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- Abundance dynamics of a new, endemic batoid from Brazil: The Lutz’s stingray, Hypanus berthalutzaePublication . Branco-Nunes, Ilka; Niella, Yuri; Hazin, Fábio Hissa Vieira; Creio, Emmanuelly; Oliveira, Paulo Guilherme Vasconcelos de; Afonso, André SucenaThe Lutz’s stingray, Hypanus berthalutzae, is a recently identified, endemic dasyatid species which occurs in shallow habitats exclusively along most of the Brazilian coast. It is frequently caught as bycatch in different fisheries throughout the range of its distribution, but information on population trends and the impacts of fishing is still scarce. Therefore, this study aimed to describe the structure and dynamics in H. berthalutzae relative abundance off Recife, Northeast Brazil, in order to provide baseline data for species management. H. berthalutzae were caught and sampled in a fisheries-independent, standardized longline and drumline survey which used > 500,000 hooks during ∼11 years. H. berthalutzae catch composition (N = 86) was characterized by a striking predominance of females, with a female:male ratio of 16:1. Most of these individuals measured were considered to be mature. H. berthalutzae catch rates were largely reduced after hooks previously operating directly on the seafloor were suspended in the middle of the water column. Such an effect was included in a zero-inflated generalized additive model developed to assess spatiotemporal and environmental modulators of H. berthalutzae abundance, for which both month and lunar phase proved to be statistically significant predictors. A higher abundance of H. berthalutzae was observed in the rainy season, from March to July (χ2= 19.22; p < 0.001), as well as during full moon periods (Z = 2.38; p = 0.017). The effects of other candidate predictors such as sampling site or year were either nonexistent or superimposed by other more deterministic factors. The results achieved in this fully standardized survey depict the dynamics in adult H. berthalutzae which might be expected to occur in coastal habitats off northeastern Brazil, thus providing a solid contribution to improve the management of local artisanal fisheries towards the recovery and sustainability of H. berthalutzae populations across their range.
- Acclimation capability inferred by metabolic performance in two sea cucumber species from different latitudesPublication . Kühnhold, Holger; Novais, Sara C.; Alves, Luís M. F.; Kamyab, Elham; Lemos, Marco F. L.; Slater, Matthew J.; Kunzmann, AndreasThe notion that thermal specialists from tropical regions live closer to their temperature limits than temperate eurytherms, seems too generalized. Species specific differences in physiological and biochemical stress reactions are linked to key components of organism fitness, like metabolic capacity, which indicates that acclimation potential across latitudes might be highly diverse rather than simplistic. In this study the exposure of a tropical (Holothuria scabra) and a temperate (Holothuria forskali) sea cucumber species to identical cold- and warmacclimation stress was compared using the key metabolic parameters, respiration rate, enzyme activity (ETS, LDH, IDH), and energy reserve fractions (lipid, carbohydrate and protein). Results show much broader respiratory adjustments, as response to temperature change, in H. scabra (2–30 μgO2*gww−1 *h−1) compared to H. forskali (1.5–6.6 μgO2*gww−1 *h−1). Moreover, the tropical species showed clearly pronounced up and down regulation of metabolic enzymes and shifts in energy reserves, due to thermal acclimation, while the same metabolic indicators remained consistent in the temperate species. In summary, these findings indicate enhanced metabolic plasticity in H. scabra at the cost of elevated energy expenditures, which seems to favor the tropical stenotherm in terms of thermal acclimation capacity. The comparison of such holistic metabolic analyses between conspecifics and congeners, may help to predict the heterogeneous effects of global temperature changes across latitudinal gradients.
- Accumulation of chemical elements and occurrence of microplastics in small pelagic fish from a neritic environmentPublication . Silva, Joana M. da; Alves, Luís M.F.; Laranjeiro, Maria I.; Bessa, Filipa; Silva, Andreia V.; Norte, Ana C.; Lemos, Marco F.L.; Ramos, Jaime A.; Novais, Sara C.; Ceia, Filipe R.The assessment of contaminant exposure in marine organisms often focuses on the most toxic chemical elements from upper trophic level species. Information on mid-trophic level species and particularly on potentially less harmful elements is lacking. Additionally, microplastics have been considered emergent contaminants in aquatic environments which have not been extensively studied in species from mid-trophic levels in food chains. This study aims to contribute to an overall assessment of environmental impacts of such chemicals in a community of small pelagic fish in the North Atlantic. The concentrations of 16 chemical elements, rarely simultaneously quantified (including minerals, trace elements and heavy metals), and the presence of microplastics were analysed in sardines (Sardina pilchardus) and mackerels (Scomber spp. and Trachurus trachurus) sampled along the Portuguese coast. Biochemical stress assessments and stable isotope analyses were also performed. The chemical element concentrations in S. pilchardus, T. trachurus, and Scomber spp. were relatively low and lower than the levels reported for the same species in the North Atlantic and adjacent areas. No clear relationships were found between chemical elements and oxidative damage in fish. However, the concentration of several chemical elements showed differences among species, being related with the species’ habitat use, trophic niches, and specific feeding strategies. The presence of plastic pieces in the stomachs of 29% of the sampled fishes is particularly concerning, as these small pelagic fish from mid-trophic levels compose a significant part of the diet of humans and other top predators. This study highlights the importance of multidisciplinary approaches focusing on the individual, including position data, stable isotopes, and oxidative stress biomarkers as complementary tools in contamination assessment of the marine mid-trophic levels in food chains.
