Anthropogenic carbon dioxide emissions (CO2) have led to climate change and marine acidification, with an estimated decrease in ocean surface pH of 0.3-0.4 units by the end of the current century. Chemical pollution also contributes to biodiversity loss in marine environments. This issue is particularly critical in areas under pressure from shipping activities, where the introduction of new antifouling system formulations poses a major threat to non-target species. The biocide DCOIT is the most widely used alternative to organotin compounds due to its rapid degradation in seawater. The toxicity of waterborne DCOIT to marine organisms has been documented, but sediment-bound effects are limited to apical responses and pH scenarios corresponding to current levels. In this study, we determine in a combined way, the toxicity of DCOIT under marine acidification scenarios assessing biomarker responses in the burrowing amphipod Tiburonella viscana as a parameter of sublethal effects in solid phase exposures. Environmental relevant concentrations of DCOIT caused inhibition of the enzyme glutathione S-transferases (GST), changed acetylcholinesterase-like activity (AChE), and increased DNA damage at pHs of 7.7 and 7.4. For lipid peroxidation (LPO), increased levels caused by DCOIT were found for both control (8.1) and intermediate (7.7) conditions of pH. Our data provides evidence of oxidative and genotoxic effects induced by DCOIT, with activation of detoxification and defense mechanisms in T. viscana. These results are important for ecological risk assessment and managing of antifouling paint biocides in multiple stressors scenarios.

Background: The present study reports the protective effect conferred by scavenging mitochondrial oxidative stress (mtOS) in 5-fluorouracil (5-FU)-induced renal injury. Methods: 5-FU renal toxicity model was created by administering 5-FU (12 mg/kg b.w. intraperitoneally [i.p.], for 4 days) to male BALB/c mice. The protective effect of mitochondria-targeted antioxidant (MTA), Mito-TEMPO coadministered at a dosage of 0.1 mg/kg b.w. i.p., was established in terms of levels/expressions of renal injury markers, histopathological alterations, oxidative DNA damage, proinflammatory markers, mtOS, mitochondrial dysfunction, and modulation of apoptotic proteins and apoptotic cell death. Results: A significant rise in the levels of serum urea, uric acid, and creatinine was noted after 5-FU administration to the animals. Immunohistochemical and ELISA findings demonstrated significant decrease in podocin and conversely a significant increase in neutrophil gelatinase-associated lipocalin (NGAL) expression after 5-FU challenge. The histopathological analysis further revealed Bowman's capsule dilation, glomerular condensation, and vacuolar degeneration. Mito-TEMPO treatment significantly lowered renal injury markers, reversed the expressions of podocin and NGAL to normal, and restored normal histoarchitecture of renal tissue. Mitochondrial reactive oxygen species (mtROS), mtLPO, activity of mitochondrial enzyme complexes, and mitochondrial antioxidant defense status were significantly improved in Mito-TEMPO protected group as compared to the 5-FU group. Further, significantly decreased expression of 8-OHdG, reduction in apoptotic cell death, and modulation of apoptotic proteins Bax, Bcl-2, and caspase-3 were noted in Mito-TEMPO protected group, indicating its protective effect against 5-FU-induced renal injury. Conclusion: The approach of targeting mtOS using MTA, Mito-TEMPO, may prove as safe adjuvant in alleviating renal toxicity during 5-FU chemotherapy.

