Lipids as biomarkers to assess the nutritional and physiological status of two diadromous fish (Anguilla anguilla and Chelon auratus) at early life stages in a temperate macrotidal estuary.

Estuaries are considered as key habitats for the early life stages of fish. However, in the face of massive destruction of many estuarine intertidal areas, management and conservation measures are needed. Fish condition indicators may be used as a proxy of habitat quality and provide valuable information for management of coastal areas. In this study, the larvae of golden mullet (Chelon auratus) and European glass eels (Anguilla anguilla) were sampled in three sites of the Gironde Estuary. Different lipid classes and fatty acids were quantified: phospholipids (globally, phosphatidylethanolamine and phosphatidylcholine), triglycerides, omega-3 (particularly docosahexaenoic and eicosapentaenoic acids), omega-6 and C18:1. These biomarkers provide information on the nutritional status of the larvae as well as on prey availability and larvae diet between sites. One site significantly differed from the others as it seemed to offer abundant and better-quality prey. The very high levels of omega-3 contained in mullet larvae suggested that this site provided a high amount of diatoms. However, the mullet larvae that colonized this site also showed physiological stress that could be explained by exposure to pollutants through their prey. This work constitutes an essential baseline for developing biomarkers to assess the quality of habitats in a global change context.

Comparing biomarker responses during thermal acclimation: A lethal vs non-lethal approach in a tropical reef clownfish.

Knowledge of thermal stress biology for most tropical fish species in reef ecosystems under climate change is still quite limited. Thus, the objective of this study was to measure the time-course changes of thermal stress biomarkers in the commercially exploited coral reef fish Amphiprion ocellaris, during a laboratory simulated event of increased temperature. Heat shock protein 70kDa (Hsp70) and total ubiquitin (Ub) were determined in the muscle (lethal method) and in the fin (non-lethal alternative method) under two temperature treatments (control - 26°C and elevated temperature - 30°C) throughout one month with weekly samplings. Results suggest that biomarker basal levels are tissue-specific and influence the degree of response under temperature exposure. Responses were highly inducible in the muscle but not in fin tissue, indicating that the latter is not reliable for monitoring purposes. Thermal stress was observed in the muscle after one week of exposure (both biomarkers increased significantly) and Ub levels then decreased, suggesting the animals were able to acclimate by maintaining high levels of Hsp70 and through an effective protein turnover. In addition, the results show that mortality rates did not differ between treatments. This indicates that A. ocellaris is capable of displaying a plastic response to elevated temperature by adjusting the protein quality control system to protect cell functions, without decreasing survival. Thus, this coral reef fish species presents a significant acclimation potential under ocean warming scenarios of +4°C. Monitoring of thermal stress through a non-lethal method, fin-clipping, although desirable proved to be inadequate for this species.

Single-trial properties of place cells in control and CA1 NMDA receptor subunit 1-KO mice.

The NMDA receptor plays a key role in synaptic plasticity and its disruption leads to impaired spatial representation in the CA1 area of the hippocampus, with place cells exhibiting larger place fields (McHugh et al., 1996). Place fields are defined by the spatial and nonspatial inputs of a given place and context, by intrinsic network processes, such as phase precession, but also by the matching of these inputs to a pre-existing spatial representation. Larger place fields may be a consequence of spatially widened firing upon a single crossing of a place field, or of increased variability in place field positions across traversals. We addressed this question by monitoring CA1 place cell activity, with tetrodes, in control and KO mice lacking the NMDA receptor in this region. In individual crossings of the field, we found no difference between genotypes in place field size; the larger, overall place field size turns out to be a consequence of jitter across trials. We suggest that this jitter reflects a deficit in the matching of current spatial inputs to the stored spatial representation of the track. This is supported by the finding that deficits in place field size and spatial information are rescued by extensive exposure of the mouse to the track, which may echo an increased influence of memory retrieval processes in CA3 on firing in CA1.

Global patterns and predictors of fish species richness in estuaries.

