Behavior of diclofenac from contaminated fish after cooking and in vitro digestion.

BACKGROUND: Seafood consumers are widely exposed to diclofenac due to the high contamination levels often present in aquatic organisms. It is a potential risk to public health due its endocrine disruptor properties. Limited information is available about diclofenac behavior after food digestion to enable a more realistic scenario of consumer exposure. This study aimed to evaluate cooking effects on diclofenac levels, and determine diclofenac bioaccessibility by an in vitro digestion assay, using commercial fish species (seabass and white mullet) as models. The production of the main metabolite 4'-hydroxydiclofenac was also investigated. Fish hamburgers were spiked at two levels (150 and 1000 ng g-1 ) and submitted to three culinary treatments (roasting, steaming and grilling). RESULTS: The loss of water seems to increase the diclofenac levels after cooking, except in seabass with higher levels. The high bioaccessibility of diclofenac (59.1-98.3%) observed in both fish species indicates that consumers' intestines are more susceptible to absorption, which can be worrisome depending on the level of contamination. Contamination levels did not affect the diclofenac bioaccessibility in both species. Seabass, the fattest species, exhibited a higher bioaccessibility of diclofenac compared to white mullet. Overall, cooking decreased diclofenac bioaccessibility by up to 40% in seabass and 25% in white mullet. The main metabolite 4'-hydroxydiclofenac was not detected after cooking or digestion. CONCLUSION: Thus, consumption of cooked fish, preferentially grilled seabass and steamed or baked white mullet are more advisable. This study highlights the importance to consider bioaccessibility and cooking in hazard characterization studies. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Ecotoxicological responses of juvenile Sparus aurata to BDE-99 and BPA exposure: A multi-biomarker approach integrating immune, endocrine and oxidative endpoints.

Pentabromodiphenyl ether (BDE-99) and bisphenol A (BPA) are synthetic organic compounds present in several daily use products. Due to their physicochemical properties, they are ubiquitously present in aquatic ecosystems and considered highly persistent. Recent evidence has confirmed that both emerging compounds are toxic to humans and terrestrial mammals eliciting a wide range of detrimental effects at endocrine and immune levels. However, the ecotoxicological responses that they can trigger in vertebrate marine species have not yet been established. Hence, this study aimed to investigate the ecotoxicological responses of juvenile Sparus aurata upon chronic (28 days) dietary exposure to BDE-99 and BPA (alone and combined) following an integrated multi-biomarker approach that combined fitness indicators (Fulton's K and splenosomatic indexes) with endocrine [cortisol, 17ß-estradiol (E2), 11-ketotestosterone (11-KT) concentrations] and immune (peroxidase and antiprotease activities) endpoints in fish plasma, and oxidative stress [superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) activities, and lipid peroxidation (LPO)] endpoints in the fish spleen. The mixture of BDE-99 and BPA yielded the highest IBR index value in both plasma and spleen biomarkers, therefore, suggesting that the effects of these compounds are more severe when they act together. Endocrine biomarkers were the most responsive in the three contaminated treatments. Fitness indicators were not affected by the individual nor the interactive effects of BDE-99 and BPA. These findings highlight the relevance of accounting for the interactive effects of emerging chemical contaminants and integrating responses associated with distinct biological pathways when investigating their impacts on marine life, as such a multi-biomarker approach provides a broader, more realistic and adequate perspective of challenges faced by fish in a contaminated environment.

Combined effects of climate change and BDE-209 dietary exposure on the behavioural response of the white seabream, Diplodus sargus.