Silver (Ag) and cadmium (Cd) are non-essential metals that, as a result of natural processes and human activities, reach the aquatic environment where they interact with biota inducing potential toxic effects. To determine the biological effects of these metals on the endobenthic bivalve Scrobicularia plana, specimens were exposed to Ag and Cd at two concentrations, 5 and 50 μg∙L-1, for 7 days in a controlled microcosm system. The levels of the metals were measured in the seawater, sediments and clam tissues. The possible toxic biological effects of Ag and Cd were studied using a battery of biochemical biomarkers that are responsive to oxidative stress: superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST) activities, and metallothioneins (MTs) and lipid peroxidation (LPO) levels. Since both metals have been linked to oxidative stress, redox modifications to proteins were studied by differential isotopic labelling of the oxidised and reduced forms of cysteines (Cys). An accumulation of metals was observed in the digestive gland and gills following exposure, together with the activation of enzyme activities (SOD for the Cd exposure; SOD, CAT, GST, and GR for the Ag exposure). The MT and LPO levels (after individual exposure to Ag and Cd) increased, which suggests the existence of antioxidant and detoxification processes to mitigate the toxic oxidative effects of both metals. The redox proteomic analysis identified 771 Cys-containing peptides (out of 514 proteins), of which 195 and 226 changed after exposure to Ag and Cd, respectively. Bioinformatics analysis showed that exposure to metal affects relevant functional pathways and biological processes in S. plana, such as: "cellular respiration" (Ag), "metabolism of amino acids" and "synthesis and degradation of proteins" (Ag and Cd), "carbohydrate metabolism" and "oxidative stress" (Cd). The proteomic approach implemented here is a powerful complement to conventional biochemical biomarkers, since it evaluates changes at the protein level in a high-throughput unbiased manner, thus providing a general appraisal of the biological responses altered by exposure to the contaminants.

Once in the marine environment, fishing nets and cables undergo weathering, breaking down into micro and nano-size particles and leaching plastic additives, which negatively affect marine biota. This study aims to unravel the ecotoxicological impact of different concentrations of leachate obtained from abandoned or lost fishing nets and cables in the mussel Mytilus galloprovincialis under long-term exposure (28 days). Biochemical biomarkers linked to antioxidant defense system, xenobiotic biotransformation, oxidative damage, genotoxicity, and neurotoxicity were evaluated in different mussel tissues. The chemical nature of the fishing nets and cables and the chemical composition of the leachate were assessed and metals, plasticizers, UV stabilizers, flame retardants, antioxidants, dyes, flavoring agents, preservatives, intermediates and photo initiators were detected. The leachate severely affected the antioxidant and biotransformation systems in mussels' tissues. Following exposure to 1 mg·L-1 of leachate, mussels' defense system was enhanced to prevent oxidative damage. In contrast, in mussels exposed to 10 and 100 mg·L-1 of leachate, defenses failed to overcome pro-oxidant molecules, resulting in genotoxicity and oxidative damage. Principal component analysis (PCA) and Weight of Evidence (WOE) evaluation confirmed that mussels were significantly affected by the leachate being the hazard of the leachate concentrations of 10 mg·L-1 ranked as major, while 1 and 100 mg·L-1 was moderate. These results highlighted that the leachate from fishing nets and cables can be a threat to the heath of the mussel M. galloprovincialis.

The increasing release of engineered nanoparticles (ENPs) in aquatic ecosystems stresses the need for stringent investigations of nanoparticle mixture toxicity towards aquatic organisms. Here, the individual and combined immunotoxicity of two of the most consumed ENPs, the ZnO and the TiO2 ones, was investigated on rainbow trout juveniles (Oncorhynchus mykiss). Fish were exposed to environmentally realistic concentrations (21 and 210 µg L-1 for the ZnO and 210 µg L-1 for the TiO2) for 28 days, and then challenged with the pathogenic bacterium, Aeromonas salmonicida achromogenes. Antioxidant and innate immune markers were assessed before and after the bacterial infection. None of the experimental conditions affected the basal activity of the studied innate immune markers and the redox balance. However, following the bacterial infection, the expression of genes coding for pro and anti-inflammatory cytokines (il1β and il10), as well as innate immune compounds (mpo) were significantly reduced in fish exposed to the mixture. Conversely, exposure to ZnO NPs alone seemed to stimulate the immune response by enhancing the expression of the IgM and c3 genes for instance. Overall, our results suggest that even though the tested ENPs at their environmental concentration do not strongly affect basal immune functions, their mixture may alter the development of the immune response when the organism is exposed to a pathogen by interfering with the inflammatory response.