1. Knowledge of global patterns of biodiversity and regulating variables is indispensable to develop predictive models. 2. The present study used predictive modelling approaches to investigate hypotheses that explain the variation in fish species richness between estuaries over a worldwide spatial extent. Ultimately, such models will allow assessment of future changes in ecosystem structure and function as a result of environmental changes. 3. A comprehensive worldwide data base was compiled of the fish assemblage composition and environmental characteristics of estuaries. Generalized Linear Models were used to quantify how variation in species richness among estuaries is related to historical events, energy dynamics and ecosystem characteristics, while controlling for sampling effects. 4. At the global extent, species richness differed among marine biogeographic realms and continents and increased with mean sea surface temperature, terrestrial net primary productivity and the stability of connectivity with a marine ecosystem (open vs. temporarily open estuaries). At a smaller extent (within a marine biogeographic realm or continent), other characteristics were also important in predicting variation in species richness, with species richness increasing with estuary area and continental shelf width. 5. The results suggest that species richness in an estuary is defined by predictors that are spatially hierarchical. Over the largest spatial extents, species richness is influenced by the broader distributions and habitat use patterns of marine and freshwater species that can colonize estuaries, which are in turn governed by history contingency, energy dynamics and productivity variables. Species richness is also influenced by more regional and local parameters that can further affect the process of community colonization in an estuary including the connectivity of the estuary with the adjacent marine habitat, and, over smaller spatial extents, the size of these habitats. In summary, patterns of species richness in estuaries across large spatial extents seem to reflect from global to local processes acting on community colonization. The importance of considering spatial extent, sampling effects and of combining history and contemporary environmental characteristics when exploring biodiversity is highlighted.

Relationships between marine litter and type of coastal area, in Northeast Atlantic sandy beaches.

Marine litter represents a threat to the marine environment, being estimated that around eight million items are discarded daily in the ocean. Monitoring marine debris became a relevant topic of research as marine litter is one of the descriptors of the Marine Strategy Framework Directive, for European Union's member states. Nevertheless, the patterns and processes governing the disposal of waste in coastal areas are still not clear. Our study relates characteristics of eleven coastal areas in Portugal (urbanization, slope, distance to an estuary, length, and type of substrate) to the type and abundance of marine litter found. A total of 7743 items were identified, with the main types of litter found being plastic (71.2%), paper (16.3%), and sanitary waste (9.1%). A clear spatial distribution pattern was observed, with more litter items recorded in the zone corresponding to the high tide line (2.3 items m-2). It was also verified that both beaches and seasons influenced the amount of litter found. Plastic, the dominant marine litter group, was abundant on the vast majority of beaches. It was possible to identify litter with land and sea origins. The litter with land origin came mainly from sanitary and sewage-related waste while the litter with marine origin came mainly from fisheries, including aquaculture.

Neuroactive pharmaceuticals in estuaries: Occurrence and tissue-specific bioaccumulation in multiple fish species.

Contamination of surface waters by pharmaceuticals is an emerging problem globally. This is because the increased access and use of pharmaceuticals by a growing world population lead to environmental contamination, threatening non-target species in their natural environment. Of particular concern are neuroactive pharmaceuticals, which are known to bioaccumulate in fish and impact a variety of individual processes such as fish reproduction or behaviour, which can have ecological impacts and compromise fish populations. In this work, we investigate the occurrence and bioaccumulation of 33 neuroactive pharmaceuticals in brain, muscle and liver tissues of multiple fish species collected in four different estuaries (Douro, Tejo, Sado and Mira). In total, 28 neuroactive pharmaceuticals were detected in water and 13 in fish tissues, with individual pharmaceuticals reaching maximum concentrations of 1590 ng/L and 207 ng/g ww, respectively. The neuroactive pharmaceuticals with the highest levels and highest frequency of detection in the water samples were psychostimulants, antidepressants, opioids and anxiolytics, whereas in fish tissues, antiepileptics, psychostimulants, anxiolytics and antidepressants showed highest concentrations. Bioaccumulation was ubiquitous, occurring in all seven estuarine and marine fish species. Notably, neuroactive compounds were detected in every water and fish brain samples, and in 95% of fish liver and muscle tissues. Despite variations in pharmaceutical occurrence among estuaries, bioaccumulation patterns were consistent among estuarine systems, with generally higher bioaccumulation in fish brain followed by liver and muscle. Moreover, no link between bioaccumulation and compounds' lipophilicity, species habitat use patterns or trophic levels was observed. Overall, this work highlights the occurrence of a highly diverse suite of neuroactive pharmaceuticals and their pervasiveness in waters and fish from estuarine systems with contrasting hydromorphology and urban development and emphasizes the urgent need for toxicity assessment of these compounds in natural ecosystems, linked to internalized body concentration in non-target species.

