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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., This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the project BLUESHARKER (PTDC/CTAAMB/29136/2017), co-financed by COMPETE2020 (POCI-01-0145-FEDER-029136), the Strategic Project granted to MARE – Marine and Environmental Sciences Centre (UIDB/04292/2020 and UIDP/04292/2020), the project granted to the Associate Laboratory ARNET (LA/P/0069/2020), and the grant awarded to Luís Alves (SFRH/BD/122082/2016). Vanessa F. Fonseca was supported by FCT, via research contract 2021.00244.CEECIND. Sara Novais is funded by national funds (OE), through FCT, in the scope of the framework contract foreseen in the numbers 4, 5, and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19.

The growth of human populations has been driving an unprecedent and widespread increase in marine traffic, posing a real threat to marine biodiversity. Even though we are now aware of the negative effects of shipping noise exposure on fish, information about the impact on their early life stages continues to lack. Meagre (Argyrosomus regius) is a vocal fish that uses estuaries with high levels of anthropogenic noise pollution as both breeding areas and nurseries. Here, the effects of boat noise exposure on the development and survival of meagre larvae were studied. Embryos and larvae were exposed to either noise (boat noise playback) or control treatments (coils producing a similar electric field to the speakers) and hatching rate, survival rate, morphometric traits and stress-related biomarkers, at hatching and at 2 days-post-hatching (dph) were analyzed. Results showed no conclusive effects of the impact of boat noise playback, even though there was an increased lipid droplet consumption and a decrease in body depth at 2dph larvae under this stressor. The assessment of oxidative stress and energy metabolism-related biomarkers at hatching showed a marginal decrease in superoxide dismutase (SOD) activity and no changes in DNA damage or electron transport system activity (ETS), although it cannot be disregarded that those effects could only be visible at later stages of larval development. Whether these morphological and developmental results have implications in later stages remains to be investigated. Further studies with longer exposure and wild meagre could help deepen this knowledge and provide a better understanding of how anthropogenic noise can impact meagre early stages Fundação para a Ciência e Tecnologia - FCT info:eu-repo/semantics/publishedVersion

Acknowledgments Financial support was given by FEDER through the project ReNATURE- Valorization of the Natural Endogenous Resources of the Centro Region (Centro 2020; Centro-01-0145-FEDER-000007) and SmarBioR- Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate (Centro 2020; Centro-01-0145FEDER-000018), co-funded by Centro 2020 Program, Portugal 2020, European Union, through the European Regional Development Fund. Authors also acknowledge FCT for the grant attributed to João Rosa (grant SFRH/BD/102008/2014) and to Sara Leston (grant SFRH/BPD/ 91828/2012). The funding source did not interfere with the study design, or in the collection, analysis or interpretation of the data. It also had no involvement in the writing of the report and in the decision to submit the present manuscript for publication. Background: The demand for fish and fish products is now higher than ever. However, several problems such as nutrient loading or excessive use of resources can be associated with the intensification of aquaculture systems. Integrated multitrophic aquaculture systems (IMTAs) refer to the co-culture of different species belonging to different trophic levels, and offer a sustainable approach to aquaculture development. In these systems, organic and inorganic extractive species will feed on other species waste or on uneaten feed nutrients, acting as bioremediators. Scope and approach: The extractive capacity that these organisms have to take up nutrients from the water also means they will accumulate chemicals that are often administered in intensive productions. The present review describes a vast number of substances that can be found in IMTAs, either intentionally administered or resulting from contamination, and subsequently accumulated in species reared afterwards in these systems. The presence of such chemicals in organisms produced in IMTAs raises several food safety and human health concerns, which need to be addressed. Key findings and conclusions: Although IMTAs still face many challenges in terms of large scale production, legislations are not yet ready to comprise co-cultivation of multiple species in proximity. Also, maximum residue limits already existent for fish must be set for other organisms also produced in IMTAs in order to protect consumer's health. An increase in extractive species consumption (e.g. seaweeds) has been noticed during the past few years, and as IMTAs gain importance as a sustainable production method, food safety issues must be tackled. info:eu-repo/semantics/publishedVersion

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., This study had the support of Fundaçao ˜ para a Ciˆencia e a Tecnologia (FCT) through the project BLUESHARKER (PTDC/CTAAMB/29136/ 2017), co-financed by COMPETE2020 (POCI-01-0145-FEDER-029136), the grant to MARE – Marine and Environmental Sciences Centre (UIDB/ 04292/2020 and UIDP/04292/2020) and the Associate Laboratory ARNET (LA/P/0069/2020), the grant awarded to Luís Alves (SFRH/BD/ 122082/2016), and the transitory norm contract to Filipe Ceia (DL57/ 2016/CP1370/CT90). Sara Novais is funded by national funds (OE), through FCT, in the scope of the framework contract foreseen in the numbers 4, 5, and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19

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., This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2020, project MARINE INVADERS—The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI-01-0145-FEDER-031144), and grant awarded to Carla Silva (SFRH/BD/87030/2012), and was also partially funded by Portuguese National Funds, FEDER funds within the PT2020 Partnership Agreement and Compete 2020-Programa Operacional Factores de Competitividade, and by the Portuguese Foundation for Science and Technology (FCT) through the strategic programme (UIDB/50017/2020 + UIDP/50017/2020). The project was also partially funded by the European Union through EASME Blue Labs project AMALIA—Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419) and Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund.

Funding: This study had the support of the Fundação para a Ciência e a Tecnologia (FCT) Strategic Projects UID/MAR/04292/2020 and UIDB/50006/2020 granted to MARE and REQUIMTE, respectively; the grant awarded to A.P.J. (2020.08202.BD); and project MARINE INVADERS (POCI-01-0145-FEDER-031144). The authors also wish to acknowledge the support of the European Union through EASME Blue Labs project AMALIA, Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419). The project was also partially funded by RDandT co-promotion project ORCHESTRA (POCI-01-0247-FEDER-070155) through COMPETE, Portugal 2020 and FCT, and COSMOS (MAR-04.03.01-FEAMP-0370) through GAL PESCA OESTE and MAR2020 in the framework of PORTUGAL2020 and the European Maritime and Fisheries Fund. Acne vulgaris (AV) is a chronic skin disease of the pilosebaceous unit affecting both adolescents and adults. Its pathophysiology includes processes of inflammation, increased keratinization, sebum production, hormonal dysregulation, and bacterial Cutibacterium acnes proliferation. Common AV has been treated with antibiotics since the 1960s, but strain resistance has emerged and is of paramount concern. Macroalgae are known producers of substances with bioactive properties, including anti-viral, antibacterial, antioxidant, and anti-inflammatory properties, among several others. In particular, red algae are rich in bioactive compounds such as polysaccharides, phenolic compounds, lipids, sterols, alkaloids, and terpenoids, conferring them antioxidant, antimicrobial, and anti-inflammatory activities, among others. Thus, the exploration of compounds from marine resources can be an appealing approach to discover new treatment options against AV. The aim of this work is to provide an overview of the current knowledge of the potentialities of red macroalgae in the treatment of AV by reviewing the main therapeutic targets of this disease, and then the existence of compounds or extracts with bioactive properties against them. info:eu-repo/semantics/publishedVersion

Argan oil is rich in long-chain unsaturated fatty acids (FA), mostly oleic and linoleic, and natural antioxidants. This study addresses the production of low-calorie structured lipids by acidolysis reaction, in a solvent-free system, between caprylic (C8:0, system I) or capric (C10:0, system II) acids and argan oil, used as triacylglycerol (TAG) source. Three commercial immobilized lipases were tested: Novozym® 435, Lipozyme® TL IM, and Lipozyme® RM IM. Higher incorporation degree (ID) was achieved when C10:0 was used as acyl donor, for all the lipases tested. Lipozyme® RM IM yielded the highest ID for both systems (28.9 ± 0.05 mol% C10:0, and 11.4 ± 2.2 mol% C8:0), being the only catalyst able to incorporate C8:0 under the reaction conditions for biocatalyst screening (molar ratio 2:1 FA/TAG and 55 °C). The optimal conditions for Lipozyme® RM IM in system II were found by response surface methodology (66 °C, molar ratio FA/TAG of 4:1), enabling to reach an ID of 40.9 mol% of C10:0. Operational stability of Lipozyme® RM IM in system II was also evaluated under optimal conditions, after eight consecutive 24 h-batches, with biocatalyst rehydration between cycles. The biocatalyst presented a half-life time of 103 h.