The synthetic hormone 17α ethinyl estradiol (EE2) is a molecule widely used in female contraceptives and recognized as a contaminant of attention (Watch List) in the European Union due to its high consumption, endocrine effects and occurrence in aquatic environments. Its main source of introduction is domestic sewage where it can be associated with other contaminants such as microplastics (MPs). Due to their characteristics, they can combine with each other and exacerbate their isolated effects on biota. This study evaluated the combined effects of microplastics (MPs) and 17α ethinylestradiol (EE2) on two tropical estuarine invertebrate species: Crassostrea gasar and Ucides cordatus. Polyethylene particles were spiked with EE2 and organisms were exposed to three treatments, categorized into three groups: control group (C), virgin microplastics (MPs), and spiked microplastics with EE2 (MPEs). All treatments were evaluated after 3 and 7 days of exposure. Oysters exhibited changes in phase 2 enzymes and the antioxidant system, oxidative stress in the gills, and reduced lysosomal membrane stability after exposure to MPs and MPEs. Crabs exposed to MPs and MPEs after seven days showed changes in phase 1 enzymes in the gills and changes in phases 1 and 2 enzymes in the hepatopancreas, such as disturbed cellular health. The combined effects of microplastics and EE2 increased the toxicity experienced by organisms, which may trigger effects at higher levels of biological organization, leading to ecological disturbances in tropical coastal ecosystems.

In the last few decades, there has been a growing interest in understanding the behavior of personal care products (PCPs) in the aquatic environment. In this regard, the aim of this study is to estimate the accumulation and effects of four PCPs within the clam Ruditapes philippinarum. The PCPs selected were triclosan, OTNE, benzophenone-3, and octocrylene. A progressive uptake was observed and maximum concentrations in tissues were reached at the end of the exposure phase, up to levels of 0.68 µg g-1, 24 µg g-1, 0.81 µg g-1, and 1.52 µg g-1 for OTNE, BP-3, OC, and TCS, respectively. After the PCP post-exposure period, the removal percentages were higher than 65%. The estimated logarithm bioconcentration factor ranged from 3.34 to 2.93, in concordance with the lipophobicity of each substance. No lethal effects were found although significant changes were observed for ethoxyresorufin O-demethylase activity, glutathione S-transferase activity, lipid peroxidation, and DNA damage.

Contaminants of emerging concern (CECs) present a new threat to the marine environment, and it is vital to understand the interactions and possible toxicity of CEC mixtures once they reach the ocean. CECs-such as metal nanoparticles, nanoplastics, and pharmaceuticals-are groups of contaminants some of which have been individually evaluated, though their interactions as mixtures are still not fully understood. To ensure a healthy and prosperous future generation, successful reproduction is key: however, if hindered, population dynamics may be at danger leading to a negative impact on biodiversity. This study aimed to understand the effects of silver (20 nm nAg, 10 μg/L), polystyrene nanoparticles (50 nm nPS, 10 μg/L), and 5-fluorouracil (5FU, 10 ng/L) individually and as a mixture (10 μg/L of nPS + 10 μg/L of nAg +10 ng/L of 5FU) in the gonads of Mytilus galloprovincialis. A multibiomarker approach, namely the antioxidant defence system (ADS; superoxide dismutase, catalase, glutathione peroxidases, glutathione - S - transferases activities), and oxidative damage (OD; lipid peroxidation) were analysed in the gonads of mussels. All exposure treatments after 3 days led to an increase of enzymatic activity, followed by an inhibition after 14 and 21 days. Thus, ADS was overwhelmed due to the generation of ROS, resulting in OD, except for nPS exposed mussels. The OD in Mix exposed mussels increased exponentially by 57-fold. When CEC mixtures interact, they are potentially more hazardous than their individual components, posing a major threat to marine species. To understand synergistic and antagonistic interactions, a model was applied, and antagonistic interactions were observed in evaluated biomarkers at all time-points, apart from a synergistic interaction at day 3 relative to LPO. Results indicate that the effects observed in Mix-exposed mussel gonads are mainly due to the interaction of nAg and 5FU but not nPS.

Recent evidences highlight role of mitochondria in the development of 5-fluorouracil (5-FU)-induced intestinal toxicity. Mitochondria-targeted antioxidants are well-known for their protective effects in mitochondrial oxidative stress- mediated diseases. In the present study, we investigated protective effect of Mito-TEMPO in 5-FU-induced intestinal toxicity.