Decabromodiphenyl-ether (BDE-209) is a persistent organic pollutant ubiquitously found in marine environments worldwide. Even though this emerging chemical contaminant is described as highly toxic, bioaccumulative and biomagnifiable, limited studies have addressed the ecotoxicological implications associated with its exposure in non-target marine organisms, particularly from a behavioural standpoint. Alongside, seawater acidification and warming have been intensifying their impacts on marine ecosystems over the years, compromising species welfare and survival. BDE-209 exposure as well as seawater acidification and warming are known to affect fish behaviour, but information regarding their interactive effects is not available. In this study, long-term effects of BDE-209 contamination, seawater acidification and warming were studied on different behavioural traits of Diplodus sargus juveniles. Our results showed that D. sargus exhibited a marked sensitivity in all the behaviour responses after dietary exposure to BDE-209. Fish exposed to BDE-209 alone revealed lower awareness of a risky situation, increased activity, less time spent within the shoal, and reversed lateralization when compared to fish from the Control treatment. However, when acidification and/or warming were added to the equation, behavioural patterns were overall altered. Fish exposed to acidification alone exhibited increased anxiety, being less active, spending more time within the shoal, while presenting a reversed lateralization. Finally, fish exposed to warming alone were more anxious and spent more time within the shoal compared to those of the Control treatment. These novel findings not only confirm the neurotoxicological attributes of brominated flame retardants (like BDE-209), but also highlight the relevance of accounting for the effects of abiotic variables (e.g. pH and seawater temperature) when investigating the impacts of environmental contaminants on marine life.

Linking chemical exposure and fish metabolome: Discovering new biomarkers of environmental exposure of Argyrosomus regius to the antidepressant venlafaxine.

In this study, a non-target metabolomic approach was used to investigate changes in the metabolome of juvenile meagre (Argyrosomus regius) exposed to venlafaxine (20 µg/L). A total of 24, 22 and 8 endogenous metabolites tentatively identified in liver, brain and plasma, respectively, were significantly changed in venlafaxine exposed meagre, showing tissue-dependent variations in the metabolic profile. The amino acids tryptophan, tyrosine and phenylalanine, which are related to the synthesis, availability, and expression of neurotransmitters (e.g., serotonin, dopamine, epinephrine), showed to be dysregulated by venlafaxine exposure. A high impact was observed in fish brain metabolome that showed a trend of up-regulation for most of the tentatively identified metabolites. In conclusion, the identification of possible biomarkers of exposure in fish metabolome to environmental stressors such as venlafaxine is crucial to assess early signal changes at molecular level, enabling the prevention of deleterious effects at the organism and population levels.

Seasonal occurrence and risk assessment of endocrine-disrupting compounds in Tagus estuary biota (NE Atlantic Ocean coast).

Estuaries are continually threatened by anthropogenic pressures, consequently, a large group of contaminants harmful to human health affects the aquatic biota; therefore, it is necessary to monitor their quality. This study deals with the determination of a large group of compounds representing different endocrine-disrupting compounds (EDCs) classes [21 pesticides, 4 polycyclic musk fragrances, 4 UV-filters, 7 bisphenols, 6 polybrominated diphenyl ethers (PBDEs) and 8 of their methoxylated (MeO-BDEs)] in several estuarine species (fish, bivalves, crustaceans, earthworm, and macroalgae) collected seasonally along one year in two distinct areas of Tagus River estuary ("contaminated" vs. "clean" areas). The most abundant compounds found were galaxolide (HHCB) (81% positive samples; 0.04-74 ng/g ww), isoamyl 4-methoxycinnamate (IMC) (64%; 1.13-251 ng/g ww), alachlor (44%; 0.08-16 ng/g ww), and BDE-47 (36%; 0.06-2.26 ng/g ww). Polycyclic musks were the most frequent contaminants in fish (seabass, barbus, mullet, and sole) and macroalgae samples, while UV-filters were predominant in bivalves and crustaceans, and bisphenols in earthworms. Seasonal variation was verified for Σpesticides and Σmusks, with significantly higher levels in summer and autumn, whereas ΣUV-filters highest levels were found in spring and summer, and for ΣPBDEs statistically higher levels were registered in cold seasons (autumn and winter). Σbisphenols were significantly lower in spring than in the other seasons. In general, considering all species analysed in both areas, no statistically significant differences (p > 0.05) were verified between the two collection areas. Based on the estimated daily intake data, consumption of fish from this estuary is unlikely to be a human health concern, since the levels of contamination were below the toxicological threshold values. Overall, the data obtained in this study will allow regulatory authorities to identify and prioritize contaminants monitoring programs in estuaries, such as the case of bisphenol A, which was found, for the first time, in earthworm and clam species.