Funding: This study was supported by the Fundação para a Ciência e a Tecnologia (FCT) to MARE (UID/MAR/04292/2020) through national funds and grant to Tânia Vicente (2020.06230.BD). The authors also acknowledge the support of project ORCHESTRA—add-value to ORCHards through thE full valoriSaTion of macRoalgAe (POCI-01-0247-FEDER-070155) co-funded by FEDER—Fundo Europeu de Desenvolvimento Regional da União Europeia, Portugal 2020, through COMPETE 2020 —Programa Operacional Competitividade e Internacionalização and through FCT. Fungal phytopathogens are a growing problem all over the world; their propagation causes significant crop losses, affecting the quality of fruits and vegetables, diminishing the availability of food, leading to the loss of billions of euros every year. To control fungal diseases, the use of synthetic chemical fungicides is widely applied; these substances are, however, environmentally damaging. Marine algae, one of the richest marine sources of compounds possessing a wide range of bioactivities, present an eco-friendly alternative in the search for diverse compounds with industrial applications. The synthesis of such bioactive compounds has been recognized as part of microalgal responsiveness to stress conditions, resulting in the production of polyphenols, polysaccharides, lipophilic compounds, and terpenoids, including halogenated compounds, already described as antimicrobial agents. Furthermore, many studies, in vitro or in planta, have demonstrated the inhibitory activity of these compounds with respect to fungal phytopathogens. This review aims to gather the maximum of information addressing macroalgae extracts with potential inhibition against fungal phytopathogens, including the best inhibitory results, while presenting some already reported mechanisms of action. info:eu-repo/semantics/publishedVersion

Acknowledgements : This work was supported by the Portuguese Foundation for Science and Technology (FCT) through Strategic Project UID/MAR/04292/2019 granted to MARE – Marine and Environmental Sciences Centre and grant 2020.09455.BD awarded to Joao ˜ Reboleira. Work was also supported by VALORMAR (Mobilizing R&TD Programs, Portugal 2020) co-funded by COMPETE (POCI-01-0247-FEDER-024517), projects MACAU (MAR04.03.01-FEAMP-0128) and COSMOS (MAR2020-P04M03-1445P) funded by PO MAR2020, Portugal2020, and the European Union through the European Fund for Maritime Affairs and Fisheries (FEAMP), the Integrated Programme of SR&TD “Smart Valorisation of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, the European Union through the European Regional Development Fund (FEDER), and project ORCHESTRA (POCI-01-0247-FEDER-070155), funded by FEDER, FCT and Portugal 2020 through COMPETE 2020. Background: Seaweeds are promising substrates for biotransformation via fermentation, something that has been primarily utilized by the field of biofuels but focused less attention from other fields of research. Considering that the fermentation of abundant land resources has become an important means by which new added-value compounds can be obtained, exploring the same process for seaweeds can contribute to an effective and sustainable exploitation of marine resources. Scope and approach: In this review, recent advances demonstrating the potential behind the fermentation of seaweeds are evaluated. A breakdown of the most relevant seaweed compounds and their effect on potential bioprocesses is presented, along with pre-processing techniques that have become popular in biofuel fermentations. The applications of seaweed fermentation products in the fields of natural product research, functional foods and nutraceuticals, as well as the limitations and opportunities of seaweed fermentation are also highlighted. Key findings and conclusions: Research revealing that seaweed fermentation can be used to create novel food and nutraceutical products that demonstrate high bioactivity and sensory quality was presented. The studies included demonstrate the use of this process in algal tissues and extracts as an enhancer of antioxidant, antimicrobial, antiinflammatory and antidiabetic activities, among others. Many of the difficulties related to fermenting seaweed have been addressed by research within the field of biofuels, providing insight on the conditions and pretreatments necessary to improve seaweed fermentability. Food applications for seaweed fermentation products are still underdeveloped, but the nutritional, sensory and bioactive profiles collected so far highly encourage further developments. info:eu-repo/semantics/publishedVersion

This work focuses on understanding the action of a novel seaweed extract with anti-browning functionality in fresh-cut apples. Organic fresh-cut apples were coated by immersion in an aqueous Codium tomentosum seaweed extract (0.5 % w/v), packaged under ambient atmospheric conditions in plastic bags, and stored at 4 ˚C for 15 days. Browning-related enzymatic activities, as well as targeted gene expression related to superficial browning, were monitored immediately after coating and followed at five-day intervals, until a final storage period of 15 days. Gene expression was particularly affected one hour after coating application (day 0), with no expression registered for peroxidase (mdPOD) and phenylalanine ammonia-lyase (mdPAL) genes in the coated samples. A reduction in polyphenol oxidase expression levels was also observed. After 15 days of storage, the coated samples developed lower browning levels and presented distinctly lower activities of polyphenol oxidase and peroxidase - the oxidative enzymes predominantly involved in enzymatic browning. The observed post-coating suppression of mdPAL and mdPOD expression, and reduction in mdPPO expression, suggest that the seaweed C. tomentosum extract delays the activation of these genes, and decreases enzymatic activity, which in turn accounts for the coating's anti-browning effect.

Funding: With support from FCT/MCTES through national funds (UID/MAR/04292/2020), the European Union through EASME Blue Labs project AMALIA, Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419), project VALORMAR (Mobilizing R&TD Programs, Portugal 2020) co-funded by COMPETE (POCI-01-0247-FEDER-024517), RD&T co-promotion project ORCHESTRA (No. 70155), and SAICTPAC/0019/2015—LISBOA-01-0145-FEDER-016405 Oncologia de Precisão: Terapias e Tecnologias Inovadoras (POINT4PAC) through the European Regional Development Fund. info:eu-repo/semantics/publishedVersion

Crustaceans are amongst the most reported invaders of coastal habitats, and predatory brachyuran crabs one of the most successful marine invasive groups. They hold high tolerance to abiotic stress and their genetic, life-history, and behavioural adaptation mechanisms prompt their invasive conspicuousness. However, there is a generalized ambiguity on the specific processes that make these bioinvasions successful. No trait-related patterns have yet been found, mostly since traits attributed solely to non-indigenous crab species lack confirmation by comparison with their native counterparts and across each other. Therefore, this review discusses the available literature on fitness and plasticity of brachyuran crabs under global changes, and advances the increasing importance of addressing native versus non-native invasive species comparisons in the annals of brachyura invasion dynamics. Further on, integrative approaches are highlighted and proposed as innovative tools to disentangle trait-related tolerance, overall crab phenotypic plasticity and further adaptation. These indicator tools will advance the knowledge on crabs’ invasive potential, which ultimately affects upcoming population dynamics and ecosystem services, adding value to an effective management of coastal bioinvasions.

This study had the support of Fundação para a Ciência e Tecnologia (FCT), through project PROTEOME (PTDC/AAG-MAA/1302/2014) cofunded by COMPETE (POCI-01-0145-FEDER-016773) and the Strategic Project UID/MAR/04292/2013 granted to MARE, and the grants awarded to Luís Alves (SFRH/BD/122082/2016) and Sara Novais (SFRH/BPD/94500/2013), and also FCT and Deutscher Akademischer Austauschdienst (DAAD) program for bilateral cooperation funding Global Invaders and Stream Cukes. The authors also wish to acknowledge the Integrated Program of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020) program, Portugal 2020, European Union, through the European Regional Development Fund. The Leibniz project budget 6057 granted financial support for Holger Kühnhold. A NAM-fellowship was awarded to Lisa Indriana for a prolonged research stay in Germany. We thank Christiane Hassenrück for helpful advice concerning data analyses and interpretation. Moreover, we thank Kai Lorkowski, Nico Steinel and Christian Brandt for technical support and Sofia Esteves da Silva and Hugo Morais for great SCUBA diving support. The 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. info:eu-repo/semantics/publishedVersion

Financial support was given by FEDER through the Operational Program for Competitiveness Factors and COMPETE, through FCT and the Portuguese Foundation for Science and Technology under project PTDC/AGR/ALI/122119/2010, and projects ReNATURE - Valorization of the Natural Endogenous Resources of the Centro Region (Centro 2020, Centro-01-0145-FEDER-000007) and SmarBioR - Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate (Centro 2020, Centro-01-0145-FEDER-000018), cofunded by Centro 2020 Program, Portugal 2020, European Union, through the European Regional Development Fund. Authors also acknowledge FCT for the grant attributed to Sara Leston (grant SFRH/ BPD/91828/2012) and to João Rosa (grant SFRH/BD/102008/2014). The funding source did not interfere with the study design, or in the collection, analysis or interpretation of the data. It also had no involvement in the writing of the report and in the decision to submit the present manuscript for publication. Concerns about antibiotic use led to stricter legislations and overall better practices in aquaculture production. However, depletion time periods in tissues vary greatly with different antimicrobials, fish species, or experimental setup. In the present work, five drugs (sulfadiazine (SDZ), trimethoprim (TRI), flumequine (FLU), oxolinic acid (OXO), and oxytetracycline (OTC)) were incorporated into medicated feed, and their retention in European seabass muscle tissues assessed. Juveniles were placed in 300 L tanks at 18 °C and were manually fed with medicated feeds for a 7 day period (two concentrations per antimicrobial, ranging from 6 to 220 mg kg−1 , which were based on previous studies on the occurrence of these antimicrobials). Residues were analyzed through a multi-class quantification method (UHPLC-MS/MS). Data on residues concentration through time followed a one-compartment model, with TRI concentrations above the established Maximum Residue Level (MRL) throughout the experiment. Similarly, OTC concentrations at the highest dose were also above 100 μg kg−1 up to 14 days after the medication period. Results obtained for TRI may indicate the presence of a dose dependency for this antimicrobial. FLU concentrations in muscle samples were the lowest through time (Cday7 of 279.70 and 386.63 μg kg−1 , for prophylactic and therapeutic treatments, respectively). Half-life values of 14.37, 10.87, 5.36, 7.20, and 27.22 h (prophylactic treatment), and 20.95, 8.41, 5.61, 11.22, and 17.99 h (therapeutic treatment), were determined for SDZ, TRI, FLU, OXO, and OTC, respectively. Withdrawal times of 0, 2, and 5 days were determined for FLU, OXO, and SDZ, but sampling times for OTC and TRI should be longer, since antimicrobial concentrations were above the MRL 14 days after the end of the feeding period with medicated feed. Therefore, special attention should be given since they are the most used antibiotics in aquaculture and European seabass plays a major part in human nutrition in the Mediterranean region. info:eu-repo/semantics/publishedVersion