Assessment of contaminants in blue sharks from the Northeast Atlantic: Profiles, accumulation dynamics, and risks for human consumers.

Chemical pollution is a major threat to marine ecosystems, and top predators such as most shark species are extremely vulnerable to being exposed and accumulating contaminants such as metals and persistent organic pollutants (POPs). This work aimed to study the degree, composition, and the sources of contamination in the blue shark (Prionace glauca) inhabiting the Northeast Atlantic, as well as the potential risk faced by human consumers. A total of 60 sharks were sampled in situ aboard fishing vessels, and the concentrations of a set of metals and POPs were analysed in various tissues and complemented with stable isotope analyses. High levels of contaminants were found in most sharks sampled. The concentrations of most metals were higher in the muscle when compared with the liver. Regarding the dangers to consumers posed by the concentrations of arsenic (As), mercury (Hg), and lead (Pb), over 75% of the sharks presented muscle concentrations of at least one contaminant above the legal limits for human consumption, and a risk assessment determined that consumption of meat of these sharks exceeding 0.07 Kg per week could potentially expose human consumers to dangerous amounts of methylmercury (MeHg). Additionally, the assessment of single contaminants may lead to an underestimation of the risk for the human health. Finally, the overall accumulation of contaminants seems to be mostly influenced by the sharks' geographical distribution, rather than sex, size, or trophic level of their prey.

Decoupling carry-over effects from environment in fish nursery grounds.

Life-history trait expression not only depends on the current environmental constraints, but also on the past ones that shaped traits expressed earlier in life. Such an effect, named carry-over, can occur in fish nursery grounds when juvenile performances after settlement are influenced by their larval traits in combination with conditions experienced in nurseries. To date, the impacts of environmental and human stressors on post-settlement traits have been assessed, but independently from larval traits, so that the contributions of environmental versus carry-over constraints remain unquantified. Here, we used a reconstructive approach based on otolith microstructure to investigate how carry-over and environment affect life-history traits of the European seabass, Dicentrarchus labrax. In the northeast Atlantic Ocean, seabass juveniles were collected in six French estuarine nursery areas with contrasted environmental conditions (water temperature, salinity, food availability, and anthropogenic impacts), and five of their life-history traits across ontogenetic stages were measured (pelagic growth, larval duration, size at settlement, post-settlement growth and body condition). Piecewise structural equation model emphasized the strong co-variation of larval traits in response to food availability and temperature in the pelagic environment, stressing that fast growing larvae are characterized by shorter pelagic larval duration, but larger size at recruitment. However, the magnitude of carry-over effects greatly varied between traits, revealing that larval trait impacts on post-settlement traits remained minor as compared to the nursery environment. In estuarine nurseries, our findings suggest that resource allocation results from a trade-off between somatic growth and energy storage. Fish juveniles exposed to anthropogenic stress or risk of food limitation tended to predominantly invest in storage, whereas individuals in favourable conditions allocated their resources in somatic growth. These findings highlight the importance of heterogeneity in pelagic and nursery environments in understanding trait variations and population dynamic of estuarine dependent fish.

Evidence of contamination-associated damage in blue sharks (Prionace glauca) from the Northeast Atlantic.