Lack of detrimental effects of ocean acidification and warming on proximate composition, fitness and energy budget of juvenile Senegalese sole (Solea senegalensis).

Rising levels of atmospheric carbon dioxide (CO2) are driving ocean warming and acidification, which may negatively affect the nutritional quality and physiological performance of commercially important fish species. Thus, this study aimed to evaluate the effects of ocean acidification (OA; ΔpH = -0.3 units equivalent to ΔpCO2 ~ +600 µatm) and warming (OW; ΔT = +4 °C) (and combined, OAW) on the proximate composition, fitness and energy budget of juvenile Senegalese sole (Solea senegalensis). After an exposure period of 75 days, growth (G), metabolism (R) and excretion (faecal, F and nitrogenous losses, U) were assessed to calculate the energy intake (C). Biometric and viscera weight data were also registered to determine animal fitness. Overall, the proximate composition and gross energy were not significantly affected by acidification and warming (alone or in combination). Weight gain, maximum and standard metabolic rates (MMR and SMR, respectively), aerobic scope (AS) and C were significantly higher in fish subjected to OA, OW and OAW than in CTR conditions. Furthermore, the highest relative growth rates (RGR), specific growth rates in terms of wet weight (SGRw) and protein (SGRp), as well as feed efficiencies (FE) occurred in fish submitted to OW and OAW. On the other hand, fish exposed to CTR conditions had significantly higher feed conversion ratio (FCR) and ammonia excretion rate (AER) than those exposed to simulated stressors. Regarding energy distribution, the highest fraction was generally allocated to growth (48-63 %), followed by excretion through faeces (36-51 %), respiration (approximately 1 %) and ammonia excretion (0.1-0.2 %) in all treatments. Therefore, ocean warming and acidification can trigger physiological responses in juvenile Senegalese sole, particularly in their energy budget, which can affect the energy flow and allocation of its population. However, and in general, this species seems highly resilient to these predicted ocean climate change stressors.

Physicochemical properties of iodine and selenium biofortified Sparus aurata and Cyprinus carpio during frozen storage.

Fish biofortification with natural ingredients like iodine-rich macroalgae and selenized-yeast is an excellent strategy to enhance the nutritional quality of farmed fish. This study aimed to assess the effect of frozen storage during 12-months on physicochemical quality of biofortified seabream (Sparus aurata) and carp (Cyprinus carpio). Frozen storage reduced iodine content in biofortified seabream fillets (17%), as well as selenium content in biofortified carp fillets (24%). Yet, biofortified fillets still presented enhanced iodine and selenium contents at the end of the storage period. Increased lipid oxidation (3.45 mg MDA kg-1 for seabream and 2.41 mg MDA kg-1 for carp) and decreased water holding capacity (23-29% for seabream and 14-23% for carp) was observed during storage, whereas major changes in colour and texture occurred after 45 days (seabream) and 225 days (carp) of storage. In general, biofortified fish fillets maintained their nutritional value and quality after 360 days of frozen storage.

Impacts of ocean warming and acidification on the energy budget of three commercially important fish species.

A mechanistic model based on Dynamic Energy Budget (DEB) theory was developed to predict the combined effects of ocean warming, acidification and decreased food availability on growth and reproduction of three commercially important marine fish species: white seabream (Diplodus sargus), zebra seabream (Diplodus cervinus) and Senegalese sole (Solea senegalensis). Model simulations used a parameter set for each species, estimated by the Add-my-Pet method using data from laboratory experiments complemented with bibliographic sources. An acidification stress factor was added as a modifier of the somatic maintenance costs and estimated for each species to quantify the effect of a decrease in pH from 8.0 to 7.4 (white seabream) or 7.7 (zebra seabream and Senegalese sole). The model was used to project total length of individuals along their usual lifespan and number of eggs produced by an adult individual within one year, under different climate change scenarios for the end of the 21st century. For the Intergovernmental Panel on Climate Change SSP5-8.5, ocean warming led to higher growth rates during the first years of development, as well as an increase of 32-34% in egg production, for the three species. Ocean acidification contributed to reduced growth for white seabream and Senegalese sole and a small increase for zebra seabream, as well as a decrease in egg production of 48-52% and 14-33% for white seabream and Senegalese sole, respectively, and an increase of 4-5% for zebra seabream. The combined effect of ocean warming and acidification is strongly dependent on the decrease of food availability, which leads to significant reduction in growth and egg production. This is the first study to assess the combined effects of ocean warming and acidification using DEB models on fish, therefore, further research is needed for a better understanding of these climate change-related effects among different taxonomic groups and species.

