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- 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.
- Seasonal Variation in Proximate Composition and Fatty Acid Profile of Grey Triggerfish Balistes capriscus Captured Along the Coast of PortugalPublication . Simões, Tiago; Carvalho, João; Sousa, Ana; Lemos, Marco F. L.; Gil, Maria M.; Pedrosa, Rui; Tecelão, CarlaProximate composition (ash, moisture, total protein, and fat contents) and fatty acid profile of Balistes capriscus (grey triggerfish) were assessed over a 6-months period (April to September, 2011) in animals captured along the coast of Peniche (Portugal). High protein (18.9% to 21.4%) and low lipid (0.5% to 0.8%) contents were determined. The highest lipid level was found in June-captured animals. High proportions of n-3 polyunsaturated fatty acids (n-3 PUFA) were observed (53.87 ± 2.00%) together with substantial quantities of oleic acid (27.58 ± 1.09%) and palmitic acid (16.52 ± 0.93%). Docosahexaenoic acid accounted for 77% of the total PUFA and was 7 times more abundant than eicosapentaenoic acid, and the n-3/n-6 ratio was 7.2. Gender did not influence lipid levels and the fatty acid profile, but seasonal variations were observed for PUFA and monounsaturated fatty acid contents. Overall, the nutritional properties of this underexploited species may be comparable to those of other lean fish species with higher economic value. Practical Application: Balistes capriscus is an abundant species in the Atlantic and underexploited in the Eastern Atlantic coast, with very few studies reported. This study was aimed to add commercial value to the species through its nutritional evaluation. B. capriscus is characterized by high protein and low fat contents. High proportions of n-3 PUFA, mainly docosahexaenoic acid, were noted for the grey triggerfish. Nutritional properties of this species may be comparable to those of other lean fish species.
- Metabolic Profile of Four Selected Cathinones in Microsome Incubations: Identification of Phase I and II Metabolites by Liquid Chromatography High Resolution Mass SpectrometryPublication . Lopes, Beatriz T.; Caldeira, Maria João; Gaspar, Helena; Antunes, Alexandra M. M.Consumption of synthetic cathinones, the second largest class of new psychoactive substances (NPS) reported worldwide, represents a serious public health risk. One of the biggest challenges created by the rapid spread of NPS on the illegal drug market is the discovery of selective biomarkers for their detection in biological matrices, which is only possible through the study of their metabolic profile. The synthetic cathinones 4′-methyl-N,N-dimethylcathinone (4-MDMC), 4′-methyl-N,N-diethylcathinone (4-MDEC), 4′-chloro-α-pyrrolidinovalerophenone (4Cl-PVP), and 4′-chloroethylcathinone (4-CEC) are NPS recently seized in Europe, and, with the exception of 4-CEC, no metabolism study was reported for these cathinones. With the ultimate goal of overcoming this gap, these cathinones were incubated in vitro in human and rat liver microsomes in the presence of Phase I and II (glucuronidation) co-factors, using α-pyrrolidinovalerophenone (α-PVP) as positive control. The metabolite identification was performed by liquid chromatography coupled to tandem high resolution mass spectrometry (LC-HRMS/MS). This allowed the identification of multiple Phase I and glucuronide metabolites of the selected cathinones. Additionally, a new glucuronide conjugate, derived from the recreational drug α-PVP, was herein identified for the first time. Importantly, we have demonstrated that 4-MDMC and 4-MDEC can act as prodrugs of the controlled substances 4-MMC and 4-MEC, respectively. The metabolites herein identified are expected to play an important role not only by acting as potential selective biomarkers of the intake of the synthetic cathinones selected for this study but also to understand their potential adverse effects and link these causative agents to toxicities, thereby helping in the treatment of non-fatal intoxications.
- Assessment of the Levels of Pollution and of Their Risks by Radioactivity and Trace Metals on Marine Edible Fish and Crustaceans at the Bay of Bengal (Chattogram, Bangladesh)Publication . Biswas, Krishna Prasad; Hossain, Shahadat; Deb, Nipa; Bhuian, A.K.M. Saiful Islam; Gonçalves, Sílvia C.; Hossain, Shahadat; Hossen, Mohammad BelalMarine environmental pollution is a longstanding global problem and has a particular impact on the Bay of Bengal. Effluent from different sources directly enters rivers of the region and eventually flows into the Bay of Bengal. This effluent may contain radioactive materials and trace metals and pose a serious threat to the coastal environment, in addition to aquatic ecosystems. Using gamma spectrometry and atomic absorption spectrometry, a comprehensive study was carried out on the radioactivity (226Ra, 232Th, 40K, and 137Cs) and trace metal (Cd, Pb, Zn, Cu, Ni, Fe, Mn, and Cr) concentrations, respectively, in fish and crustacean species collected from the coastal belt of the Bay of Bengal (Chattogram, Bangladesh). The analysis showed a noticeable increment in the levels of different radioactive pollutants in the marine samples, although the consumption of the studied fish and crustacean species should be considered safe for human health. Anthropogenic radionu-clide (137Cs) was not detected in any sample. Furthermore, the metal concentrations of a small num-ber of trace elements (Pb, Cd, Cr) were found to be higher in most of the samples, which indicates aquatic fauna are subject to pollution. The estimated daily intake (EDI), target hazard quotient (THQ), hazard index (HI), and target cancer risk (TR) were calculated and compared with the per-missible safety limits. It was found that consuming the seafood from the Bay of Bengal may cause adverse health impacts if consumption and/or means of pollution are not controlled.