Top predators such as most shark species are extremely vulnerable to amassing high concentrations of contaminants, but not much is known about the effects that the contaminant body burden imparts on these animals. Species like the blue shark (Prionace glauca) are very relevant in this regard, as they have high ecological and socioeconomic value, and have the potential to act as bioindicators of pollution. This work aimed to assess if differences in contaminant body burden found in blue sharks from the Northeast Atlantic would translate into differences in stress responses. Biochemical responses related to detoxification and oxidative stress, and histological alterations were assessed in the liver and gills of 60 blue sharks previously found to have zone-related contamination differences. Similar zone-related differences were found in biomarker responses, with the sharks from the most contaminated zone exhibiting more pronounced responses. Additionally, strong positive correlations were found between contaminants (i.e., As, PCBs, and PBDEs) and relevant biomarkers (e.g., damaged DNA and protective histological alterations). The present results are indicative of the potential that this species and these tools have to be used to monitor pollution in different areas of the Atlantic.

Bioconcentration of neuroactive pharmaceuticals in fish: Relation to lipophilicity, experimental design and toxicity in the aquatic environment.

Uptake of contaminants is linked to their toxicity and is usually estimated through their lipophilicity (logKow). Here, we review current literature regarding bioconcentration, i.e. uptake of contaminants from the external environment only, and the effects of exposure to neuroactive pharmaceuticals in fish. We aim to determine if lipophilicity is a suitable predictor of bioconcentration of these compounds in fish, to identify major drivers of bioconcentration and explore the link between bioconcentration potential and toxicity, focusing on survival, growth, condition, behaviour and reproduction endpoints. Additionally, we compare concentrations known to elicit significant effects in fish with current environmental concentrations, identifying exposure risk in ecosystems. The majority of studies have focused on antidepressants, mainly fluoxetine, and encompasses mostly freshwater species. Few studies determined pharmaceuticals bioconcentration, and even a smaller portion combined bioconcentration with other toxicity endpoints. Results show that lipophilicity isn't a good predictor of neuroactive pharmaceuticals' bioconcentration in fish, which in turn is highly influenced by experimental parameters, including abiotic conditions, species and life-stage. The need for increased standardization of experimental settings is key towards improving accuracy of environmental risk assessments and application in future regulatory schemes. Still, increased fish lethality was linked to increased bioconcentration, yet no other correlations were observed when considering effects on growth, condition, behaviour or reproduction, likely as a result of insufficient and variable data. In the context of current environmental concentrations, several neuroactive pharmaceuticals were found to be potentially threatening, while data on occurrence is lacking for some compounds, particularly in brackish/marine systems. Specifically, nine compounds (fluoxetine, citalopram, sertraline, amitriptyline, venlafaxine, clozapine, carbamazepine, metamfetamine and oxazepam) were found at concentrations either above or critically close to minimum response concentrations, thus likely to affect fish in freshwater and brackish or marine environments, which supports further exploration in risk management strategies and monitoring programs in aquatic environments.

Elasmobranchs as bioindicators of pollution in the marine environment.

Bioindicator species are increasingly valuable in environmental pollution monitoring, and elasmobranch species include many suitable candidates for that role. By measuring contaminants and employing biomarkers of effect in relevant elasmobranch species, scientists may gain important insights about the impacts of pollution in marine ecosystems. This review compiles biomarkers applied in elasmobranchs to assess the effect of pollutants (e.g., metals, persistent organic pollutants, and plastics), and the environmental changes induced by anthropogenic activities (e.g., shifts in marine temperature, pH, and oxygenation). Over 30 biomarkers measured in more than 12 species were examined, including biotransformation biomarkers (e.g., cytochrome P450 1A), oxidative stress-related biomarkers (e.g., superoxide anion, lipid peroxidation, catalase, and vitamins), stress proteins (e.g., heat shock protein 70), reproductive and endocrine biomarkers (e.g., vitellogenin), osmoregulation biomarkers (e.g., trimethylamine N-oxide, Na+/K+-ATPase, and plasma ions), energetic and neurotoxic biomarkers (e.g., lactate dehydrogenase, lactate, and cholinesterases), and histopathological and morphologic biomarkers (e.g., tissue lesions and gross indices).