Effects of steaming on health-valuable nutrients from fortified farmed fish: Gilthead seabream (Sparus aurata) and common carp (Cyprinus carpio) as case studies.

Fish fortification with iodine-rich macroalgae (Laminaria digitata) and Selenium-rich yeast is expected to promote nutritional added value of this crucial food item, contributing to a healthy and balanced diet for consumers. However, it is not known if steaming can affect these nutrient levels in fortified fish. The present study evaluates the effect of steaming on nutrients contents in fortified farmed gilthead seabream (Sparus aurata) and common carp (Cyprinus carpio). Fortified seabream presented enhanced I, Se and Fe contents, whereas fortified carp presented enhanced I, Se and Zn contents. Steaming resulted in increased I and Se contents in fortified seabream, and increased Fe and Zn levels in fortified carp, with higher elements true retention values (TRVs >90%). The consumption of 150 g of steamed fortified seabream contributes to a significant daily intake (DI) of I (up to 12%) and Se (up to >100%). On the other hand, steamed fortified carp contributes to 19-23% of I DI and 30%-71% of Se DI. These results demonstrate that steaming is a healthy cooking method, maintaining the enhanced nutritional quality of fortified fish. Moreover, the present fortification strategy is a promising solution to develop high-quality farmed fish products to overcome nutritional deficiencies.

Green tea infusion reduces mercury bioaccessibility and dietary exposure from raw and cooked fish.

Human exposure to mercury (Hg) and methylmercury (MeHg) through the ingestion of seafood raises human health-related concerns. In contrast, green tea has health benefits and its consumption potentially reduces bioaccessibility of dietary Hg. The present study aimed to assess the effect of green tea in total mercury (THg) and MeHg bioaccessibility in raw and cooked marine fish species commonly having high Hg levels. Preliminary results demonstrated that significantly higher reductions of bioaccessible THg were attained after the co-ingestion of green tea infusion (1 cup or more) in the oral and intestinal phases. Overall, the present findings clearly show that the co-ingestion of green tea along with seafood grilling strongly reduces THg and MeHg bioaccessibility in all fish species and consequently diminishes the probability of exceeding MeHg provisional tolerable weekly intakes through the consumption of these species with high Hg levels. Such results point out the need to better understand the beneficial/preventive role of green tea infusions and other food processing techniques in bioaccessibility reduction of other chemical contaminants present in food products. Such information is certainly useful to help consumers to wisely select their food, and to enable food safety authorities to integrate such information in risk assessment.

Insights on the metabolization of the antidepressant venlafaxine by meagre (Argyrosomus regius) using a combined target and suspect screening approach.

Bioaccumulation of pharmaceuticals in fish exposed to contaminated water can be shaped by their capability to metabolize these xenobiotics, affecting their toxicity and animal welfare. In this study the in vivo metabolization of the antidepressant venlafaxine by the juvenile marine fish meagre (Argyrosomus regius) was evaluated using a combined target and suspect screening analytical approach. Thirteen venlafaxine metabolites were identified, namely N-desmethylvenlafaxine and N,N-didesmethylvenlafaxine, which were unequivocally identified using analytical standards, and 11 more tentatively identified by suspect screening analysis, including two Phase II metabolites formed by amino acid conjugation. All of them were detected in the liver, while in plasma and brain only 9 and 6 metabolites, respectively, were detected. Based on these findings, for the first time, a tentative metabolization pathway of venlafaxine by A. regius is proposed. Contrarily to what happen in humans, N-demethylation was identified as the main route of metabolization of venlafaxine by fish. Our findings highlight species-specificity in the metabolization of venlafaxine and allow a better understanding of venlafaxine's toxicokinetic in fish. These results emphasize the need to investigate the biotransformation of xenobiotics by non-target organisms to have an integrated overview of their environmental exposure and to improve future evaluations of environmental risk assessment.