- Acute Stress and an Electrolyte-Imbalanced Diet, but Not Chronic Hypoxia, Increase Oxidative Stress and Hamper Innate Immune Status in a Rainbow Trout (Oncorhynchus mykiss) Isogenic LinePublication . Magnoni, Leonardo J.; Novais, Sara C.; Eding, Ep; Leguen, Isabelle; Lemos, Marco F. L.; Ozório, Rodrigo O. A.; Geurden, Inge; Prunet, Patrick; Schrama, Johan W.In aquaculture, fish may be exposed to sub-optimal rearing conditions, which generate a stress response if full adaptation is not displayed. However, our current knowledge of several coexisting factors that may give rise to a stress response is limited, in particular when both chronic and acute stressors are involved. This study investigated changes in metabolic parameters, oxidative stress and innate immune markers in a rainbow trout (Oncorhynchus mykiss) isogenic line exposed to a combination of dietary (electrolyteimbalanced diet, DEB 700 mEq Kg−1 ) and environmental (hypoxia, 4.5 mg O2 L −1 ) challenges and their respective controls (electrolyte-balanced diet, DEB 200 mEq Kg−1 and normoxia, 7.9 or mg O2 L −1 ) for 49 days. At the end of this period, fish were sampled or subjected to an acute stressor (2 min of handling/confinement) and then sampled. Feeding trout an electrolyte-imbalanced diet produced a reduction in blood pH, as well as increases in cortisol levels, hepato-somatic index (HSI) and total energy content in the liver. The ratio between the lactate dehydrogenase (LDH) and isocitrate dehydrogenase (IDH) activities decreased in the liver of trout fed the DEB 700 diet, but increased in the heart, suggesting a different modulation of metabolic capacity by the dietary challenge. Several markers of oxidative stress in the liver of trout, mainly related to the glutathione antioxidant system, were altered when fed the electrolyteimbalanced diet. The dietary challenge was also associated with a decrease in the alternative complement pathway activity (ACH50) in plasma, suggesting an impaired innate immune status in that group. Trout subjected to the acute stressor displayed reduced blood pH values, higher plasma cortisol levels as well as increased levels of metabolic markers associated with oxidative stress in the liver. An interaction between diet and acute stressor was detected for oxidative stress markers in the liver of trout, showing that the chronic electrolyte-imbalance impairs the response of rainbow trout to handling/confinement. However, trout reared under chronic hypoxia only displayed changes in parameters related to energy use in both liver and heart. Taken together, these results suggest that trout displays an adaptative response to chronic hypoxia. Conversely, the dietary challenge profoundly affected fish homeostasis, resulting in an impaired physiological response leading to stress, which then placed constraints on a subsequent acute challenge.
- Adding value to bycatch fish species captured in the Portuguese coast—development of new food productsPublication . Silva, Frederica; Duarte, Ana M.; Mendes, Susana; Borges, Patrícia; Magalhães, Elisabete; Pinto, Filipa R.; Barroso, Sonia; Neves, Ana; Sequeira, Vera; Vieira, Ana Rita; Magalhães, Maria Filomena; Rebelo, Rui; Assis, Carlos; Gordo, Leonel SerranoWe live in a world of limited biological resources and ecosystems, which are essential to feed people. Consequently, diversifying target species and considering full exploitation are essential for fishery sustainability. The present study focuses on the valorization of three low commercial value fish species (blue jack mackerel, Trachurus picturatus; black seabream, Spondyliosoma cantharus; and piper gurnard, Trigla lyra) and of two unexploited species (comber, Serranus cabrilla and boarfish, Capros aper) through the development of marine-based food products with added value. A preliminary inquiry with 155 consumers from Região de Lisboa e Vale do Tejo (Center of Portugal) was conducted to assess fish consumption, the applicability of fish product innovation, and the importance of valorizing discarded fish. Five products (black seabream ceviche, smoked blue jack mackerel pâté, dehydrated piper gurnard, fried boarfish, and comber pastries) were developed and investigated for their sensory characteristics and consumer liking by hedonic tests to 90 consumers. The most important descriptors were identified for each product (texture, flavor, color, and appearance). Comber pastries had the highest purchase intention (88%), followed by black seabream ceviche (85%) and blue jack mackerel pâté (76%). Sensory evaluations showed a clear tendency of consumers to accept reformulated products, with the introduction of the low-value and unexploited species under study.