Does Predation Exacerbate the Risk of Endosymbiont Loss in Heat Stressed Hermatypic Corals? Molecular Cues Provide Insights Into Species-Specific Health Outcomes in a Multi-Stressor Ocean.

Ocean warming has been a major driver of coral reef bleaching and mass mortality. Coupled to other biotic pressures, corals' ability for acclimatization and adaptation may become compromised. Here, we tested the combined effects of warming scenarios (26, 30, and 32°C) and predation (wound vs. no wound) in coral health condition (paleness, bleaching, and mortality), cellular stress responses (heat shock protein 70 kDa Hsp70, total ubiquitin Ub, and total antioxidant capacity TAC), and physiological state (integrated biomarker response index, IBR) of seven Scleractinian coral species, after being exposed for 60 days. Results show that although temperature was the main factor driving coral health condition, thermotolerant species (Galaxea fascicularis, Psammocora contigua, and Turbinaria reniformis) displayed increased paleness, bleaching, and mortality in predation treatments at high temperature, whereas thermosensitive species (Acropora tenuis, Echinopora lamellosa, and Montipora capricornis brown and green morphotypes) all died at 32°C, regardless of predation condition. At the molecular level, results show that there were significant main and interactive effects of species, temperature, and predation in the biomarkers assessed. Temperature affected Hsp70, Ub, and TAC, evidencing the role of protein folding and turnover, as well as reactive oxygen species scavenging in heat stress management. Predation increased Hsp70 and Ub, suggesting the activation of the pro-phenoloxidase system and cytokine activity, whereas the combination of both stressors mainly affected TAC during moderate stress and Ub under severe stress, suggesting that redox balance and defense of homeostasis are crucial in tissue repair at high temperature. IBR levels showed an increasing trend at 32°C in predated coral fragments (although non-significant). We conclude that coral responses to the combination of high temperature and predation pressure display high inter-species variability, but these stressors may pose a higher risk of endosymbiont loss, depending on species physiology and stress intensity.

Selecting statistical models and variable combinations for optimal classification using otolith microchemistry.

Reliable assessment of fish origin is of critical importance for exploited species, since nursery areas must be identified and protected to maintain recruitment to the adult stock. During the last two decades, otolith chemical signatures (or "fingerprints") have been increasingly used as tools to discriminate between coastal habitats. However, correct assessment of fish origin from otolith fingerprints depends on various environmental and methodological parameters, including the choice of the statistical method used to assign fish to unknown origin. Among the available methods of classification, Linear Discriminant Analysis (LDA) is the most frequently used, although it assumes data are multivariate normal with homogeneous within-group dispersions, conditions that are not always met by otolith chemical data, even after transformation. Other less constrained classification methods are available, but there is a current lack of comparative analysis in applications to otolith microchemistry. Here, we assessed stock identification accuracy for four classification methods (LDA, Quadratic Discriminant Analysis [QDA], Random Forests [RF], and Artificial Neural Networks [ANN]), through the use of three distinct data sets. In each case, all possible combinations of chemical elements were examined to identify the elements to be used for optimal accuracy in fish assignment to their actual origin. Our study shows that accuracy differs according to the model and the number of elements considered. Best combinations did not include all the elements measured, and it was not possible to define an ad hoc multielement combination for accurate site discrimination. Among all the models tested, RF and ANN performed best, especially for complex data sets (e.g., with numerous fish species and/or chemical elements involved). However, for these data, RF was less time-consuming and more interpretable than ANN, and far more efficient and less demanding in terms of assumptions than LDA or QDA. Therefore, when LDA and QDA assumptions cannot be reached, the use of machine learning methods, such as RF, should be preferred for stock assessment and nursery identification based on otolith microchemistry, especially when data set include multispecific otolith signatures and/or many chemical elements.