Enriched feeds with iodine and selenium from natural and sustainable sources to modulate farmed gilthead seabream (Sparus aurata) and common carp (Cyprinus carpio) fillets elemental nutritional value.

Developing tailor-made fortified farmed fish is a promising solution to overcome nutritional deficiencies and increase consumer confidence in these products. This study evaluated the supplementation of three fortified diets with I-rich seaweed and selenised-yeast on essential and toxic elements levels in gilthead seabream (Sparus aurata) and common carp (Cyprinus carpio). Fortified diets resulted in increased I, Se and Fe in fish muscle. Biofortified seabream and carp revealed lower Cu and Br. The reduction of fishmeal and fish oil in fortified diets resulted in lower Hg and Cd in seabream muscle. Contrarily, fortified diets increased As and Hg in carp fillets. The consumption of 150 g of fortified seabream enabled a significantly higher contribution to the daily recommended intake (DRI) of I (10%) and Se (76%) than non-fortified fish, whereas fortified carp fulfilled 23% of I DRI and 91% of Se DRI. Moreover, the exposure to Pb decreased with the consumption of biofortified seabream (23-82% BMDL01) and carp (26-92% BMDL01). These results support the strategy of developing eco-innovative biofortified farmed fish using sustainable, natural, safe and high-quality ingredients in feeds, to enable consumers to overcome nutritional deficiencies without significantly increased feed costs.

Bioaccessibility of polybrominated diphenyl ethers and their methoxylated metabolites in cooked seafood after using a multi-compartment in vitro digestion model.

Polybrominated diphenyl ethers (PBDEs) comprise a major class of brominated flame retardants and are well-known endocrine disruptors. The dietary route, through contaminated seafood consumption, is a main contributor to human exposure. Hence, the aim of this work was to provide thorough information on the dietary pathway of PBDEs and their methoxylated metabolites (MeO-PBDEs) after consumption of contaminated cooked seafood. The analyses were performed by gas chromatography (tandem) mass spectrometry using environmental-friendly extractive methods validated for fish and samples from several digestion segments with limits of detection at the pictogram level (per gram or milliliter of sample). Selected fish species were artificially contaminated and cooked using common household practices (steamed, grilled and microwaved), resulting in considerable loss of pollutants (up to 32% loss), with significant differences between cooking methods and species. Finally, an in vitro method that simulates four human adult digestion steps (oral, gastric digestion, small and large intestinal digestion) was applied to raw and cooked fish. Bioaccessibility of PBDEs and MeO-PBDEs in small intestinal was low (below 24%), pointing to a heavy impact in gut microbiota. Nevertheless, gut microbiota was able to reduce the amounts of targeted contaminants (up to 82%) in the large intestine. The results achieved herein are of great value to predict both amounts and nature of PBDEs and MeO-PBDEs that seafood consumers may be exposed after the ingestion of contaminated food as to ascertain more accurately the impact on human and environmental health.

Does the addition of ingredients affect mercury and cadmium bioaccessibility in seafood-based meals?