Anthropogenic noise is a growing threat to marine organisms, including fish. Yet very few studies have addressed the impact of anthropogenic noise on fish reproduction, especially in situ. In this study, we investigated the impacts of boat noise exposure in the reproductive success of wild Lusitanian toadfish (Halobatrachus didactylus), a species that relies on advertisement calls for mate attraction, using behavioural, physiological and reproductive endpoints. Two sets of artificial nests were deployed in the Tagus estuary and exposed to either ambient sound or boat noise during their breeding season. Toadfish males spontaneously used these nests to breed. We inspected nests for occupation and the presence of eggs in six spring low tides (in two years) and assessed male vocal activity and stress responses. Boat noise did not affect nest occupation by males but impacted reproductive success by decreasing the likelihood of receiving eggs, decreasing the number of live eggs and increasing the number of dead eggs, compared to control males. Treatment males also showed depressed vocal activity and slightly higher cortisol levels. The assessment of oxidative stress and energy metabolism-related biomarkers revealed no oxidative damage in noise exposed males despite having lower antioxidant responses and pointed towards a decrease in the activity levels of energy metabolism-related biomarkers. These results suggest that males exposed to boat noise depressed their metabolism and their activity (such as parental care and mate attraction) to cope with an acoustic stressor, consistent with a freezing defensive response/behaviour. Together, our study demonstrates that boat noise has severe impacts on reproductive fitness in Lusitanian toadfish. We argue that, at least fishes that cannot easily avoid noise sources due to their dependence on specific spawning sites, may incur in significant direct fitness costs due to chronic noise exposure. Fundação para a Ciência e Tecnologia - FCT info:eu-repo/semantics/publishedVersion

Acknowledgements : This study had the support of Fundaçao ˜ para a Ciˆencia e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2020 granted to MARE – Marine and Environmental Sciences Centre, project BLUESHARKER (PTDC/CTA-AMB/29136/2017), co-financed by COMPETE2020 (POCI-01-0145-FEDER-029136), and grant awarded to Luís Alves (SFRH/BD/122082/2016). Sara Novais is funded by national funds (OE), through FCT – Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. Authors would also like to acknowledge the fishermen enabling access to the samples, and all the members of Lemos Lab research group that helped in the laboratory during the dissection and sampling. One of the most landed sharks in Portuguese fisheries is the lesser-spotted dogfish (Scyliorhinus canicula), which is ever-present in Portuguese fish markets and consumed as cheap fish protein source. The focus of this study was to evaluate element contamination in consumed tissues of Atlantic S. canicula, with the intent of safeguarding possible public health issues. A total of 74 specimens were analysed for metals and metalloids in the deeper white muscle and skin. Arsenic, zinc, iron and aluminium were the elements with higher mean values. There was a tendency for higher levels in the skin, with differences between life-stage and gender. Many individuals surpassed stipulated guideline limits for mercury and arsenic, posing a risk for human consumption (according to the health risk assessment performed for the average Portuguese fish consumption) or even for use in feed production. Besides the public health concern, this study also evidences troubling signs on marine contamination status. info:eu-repo/semantics/publishedVersion

This study was supported by UID/MAR/04292/2020 with funding from FCT/MCTES through national funds, and by the grant awarded to Rafael Félix (SFRH/BD/139763/2018). The authors also wish to acknowledge the support of the European Union through the EASME Blue Labs project AMALIA, Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419), project VALORMAR (Mobilizing R&TD Programs, Portugal 2020), co-funded by COMPETE (POCI-01-0247-FEDER-024517), the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018) and RD&T co-promotion project ORCHESTRA (nº 70155), co-funded by Centro 2020 program, Portugal 2020, European Union, European Regional Development Fund, and SAICTPAC/0019/2015—LISBOA01-0145-FEDER-016405 Oncologia de Precisão: Terapias e Tecnologias Inovadoras (POINT4PAC), through the European Regional Development Fund. The invasive macroalga Grateloupia turuturu is known to contain a diversity of bioactive compounds with different potentialities. Among them are compounds with relevant bioactivities for cosmetics. Considering this, this study aimed to screen bioactivities with cosmeceutical potential, namely, antioxidant, UV absorbance, anti-enzymatic, antimicrobial, and anti-inflammatory activities, as well as photoprotection potential. Extractions with higher concentrations of ethanol resulted in extracts with higher antioxidant activities, while for the anti-enzymatic activity, high inhibition percentages were obtained for elastase and hyaluronidase with almost all extracts. Regarding the antimicrobial activity, all extracts showed to be active against E. coli, S. aureus, and C. albicans. Extracts produced with higher percentages of ethanol were more effective against E. coli and with lower percentages against the other two microorganisms. Several concentrations of each extract were found to be safe for fibroblasts, but no photoprotection capacity was observed. However, one of the aqueous extracts was responsible for reducing around 40% of the nitric oxide production on macrophages, showing its anti-inflammatory potential. This work highlights G. turuturu’s potential in the cosmeceutical field, contributing to the further development of natural formulations for skin protection. info:eu-repo/semantics/publishedVersion

Acknowledgements : We would like to thank the Instituto da Conservação da Natureza e Florestas (ICNF) for permits and logistical support (lodging) to conduct this work. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed and all necessary approvals have been obtained from the ICNF. Special thanks to the wardens, Paulo Crisostomo ´ and Eduardo Mourato (Reserva Natural das Berlengas). We would like to thank Jos´e Seco for valuable inputs during earlier drafts of the manuscript. LMFA and JMP acknowledge their grants (SFRH/BD/122082/2016, SFRH/BD/123499/2016, respectively) attributed by the Foundation for Science and Technology (FCT; Portugal). This study benefited from the strategic program of MARE, financed by FCT (MARE - UID/MAR/04292/2020) and the transitory norm contracts DL57/2016 to FRC (DL57/2016/CP1370/ CT90), ACN (DL57/2016/CP1370/CT89), VHP (SFRH/BPD/85024/2012) and SCN (SFRH/BPD/94500/2013). Financial support provided by FCT and MARE through a PhD grant attributed to MIL (UI/BD/150956/2021) and the project ECOLIND – Seabirds and fish as ecological indicators of contaminants and health in the North Atlantic relevant to ocean resources assessment (MFM/2017/0003) Multidisciplinary approaches are essential to diligently assess environmental health status of ecosystems. In this study, year-round chemical elements’ exposure and impacts were assessed on the wide-ranging Cory’s shearwater Calonectris borealis breeding in Berlenga Island, offshore Portugal, North Atlantic Ocean. The aim was to identify potential contamination and oxidative stress sources associated with trophic ecology, habitat and spatial use, and foraging patterns. A set of 20 chemical elements were quantified, along with oxidative stress biomarkers, stable isotope analyses, and GPS tracking data. Birds presented higher accumulation to some nonessential elements along the year (i.e. arsenic, As; cadmium, Cd; mercury, Hg; lead, Pb; and strontium, Sr), in which concentrations were similar or surpassed other procellariform seabird populations all over the world. No significant differences were found for any of the elements between different periods within the breeding season, with exception of Hg. However, a Principal Component Analysis taking into consideration a group of elements showed differences between pre-laying and chick-rearing periods, with overall higher concentrations in the former. Individuals spending more time engaging in an intensive search for food, and in more coastal environments, presented overall higher element concentrations, and particularly Hg. Contrary to expectations, no relationships were found between chemical elements and oxidative stress. On the other hand, spatial use and foraging patterns of Cory’s shearwaters influenced their oxidative stress responses. Our results highlight the need for multidisciplinary approaches to deepen understanding of the large-scale vulnerability of bioindicators such as seabirds and, by extension, the overall environmental health of ecosystems in which they rely. info:eu-repo/semantics/publishedVersion

This study was funded by Fundaçao ˜ para a Ciˆencia e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2019, project MARINE INVADERS – The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI01-0145-FEDER-031144), and the grants awarded to Carla Silva (SFRH/ BD/87030/2012). The project was also partially funded by European Union through EASME Blue Labs project AMALIA—Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419), and the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145- FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. Authors are also thankful to all the team members for the help during field work Biological invasions represent a threat to ecosystems, through competition and habitat destruction, which may result in significant changes of the invaded community. Asparagopsis armata Harvey 1855 is a red macroalgae (Rodophyta) globally recognized as an invasive species. It is found from the intertidal to shallow subtidal areas, on rock or epiphytic, forming natural vegetation belts on exposed coasts. This study evaluated the variations on native intertidal seaweed and macroinvertebrate assemblages inhabiting rock pools with and without the presence of the invasive macroalgae A. armata. To achieve this, manipulation experiments on Atlantic (Portugal) rock pools were done. Three rock pools were maintained without A. armata by manual removal of macroalgae, and three others were not experimentally manipulated during the study period and A. armata was freely present. In this study the variations between different rock pools were assessed. Results showed different patterns in the macroalgae composition of assemblages but not for the macrobenthic communities. Ellisolandia elongata (J. Ellis & Solander) K.R. Hind & G.W. Saunders 2013 was the main algal species affected by the invasion of A. armata. Invaded pools tended to show less macroalgal species richness, showing a more constant and conservative structure, with lower variation of its taxonomic composition than the pools not containing A. armata, where the variability between samples was always higher. Despite the importance of the achieved results, further data based on observation of long-term series are needed, in order to further understand more severe effects of the invader A. armata on native macroalgal assemblage. info:eu-repo/semantics/publishedVersion

