and respective byproducts. Final analysis included TAC values for comparison between fruits and their byproducts and also the influence of the extraction conditions on the TAC levels. At the end, the students presented their findings in a scientific poster and in a postlaboratory quiz. This laboratorial internship has been carried out since 2012 and was designed to be performed during a week, 7 h per day, in groups of two young students antioxidant power (FRAP) and the total phenolic content (TPC) assays. First, the students were introduced to solid−liquid extraction as a conventional technique to extract antioxidants from the selected matrices. In order to optimize the extraction yield, different solvents, temperatures, and extraction times were used. Then, the students developed skills on TPC and FRAP assays by performing calibration curves using standard antioxidants, namely, gallic acid and ascorbic acid, prior to the measurement of the TAC of fruits (apple and orange) and respective byproducts. Final analysis included TAC values for comparison between fruits and their byproducts and also the influence of the extraction conditions on the TAC levels. At the end, the students presented their findings in a scientific poster and in a postlaboratory quiz. This laboratorial internship has been carried out since 2012 and was designed to be performed during a week, 7 h per day, in groups of two young students.
- Involvement of the Iron-Regulated Loci hts and fhuC in Biofilm Formation and Survival of Staphylococcus epidermidis within the HostPublication . Oliveira, Fernando; Lima, Tânia; Correia, Alexandra; Silva, Ana Margarida; Soares, Cristina; Morais, Simone; Weißelberg, Samira; Vilanova, Manuel; Rohde, Holger; Cerca, NunoStaphylococcus epidermidis is a major nosocomial pathogen with a remarkable ability to persist on indwelling medical devices through biofilm formation. Nevertheless, it remains intriguing how this process is efficiently achieved under the host’s harsh conditions, where the availability of nutrients, such as essential metals, is scarce. Following our previous identification of two iron-regulated loci putatively involved in iron transport, hts and fhuC, we assessed here their individual contribution to both bacterial physiology and interaction with host immune cells. Single deletions of the hts and fhuC loci led to marked changes in the cell iron content, which were partly detrimental for planktonic growth and strongly affected biofilm formation under iron-restricted conditions. Deletion of each of these two loci did not lead to major changes in S. epidermidis survival within human macrophages or in an ex vivo human blood model of bloodstream infection. However, the lack of either hts or fhuC loci significantly impaired bacterial survival in vivo in a murine model of bacteremia. Collectively, this study establishes, for the first time, the pivotal role of the iron-regulated loci hts and fhuC in S. epidermidis biofilm formation and survival within the host, providing relevant information for the development of new targeted therapeutics against this pathogen
- 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.