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- Potential of Marine Biomolecules: Advances in Extraction and Applications of Proteins, Polysaccharides, and Antioxidant CompoundsPublication . Sousa, Gabriela; Ferreira-Dias, Suzana; Tecelão, Carla; Alves, Vítor D.Oceans are increasingly viewed as a new frontier for economic development, contributing to the bridge between food industry, sea bioeconomy, and health. Nowadays, oceans are under attention as a strategy for creating opportunities and driving innovation, and their vital importance will become even more evident in the years to come. Therefore, it is crucial to study oceans under a holistic approach, taking the maximum value of their abundant resources in a sustainable way. As such, blue bioeconomy is the path forward, since it is a development strategy that meets the economic potential without compromising the environmental health. A special look needs to be taken at the underutilized resources and by-products, which hold unexploited value. For instance, green macroalgae are widespread marine macroalgae that lack industry applications, despite being rich in biopolymers (polysaccharides) and antioxidants. Moreover, fish by-products are also rich sources of biopolymers, mostly proteins. Thus, among other potential uses, raw materials could be explored to produce functional edible coatings under a blue bioeconomy approach. A detailed characterization of raw materials is the first step for the development of value-added products. These topics will be addressed in this review.
- Broodstock conditioning of the Portuguese oyster (Crassostrea angulata, Lamarck, 1819): influence of different dietsPublication . Anjos, Catarina; Baptista, Teresa; Joaquim, Sandra; Mendes, Susana; Matias, Ana Margarete; Moura, Paula; Simões, Tiago; Matias, DomitíliaThe Portuguese oyster Crassostrea angulata shows great potential in oyster farming. The conservation of pure populations of this species is important for production diversification and biodiversity preservation. In this way, the zootechnological development for seed hatchery production is extremely important. Broodstock conditioning is a key step in the process of rearing bivalves in a hatchery. Many factors regulate the reproductive cycle, being food one of the most important ones. To evaluate the effect of different diets on C. angulata reproductive performance, broodstock were conditioned with different food regimes formulated fundamentally by flagellates (Diet 1 – Pavlova lutheri and Isochrysis galbana clone T-ISO; Diet 2 – P. lutheri, T-ISO and Skeletonema costatum) and constituted fundamentally by diatoms (Diet 3 – S. costatum and Chaetoceros calcitrans; Diet 4 – P. lutheri, S. costatum and C. calcitrans). During conditioning, samples of oysters were collected to evaluate condition index, gonadal development and biochemical composition. At the end of the conditioning period, oysters were induced to spawn to evaluate reproductive output (fecundity, fertilization rate and D-larvae development). The diets had an impact on the gametogenesis process, energy storage and reproductive output performance, being the best results those obtained in broodstock fed with the diatoms-predominant diets. However, those fed with diets majority flagellates had an unsuccessful performance. Holistic approaches incorporating all results in this study reveal and reinforce the idea that the diatom species used presented the nutritional requirements to C. angulata broodstock, being essential in the conditioning phase.
- Anti-Hepatocellular Carcinoma (HepG2) Activities of Monoterpene Hydroxy Lactones Isolated from the Marine Microalga Tisochrysis LuteaPublication . Gangadhar, Katkam N.; Rodrigues, Maria João; Pereira, Hugo; Gaspar, Helena; Malcata, F. Xavier; Barreira, Luísa; Varela, JoãoTisochrysis lutea is a marine haptophyte rich in omega-3 polyunsaturated fatty acids (e.g., docosahexaenoic acid (DHA)) and carotenoids (e.g., fucoxanthin). Because of the nutraceutical applications of these compounds, this microalga is being used in aquaculture to feed oyster and shrimp larvae. In our earlier report, T. lutea organic crude extracts exhibited in vitro cytotoxic activity against human hepatocarcinoma (HepG2) cells. However, so far, the compound(s) accountable for the observed bioactivity have not been identified. Therefore, the aim of this study was to isolate and identify the chemical component(s) responsible for the bioactivity observed. Bioassay-guided fractionation through a combination of silica-gel column chromatography, followed by preparative thin layer chromatography (PTLC), led to the isolation of two diastereomers of a monoterpenoid lactone, namely, loliolide (1) and epi-loliolide (2), isolated for the first time in this species. The structural elucidation of both compounds was carried out by GC-MS and 1D (1 H and13 C APT) and 2D (COSY, HMBC, HSQC-ed, and NOESY) NMR analysis. Both compounds significantly reduced the viability of HepG2 cells and were considerably less toxic towards a non-tumoral murine stromal (S17) cell line, although epi-loliolide was found to be more active than loliolide. © 2020 by the authors.