Despite the bioaccessibility of nutrients and contaminants present in individual seafood products has been thoroughly studied, information is extremely limited in what concerns complete seafood-based meals, where interactions between ingredients may occur. Hence, this study aimed to evaluate the effect of different ingredients and cooking processes in mercury (Hg) and cadmium (Cd) bioaccessibility in complete meals of tuna (Thunnus spp.) and edible crab (Cancer pagurus), respectively. The addition of ingredients/side dishes decreased Hg levels in cooked tuna meals, but increased Hg bioaccessibility (up to 31% of bioaccessible Hg in complete meals, against 13.5% in stewed tuna alone). Cd levels in edible crab meals were significantly decreased by the addition of ingredients (~36% and ~65% decrease in boiled crab and paté, respectively), but its' bioaccessibility was not significantly affected (>94% in all cases). Results showed that the weekly consumption of 2 complete tuna meals does not exceed MeHg tolerable weekly intake (TWI), whereas Cd's TWI is largely surpassed with the consumption of 50 g/week of edible crab meals. This highlights the importance of determining contaminant levels and bioaccessibility in a whole seafood-based meal context, as such approach enables a more realistic assessment of the risks that seafood can pose to consumers.

Paralytic Shellfish Toxins and Ocean Warming: Bioaccumulation and Ecotoxicological Responses in Juvenile Gilthead Seabream (Sparus aurata).

Warmer seawater temperatures are expected to increase harmful algal blooms (HABs) occurrence, intensity, and distribution. Yet, the potential interactions between abiotic stressors and HABs are still poorly understood from ecological and seafood safety perspectives. The present study aimed to investigate, for the first time, the bioaccumulation/depuration mechanisms and ecotoxicological responses of juvenile gilthead seabream (Sparus aurata) exposed to paralytic shellfish toxins (PST) under different temperatures (18, 21, 24 °C). PST were detected in fish at the peak of the exposure period (day five, 0.22 µg g-1 N-sulfocarbamoylGonyautoxin-1-2 (C1 and C2), 0.08 µg g-1 Decarbamoylsaxitoxin (dcSTX) and 0.18 µg g-1 Gonyautoxin-5 (B1)), being rapidly eliminated (within the first 24 h of depuration), regardless of exposure temperature. Increased temperatures led to significantly higher PST contamination (275 µg STX eq. kg-1). During the trial, fish antioxidant enzyme activities (superoxide dismutase, SOD; catalase, CAT; glutathione S-transferase, GST) in both muscle and viscera were affected by temperature, whereas a significant induction of heat shock proteins (HSP70), Ubiquitin (Ub) activity (viscera), and lipid peroxidation (LPO; muscle) was observed under the combination of warming and PST exposure. The differential bioaccumulation and biomarker responses observed highlight the need to further understand the interactive effects between PST and abiotic stressors, to better estimate climate change impacts on HABs events, and to develop mitigation strategies to overcome the potential risks associated with seafood consumption.

Bioaccumulation and ecotoxicological responses of juvenile white seabream (Diplodus sargus) exposed to triclosan, warming and acidification.

Triclosan (TCS) is a synthetic microbial compound widely used in the formulation of various personal care products. Its frequent detection in marine ecosystems, along with its physical and chemical properties, suggest that TCS can be highly persistent, being easily bioaccumulated by biota and, therefore, eliciting various toxicological responses. Yet, TCS's mechanisms of bioaccumulation and toxicity still deserve further research, particularly focusing on the interactive effects with climate change-related stressors (e.g. warming and acidification), as both TCS chemical behaviour and marine species metabolism/physiology can be strongly influenced by the surrounding abiotic conditions. Hence, the aim of this study was to assess TCS bioaccumulation and ecotoxicological effects (i.e. animal fitness indexes, antioxidant activity, protein chaperoning and degradation, neurotoxicity and endocrine disruption) in three tissues (i.e. brain, liver and muscle) of juvenile Diplodus sargus exposed to the interactive effects of TCS dietary exposure (15.9 µg kg-1 dw), seawater warming (ΔTºC = +5 °C) and acidification (ΔpCO2 âˆ¼ +1000 µatm, equivalent to ΔpH = -0.4 units). Muscle was the primary organ of TCS bioaccumulation, and climate change stressors, particularly warming, significantly reduced TCS bioaccumulation in all fish tissues. Furthermore, the negative ecotoxicological responses elicited by TCS were significantly altered by the co-exposure to acidification and/or warming, through either the enhancement (e.g. vitellogenin content) or counteraction/inhibition (e.g. heat shock proteins HSP70/HSC70 content) of molecular biomarker responses, with the combination of TCS plus acidification resulting in more severe alterations. Thus, the distinct patterns of TCS tissue bioaccumulation and ecotoxicological responses induced by the different scenarios emphasized the need to further understand the interactive effects between pollutants and abiotic conditions, as such knowledge enables a better estimation and mitigation of the toxicological impacts of climate change in marine ecosystems.