Distribution and structure of the upper sublittoral macrobenthic communities of Tróia sand beaches (Setúbal, Portugal) and their relationship with environmental factors.

The present study dealt with the spatial and temporal variability of the distribution of the upper sublittoral benthic macrofauna of the Tróia peninsula sand beaches and its relationship with abiotic environmental factors. The existence of a relationship between the data set of macrobenthic species distribution and community structure in the Sado estuary (432 individual samples) and the environmental factors in analysis was investigated. Morpho-sedimentary data analysis revealed an environmental gradient, from the marine margin (exposed marine environment) to the estuarine margin (sheltered estuarine environment). Benthic macrofauna analysis showed a gradient of increasing number of individuals, species richness and diversity from the marine margin (Exposed) to the estuarine margin (Sheltered). Canonical Correspondence Analysis showed the dominant patterns in the community structure to be explained by the environmental factors considered, the most important, of which in influencing the spatial and temporal pattern, being beach slope, organic matter and calcium carbonate contents. The structure of the sandy beach communities studied showed a clear dominance of the spatial patterns over the seasonal ones. Four assemblages were defined-(1) an assemblage dominated by Angulus tenuis, on the marine margin of the peninsula; (2) an assemblage dominated by Euclymene sp. and Apseudes latreillei, on the sea-estuary transition area and related to the presence of a Zostera spp. meadow; (3) an assemblage dominated by Glycera sp. and Scoloplos armiger, on the sea-estuary transition area; (4) an assemblage dominated by Notomastus latericeus, Nassarius reticulatus and Cyathura carinata, on the estuarine margin.

Depressed, hypertense and sore: Long-term effects of fluoxetine, propranolol and diclofenac exposure in a top predator fish.

Pharmaceutical compounds are continuously released into the aquatic environment, resulting in their ubiquitous presence in many estuarine and coastal systems. As pharmaceuticals are designed to produce effects at very low concentrations and target specific evolutionary conserved pathways, there are growing concerns over their potential deleterious effects to the environment and specifically to aquatic organisms, namely in early life-stages. In this context, the long-term effects of exposure of juvenile meagre Argyrosomus regius to three different pharmaceuticals were investigated. Fish were exposed to environmental concentrations of one of three major used pharmaceuticals: the antidepressant fluoxetine (0.3 and 3 µg/L for 15 days), the anti-hypertensive propranolol and the non-steroidal anti-inflammatory agent diclofenac (0.3 and 15 µg/L for 30 days). Pharmaceuticals bioconcentration in fish muscle was examined, along with biomarkers in different tissues related with antioxidant and biotransformation responses (catalase, superoxide dismutase, ethoxyresorufin-O-deethylase and glutathione S-transferase), energetic metabolism (lactate dehydrogenase, isocitrate dehydrogenase and electron transport system activities), neurotransmission (acetylcholinesterase activity) and oxidative damage (DNA damage and lipid peroxidation levels). Overall, each pharmaceutical had different potential for bioconcentration in the muscle (FLX > PROP > DCF) and induced different biological responses: fluoxetine was the most toxic compound to juvenile meagre, affecting fish growth, triggering antioxidant defense responses, inhibiting detoxification mechanisms and increasing lipid peroxidation and DNA damage in the liver; propranolol exposure increased DNA damage and decreased aerobic metabolism in fish muscle; and diclofenac showed no potential to bioconcentrate, yet it affected fish metabolism by increasing cellular energy consumption in the muscle and consequently reducing fish net energy budget. The diverse response patterns evidence the need for future research focused on pharmaceuticals with different modes of action and their exposure effects on organismal physiological mechanisms and homeostatic status. Ultimately, the combination of sub-individual and individual responses is key for ecologically relevant assessments of pharmaceutical toxicity.

Synergistic Effects of Climate Change and Marine Pollution: An Overlooked Interaction in Coastal and Estuarine Areas.