Mito-TEMPO (0.1 mg/kg b.w.) was administered intraperitoneally to male BALB/c mice for 7 days, followed by co-administration of 5-FU for next 4 days (intraperitoneal 12 mg/kg b.w.). Protective effect of Mito-TEMPO on intestinal toxicity was assessed in terms of histopathological alterations, modulation in inflammatory markers, apoptotic cell death, expression of 8-OhDG, mitochondrial functional status and oxidative stress.

5-FU administered animals showed altered intestinal histoarchitecture wherein a shortening and atrophy of the villi was observed. The crypts were disorganized and inflammatory cell infiltration was noted. Mito-TEMPO pre-protected animals demonstrated improved histoarchitecture with normalization of villus height, better organized crypts and reduced inflammatory cell infiltration. The inflammatory markers and myeloperoxidase activity were normalized in mito-TEMPO protected group. A significant reduction in intestinal apoptotic cell death and expression of 8-OhDG was also observed in mito-TEMPO group as compared to 5-FU group. Further, mtROS, mtLPO and mitochondrial antioxidant defense status were improved by mito-TEMPO.

Mito-TEMPO exerted significant protective effect against 5-FU-induced intestinal toxicity. Therefore, it may be used as an adjuvant in 5-FU chemotherapy.

Mercury (Hg) is a toxic environmental contaminant associated with oxidative stress in freshwater fish. A known antagonist to Hg, selenium (Se), may reduce the toxic effects of Hg. In this study, the relation among Se, methylmercury (MeHg), inorganic mercury (IHg), total mercury (THg), and the expression of biomarkers of oxidative stress and metal regulation in livers of northern pike were examined. Livers from northern pike were collected from 12 lakes in Isle Royale National Park, Pictured Rocks National Lakeshore, Sleeping Bear Dunes National Lakeshore, and Voyageurs National Park. The concentrations of MeHg, THg, and Se were measured in liver tissue, and the expression of superoxide dismutase (sod), catalase (cat), glutathione s-transferase (gst), and metallothionein (mt) was assessed. There was a positive relationship between the concentrations of THg and Se, with a Hg:Se molar ratio less than one in all livers examined. There was no significant relation between sod, cat, gst, or mt expression and Hg:Se molar ratios. cat and sod expression were significantly related to increases in percent MeHg, relative to THg; however, gst and mt expression were not significantly altered. This suggests that incorporating biomarkers containing Se may be a better indicator than non-selenium-containing proteins of assessing the long-term effect of Hg and the interactions between Hg and Se in the livers of fish, such as northern pike, especially when molar concentrations of Se are greater than Hg.

Human exposure to multiple xenobiotics, over various developmental windows, results in adverse health effects arising from these concomitant exposures. Humans are widely exposed to bisphenol A, and acetaminophen is the most commonly used over-the-counter drug worldwide. Bisphenol A is a well-recognized male reproductive toxicant, and increasing evidence suggests that acetaminophen is also detrimental to the male reproductive system. The recent recognition of male reproductive system dysfunction in conditions of suboptimal reproductive outcomes makes it crucial to investigate the contributions of toxicant exposures to infertility and sub-fertility. We aimed to identify toxicity in the male reproductive system at the mitochondrial level in response to co-exposure to bisphenol A and acetaminophen, and we investigated whether melatonin ameliorated this toxicity.

Male Wistar rats were divided into six groups (n=10 each): a control group and groups that received melatonin, bisphenol A, acetaminophen, bisphenol A and acetaminophen, and bisphenol A and acetaminophen with melatonin treatment.

Significantly higher lipid peroxidation was observed in the testicular mitochondria and sperm in the treatment groups than in the control group. Levels of glutathione and the activities of catalase, glutathione peroxidase, glutathione reductase, and manganese superoxide dismutase decreased significantly in response to the toxicant treatments. Likewise, the toxicant treatments significantly decreased the sperm count and motility, while significantly increasing sperm mortality. Melatonin mitigated the adverse effects of bisphenol A and acetaminophen.

Co-exposure to bisphenol A and acetaminophen elevated oxidative stress in the testicular mitochondria, and this effect was alleviated by melatonin.