Living in a multi-stressors environment: An integrated biomarker approach to assess the ecotoxicological response of meagre (Argyrosomus regius) to venlafaxine, warming and acidification.

Pharmaceuticals, such as the antidepressant venlafaxine (VFX), have been frequently detected in coastal waters and marine biota, and there is a growing body of evidence that these pollutants can be toxic to non-target marine biota, even at low concentrations. Alongside, climate change effects (e.g. warming and acidification) can also affect marine species' physiological fitness and, consequently, compromising their ability to cope with the presence of pollutants. Yet, information regarding interactive effects between pollutants and climate change-related stressors is still scarce. Within this context, the present study aims to assess the differential ecotoxicological responses (antioxidant activity, heat shock response, protein degradation, endocrine disruption and neurotoxicity) of juvenile fish (Argyrosomus regius) tissues (muscle, gills, liver and brain) exposed to VFX (via water or feed), as well as to the interactive effects of warming (ΔT °C = +5 °C) and acidification (ΔpCO2 ~ +1000 µatm, equivalent to ΔpH = -0.4 units), using an integrated multi-biomarker response (IBR) approach. Overall, results showed that VFX toxicity was strongly influenced by the uptake pathway, as well as by warming and acidification. More significant changes (e.g. increases surpassing 100% in lipid peroxidation, LPO, heat shock response protein content, HSP70/HSC70, and total ubiquitin content, Ub,) and higher IBR index values were observed when VFX exposure occurred via water (i.e. average IBR = 19, against 17 in VFX-feed treatment). The co-exposure to climate change-related stressors either enhanced (e.g. glutathione S-transferases activity (GST) in fish muscle was further increased by warming) or attenuated the changes elicited by VFX (e.g. vitellogenin, VTG, liver content increased with VFX feed exposure acting alone, but not when co-exposed with acidification). Yet, increased stress severity was observed when the three stressors acted simultaneously, particularly in fish exposed to VFX via water (i.e. average IBR = 21). Hence, the distinct fish tissues responses elicited by the different scenarios emphasized the relevance of performing multi-stressors ecotoxicological studies, as such approach enables a better estimation of the environmental hazards posed by pollutants in a changing ocean and, consequently, the development of strategies to mitigate them.

Integrated multi-biomarker responses of juvenile seabass to diclofenac, warming and acidification co-exposure.

Pharmaceutical drugs, such as diclofenac (DCF), are frequently detected in the marine environment, and recent evidence has pointed out their toxicity to non-target marine biota. Concomitantly, altered environmental conditions associated with climate change (e.g. warming and acidification) can also affect the physiology of marine organisms. Yet, the underlying interactions between these environmental stressors (pharmaceutical exposure and climate change-related stressors) still require a deeper understanding. Comprehending the influence of abiotic variables on chemical contaminants' toxicological attributes provides a broader view of the ecological consequences of climate change. Hence, the aim of this study was to assess the ecotoxicological responses of juvenile seabass Dicenthrachus labrax under the co-exposure to DCF (from dietary sources, 500 ±â€¯36 ng kg-1 dw), warming (ΔTºC = +5 °C) and acidification (ΔpCO2 ∼1000 µatm, equivalent to ΔpH = -0.4 units), using an "Integrated Biomarker Response" (IBR) approach. Fish were exposed to these three stressors, acting alone or combined, for 28 days in a full cross-factorial design, and blood, brain, liver and muscle tissues were subsequently collected in order to evaluate: i) animal/organ fitness; ii) hematological parameters and iii) molecular biomarkers. Results not only confirmed the toxicological attributes of dietary exposure to DCF in marine fish species at the tissue (e.g. lower HSI), cellular (e.g. increased ENAs and lower erythrocytes viability) and molecular levels (e.g. increased oxidative stress, protein degradation, AChE activity and VTG synthesis), but also showed that such attributes are altered by warming and acidification. Hence, while acidification and/or warming enhanced some effects of DCF exposure (e.g. by further lowering erythrocyte viability, and increasing brain GST activity and Ub synthesis in muscle), the co-exposure to these abiotic stressors also resulted in a reversion/inhibition of some molecular responses (e.g. lower CAT and SOD inhibition and VTG synthesis). IBRs evidenced that an overall higher degree of stress (i.e. high IBR index) was associated with DCF and warming co-exposure, while the effects of acidification were less evident. The distinct responses observed when DCF acted alone or the animals were co-exposed to the drug together with warming and acidification not only highlighted the relevance of considering the interactions between multiple environmental stressors in ecotoxicological studies, but also suggested that the toxicity of pharmaceuticals can be aggravated by climate change-related stressors (particularly warming), thus, posing additional biological challenges to marine fish populations.