- Adding value to stalked barnacles from Berlengas Nature Reserve (Portugal) by the development of a new foodPublication . Pinheiro, Joaquina; Fernandes, Wilson; Sá, Hugo; Bernardino, Raul; Leandro, Sérgio Miguel; Ganhão, RuiIn this work, a new and natural food product, stalked barnacle (Pollicipes Pollicipes) pâté enriched with blackberry, was developed to valorise the rejected stalked barnacle. To evaluate the addition of blackberry fruits (Rubus ulmifolius Schott.) as a natural preservative on pâté quality, four pâté sample groups were considered: a negative control without a synthetic additive (CTR), a positive control with butylated hydroxytoluene (BHT), a group with blackberry extract (blackberry), and a group with a mixture of BHT and blackberry (blackberry + BHT). In addition, the effect of pasteurization (80 °C for 30 min) versus sterilization (121 °C for 30 min) on the pâté quality were evaluated. The bioactive evaluation expressed by the total phenolic content (TPC) and antiradical activity by the DPPH radical scavenging associated with oxidative stability determined by thiobarbituric acid reacting substances (TBARS) were performed. The impact of packaging opening followed by refrigerated storage (4 ± 1 °C) simulating the consumer behavior at home, was assessed. All the heat-treated stalked barnacle pâté samples were found microbially safe with an interesting content of total phenolic, the highest ones being the CTR and Blackberry + BHT, ranging from 58.79 to 55.38 mg GAE/100 g. After the package opening of the barnacle pâté sample of Blackberry + BHT, it revealed a superior inhibition of TBARS (0.46 mg MDA/100 g) after 7 days at refrigerated storage compared to the other samples. These results state the efficacy of blackberry in minimizing the lipid oxidation of stalked barnacle pâte. This study showed the potential for rejected stalked barnacle to be valorised and improve the sustainability of resources.
- Adjunctive role of Calyptranthes tricona extract with probiotic Kluyveromyces marxianus on colorectal adenocarcinoma Caco-2 cellsPublication . Kich, Débora Mara; Bitencourt, Shanna; Faleiro, Dalana; Immich, Sheila Mariele; Marmitt, Diorge Jonatas; Baldasso, Tamara; Haute, Gabriela Viegas; Oliveira, Jarbas Rodrigues de; Laufer, Stefan; Pedrosa, Rui; Souza, Claucia Fernanda Volken de; Goettert, Márcia InêsThe composition of microbiota may influence the development of colorectal cancer (CRC). In addition, probiotics can decrease the chance of developing cancer or its progress. For that reason, it is encouraging to assess the influence of plant extracts as adjuvants of the health-promoting effects of probiotics. Thus, this study aimed to investigate the in vitro beneficial properties of Calyptranthes tricona leaf ethanol extract in association or not with the lactic yeast Kluyveromyces marxianus on colon adenocarcinoma Caco-2 cells. C. tricona extract inhibited the in vitro p38α MAPK activity and exhibited immunomodulation on isolated human lymphocytes. Further, the extract did not induce cytotoxicity towards K. marxianus or Caco-2 cells; leading to an increased yeast adhesion to cells in a dose-dependent manner. In conclusion, this preliminary study demonstrates that C. tricona extract has effects on enzyme inhibition and immune function. In addition, when associated with K. marxianus, C. tricona may possess beneficial properties for application as enhancer of probiotic’s protective role on CRC cells. However, further studies are necessary in order to elucidate the mechanisms involved.