Environmental pollution increases due to anthropogenic activities. Toxic chemicals in the environment affect the health of aquatic organisms. Tributyltin (TBT) is a toxic chemical widely used as an antifouling paint on boats, hulls, and ships. The toxic effect of TBT is well documented in aquatic organisms; however, little is known about the effects of TBT on DNA lesions in shellfish. The American oyster (Crassostrea virginica, an edible and commercially important species) is an ideal marine mollusk to examine the effects of TBT exposure on DNA lesions and oxidative/nitrative stress. In this study, we investigated the effects of TBT on 8'-hydroxy-2'-deoxyguanosine (8-OHdG, a biomarker of pro-mutagenic DNA lesion), double-stranded DNA (dsDNA), dinitrophenyl protein (DNP, a biomarker on reactive oxygen species, ROS), 3-nitrotyrosine protein (NTP, a biomarker of reactive nitrogen species, RNS), catalase (CAT, an antioxidant), and acetylcholinesterase (AChE, a cholinergic enzyme) expressions in the gills and digestive glands of oysters. We also analyzed extrapallial (EF) fluid conditions. Immunohistochemical and qRT-PCR results showed that TBT exposure significantly increased 8-OHdG, dsDNA, DNP, NTP, and CAT mRNA and/or protein expressions in the gills and digestive glands. However, AChE mRNA and protein expressions, and EP fluid pH and protein concentrations were decreased in TBT-exposed oysters. Taken together, these results suggest that antifouling biocide-induced production of ROS/RNS results in DNA damage, which may lead to decreased cellular functions in oysters. To the best of our knowledge, the present study provides the first molecular/biochemical evidence that TBT exposure results in oxidative/nitrative stress and DNA lesions in oysters.

Selenium (Se) is a vital trace element for many living organisms inclusive of aquatic species. Although the antagonistic action of this element against other pollutants has been previously described for mammals and birds, limited information on the join effects in bivalves is available. To this end, bivalves of the species Scrobicularia plana were exposed to Se and Cd individually and jointly. Digestive glands were analysed to determine dose-dependent effects, the potential influence of Se on Cd bioaccumulationas well as the possible recover of the oxidative stress and metabolic alterations induced by Cd. Selenium co-exposure decreased the accumulation of Cd at low concentrations. Cd exposure significantly altered the metabolome of clams such as aminoacyltRNA biosynthesis, glycerophospholipid and amino acid metabolism, while Se co-exposure ameliorated several altered metabolites such asLysoPC (14:0), LysoPE (20:4), LysoPE (22:6), PE (14:0/18:0), PE (20:3/18:4) andpropionyl-l-carnitine.Additionally, Se seems to be able to regulate the redox status of the digestive gland of clams preventing the induction of oxidativedamage in this organ. This study shows the potential Se antagonism against Cd toxicity in S. plana and the importance to study join effects of pollutants to understand the mechanism underlined the effects.

Domestic sewage is an important source of pollutants in aquatic ecosystems and includes both microplastics (MPs) and pharmaceuticals and personal care products (PPCPs). This study sought to assess the biological effects of the interaction between plastic particles and the antibacterial agent triclosan (TCS). The study relied on the swamp ghost crab Ucides cordatus as a model. Herein polyethylene particles were contaminated with triclosan solution. Triclosan concentrations in the particles were then chemically analyzed. Swamp ghost crab specimens were exposed to experimental compounds (a control, microplastics, and microplastics with triclosan) for 7 days. Samplings were performed on days 3 (T3) and 7 (T7). Gill, hepatopancreas, muscle and hemolymph tissue samples were collected from the animals to evaluate the biomarkers ethoxyresorufin O-deethylase (EROD), dibenzylfluorescein dealkylase (DBF), glutathione S-transferase (GST), glutathione peroxidase (GPx), reduced glutathione (GSH), lipid peroxidation (LPO), DNA strands break (DNA damage), cholinesterase (ChE) through protein levels and neutral red retention time (NRRT). Water, organism, and microplastic samples were collected at the end of the assay for post-exposure chemical analyses. Triclosan was detected in the water and crab tissue samples, results which indicate that microplastics serve as triclosan carriers. Effects on the gills of organisms exposed to triclosan-spiked microplastics were observed as altered biomarker results (EROD, GST, GPx, GSH, LPO, DNA damage and NRRT). The effects were more closely associated with microplastic contaminated with triclosan exposure than with microplastic exposure, since animals exposed only to microplastics did not experience significant effects. Our results show that microplastics may be important carriers of substances of emerging interest in marine environments in that they contaminate environmental matrices and have adverse effects on organisms exposed to these stressors.