Funding : This study was supported by FCT/MCTES through funding to MARE (UID/MAR/04292/2020) and by the grant awarded to Rafael Félix (SFRH/ BD/139763/2018). The authors also wish to acknowledge the support of the European Union through EASME Blue Labs project AMALIA (EASME/EMFF/2016/1.2.1.4/03/SI2.750419), project VALORMAR (Mobilizing R&TD Programs, Portugal 2020) cofunded by COMPETE (POCI-01-0247-FEDER-024517), the Integrated Programme of SR&TD SmartBioR (reference Centro-01-0145-FEDER-000018), and RD&T copromotion project ORCHESTRA (n◦ 70155) cofunded by Centro 2020 program, Portugal 2020, European Union, European Regional Development Fund, and SAICTPAC/0019/2015—LISBOA-01-0145-FEDER POINT4PAC through the European Regional Development Fund, and MACAU (MAR-04.03.01-FEAMP-0128) and COSMOS (MAR04.03.01-FEAMP-0370) cofounded by GAL PESCA OESTE and MAR2020 in the framework of PORTUGAL2020 through the European Maritime and Fisheries Fund. Marine biomass is a resource with great potential to fuel a circular bioeconomy addressing the current economic and environmental world crisis, while seaweeds are known to be a great source of bioactive compounds with potential use for human and animal nutrition, novel natural preservatives, phytopharmaceuticals, cosmeceuticals, biopharmaceuticals, among others. Asparagopsis armata (Harvey, 1855) is a seaweed with high biomass production and exquisite biotechnological potential due to a myriad of produced secondary metabolites. Furthermore, it is one of the most aggressive invasive seaweed species becoming an economic and ecological problem in Europe. Simultaneously, it is a resource with market demand for its applications in cosmetics and, more recently, for its potential anti-methanogenic activity in ruminants. There is an appreciable amount of literature concerning the chemistry and the bioactivity of this seaweed species (and particularly its gametophyte, the focus of this review), as well as multiple products in the cosmetics market, but all this information is scattered and therefore poorly used. To potentiate the bioindustry of A. armata, the available scientific knowledge is here reviewed and an overview of the market for this species is comprehensively provided. Thus, the chemical composition of A. armata (primary metabolism, polysaccharide characteristics, amino acid profile, and nonhalogenated and halogenated secondary metabolites), its bioactivities (antibacterial, antifungal, antiviral, antiprotozoal, antioxidant, antitumor, anti-inflammatory, among others) and the existent, marketed extracts (cosmetic ingredients) are thoroughly compiled and discussed herein. info:eu-repo/semantics/publishedVersion

Acknowledgments : We are thankful to the Air Force Base No. 6 of Montijo (Portugal) for allowing the field study in their military establishment. We are also grateful to Marilia Freire and Daniel Alves for the valuable assistance during field work. This study was funded by the Science and Technology Foundation, Portugal, project PTDC/BIA-BMA/29662/2017; strategic projects UID/MAR/04292/2020 to A.F. and M.C.P.A. by MARE and UID/BIA/00329/2020 to P.J.F. by cE3c; grant SFRH/BD/115562/2016 to M.V. Marine traffic is themost common and chronic source of ocean noise pollution. Despite the evidence of detrimental effects of noise exposure on fish, knowledge about the effects on the critical early life stages - embryos and larvae - is still scarce. Here, we take a natural habitat-based approach to examine potential impacts of boat noise exposure in early life stages in a wild fish population of the Lusitanian toadfish (Halobatrachus didactylus). In-situ experiments were carried out in the Tagus estuary, an estuary with significant commercial and recreational boat traffic. Nestswith eggswere exposed to either ambient (control) or boat noise (treatment), for 1 fortnight. Eggs were photographed before being assigned to each treatment, and after exposure, to count number of eggs and/or larvae to assess survival, and sampled to study development and oxidative stress and energy metabolism-related biomarkers. Data concerns 4 sampling periods (fortnights) from 2 years. Results indicate that offspring survival did not differ between treatments, but boat noise induced a detrimental effect on embryos and larvae stress response, and on larvae development. Embryos showed reduced levels of electron transport system (ETS), an energy metabolism-related biomarker, while larvae showed higher overall stress responses, with increased levels of superoxide dismutase (SOD) and DNA damage (oxidative stress related responses), ETS, and reduced growth. With this study,we provided the first evidence of detrimental effects of boat noise exposure on fish development in the field and on stress biomarker responses. If these critical early stages are not able to compensate and/or acclimate to the noise stress later in the ontogeny, then anthropogenic noise has the potential to severely affect this and likely other marine fishes, with further consequences for populations resilience and dynamics. info:eu-repo/semantics/publishedVersion

The authors would like to thank Fundação para a Ciência e a Tecnologia (FCT) for funding the research via project grants PTDC/MAR-EST/3048/2014 (BIOPHARMA), PTDC/CTAAMB/30056/2017 (OPTOX), UIDB/04292/2020 and UID/MULTI/04046/2019. Work was also funded by the Integrated Programme of SR&TD SmartBioR (reference Centro-01-0145-FEDER000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. B. Duarte and V. F. Fonseca were supported by researcher contracts (CEECIND/00511/2017 and DL57/2016/CP1479/CT0024). Worldwide anthropogenic activities result in the production and release of potentially damaging toxic pollutants into ecosystems, thereby jeopardizing their health and continuity. Research studies and biomonitoring programs attend to this emerging problematic by applying and developing statistically relevant indexes that integrate complex biomarker response data to provide a holistic approach, reflecting toxically induced alterations at the organism or population level. Ultimately, indexes allow simple result communications, enhancing policy makers understanding, and contributing to better resource and environmental managing policies. In this study three indexes, the integrated biomarker response index (IBR), the bioeffects assessment index (BAI) and principal components analysis (PCA), were evaluated for their sensitivity in revealing toxically induced stress patterns in cells of the diatom Phaeodactylum tricornutum under contaminant exposure. The set of biomarkers selected for index construction comprised the anti-oxidant enzymes APX, CAT and SOD, and the lipid peroxidation marker TBARS. Several significant correlations with the applied concentration gradients were noticed for all indexes, although IBR excelled for its reliability in delivering statistically significant dose-response patterns for four out of the five tested compounds. info:eu-repo/semantics/publishedVersion

Invasive macroalgae represent one of the major threats to marine biodiversity, ecosystem functioning and structure, as well as being important drivers of ecosystem services depletion. Many such species have become well established along the west coast of the Iberian Peninsula. However, the lack of information about the distribution of the invaders and the factors determining their occurrence make bioinvasions a difficult issue to manage. Such information is key to enabling the design and implementation of effective management plans. The present study aimed to map the current probability of presence of six invasive macroalgae: Grateloupia turuturu, Asparagopsis armata, Colpomenia peregrina, Sargassum muticum, Undaria pinnatifida, and Codium fragile ssp. fragile. For this purpose, an extensive field survey was carried out along the coast of the north-western Iberian Peninsula. Species distribution models (SDMs) were then used to map the presence probability of these invasive species throughout the study region on the basis of environmental and anthropogenic predictor variables. The southern Galician rias were identified as the main hotspots of macroalgal invasion, with a high probability of occurrence for most of the species considered. Conversely, the probability of presence on the Portuguese coast was generally low. Physico-chemical variables were the most important factors for predicting the distribution of invasive macroalgae contributing between 57.27 and 85.24% to the ensemble models. However, anthropogenic factors (including size of vessels, number of shipping lines, distance from ports, population density, etc.) considerably improved the estimates of the probability of occurrence for most of the target species. This study is one of the few to include anthropogenic factors in SDMs for invasive macroalgae. The findings suggest that management actions aimed at controlling these species should strengthen control and surveillance at ports, particularly in southern Galician rias. Early detection should be of main concern for risk assessment plans on the Portuguese coast., A. B. was supported by the Axencia Galega de Innovación (GAIN), Xunta de Galicia (grant ED481B 2018/68). This work was supported by the European Union through EASME Blue Labs project AMALIA—Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/ 1.2.1.4/03/SI2.750419). The authors acknowledge financial support provided by the Portuguese Foundation for Science and Technology (FCT) through Strategic Project UID/MAR/04292/2019 granted to MARE – Marine and Environmental Sciences Centre, through MARINE INVADERS Project (PTDC/BIA-CBI/31144/2017), co-financed by COMPETE (POCI-01-0145-FEDER-031144), and the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145- FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund.

