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- Temporal changes in macrofauna as response indicator to potential human pressures on sandy beachesPublication . Bessa, Filipa; Gonçalves, Sílvia C.; Franco, João N.; André, José N.; Cunha, Pedro P.; Marques, João CarlosSandy beaches are natural dynamic ecosystems, which are becoming worldwide increasingly disturbed by intensive human direct use, coastal development and erosive evolution. In this study, we have examined whether ten years of potential increased human pressures have resulted in significant changes in the macrofaunal assemblages’ structure and composition of two mesotidal sandy beaches (Cabedelo – urban beach, and Quiaios – rural beach) on the European Atlantic coast (Portugal). Seasonal macrofauna collections were performed at both beaches in two different periods, one in 1999–2000 and another in 2010–2011. The physical variables did not change significantly in both beaches throughout the studied periods, however, the urban beach was subject to an increase of human pressures (tourism and shoreline modifications) over the 10-years interval considered when compared with the rural beach. The univariate community descriptors (total density, species richness and diversity) did not differ significantly among periods for the rural beach. In contrast, temporal differences were found at the most urbanised beach, principally regarding the abundances of the amphipod Talitrus saltator and the isopod Tylos europaeus, two of the most abundant species at both beaches. PERMANOVA tests enhanced these temporal variations and the SIMPER analysis attributed to these species the main differences found between periods in this beach. Since the physical environment was similar in both periods, the ecological changes were most likely attributed to the increased human pressures observed at the urban beach. Nevertheless, this study highlights the need of further robust and effective impact assessments and long-term studies to better discern between natural and human induced changes on sandy beaches.
- Preparation of Triacylglycerols Rich in Omega-3 Fatty Acids from Sardine Oil Using a Rhizomucor miehei Lipase: Focus in the EPA/DHA RatioPublication . Bispo, Paulo; Batista, Irineu; Bernardino, Raul J.; Bandarra, Narcisa MariaThe increasing evidence on the differential biochemical effects of eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) raises the need of n-3 highly unsaturated fatty acid concentrates with different amounts of these fatty acids. In the present work, physicochemical and enzymatic techniques were combined to obtain acylglycerols, mainly triacylglycerols (TAG), rich in n-3 fatty acids. Sardine oil was obtained by washing sardine (Sardina pilchardus) mince with a NaHCO3 solution, hydrolyzed in a KOH-ethanol solution, and concentrated with urea. The esterification reaction was performed in the stoichiometric proportion of substrates for re-esterification to TAG, with 10 % level of Rhizomucor miehei lipase based on the weight of substrates, without any solvent, during 48 h. This procedure led to approximately 88 % of acylglycerols, where more than 66 % were TAG and the concentration of n-3 fatty acids was higher than 60 %, the EPA and DHA ratio (EPA/DHA) was 4:1. The content of DHA in the unesterifed fraction (free fatty acids) increased from 20 to 54 %, while the EPA level in the same fraction decreased from 33 to 12.5 % (EPA/DHA ratio ≈1:4). Computational methods (density functional theory calculations) have been carried out at the B3LYP/6-31G(d,p) level to explain some of the experimental results.
- Application of optimal experimental design concept to improve the estimation of model parameters in microbial thermal inactivation kineticsPublication . Gil, Maria M.; Miller, Fátima A.; Silva, Cristina L. M.; Brandão, Teresa R. S.The estimation of model parameters with high precision is of major importance in mathematical predictions. If a mathematical model is properly chosen and if the primary objective is to improve parameter estimation, underlying statistical theories can be applied. Precision increases with the number of experimental points. However, and in many situations, maximum precision is attained when sampling consists of replicates of specific experimental points. Experimental conditions can be optimized using the D-optimal design concept based on minimization of the generalized variance of the parameter estimates. The objective of this work was to use this methodology for the design of experiments for microbial inactivation processes described by a Gompertz-based model under isothermal and non-isothermal conditions. The application of D-optimal design concept considerably improved parameters precision, when compared to the commonly used heuristic designs.