The assessment of contaminants of emerging concern, alone and in mixtures, and their effects on marine biota requires attention. 5-Fluorouracil is a cytostatic category 3 anti-cancer medication (IARC) that is used to treat a variety of cancers, including colon, pancreatic, and breast cancer. In the presence of other pollutants, this pharmaceutical can interact and form mixtures of contaminants, such as adhering to plastics and interaction with metal nanoparticles. This study aimed to comprehend the effects of 5-Fluorouracil (5FU; 10 ng/L) and a mixture of emerging contaminants (Mix): silver nanoparticles (nAg; 20 nm; 10 μg/L), polystyrene nanoparticles (nPS; 50 nm; 10 μg/L) and 5FU (10 ng/L), in an in vivo (21 days) exposure of the mussel Mytilus galloprovincialis. A multibiomarker approach namely genotoxicity, the antioxidant defence system (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx), glutathione - S - transferases (GST) activities), and oxidative damage (LPO) was used to assess the effects in gills and digestive gland of mussels. Both treatments cause genotoxicity in mussel's haemolymph, and antagonism between contaminants was observed in the Mix. Genotoxicity observed confirms 5FU's mode of action (MoA) by DNA damage. The antioxidant defence system of mussels exposed to 5FU kicked in and counter balanced ROS generated during the exposure, though the same was not seen in Mix-exposed mussels. Mussels were able to withstand the effects of the single compound but not the effects of the Mix. For oxidative stress and damage, the interactions of the components of the mixture have a synergistic effect.

Pharmaceutical drugs in the aquatic medium may pose significant risk to non-target organisms. In this study, the potential toxicity of a mixture of three compounds commonly detected in marine waters (ibuprofen, ciprofloxacin and flumequine) was assessed, by studying bioaccumulation, oxidative stress and neurotoxicity parameters (catalase CAT, superoxide dismutase SOD, glutathione reductase GR, glutathione S-transferase GST, lipid peroxidation LPO, glutathione peroxidase GPX, metallothionein MT and acetylcholinesterase AChE) in the clam Scrobicularia plana. Temporal evolution of selected endpoints was evaluated throughout an exposure period (1, 7 and 21 days) followed by a depuration phase. The accumulation of all drugs was fast, however clams showed the ability to control the internal content of drugs, keeping their concentration constant throughout the exposure and reducing their content after 7 days of depuration. The induction of biochemical alterations (SOD, CAT, LPO, MT, AChE) was observed in gills and digestive gland probably related to an imbalance in the redox state of clams as a consequence of the exposure to the drug mixture. These alterations were also maintained at the end of the depuration week when the high levels of SOD, CAT, GST and LPO indicated the persistence of oxidative stress and damage to lipids despite the fact that clams were no longer exposed to the mixture.

The high consumption and subsequent input of antibacterial compounds in marine ecosystems has become a worldwide problem. Their continuous presence in these ecosystems allows a direct interaction with aquatic organisms and can cause negative effects over time. The objective of the present study was to evaluate the effects of exposure to three antibacterial compounds of high consumption and presence in marine ecosystems (Ciprofloxacin CIP, Sulfadiazine SULF and Trimethoprim TRIM) on the physiology of the gilthead sea bream, Sparus aurata. Plasma parameters, enzymatic biomarkers of oxidative stress and damage and expression of genes related to stress and growth were assessed in exposed S. aurata specimens. For this purpose, sea bream specimens were exposed to individual compounds at concentrations of 5.2 ± 2.1 μg L^-1 for CIP, 3.8 ± 2.7 μg L^-1 for SULF and 25.7 ± 10.8 μg L^-1 for TRIM during 21 days. Exposure to CIP up-regulated transcription of genes associated with the hypothalamic-pituitary-thyroid (HPT) (thyrotropin-releasing hormone, trh) and hypothalamic-pituitary-interrenal (HPI) axes (corticotropin-releasing hormone-binding protein, crhbp) in the brain, as well as altering several hepatic stress biomarkers (catalase, CAT; glutathione reductase, GR; and lipid peroxidation, LPO). Similar alterations at the hepatic level were observed after exposure to TRIM. Overall, our study indicates that S. aurata is vulnerable to environmentally relevant concentrations of CIP and TRIM and that their exposure could lead to a stress situation, altering the activity of antioxidant defense mechanisms as well as the activity of HPT and HPI axes.