- Adverse outcome pathways induced by 3,4‑dimethylmethcathinone and 4‑methylmethcathinone in differentiated human SH‑SY5Y neuronal cellsPublication . Soares, Jorge; Costa, Vera Marisa; Gaspar, Helena; Santos, Susana; Bastos, Maria de LourdesCathinones (β-keto amphetamines), widely abused in recreational settings, have been shown similar or even worse toxicological profile than classical amphetamines. In the present study, the cytotoxicity of two β-keto amphetamines [3,4-dimethylmethcathinone (3,4-DMMC) and 4-methylmethcathinone (4-MMC)], was evaluated in differentiated dopaminergic SH-SY5Y cells in comparison to methamphetamine (METH). MTT reduction and NR uptake assays revealed that both cathinones and METH induced cytotoxicity in a concentration- and time-dependent manner. Pre-treatment with trolox (antioxidant) partially prevented the cytotoxicity induced by all tested drugs, while N-acetyl-l-cysteine (NAC; antioxidant and glutathione precursor) and GBR 12909 dopamine transporter inhibitor) partially prevented the cytotoxicity induced by cathinones, as evaluated by the MTT reduction assay. Unlike METH, cathinones induced oxidative stress evidenced by the increase on intracellular levels of reactive oxygen species (ROS), and also by the decrease of intracellular glutathione levels. Trolox prevented, partially but significantly, the ROS generation elicited by cathinones, while NAC inhibited it completely. All tested drugs induced mitochondrial dysfunction, since they led to mitochondrial membrane depolarization and to intracellular ATP depletion. Activation of caspase-3, indicative of apoptosis, was seen both for cathinones and METH, and confirmed by annexin V and propidium iodide positive staining. Autophagy was also activated by all drugs tested. Pre-incubation with bafilomycin A1, an inhibitor of the vacuolar H+- ATPase, only protected against the cytotoxicity induced by METH, which indicates dissimilar toxicological pathways for the tested drugs. In conclusion, the mitochondrial impairment and oxidative stress observed for the tested cathinones may be key factors for their neurotoxicity, but different outcome pathways seem to be involved in the adverse effects, when compared to METH.
- Age and growth of Brauer's lanternfish Gymnoscopelus braueri and rhombic lanternfish Krefftichthys anderssoni (Family Myctophidae) in the Scotia Sea, Southern OceanPublication . Saunders, Ryan A.; Lourenço, Sílvia; Vieira, Rui P.; Collins, Martin A.; Assis, Carlos A.; Xavier, Jose C.This study examines age and growth of Brauer's lanternfish Gymnoscopelus braueri and rhombic lanternfish Krefftichthys anderssoni from the Scotia Sea in the Southern Ocean, through the analysis of annual growth increments deposited on sagittal otoliths. Otolith pairs from 177 G. braueri and 118 K. anderssoni were collected in different seasons from the region between 2004 and 2009. Otolith‐edge analysis suggested a seasonal change in opaque and hyaline depositions, indicative of an annual growth pattern, although variation within the populations of both species was apparent. Age estimates varied from 1 to 6 years for G. braueri (40 to 139 mm standard length; L s) and from 0 to 2 years for K. anderssoni (26 to 70 mm L s). Length‐at‐age data were broadly consistent with population cohort parameters identified in concurrent length‐frequency data from the region for both species. The estimated values of von Bertalanffy growth curves for G. braueri were L∞ = 133.22 mm, k = 0.29 year−1 and t 0 = − 0.21 year and the values for K. anderssoni were L∞ = 68.60 mm, k = 0.71 year−1 and t 0 = − 0.49 year. There were no significant (P > 0.05) differences in growth between sexes for either species, suggesting that males and females have similar growth and development trajectories in the Scotia Sea. A positive allometric relationship between Ls and wet mass was found for each species, as well as a significant (P < 0.0001) linear relationship between otolith size and L s. Growth performance (Ф ′) was similar between the two species and congruent with other myctophid species across the Southern Ocean. This study provides important parameters for future Southern Ocean ecosystem studies in a resource management context.
- Algae from Portuguese Coast Presented High Cytotoxicity and Antiproliferative Effects on an In vitro Model of Human Colorectal CancerPublication . Alves, Celso; Pinteus, Susete; Rodrigues, Ana; Horta, André; Pedrosa, RuiBackground: The marine environment has shown to be an interesting source of new antitumor agents, representing an important tool in cancer research. Objective: The aim of this study was to evaluate the antitumor activities of 12 algae from Peniche coast (Portugal) on an in vitro model of human colorectal cancer (Caco‑2 cells). Materials and Methods: The antitumor potential was accessed by evaluating Caco‑2 cell’s viability and proliferation through the 3‑[4, 5‑dimethylthiazol‑2‑yl]‑2, 5‑diphenyl tetrazolium bromide and calcein‑AM methods. Results: The dichloromethane extracts of Asparagopsis armata and Sphaerococcus coronopifolius induced the highest decrease on cell’s viability (1 mg/mL; 24 h), 98.96% ± 0.39% and 98.08% ± 0.89%, respectively, followed by the methanolic extracts of S. coronopifolius (96.47% ± 1.26%) and A. armata (92.68% ± 1.17%). Regarding cell proliferation, the highest decrease of Caco‑2 cell’s proliferation (1 mg/mL; 24 h) was induced by the dichloromethane extract of A. armata (100% ± 0.48%), S. coronopifolius (99.04 ± 0.51%), and Plocamium cartilagineum (95.05% ± 1.19%). The highest potency was shown by the dichloromethane extract of S. coronopifolius in both, cytotoxicity and antiproliferative tests, with an IC50 of 21.3 and 36.5 µg/mL, respectively. Conclusion: The extracts of A. armata and S. coronopifolius are promising sources of new bioactive molecules with application in cancer therapeutics.