- Impacts of the Invasive Seaweed Asparagopsis armata Exudate on Energetic Metabolism of Rock Pool InvertebratesPublication . Silva, Carla O.; Novais, Sara C.; Soares, Amadeu M. V. M.; Barata, Carlos; Lemos, Marco F. L.The marine red algae Asparagopsis armata is an invasive species gaining competitive advantage by releasing large amounts of toxic compounds to the surrounding invaded area. The main objective of this study was to evaluate the effects of this invasive seaweed on marine invertebrates by exposing the common prawn Palaemon elegans and the marine snail Gibbula umbilicalis to the exudate of this seaweed. The seaweed was collected and placed in a tank for 12 h in the dark in a 1:10 ratio. Afterwards the seawater medium containing the released secondary metabolites was collected for further testing. Lethal and sublethal effects of A. armata were investigated. Biochemical biomarker responses associated with energy metabolism (lactate dehydrogenase, LDH; electron transport system activity, ETS; lipid, protein and carbohydrate content) were analysed. The biomarker responses showed physiological status impairment of invertebrates after exposure to low concentrations of this algal exudate. The highest concentrations of exudate significantly increased lipid content in both organisms. In the shrimp, protein content, ETS, and LDH were also significantly increased. By contrast, these parameters were significantly decreased in G. umbilicalis. A behavioural impairment was also observed in G. umbilicalis exposed to A. armata exudate, reducing feeding consumption. These results represent an important step in the research of natural toxic exudates released to the environment and prospective effects of this seaweed in invaded communities under increasing global change scenarios.
- Impact of trematodes on the population structure and shell shape of the estuarine mud snail Hydrobia ulvae from a Southern European estuary.pdfPublication . Bordalo, Maria D.; Ferreira, Susana M. F.; Jensen, Kurt T.; Pardal, Miguel A.The occurrence of trematodes within the gastropod Hydrobia ulvae was studied in two areas (a Zostera noltii bed and a eutrophic area) in the Mondego Estuary, Portugal. The aim was to assess trematode infections across snail age classes, as well as to investigate any influence of parasitism on the shell shape and size of these gastropods. In the Z. noltii bed, infection occurred in snails with shell height of 1.5-6 mm, but a higher prevalence was seen in intermediate size classes (3-5 mm). Infected individuals were never found among the elder gastropods. In the eutrophic area, infection was commonly observed within small individuals (1.6-2.5 mm), as large individuals were rarely found. The warm temperate climate in this coastal system may cause infected specimens to die before they can grow to larger dimensions, in contrast to Northern European populations. The frequency of snails with thin, distorted and corroded shells was higher among infected than non-infected snails. Infected individuals were frequently colonized by epibionts that may have additional detrimental effects and probably promote shell erosion. Infection with trematodes and their influence on snail size needs consideration in the evaluation of parameters related to population dynamics and population structure. If the implications of parasite infection are ignored, inadequate decisions about ecosystem management may be taken, resulting mainly from erroneous assessment of population structure, age composition and ecological longevity. The role of parasites is especially important to consider for systems under environmental stress.
- Cholinesterase activity in the caddisfly Sericostoma vittatum: Biochemical enzyme characterization and in vitro effects of insecticides and psychiatric drugsPublication . Pestana, João L. T.; Novais, Sara C.; Lemos, Marco; Soares, Amadeu M. V. M.Sericostoma vittatum is a caddisfly species, endemic to the Iberian Peninsula, proposed as a biomonitor species for lotic ecosystems. Since inhibition of cholinesterases[U+05F3] (ChE) activity has been used to evaluate the exposure of macroinvertebrates to organophosphates and carbamate pesticides, this work intended to characterize the ChE present in this species so their activity can be used as a potential biomarker of exposure. Biochemical and pharmacological properties of ChE were characterized in this caddisfly species using different substrates (acetylthiocholine iodide, propionylthiocholine iodide, and butyrylthiocholine iodide) and selective inhibitors (eserine sulfate, BW284c51, and iso-OMPA). Also, the in vitro effects of two insecticides (carbaryl and chlorantraniliprole) and two psychiatric drugs (fluoxetine and carbamazepine) on ChE activity were investigated. The results suggest that S. vittatum possess mainly AChE able to hydrolyze both substrates acetylthiocholine and propionylthiocholine since: (1) it hydrolyzes the substrate acetylthiocholine and propionylcholine at similar rates and butyrylthiocholine at a much lower rate; (2) it is highly sensitive to eserine sulfate and BW284c51, but not to iso-OMPA; and (3) its activity is inhibited by excess of substrate, a characteristic of typical AChE. in vitro inhibitions were observed only for carbaryl exposure while exposure to chlorantraniliprole and to relevant environmental concentrations of psychiatric drugs did not cause any significant effect on AChE activity. This study suggests that AChE activity in caddisflies can indeed be used to discriminate the effects of specific insecticides in monitoring programs. The use of non-target species such as caddisflies in ecotoxicological research in lotic ecosystems is also discussed.