Coastal areas have been increasingly affected by human activities, marine pollution and climate change are among the most important pressures affecting these environments. Human-induced pressures occur in a cumulative way and generate additive, antagonistic or synergistic effects. Knowledge on synergistic effects is crucial to coastal zone management, since they may imply a change in human uses of these systems, as well as dedicated action plans in order to reduce hazards and environmental risks. In this work, we provide an overview of the available literature on synergistic effects between climate change and chemical pollution, and discuss current knowledge, methodological approaches, and research gaps and needs. Interactions between these two pressures may be climate change dominant (climate change leads to an increase in contaminant exposure or toxicity) or contaminant-dominant (chemical exposure leads to an increase in climate change susceptibility), but the mechanistic drivers of such processes are not well known. Results from a few meta-analyses studies and reviews showed that synergistic interactions tend to be more frequent compared to additive and antagonistic ones. However, most of the studies are individual-based and assess the cumulative effects of a few contaminants individually in laboratory settings together with few climate variables, particularly temperature and pH. Nevertheless, a wide diversity of contaminants have already been individually tested, spanning from metals, persistent organic pollutants and, more recently, emergent pollutants. Population and community based approaches are less frequent but have generated very interesting and more holistic perspectives. Methodological approaches are quite diverse, from laboratory studies to mesocosm and field studies, or based on statistical or modelling tools, each with their own potential and limitations. More holistic comparisons integrating several pressures and their combinations and a multitude of habitats, taxa, life-stages, among others, are needed, as well as insights from meta-analyses and systematic reviews.

Long-term exposure to increasing temperatures on scleractinian coral fragments reveals oxidative stress.

Global warming is leading to increases in tropical storms' frequency and intensity, allowing fragmentation of reef-forming coral species, but also to coral bleaching and mortality. The first level of organism's response to an environmental perturbation occurs at the cellular level. This study investigated the long-term oxidative stress on fragments of nine Indo-Pacific reef-forming coral species exposed for 60 days to increasing temperatures (30 °C and 32 °C) and compared results with control temperature (26 °C). Coral overall condition (appearance), lipid peroxidation (LPO), catalase activity (CAT), and glutathione S-transferase (GST) were assessed. The species Turbinaria reniformis, Galaxea fascicularis, and Psammocora contigua were the most resistant to heat stress, presenting no oxidative damage at 30 °C. Unlike G. fasciularis, both T. reniformis and P. contigua showed no evidence of oxidative damage at 32 °C. All remaining species' fragments died at 32 °C. Stylophora pistillata and Pocillopora damicornis were the most susceptible species to heat stress, not resisting at 30 °C.

Biomarker and behavioural responses of an estuarine fish following acute exposure to fluoxetine.

Antidepressants such as fluoxetine are frequently detected in estuaries and can have profound effects on non-target organisms by interfering with the neural system and affecting essential physiological processes and behaviours. In this context, short-term effects of fluoxetine exposure were analysed in the common goby Pomatoschistus microps, an estuarine resident fish species. Two experiments were conducted with fish exposed to: i) fluoxetine concentrations within the µg/L range for 96 h (0.1, 0.5, 10 and 100 µg/L) and ii) fluoxetine concentrations within the mg/L range for 1 h (1, 5 and 10 mg/L). Acute toxicity was assessed via multiple biomarker responses, namely: activity levels of antioxidant (superoxide dismutase and catalase) and detoxification enzymes (ethoxyresorufin O-deethylase and glutathione S-transferase); and biomarkers of effects (lipid peroxidation and DNA damage) and of neurotoxicity (acetylcholinesterase inhibition). Furthermore, behavioural responses concerning activity (active time, movement delay and number of active individuals) and feeding (number of feeding individuals) were also recorded and analysed. Acute fluoxetine exposure for 96 h (in the µg/L range) reduced antioxidant CAT activity with increasing concentrations but had no significant effect on SOD activity. Biotransformation enzymes showed bell-shaped response curves, suggesting efficient fluoxetine metabolism at concentrations up to 10 µg/L. No significant damage (LPO and DNAd) was observed at both concentration ranges (µg/L and mg/L), yet 1 h exposure to higher fluoxetine concentrations (mg/L range) inhibited acetylcholinesterase activity (up to 37%). Fluoxetine (at mg/L) also decreased the number of both feeding and active individuals (by 67%), decreased fish active time (up to 93%) and increased movement delay almost 3-fold (274%). Overall, acutely exposed P. microps were able to cope with fluoxetine toxicity at the µg/L range but higher concentrations (mg/L) affected fish cholinergic system and behavioural responses.