- Growth Performance after Agouti-Signaling Protein 1 (Asip1) Overexpression in Transgenic ZebrafishPublication . Godino-Gimeno, Alejandra; Sánchez, Elisa; Guillot, Raúl; Rocha, Ana; Angotzi, Anna Rita; Leal, Esther; Rotllant, Josep; Cerdá-Reverter, José MiguelThe melanocortin system is a key structure in the regulation of energy balance. Overexpression of inverse agonists, agouti-signaling protein (ASIP), and agouti-related protein (AGRP) results in increased food intake, linear growth, and body weight. ASIP regulates dorsal-ventral pigment polarity through melanocortin 1 receptor (MC1R) and overexpression induces obesity in mice by binding to central MC4R. Asip1 overexpression in transgenic zebrafish (asip1-Tg) enhances growth, yet experiments show fish overexpressing Asip1 do not develop obesity even under severe feeding regimes. Asip1-Tg fish do not need to eat more to grow larger and faster; thus, increased food efficiency can be observed. In addition, asip1-Tg fish reared at high density are able to grow far more than wild-type (WT) fish reared at low density, although asip1-Tg fish seem to be more sensitive to crowding stress than WT fish, thus making the melanocortin system a target for sustainable aquaculture, especially as the U.S. Food and Drug Association has recently approved transgenic fish trading.
- Bioproducts from forest biomass II. Bioactive compounds from the steam-distillation by-products of Cupressus lusitanica Mill. and Cistus ladanifer L. wastesPublication . Tavares, Cláudia S.; Martins, Alice; Miguel, M. Graça; Carvalheiro, Florbela; Duarte, Luís C.; Gameiro, José A.; Figueiredo, A. Cristina; Roseiro, Luísa B.Obtaining essential oils and hydrolates from underutilized biomass is an economic and sustainable way for production of these high added-value bioproducts. However, this process still generates large amounts of residues as the by-products obtained during distillation, which can be a concern for the environment, but also adequate substrates for other applications. Considering this fact, the waste distilled by-products remaining after steam-distillation of underutilized biomass from Cupressus lusitanica and Cistus ladanifer, were evaluated as a natural source of other high value products with biological activities, namely, phenolic compounds. Thus, the remaining extracted solid residues (ESRs) were characterized and subject to further treatments by ultrasound-assisted extraction (UAE) with ethanol and 70 % acetone, in order to prepare phenolic-rich extracts thereof: ESRs(EtOH) and ESRs(70 % Ace). Together with the distiller condensation waters (DCWs), these extracts were characterized for their phenolic content (total phenols, tannins and flavonoids). Their antioxidant activity was also evaluated by different methodologies. The phenolic profile of DCWs, ESRs(EtOH) and ESRs(70 % Ace) from both waste species was obtained by capillary zone electrophoresis (CZE) and phenolic compounds were tentatively identified. Results obtained for C. lusitanica biomass are here disclosed for the first time. Generally, all samples revealed to be rich in phenolic compounds, being C. ladanifer biomass the one with higher phenolic content. DCWs presented values of 140 mgGAE/g for C. lusitanica and 210 mg GAE/g for C. ladanifer, from which ca. 60 % were tannins. Extracts obtained with 70 % acetone were the ones with the highest results, except for the antioxidant activity by xanthine oxidase and superoxide inhibition, which was higher in DCWs. Catechins were the major compounds found for both species, but gallocatechins and gallic acid were only identified in C. ladanifer. Hydroxycinnamic acid derivatives and salicylic acid were also identified in C. ladanifer, partly justifying the anti-inflammatory effect referred for this species.
- Identification of the main quinolone resistance determinant in Campylobacter jejuni and Campylobacter coli by MAMA-DEG PCRPublication . Hormeño, Lorena; Palomo, Gonzalo; Ugarte-Ruiz, María; Porrero, M. Concepción; Borge, Carmen; Vadillo, Santiago; Píriz, Segundo; Domínguez, Lucas; Campos, Maria Jorge; Quesada, AlbertoAmong zoonotic diseases, campylobacteriosis stands out as the major bacterial infection producing human gastroenteritis. Antimicrobial therapy, only recommended in critical cases, is challenged by resistance mechanisms that should be unambiguously detected for achievement of effective treatments. Quinolone (ciprofloxacin) resistance of Campylobacter jejuni and Campylobacter coli, the2main Campylobacter detected in humans, is conferred by the mutation gyrA C-257-T, which can be genotyped by several methods that require a previous identification of the pathogen species to circumvent the sequence polymorphism of the gene. A multiplex PCR, based on degenerated oligonucleotides, has been designed for unambiguous identification of the quinolone resistance determinant in Campylobacter spp. isolates. The method was verified with 249 Campylobacter strains isolated from humans (141 isolates) and from the 3 most important animal sources for this zoonosis: poultry (34 isolates), swine (38 isolates), and cattle (36 isolates). High resistance to ciprofloxacin, MIC above 4 μg/mL, linked to the mutated genotype predicted by MAMA-DEG PCR (mismatch amplification mutation assay PCR with degenerated primers) was found frequently among isolates from the different hosts.