- Effects of cadmium and resource quality on freshwater detritus processing chains: a microcosm approach with two insect speciesPublication . Campos, Diana; Alves, Artur; Lemos, Marco F. L.; Correia, António; Soares, Amadeu M. V. M.; Pestana, João L. T.Detritus processing is vital for freshwater ecosystems that depend on the leaf litter from riparian vegetation and is mediated by microorganisms and aquatic invertebrates. Shredder invertebrates transform coarse particulate organic matter into fine particulate organic matter used as food by collector species. Direct and indirect effects of contaminants can impair detritus processing and thus affect the functioning of these ecosystems. Here, we assessed the combined effects of a toxic metal (cadmium) and resource quality (leaf species) on detritus processing and shredder-collector interactions. We considered two types of leaves, alder and eucalyptus that were microbially conditioned under different Cd concentrations in the laboratory. The microbial communities present on leaves were analyzed by Denaturing Gradient Gel Electrophoresis (DGGE), and we also measured microbial respiration rates. Sericostoma vittatum (a caddisfly shredder) and Chironomus riparius (a midge collector) were also exposed to Cd and allowed to consume the corresponding alder or eucalyptus leaves. We evaluated C. riparius growth and leaf mass loss in multispecies microcosms. Cadmium exposure affected leaf conditioning and fungal diversity on both leaf species, as assessed by DGGE. Cadmium exposure also affected the mass loss of alder leaves by reductions in detritivore feeding, and impaired C. riparius growth. Chironomus riparius consumed alder leaf discs in the absence of shredders, but S. vittatum appear to promote C. riparius growth in treatments containing eucalyptus. These results show that indirect effects of contaminants along detritus-processing chains can occur through effects on shredder-collector interactions such as facilitation but they also depend on the nutritional quality of detritus and on sensitivity and feeding plasticity of detritivore species.
- Human milk fat substitutes: Advances and constraints of enzyme‐catalyzed productionPublication . Ferreira-Dias, Suzana; Tecelão, CarlaThe production of triacylglycerols (TAGs) with similar structures to the human milk fat TAGs is only possible using sn-1,3 regioselective lipases as biocatalysts. The advances on the processes currently performed for the production of human milk fat substitutes (HMFS), namely the biocatalysts used, the best operation conditions, reactor types and operation modes, as well as the major drawbacks of these enzymatic systems, are addressed in this article.
- C2-Domain Abscisic Acid-Related Proteins Mediate the Interaction of PYR/PYL/RCAR Abscisic Acid Receptors with the Plasma Membrane and Regulate Abscisic Acid Sensitivity in ArabidopsisPublication . Rodriguez, Lesia; Gonzalez-Guzman, Miguel; Diaz, Maira; Rodrigues, Américo; Izquierdo-Garcia, Ana C.; Peirats-Llobet, Marta; Fernandez, Maria A.; Antoni, Regina; Fernandez, Daniel; Marquez, Jose A.; Mulet, Jose M.; Albert, Armando; Rodriguez, Pedro L.Membrane-delimited abscisic acid (ABA) signal transduction plays a critical role in early ABA signaling, but the molecular mechanisms linking core signaling components to the plasma membrane are unclear. We show that transient calcium-dependent interactions of PYR/PYL ABA receptors with membranes are mediated through a 10-member family of C2-domain ABA-related (CAR) proteins in Arabidopsis thaliana. Specifically, we found that PYL4 interacted in an ABA independent manner with CAR1 in both the plasma membrane and nucleus of plant cells. CAR1 belongs to a plant specific gene family encoding CAR1 to CAR10 proteins, and bimolecular fluorescence complementation and coimmunoprecipitation assays showed that PYL4-CAR1 as well as other PYR/PYL-CAR pairs interacted in plant cells. The crystal structure of CAR4 was solved, which revealed that, in addition to a classical calcium-dependent lipid binding C2 domain, a specific CAR signature is likely responsible for the interaction with PYR/PYL receptors and their recruitment to phospholipid vesicles. This interaction is relevant for PYR/PYL function and ABA signaling, since different car triple mutants affected in CAR1, CAR4, CAR5, andCAR9 genes showed reduced sensitivity to ABA in seedling establishment and root growth assays. In summary, we identified PYR/PYL-interacting partners that mediate a transient Ca2+-dependent interaction with phospholipid vesicles, which affects PYR/PYL subcellular localization and positively regulates ABA signaling.
- The golden mackerel: first report of a xanthic Mediterranean horse mackerel Trachurus mediterraneus (Steindachner, 1868) from Sines, PortugalPublication . Vasco-Rodrigues, Nuno; Santos, Simão; Espírito-Santo, Cristina; Almada, FredericoIn this study, a single xanthic case of the Mediterranean horse mackerel Trachurus mediterraneus is reported. The individual, with a total lenght (TL) of 306 mm, was captured off Sines (Portugal) in March 2018, by a local fishing boat, using a seine net. This represents the first record of xanthism for this species and for any species of mackerel. The specimen is described here and compared with those of normal pigmentation. Considerations about the paucity of records for this or similar species are presented here, and the possibility of this condition being more frequent than previously thought is discussed.