Ecotoxicity of the lipid-lowering drug bezafibrate on the bioenergetics and lipid metabolism of the diatom Phaeodactylum tricornutum.

Pharmaceutical residues impose a new and emerging threat to the marine environment and its biota. In most countries, ecotoxicity tests are not required for all pharmaceutical residues classes and, even when mandatory, these tests are not performed using marine primary producers such as diatoms. These microalgae are among the most abundant class of primary producers in the marine realm and key players in the marine trophic web. Blood-lipid-lowering agents such as bezafibrate and its derivatives are among the most prescribed drugs and most frequently found human pharmaceuticals in aquatic environments. The present study aims to investigate the bezafibrate ecotoxicity and its effects on primary productivity and lipid metabolism, at environmentally relevant concentrations, using the model diatom Phaeodactylum tricornutum. Under controlled conditions, diatom cultures were exposed to bezafibrate at 0, 3, 6, 30 and 60 µg L-1, representing concentrations that can be found in the vicinity of discharges of wastewater treatment plants. High bezafibrate concentrations increased cell density and are suggested to promote a shift from autotrophic to mixotrophic metabolism, with diatoms using light energy generated redox potential to breakdown bezafibrate as carbon source. This was supported by an evident increase in cell density coupled with an impairment of the thylakoid electron transport and consequent photosynthetic activity reduction. In agreement, the concentrations of plastidial marker fatty acids showed negative correlations and Canonical Analysis of Principal coordinates of the relative abundances of fatty acid and photochemical data allowed the separation of controls and cells exposed to bezafibrate with high classification efficiency, namely for photochemical traits, suggesting their validity as suitable biomarkers of bezafibrate exposure. Further evaluations of the occurrence of a metabolic shift in diatoms due to exposure to bezafibrate is paramount, as ultimately it may reduce O2 generation and CO2 fixation in aquatic ecosystems with ensuing consequences for neighboring heterotrophic organisms.

Biochemical impacts in adult and juvenile farmed European seabass and gilthead seabream from semi-intensive aquaculture of southern European estuarine systems.

The nutritional value and developmental variations of cultured fish were assessed for European seabass and gilthead seabream specimens reared in semi-intensive aquaculture systems in two Portuguese estuaries. Quantification of total protein and of carbohydrate and fatty acid profiles was carried out to determine differences between the composition of the same species in two development stages reared in four distinct farms. A significant influence of the rearing site on the nutritional composition of the same species was found for adult European seabass regarding saturated, monounsaturated and highly unsaturated fatty acids contents, both between estuaries and within each estuary. In gilthead seabream, saturated, monounsaturated, polyunsaturated and highly unsaturated fatty acids content were also influenced by the rearing site. Carbohydrate analysis showed a significant influence of the rearing site on free sugar and polysaccharide content in fish of both species, and there was no influence on the species' protein content. Differences in fatty acid and carbohydrate content among juvenile and adult stages were found for all the groups studied. The present study supported the existing evidence that semi-intensive rearing systems are subjected to the variability of extrinsic factors in the rearing sites, influencing the nutritional value of the same species, namely regarding lipid and carbohydrate profiles, depending on the production site. From a consumer's perspective, such differences may come as a disadvantage of the rearing method, as it is expected for a product to provide equal nutritional properties and benefits regardless its origin, especially within the same country.