Antidepressants in a changing ocean: Venlafaxine uptake and elimination in juvenile fish (Argyrosomus regius) exposed to warming and acidification conditions.

The presence of antidepressants, such as venlafaxine (VFX), in marine ecosystems is increasing, thus, potentially posing ecological and human health risks. The inherent mechanisms of VFX uptake and elimination still require further understanding, particularly accounting for the impact of climate change-related stressors, such as warming and acidification. Hence, the present work aimed to investigate, for the first time, the effects of increased seawater temperature (ΔT°C = +5 °C) and pCO2 levels (ΔpCO2 ∼1000 µatm, equivalent to ΔpH = -0.4 units) on the uptake and elimination of VFX in biological tissues (muscle, liver, brain) and plasma of juvenile meagre (Argyrosomus regius) exposed to VFX through two different exposure pathways (via water, i.e. [VFX ] ∼20 µg L-1, and via feed, i.e. [VFX] ∼160 µg kg-1 dry weight, dw). Overall, results showed that VFX can be uptaken by fish through both water and diet. Fish liver exhibited the highest VFX concentration (126.7 ±â€¯86.5 µg kg-1 and 6786.4 ±â€¯1176.7 µg kg-1 via feed and water exposures, respectively), as well as the highest tissue:plasma concentration ratio, followed in this order by brain and muscle, regardless of exposure route. Both warming and acidification decreased VFX uptake in liver, although VFX uptake in brain was favoured under warming conditions. Conversely, VFX elimination in liver was impaired by both stressors, particularly when acting simultaneously. The distinct patterns of VFX uptake and elimination observed in the different scenarios calls for a better understanding of the effects of exposure route and abiotic conditions on emerging contaminants' toxicokinetics.

Effects of steaming on contaminants of emerging concern levels in seafood.

Seafood consumption is a major route for human exposure to environmental contaminants of emerging concern (CeCs). However, toxicological information about the presence of CeCs in seafood is still insufficient, especially considering the effect of cooking procedures on contaminant levels. This study is one among a few who evaluated the effect of steaming on the levels of different CeCs (toxic elements, PFCs, PAHs, musk fragrances and UV-filters) in commercially relevant seafood in Europe, and estimate the potential risks associated with its consumption for consumers. In most cases, an increase in contaminant levels was observed after steaming, though varying according to contaminant and seafood species (e.g. iAs, perfluorobutanoate, dibenzo(ah)anthracene in Mytilus edulis, HHCB-Lactone in Solea sp., 2-Ethylhexyl salicylate in Lophius piscatorius). Furthermore, the increase in some CeCs, like Pb, MeHg, iAs, Cd and carcinogenic PAHs, in seafood after steaming reveals that adverse health effects can never be excluded, regardless contaminants concentration. However, the risk of adverse effects can vary. The drastic changes induced by steaming suggest that the effect of cooking should be integrated in food risk assessment, as well as accounted in CeCs regulations and recommendations issued by food safety authorities, in order to avoid over/underestimation of risks for consumer health.