Funding: This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2020 grants to MARE, project MARINE INVADERS—The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI-01-0145-FEDER-031144), and grant awarded to Lénia Rato (SFRH/BD/138492/2018). The project was also partially funded by “Global Invaders—Global trends among valued aquatic invertebrate species: competitive advantages across different latitudes” funded by FCT and DAAD- Deutscher Akademischer Austauschdienst. Sara Novais is funded by national funds (OE), through FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of 29 August, changed by Law 57/2017, of 19 July. Sulfoxaflor is an insecticide belonging to the recent sulfoximine class, acting as a nicotinic acetylcholine receptor (nAChRs) agonist. There are few studies regarding sulfoxaflor’s toxicity to non-target organisms. The present study aimed to investigate the acute and sub-lethal effects of sulfoxaflor on Carcinus maenas by addressing survival, behaviour (feed intake and motricity), and neuromuscular, detoxification and oxidative stress, and energy metabolism biomarkers. Adult male green crabs were exposed to sulfoxaflor for 96 h and an LC50 of 2.88 mg L􀀀1 was estimated. All biomarker endpoints were sampled after three (T3) and seven (T7) days of exposure and behavioural endpoints were addressed at T3 and day six (T6). Sulfoxaflor affected the feed intake and motricity of C. maenas at T6. From the integrated analysis of endpoints, with the increase in concentrations of sulfoxaflor, after seven days, one can notice a lower detoxification capacity (lower GST), higher LPO levels and effects on behaviour (higher motricity effects and lower feed intake). This integrated approach proved to be valuable in understanding the negative impacts of sulfoxaflor on green crabs, while contributing to the knowledge of this pesticide toxicity to non-target coastal invertebrates. info:eu-repo/semantics/publishedVersion

Funding: This research was funded by the Ministry of Science and Higher Education of the Russian Federation, project number FZWM-2021-0016, and Fundação para a Ciência e a Tecnologia through the Strategic Project UID/MAR/04292/2020. Polysaccharides are complex macromolecules long regarded as energetic storage resources or as components of plant and fungal cell walls. They have also been described as plant mucilages or microbial exopolysaccharides. The development of glycosciences has led to a partial and difficult deciphering of their other biological functions in living organisms. The objectives of glycobiochemistry and glycobiology are currently to correlate some structural features of polysaccharides with some biological responses in the producing organisms or in another one. In this context, the literature focusing on bioactive polysaccharides has increased exponentially during the last two decades, being sometimes very optimistic for some new applications of bioactive polysaccharides, notably in the medical field. Therefore, this review aims to examine bioactive polysaccharide, taking a critical look of the different biological activities reported by authors and the reality of the market. It focuses also on the chemical, biochemical, enzymatic, and physical modifications of these biopolymers to optimize their potential as bioactive agents. info:eu-repo/semantics/publishedVersion

Present demographic trends suggest a rise in the contributions of human pharmaceuticals into coastal ecosystems, underpinning an increasing demand to evaluate the ecotoxicological effects and implications of drug residues in marine risk assessments. Propranolol, a non-selective &beta, adrenoceptor blocker, is used worldwide to treat high blood pressure conditions and other related cardiovascular conditions. Although diatoms lack &beta, adrenoceptors, this microalgal group presents receptor-like kinases and proteins with a functional analogy to the animal receptors and that can be targeted by propranolol. In the present work, the authors evaluated the effect of this non-selective &beta, adrenoceptor blocker in diatom cells using P. tricornutum as a model organism, to evaluate the potential effect of this compound in cell physiology (growth, lipids and energy metabolism and oxidative stress) and its potential relevance for marine ecosystems. Propranolol exposure leads to a significant reduction in diatom cell growth, more evident in the highest concentrations tested. This is likely due to the observed impairment of the main primary photochemistry processes and the enhancement of the mitochondrial respiratory activity. More specifically, propranolol decreased the energy transduction from photosystem II (PSII) to the electron transport chain, leading to an increase in oxidative stress levels. Cells exposed to propranolol also exhibited high-dissipated energy flux, indicating that this excessive energy is efficiently diverted, to some extent, from the photosystems, acting to prevent irreversible photoinhibition. As energy production is impaired at the PSII donor side, preventing energy production through the electron transport chain, diatoms appear to be consuming storage lipids as an energy backup system, to maintain essential cellular functions. This consumption will be attained by an increase in respiratory activity. Considering the primary oxygen production and consumption pathways, propranolol showed a significant reduction of the autotrophic O2 production and an increase in the heterotrophic mitochondrial respiration. Both mechanisms can have negative effects on marine trophic webs, due to a decrease in the energetic input from marine primary producers and a simultaneous oxygen production decrease for heterotrophic species. In ecotoxicological terms, bio-optical and fatty acid data appear as highly efficient tools for ecotoxicity assessment, with an overall high degree of classification when these traits are used to build a toxicological profile, instead of individually assessed.

Glyphosate is the main active component of the commercial formulation Roundup&reg, the most widely used chemical herbicide worldwide. However, its potential high toxicity to the environment and throughout trophic webs has come under increasing scrutiny. The present study aims to investigate the application of bio-optical techniques and their correlation to physiological and biochemical processes, including primary productivity, oxidative stress, energy balance, and alterations in pigment and lipid composition in Phaeodactylum tricornutum, a representative species of marine diatoms, using the case study of its response to the herbicide glyphosate-based Roundup&reg, formulation, at environmentally relevant concentrations. Cultures were exposed to the herbicide formulation representing effective glyphosate concentrations of 0, 10, 50, 100, 250, and 500 &mu, g L&minus, 1. Results showed that high concentrations decreased cell density, furthermore, the inhibition of photosynthetic activity was not only caused by the impairment of electron transport in the thylakoids, but also by a decrease of antioxidant capacity and increased lipid peroxidation. Nevertheless, concentrations of one of the plastidial marker fatty acids had a positive correlation with the highest concentration as well as an increase in total protein. Cell energy allocation also increased with concentration, relative to control and the lowest concentration, although culture growth was inhibited. Pigment composition and fatty acid profiles proved to be efficient biomarkers for the highest glyphosate-based herbicide concentrations, while bio-optical data separated controls from intermediate concentrations and high concentrations.

The red seaweed Asparagopsis armata exhibits a strong invasive behaviour and is included in the list of the “Worst invasive alien species threatening biodiversity in Europe”. This seaweed has been shown to produce a large diversity of halogenated compounds with effective biological effects, deeply affecting rockpool species. Therefore, the present study aimed to investigate the biochemical responses to sublethal concentrations of Asparagopsis armata exudate on two coastal organisms, the marine snail Gibbula umbilicalis and the rockpool shrimp Palaemon elegans. Antioxidant defences superoxide dismutase (SOD) and glutathione-S-transferase (GST), oxidative damage endpoints lipid peroxidation (LPO) and DNA damage, the neuronal parameter acetylcholinesterase (AChE), as well as the fatty acid profile were evaluated. Results revealed different metabolic responses between species, indicating that A. armata exudate affected the organisms through different pathways. Despite previous studies indicating that the exudate effected G. umbilicalis’ survival and behaviour, this does not seem to result from oxidative stress or addressed neurotoxicity. On the other hand, for P. elegans, an inhibition of AChE and the decrease of antioxidant capacity concomitant with the increase of LPO, suggests neurotoxicity and oxidative stress as mechanisms of exudate toxicity for this species. For fatty acids, there were different profile changes between species, also more pronounced for P. elegans with a general increase in PUFA with exudate exposure, which commonly means a defence mechanism protecting from membrane disruption. Nonetheless, the omega-3 PUFAs ARA and DPA were increased in both invertebrates, indicating a common mechanism regulation of inflammation and immunity responses to this stress. This work provides further insight on the mechanisms of invertebrate response and tolerance to an expanding coastal environmental stress as is the marine invader A. armata., This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2013, project MARINE INVADERS – The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI‐01‐0145‐FEDER‐031144), and grant awarded to Carla Silva (SFRH/BD/87030/2012). The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro‐01‐0145‐FEDER‐000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. Authors also thank Sofia Silva (MARE, Portugal) for help in laboratorial work.

The red seaweed Asparagopsis armata exhibits a strong invasive behaviour and is included in the list of the “Worst invasive alien species threatening biodiversity in Europe”. This seaweed has been shown to produce a large diversity of halogenated compounds with effective biological effects, deeply affecting rockpool species. Therefore, the present study aimed to investigate the biochemical responses to sublethal concentrations of Asparagopsis armata exudate on two coastal organisms, the marine snail Gibbula umbilicalis and the rockpool shrimp Palaemon elegans. Antioxidant defences superoxide dismutase (SOD) and glutathione-S-transferase (GST), oxidative damage endpoints lipid peroxidation (LPO) and DNA damage, the neuronal parameter acetylcholinesterase (AChE), as well as the fatty acid profile were evaluated. Results revealed different metabolic responses between species, indicating that A. armata exudate affected the organisms through different pathways. Despite previous studies indicating that the exudate effected G. umbilicalis’ survival and behaviour, this does not seem to result from oxidative stress or addressed neurotoxicity. On the other hand, for P. elegans, an inhibition of AChE and the decrease of antioxidant capacity concomitant with the increase of LPO, suggests neurotoxicity and oxidative stress as mechanisms of exudate toxicity for this species. For fatty acids, there were different profile changes between species, also more pronounced for P. elegans with a general increase in PUFA with exudate exposure, which commonly means a defence mechanism protecting from membrane disruption. Nonetheless, the omega-3 PUFAs ARA and DPA were increased in both invertebrates, indicating a common mechanism regulation of inflammation and immunity responses to this stress. This work provides further insight on the mechanisms of invertebrate response and tolerance to an expanding coastal environmental stress as is the marine invader A. armata.