- Mono-specific facies of Parazoanthus axinellae in Luiz Saldanha Marine ParkPublication . Gomes-Pereira, J.N.; Vasco-Rodrigues, N.; Afonso, A.; Albuquerque, M.; Berecibar, E.; Bernardes, L.; Calado, A.; Castanheira, A.; Tojeira, I.; Tempera, Fernando; Dias, F.C.; Santos, R.S.; Campos, A.S.We report the first mono-specific facies ofParazoanthus axinellaefrom the western Portuguese coast. The habitat was recorded using a MiniRov Observer on a large outcrop at ca. 55–60 m in the Luiz Saldanha Marine Park. A method to estimate the maximum uncertainty of the ROV position is illustrated and the occurrence of the facies is discussed.
- Seaweed Pelvetia canaliculata as a Source of Bioactive Compounds for Application in Fried Pre-Coated Mackerel (Scomber scombrus) Fillets: A Functional Food ApproachPublication . Freire, Catarina D.; Antunes, Madalena; Silva, Susana F. J.; Neves, Marta; Tecelão, CarlaFatty fish, such as mackerel (Scomber scombrus), are recommended as part of a healthy diet, providing essential fatty acids (FA). Fried fish is appreciated for its attributes, including a crispy texture, golden crust, and pleasant taste. However, frying increases the fat content and the caloric value of food. This study evaluated the use of pre-frying edible coatings on mackerel fillets aiming to: (i) reduce oil absorption, (ii) minimize water loss, preserving fish succulence, and (iii) prevent fat oxidation. For this purpose, alginate- and carrageenan-based coatings were supplemented with extracts of Pelvetia canaliculata (Pc), a seaweed with high potential as a source of bioactive compounds. The fried fillets were analysed for colour, texture, moisture, ash, lipid content, and FA profile. No significant differences were observed for colour and textural parameters. Fillets coated with Pc-supplemented carrageenan showed the highest moisture (an increase of 3%) and the lowest fat content (a decrease of 7,5%) compared to the control (fried uncoated fillets). Coated fillets also exhibited reduced saturated FA and increased monounsaturated FA. In general, linoleic acid (C18:2) decreased markedly, while the values for docosahexaenoic acid (C22:6, n-3) remained stable (11–12% of total FA). Moreover, the n3/n6 ratio and atherogenic indices (AI) were improved in the coated fillets.
- Ridolfia segetum (L.) Moris (Apiaceae) from Portugal: A source of safe antioxidant and anti-inflammatory essential oilPublication . Cabral, C.; Poças Almeida, Juliana; Gonçalves, M.J.; Cavaleiro, C.; Cruz, M.T.; Salgueiro, L.Ridolfia segetum (L.) Moris essential oil was obtained by hydrodistillation and analysed by GC and GC-MS. The antioxidant properties were evaluated through the TBARS assay, and the in vitro anti-inflammatory activity was assessed through inhibition of NO production triggered by the Toll-like receptor 4 (TLR4) agonist, lipopolysaccharide (LPS), in mouse macrophages. Assessment of cell viability was made through a colorimetric assay using MTT on macrophages, keratinocytes, and hepatocytes. The main compounds are α-phellandrene (53.0-63.3%), terpinolene (11.9-8.6%), ß-phellandrene (5.5-6.0%), and dillapiol (1.9-8.0%). The oil demonstrated high antioxidant capacity in the concentrations tested (0.05-0.20. μL/mL) and significantly inhibited NO production without cytotoxicity in concentrations up to 1.25. μL/mL. The plants showed a high essential oil yield, which confers an industrial potential interest. Also, the high content of α-phellandrene (53.0-63.3%) makes it an important natural source of this compound. These results provide evidence of the therapeutic effects of this oil, and suggest that the anti-inflammatory activity in doses without cytotoxicity on mammalian cells, combined with its antioxidant potential and pleasant smell, could be of great value in the development of new phytopharmaceuticals.
- Marine Natural Products as Anticancer Agents 3.0Publication . Alves, Celso; Diederich, MarcCancer represents a major global health challenge[...]