Biological invasions represent a threat to ecosystems, through competition and habitat destruction, which may result in significant changes of the invaded community. Asparagopsis armata Harvey 1855 is a red macroalgae (Rodophyta) globally recognized as an invasive species. It is found from the intertidal to shallow subtidal areas, on rock or epiphytic, forming natural vegetation belts on exposed coasts. This study evaluated the variations on native intertidal seaweed and macroinvertebrate assemblages inhabiting rock pools with and without the presence of the invasive macroalgae A. armata. To achieve this, manipulation experiments on Atlantic (Portugal) rock pools were done. Three rock pools were maintained without A. armata by manual removal of macroalgae, and three others were not experimentally manipulated during the study period and A. armata was freely present. In this study the variations between different rock pools were assessed. Results showed different patterns in the macroalgae composition of assemblages but not for the macrobenthic communities. Ellisolandia elongata (J. Ellis & Solander) K.R. Hind & G.W. Saunders 2013 was the main algal species affected by the invasion of A. armata. Invaded pools tended to show less macroalgal species richness, showing a more constant and conservative structure, with lower variation of its taxonomic composition than the pools not containing A. armata, where the variability between samples was always higher. Despite the importance of the achieved results, further data based on observation of long-term series are needed, in order to further understand more severe effects of the invader A. armata on native macroalgal assemblage.

This study was supported by Fundação para a Ciência e a Tecnologia (FCT) and FEDER through the project BLUESHARKER (PTDC/CTAAMB/29136/2017) and the Strategic Project UID/MAR/04292/2019. The project was also funded by the Integrated Programme of SR&TD “SmartBioR” (Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. Luís M.F. Alves also wishes to L.M.F. Alves, et al. Fisheries Research 230 (2020) 105648 7 acknowledge the financial support given by FCT (SFRH/BD/122082/ 2016). This study was also supported by Oceanário de Lisboa and Oceano Azul Foundation through the project Shark Attract, winner of the first edition of the Ocean’s Conservation Fund. Portugal plays a major role in shark, skate and ray fisheries in the European Union. With the decline of these animals raising concerns amongst scientists all over the world, we set out to provide an updated assessment on elasmobranch landings in Portugal between 1986 and 2017. The dataset analysed in this study consisted of 15,521 records, each with information concerning the species (or higher taxonomic level), port, month, year, and weight of fish landed. A non-parametric change points assessment statistical technique was used to evaluate important changes in landings throughout the time series. Principal components analysis was also performed to mean values of landings per year for the most captured species, and similar analysis was conducted using fishing ports instead of year, for the same sub-set of species / taxa. According to the data, more than 143 thousand metric tons of elasmobranchs were landed in Portuguese ports between 1986 and 2017, divided by 58 taxa of sharks, skates and rays. The most landed taxa were demersal rays and skates (Raja spp.), followed by spotted dogfishes (Scyliorhinus spp.), blue shark (Prionace glauca), Portuguese dogfish (Centroscymnus coelolepis), leafscale gulper shark (Centrophorus squamosus), gulper shark (Centrophorus granulosus) and shortfin mako (Isurus oxyrinchus). Together, these seven taxa accounted for more than 75% of all landings. Several cases of misreporting were identified and discussed. Overall, landings of elasmobranchs in Portugal show marked decreases and changes in composition, with deep-sea sharks being the most influential group during the studied period. info:eu-repo/semantics/publishedVersion

This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2013, project MARINE INVADERS – The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI01-0145-FEDER-031144) and BLUESHARKER – BLUE Shark as biomonitor of Atlantic waters through an integrated biomarker assessment (PTDC/BIA-CBI/29136/2017). This work was also supported by FCT and Deutscher Akademischer Austauschdienst (DAAD) through project GLOBAL INVADERS - GLOBAL Impacts and treNds for VAlued aquatic species: competitive aDvantages across diffERent latitudeS. The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. The stay abroad was financially supported by ERASMUS+. In nature, a multitude of factors influences the fitness of an organismat a given time,whichmakes single stressor assessments far from ecologically relevant scenarios. This study focused on the effects of water temperature and predation stress on the metabolismand bodymass gain of a common intertidal rock pool fish, Gobius paganellus, addressing the following hypotheses: (1) the energy metabolism of G. paganellus under predation stress is reduced; (2) G. paganellus shows thermal compensation under heat stress; and (3) thermal stress is the dominant stressor that may override predation stress responses. Individuals were exposed to simulated predation stress and temperature increase from 20 °C to 29 °C, and both stressors combined. Physiological effects were addressed using biochemical biomarkers related with energy metabolism (isocitrate dehydrogenase, lactate dehydrogenase, energy available, energy consumption rates), oxidative stress (superoxide dismutase, catalase, DNA damage, lipid peroxidation), and biotransformation (glutathione-S-transferase). The results of this study revealed that predation stress reduced the cellular metabolism of G. paganellus, and enhanced storage of protein reserves. As hypothesized, hyperthermia decreased the aerobic mitochondrial metabolism, indicating thermal compensationmechanisms to resist against unfavourable temperatures. Hyperthermia was the dominant stressor overriding the physiological responses to predation stress. Both stressors combined might further have synergistically activated detoxification pathways, even though not strong enough to counteract lipid peroxidation and DNA damage completely. The synergistic effect of combined thermal and predation stress thus may not only increase the risk of being preyed upon, but also may indicate extra energy trade-off for the basal metabolism,which in turnmay have ecologically relevant consequences for general body functions info:eu-repo/semantics/publishedVersion

The marine red algae Asparagopsis armata is an invasive species which competitive advantage arises from the production and release of large amounts of toxic compounds to the surrounding invaded area, reducing the abundance of native species. 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 macroalgae. The seaweed was collected and placed in tanks, for 12 hours, in the dark in a 1:10 ratio. Afterwards the media containing its secondary metabolites was collected for further testing. Lethal and sublethal effects of A. armata were investigated. Biochemical biomarkers responses associated with energy metabolism (lactate dehydrogenase, LDH; electron transport system activity, ETS; content in lipids, proteins and carbohydrates) were analysed. The biomarker responses showed invertebrates’ physiological status impairment after exposure to low concentrations of this algae exudate. Highest concentrations of exudate significantly increased lipid content in both organisms. In the shrimp, protein content, ETS, and LDH were also significantly increased. On the contrary, these parameters were significantly decreased in G. umbilicalis. A behavioural impairment was also observed in G. umbilicalis exposed to A. armata exudate, with reduction in 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.

Grateloupia turuturu is an invasive macroalga on the Iberian coast, known to produce bioactive compounds with different cosmeceutical bioactivities, namely UV shielding and antioxidants. The goal of this study was to optimize the extraction procedure of main bioactivities of this species with cosmetic potential, using Response Surface Methodology. Two Box&ndash, Behnken designs were used to evaluate the effect of ethanol concentration (0&ndash, 50%), liquid-solid ratio, time, pH, and temperature on yield, UV absorbance, and antioxidant activity. Both optimizations showed a similar trend: aqueous extracts have higher yields and extracts performed with ethanol as part of the solvent have higher activities concerning UV absorbance and antioxidant activity. For all the extracts an absorption peak between 320 and 340 nm was observed. This data now allows further studies by narrowing the extracts worthful of characterization. The development of industry-friendly extraction methods allows the valorization of this invasive species, contributing for the potential creation of natural and eco-friendly products by the cosmetic industry while contributing to the restoration of affected environments.

The authors would like to thank Fundaçao para a Ciencia e a Tecnologia (FCT) for funding the research via project grants PTDC/CTAAMB/30056/2017 (OPTOX), PTDC/AAG-MAA/1302/2014 (PROTEOME), UID/MAR/04292/2019 and UID/MULTI/04046/2013. B. Duarte and V. F. Fonseca were supported by investigation contracts (CEECIND/00511/2017 and DL57/2016/CP1479/CT0024). P. Reis-Santos was supported by FCT through a postdoctoral grant (SFRH/BPD/95784/ 2013). M.T.Cabrita is also supported by a DL57 investigation contract. Work was also funded by the Integrated Programme of SR&TD SmartBioR (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. Emerging pollutants such as pharmaceuticals are continuously released to aquatic environments posing a rising threat to marine ecosystems. Yet, monitoring routines and ecotoxicity data on biota worldwide for these substances are lacking. Non-steroidal anti-inflammatory drugs are among the most prescribed and found pharmaceuticals in aquatic environments. The toxicity effects of environmentally relevant concentrations of ibuprofen on primary productivity, oxidative stress and lipid metabolism of the diatom Phaeodactylum tricornutum were assessed. Diatom cultures were exposed to 0, 0.8, 3, 40, 100 and 300 μg L-1 ibuprofen concentrations, usually found in the vicinity of wastewater treatment plants and coastal environments. Higher concentrations (100 and 300 μg L-1) had a negative impact in P. triconutum growth, inhibiting the chloroplastic energy transduction in the electron transport chain resulting in lower energy reaching the PS I (r2 = - 0.55, p < 0.05). In contrast, the mitochondrial electron transport and available energy increased (r2 = 0.68 and r2 = 0.85, p < 0.05 respectively), mostly due to enhancements in lipid and protein contents as opposed to reduction of carbohydrates. A general up-regulation of the antioxidant enzymes could contributed to alleviate oxidative stress resulting in the decrease of lipid peroxidation products (r2 = 0.77, p < 0.05). Canonical analysis of principal components was performed and successfully discriminated exposure groups, with optical data excelling in classifying samples to different ibuprofen concentrations, being potentially used as environmental indicators. Finally, the identified mild to severe effects of ibuprofen on diatoms are likely to be exacerbated by the sustained use of this drug worldwide, underpinning the urgency of evaluating the impacts of this pharmaceutical on coastal and marine trophic webs. info:eu-repo/semantics/publishedVersion

This study aimed to evaluate the effect of ethanolic extract of Pterospartum tridentatum flowers in the stability of sunflower oil. The extract was characterized regarding to its antioxidant activity by the 2,2,1-diphenyl-1-picrylhydrazyl (DPPH) scavenging method (EC50 = 76.3 ± 2.6 µg/mL) and total phenolic content (200 ± 8 mg GAE/g). Extracts were added at 500 mg/L (E1) and 1,000 mg/L (E2), and after 30 days of storage at room temperature, E2 oil showed improved quality parameters, with a reduction of 22.4%, 17.2%, and 45.6% in the values of acidity, peroxide, and p-anisidine, respectively. The extract also increased oil stability at 180 °C. After 27 hr, the acidity (0.216 ± 0.016 mg KOH/g) and the total oxidation value (TOTOX) (69.30 ± 0.26) values of E2 oil were significantly lower than the control. These results showed that P. tridentatum effectively improved the shelf-life and thermal stability of sunflower oil, being a promising source of antioxidants for edible oils processing. PRACTICAL APPLICATION: Pterospartum tridentatum ethanolic extracts increased stability of sunflower oil during storage at room temperature and at 180 °C. Oil stability increased with extract concentration, being highest for 1,000 mg/L. Therefore, P. tridentatum may be a promising source of antioxidants for edible oils processing.

This work was supported by the European Union through EASME Blue Labs project AMALIA - Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419). Further support was provided by Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2013 and project MARINE INVADERS – The impact and mechanisms of success of the invasive seaweed Asparagopsis armata on coastal environments (POCI-01-0145-FEDER-031144). The project was also partly funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund, and Operational Programme MAR2020 through project MACAU (MAR-04.03.01-FEAMP-0128). These funding bodies were not directly involved in the development of the current study. Data on the number of licences and people landing in the Berlengas Natural Park were provided by Instituto da Conservação da Natureza e das Florestas and Life Berlengas, LIFE13/NAT/PT/000458, funded by LIFE program. The authors thank J. Pascual for assistance during SCUBA diving fieldwork, A. R. Larrinaga for developing the distribution maps, and E. Pérez and A. Fuentes for their assistance in laboratory sampling Marine ecosystems are facing major anthropogenic disturbances, including loss of biodiversity, eutrophication, and biological invasions. Thus, attention has raised on marine conservation actions to preserve habitat resilience and biodiversity. Marine Protected Areas (MPAs) play an essential role in marine conservation as they are usually designated to provide marine ecosystem resilience of native communities to human-induced impacts (including non-native introductions) while contributing with positive effects on other ecosystem services. The introduction of a new species in novel marine habitats has been attributed to biotic, abiotic, and anthropogenic factors. In the present study, the effects of native functional diversity, wave exposure were studied, and for the first time, marine protection was addressed concerning the invasion success of six macroalgae in two MPAs in the NW Iberian Peninsula. The correlation between the presence/absence of some native functional groups and the invasion success of some invasive species highlighted the importance of conserving native canopy-formers. Despite local differences, wave exposure did not affect invasion success. The protection provided by both MPAs was very limited to prevent the establishment and spread of the most abundant invasive macroalgae. Therefore, stricter management plans should be implemented to ensure native ecosystem resilience within the MPAs. info:eu-repo/semantics/publishedVersion

This study was supported by UIDB/50006/2020 with funding from FCT/MCTES through national funds, by the FCT Strategic Project UID/MAR/04292/2019 granted to MARE, and by the grants awarded to Rafael Félix (SFRH/ BD/139763/2018). The authors also wish to acknowledge the support of the European Union through EASME Blue Labs project AMALIA, Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/SI2.750419), project VALORMAR (Mobilizing R&TD Programs, Portugal 2020) co-funded by COMPETE (POCI-01-0247-FEDER-024517), the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund, SAICTPAC/0019/2015—LISBOA-01-0145-FEDER-016405 Oncologia de Precisão: Terapias e Tecnologias Inovadoras (POINT4PAC), and project “Valorização dos subprodutos do processo biotecnológico de produção de esqualeno e DHA pela microalga Aurantiochytrium sp.” (AlgaValue) (ref. 17680), ANI—Agência Nacional de Inovação, S.A., FEDER—Fundo Europeu de Desenvolvimento Regional. Lipid peroxidation is a chemical reaction known to have negative impacts on living organisms’ health and on consumer products’ quality and safety. Therefore, it has been the subject of extensive scientific research concerning the possibilities to reduce it, both in vivo and in nonliving organic matrices. It can be started by a variety of oxidants, by both ROS-dependent and -independent pathways, all of them reviewed in this document. Another feature of this reaction is the capacity of lipid peroxyl radicals to react with the non-oxidized lipids, propagating the reaction even in the absence of an external trigger. Due to these specificities of lipid peroxidation, regular antioxidant strategies—although being helpful in controlling oxidative triggers—are not tailored to tackle this challenge. Thus, more suited antioxidant compounds or technologies are required and sought after by researchers, either in the fields of medicine and physiology, or in product development and biotechnology. Despite the existence of several laboratory procedures associated with the study of lipid peroxidation, a methodology to perform bioprospecting of natural products to prevent lipid peroxidation (a Lipid Peroxidation Inhibitory Potential assay, LPIP) is not yet well established. In this review, a critical look into the possibility of testing the capacity of natural products to inhibit lipid peroxidation is presented. In vitro systems used to peroxidize a lipid sample are also reviewed on the basis of lipid substrate origin, and, for each of them, procedural insights, oxidation initiation strategies, and lipid peroxidation extent monitoring are discussed. info:eu-repo/semantics/publishedVersion

Authors would like to acknowledge the financial support of the Portuguese Foundation for Science and Technology (FCT) through Strategic Project UID/MAR/04292/2019 and UID/Multi/04046/2019 granted to MARE – Marine and Environmental Sciences Centre; through Red2Discovery Project (PTDC/MAR-BIO/6149/2014), co-financed by COMPETE (POCI-01-0145-FEDER-016791); through CrossAtlantic project (PTDC/BIA-OUT/29250/2017), co-financed by COMPETE (POCI-01-0145-FEDER-029250); through Jellyfisheries (POCI-01-0145-FEDER016772) and VALORMAR (Mobilizing R&TD Programs, Portugal 2020) co-funded by COMPETE (POCI-01-0247-FEDER-024517). The authors also wish to acknowledge the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), cofunded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund, and SAICTPAC/ 0019/2015 - LISBOA-01-0145-FEDER-016405 Oncologia de Precisão: Terapias e Tecnologias Inovadoras (POINT4PAC). This work was also supported by the European Union through EASME Blue Labs project AMALIA—Algae-to-MArket Lab IdeAs (EASME/EMFF/2016/1.2.1.4/03/ SI2.750419). Susete Pinteus, Celso Alves, Joana Silva and Sónia C. Marques were financial supported by FCT grants SFRH/BD/96203/ 2013, SFRH/BD/97764/2013, SFRH/BD/103255/2014 and SFRH/BPD/ 110400/2015, respectively. Although in the last decades significant advances have been made to improve antifouling formulations, the main current options continue to be highly toxic tomarine environment, leading to an urgent need for new safer alternatives. For anti-adherence studies, barnacles and mussels are commonly the first choice for experimental purposes. However, the use of these organisms involves a series of laborious and time-consuming stages. In the present work, a new approach for testing antifouling formulations was developed under known formulations and novel proposed options. Due to their high resilience, ability of surviving in hostile environments and high abundance in different ecosystems, medusa polyps present themselves as prospect candidates for antifouling protocols. Thus, a complete protocol to test antifouling formulations using polyps is presented, while the antifouling properties of two invasive seaweeds, Asparagopsis armata and Sargassum muticum, were evaluated within this new test model framework. The use of medusa polyps as model to test antifouling substances revealed to be a reliable alternative to the conventional organisms, presenting several advantages since the protocol is less laborious, less time-consuming and reproductive. The results also show that the seaweeds A. armata and S. muticum produce compounds with anti-adherence properties being therefore potential candidates for the development of new greener antifouling formulations. info:eu-repo/semantics/publishedVersion

Funding: This study had the support of the Fundação para a Ciência e a Tecnologia (FCT) through MARE: UIDB/04292/2020, CESAM: UIDP/50017/2020+UIDB/50017/2020, FCT/MCTES through national funds, the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020, and project PROTEOME (PTDC/AAG-MAA/1302/2014) co-financed by COMPETE (POCI-01-0145-FEDER-016773). The project was also partially funded by the Integrated Program of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. Hugo R. Monteiro and João L.T. Pestana wish to acknowledge the financial support given by FCT (SFRH/BD/80988/2011, and IF/01420/2015, respectively). The potential of proteome responses as early-warning indicators of insecticide exposure was evaluated using the non-biting midge Chironomus riparius (Meigen) as the model organism. Chironomus riparius larvae were exposed to environmentally relevant concentrations of the neurotoxic pesticide spinosad to uncover molecular events that may provide insights on the long-term individual and population level consequences. The iTRAQ labeling method was performed to quantify protein abundance changes between exposed and non-exposed organisms. Data analysis revealed a general dose-dependent decrease in the abundance of globin proteins as a result of spinosad exposure. Additionally, the downregulation of actin and a larval cuticle protein was also observed after spinosad exposure, which may be related to previously determined C. riparius life-history traits impairment and biochemical responses. Present results suggest that protein profile changes can be used as early warning biomarkers of pesticide exposure and may provide a better mechanistic interpretation of the toxic response of organisms, aiding in the assessment of the ecological effects of environmental contamination. This work also contributes to the understanding of the sublethal effects of insecticides in invertebrates and their molecular targets. info:eu-repo/semantics/publishedVersion

This study had the support of Fundação para a Ciência e a Tecnologia (FCT) via UID/MAR/04292/2019 and project grant PTDC/MAR-EST/3048/2014. IAD was funded with an FCT PhD grant (SFRH/BD/138376/2018), PRS was funded with an FCT postdoctoral grant (SFRH/BPD/95784/2013) and VFF was funded with an FCT researcher contract (DL57/2016/CP1479/CT0024) at FCUL. The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. 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 systemactivities), neurotransmission (acetylcholinesterase activity) and oxidative damage (DNA damage and lipid peroxidation levels). Overall, each pharmaceutical had different potential for bioconcentration in the muscle (FLX N PROP N 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. info:eu-repo/semantics/publishedVersion

Acknowledgements : This work was funded by the project “Valorização dos subprodutos do processo biotecnológico de produção de esqualeno e DHA pela microalga Aurantiochytrium sp.” (AlgaValue) (ref. 17680), and within the scope of the project CICECO Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, and strategic program of MARE (MARE-UID/MAR/04292/2013), financed by national funds through the FCT/MCTES. Authors also thank the funding from Project AgroForWealth (CENTRO-01-0145-FEDER- 000001), funded by Centro2020, through FEDER and PT2020 and University of Aveiro and FCT/MCT for the financial support for the QOPNA research Unit (FCT UID/QUI/00062/2019) through national founds and, where applicable, co-financed by the FEDER, within the PT2020 Partnership Agreement. The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund. The microalgae Aurantiochytrium sp. is a strong alternative source of ω-3 fatty acids, including ocosahexaenoic acid (DHA). This work encompasses the optimization of SFE conditions to maximize the total extraction yield (ηTotal), DHA content (CDHA), total phenolics content (TPC), and antioxidant capacity (AOC) of the extracts produced from Aurantiochytrium sp. biomass. A full factorial experimental plan was performed, comprising three factors (pressure, temperature, and flow rate) and two levels (200−300 bar, 40−80 °C, and 6–12 gmin−1, respectively). The maximum and minimum experimental results were ηTotal = 2.1 and 13.4 wt.%, CDHA= 27.3 and 39.3 wt.%, TPC =1.19 and 2.24 mgGAE g−extract 1 , and AOC = 0.3 and 1.4 mg g− TEAC extract 1. Under the studied experimental conditions, increasing pressure up to 300 bar is the optimum to rise both ηTotal and CDHA. Temperature increase from 40 to 80 °C leads to opposing effects: it favors the concentration of phenolics in the supercritical extracts at the expenses of decreasing DHA content and total yield. Surface models were adjusted to ηTotal, CDHA and TPC data, and the goodness of the fits ranged from coefficients of determination of 0.752-0.711 (TPC) to 0.997-0.994 (CDHA). Under optimized conditions, supercritical extracts exhibited a DHA content more than 3.5-fold richer than fish oil, and 7.9-fold richer than the best alternative microalgae species (Pavlova lutheri) found in the literature. info:eu-repo/semantics/publishedVersion

Metals are persistent worldwide being harmful for diverse organisms and having complex and combined effects with other contaminants in the environment. Sea turtles accumulate these contaminants being considered good bioindicator species for marine pollution. However, very little is known on how this is affecting these charismatic animals. São Tomé and Príncipe archipelago harbours important green sea turtle (Chelonia mydas) nesting and feeding grounds. The main goal of this study was to determine metal and metalloid accumulation in the blood of females C. mydas nesting in São Tomé Island, and evaluate the possible impacts of this contamination by addressing molecular stress responses. Gene expression analysis was performed in blood targeting genes involved in detoxification/sequestration and metal transport (mt, mtf and fer), and in antioxidant and oxidative stress responses (cat, sod, gr, tdx, txrd, selp and gclc). Micronuclei analysis in blood was also addressed as a biomarker of genotoxicity. Present results showed significant correlations between different gene expressions with the metals evaluated. The best GLM models and significant relationships were found for mt expression, for which 78% of the variability was attributed to metal levels (Al, Cu, Fe, Hg, Pb and Zn), followed by micronuclei count (65% - Cr, Cu, Fe, Hg, Mn and Zn), tdx expression (52% - Cd, Fe, Mn, Pb and Se), and cat expression (52% - As, Fe, Se and Cd x Hg). Overall, this study demonstrates that these green sea turtles are trying to adapt to the oxidative stress and damage produced by metals through the increased expression of antioxidants and other protectors, which raises concerns about the impacts on these endangered organisms' fitness. Furthermore, promising biomarker candidates associated to metal stress were identified in this species that may be used in future biomonitoring studies using C. mydas' blood, allowing for a temporal follow-up of the organisms., This study had the support of Fundação para a Ciência e a Tecnologia (FCT) through the Strategic Project UID/MAR/04292/2020 and the grants awarded to Inês Morão (PD/BD/150562/2019) and Rafael Félix (SFRH/BD/139763/2018). The project was also partially funded by the Integrated Programme of SR&TD “SmartBioR” (reference Centro-01-0145-FEDER-000018) cofunded by Centro (2020) program, Portugal 2020, European Union, through the European Regional Development Fund. Sara Novais is funded by national funds (OE), through FCT – Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. The authors which to acknowledge the support of the Clear Reef Social Fund to grant awarded to Inês Morão. We are thankful to all the members of NGO Programa Tatô and research assistants (biologists Maria Branco and Yedda Oliveira) working at Jalé Beach and to Aurélio Evaristo and João Evaristo that helped us in the field under challenging conditions. We are also thankful to Marta Carmona for the help given in the treatment of the samples for cytology and micronucleus analysis.

This work had the support of Fundação para a Ciência e Tecnologia through the project ACIDLARVAE (PTDC/MAR-EST/4627/2012) and project PROTEOME (PTDC/AAG-MAA/1302/2014), co-financed by COMPETE (POCI-01-0145-FEDER-016773), grants to Ana M. Faria (SFRH/BPD/68673/2010), Cátia Silva (SFRH/BD/118195/2016), and Sara C. Novais (SFRH/BPD/94500/2013), a L'oreal award for Women in Science (2015) to Ana M. Faria, and the strategic project UID/MAR/ 04292/2013. The authors also wish to acknowledge the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01- 0145-FEDER-000018), co-funded by Centro (2020) program, Portugal 2020, European Union, through the European Regional Development Fund. Fishes are currently facing novel types of anthropogenic stressors that have never experienced in their evolutionary history, such as ocean acidification. Under these stressful conditions, energetically costly processes, such as reproduction, may be sacrificed for increased chances of survival. This trade-off does not only affect the organism itself but may result in reduced offspring fitness. In the present study, the effects of exposure to high pCO2 levels were tested on the reproductive performance of a temperate species, the two-spotted goby, Gobiusculus flavescens. Breeding pairs were kept under control (∼600 μatm, pH∼ 8.05) and high pCO2 levels (∼2300 μatm, pH∼ 7.60) conditions for a 4-month period. Additionally, oxidative stress and energy metabolism-related biomarkers were measured. Results suggest that reproductive activity is stimulated under high pCO2 levels. Parental pairs in the simulated ocean acidification conditions exhibited increased reproductive output, with 50% more clutches and 44% more eggs per clutch than pairs under control conditions. However, there was an apparent trade-off between offspring number and size, as larvae of parental pairs under high pCO2 levels hatched significantly smaller, suggesting differences in parental provisioning, which could be related to the fact that these females produce more eggs. Moreover, results support the hypothesis of different energy allocation strategies used by females under high pCO2 conditions. These changes might, ultimately, affect individual fitness and population replenishment. info:eu-repo/semantics/publishedVersion