Glutathione peroxidase (Gpx) is an important member of antioxidant enzymes, which can play a vital role in metabolizing reactive oxygen species (ROS) and in maintaining cell homeostasis. In order to study the evolutionary dynamics of gpx gene family in allotetraploid fish species, we identified a total of 14 gpx genes in common carp Cyprinus carpio, while 9 gpx genes were discovered in the diploid progenitor-like species Poropuntius huangchuchieni. Comparative genomic analysis and phylogenetic analysis revealed that the common carp gpx genes had significant expansion and were divided into five distinct subclades. Exon-intron distribution patterns and conserved motif analysis revealed highly conserved evolutionary patterns. Transcript profiles suggested that different gpx genes had specific patterns of regulation during early embryonic development. In adult tissues, gpx genes had a relatively broad expression distribution, most of which were highly expressed in the gills, intestines, and gonads. RT-qPCR studies showed that most gpx genes were downregulated during the initial cd²⁺ treatment stage. Dietary supplementation of Bacillus coagulans at different concentrations (Group 2 of 1.0 × 10⁷ cfu/g, Group 3 of 1.0 × 10⁸ cfu/g, and Group 4 of 1.0 × 10⁹ cfu/g) induced different regulatory responses of gpx subclades. This result suggested that the appropriate concentration of B. coagulans can improve gpx gene expression when exposed to heavy metal cadmium treatment, which may play a vital role in the resistance to oxidative stress and immune responses. This study has expanded our understanding of the functional evolution of the gpx gene family in common carp.

Nanoplastics (NP) (1-100 nm) are a growing global concern, and their adverse effects in marine organisms are still scarce. This study evaluated the effects of polystyrene nanoplastics (10 μg/L; 50 nm nPS) in the marine mussel Mytilus galloprovincialis after a 21 - day exposure. The hydrodynamic diameter and zeta potential of nPS were analysed, over time, in seawater and ultrapure water. A multibiomarker approach (genotoxicity (the comet assay) was assessed in mussel haemocytes, and the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)), biotransformation enzyme (glutathione - S - transferase (GST)), and oxidative damage (LPO)) was assessed in gills and digestive glands to evaluate the toxicity of nPS towards mussels. In seawater, aggregation of nPS is favoured and consequently the hydrodynamic diameter increases. Genotoxicity was highly noticeable in mussels exposed to nPS, presenting a higher % tail DNA when compared to controls. Antioxidant enzymes are overwhelmed after nPS exposure, leading to oxidative damage in both tissues. Results showed that mussel tissues are incapable of dealing with the effects that this emerging stressor pursues towards the organism. The Integrated Biomarker Response index, used to summarise the biomarkers analysed into one index, shows that nPS toxicity towards mussels are both tissue and time dependent, being that gills are the tissue most compromised.

This study investigated the sublethal effects of environmentally relevant concentrations of DCOIT on the neotropical oyster Crassostrea brasiliana. Gills and digestive glands of animals exposed to increasing concentrations of DCOIT were analyzed for biochemical, cellular, and histopathological responses. Exposure to DCOIT (0.2 to 151 μg L^-1) for 120 h triggered oxidative stress in both tissues (through the modulation of GPX, GST, GSH and GR), which led to damage of membrane lipids (increase of LPO and reduction of the NRRT). DCOIT increased histopathological pathologies in gills, such as necrosis, lymphocyte infiltration and epithelial desquamation. This study showed that short term exposure to environmental concentrations of DCOIT causes negative effects on C. brasiliana at biochemical, physiological, and histological levels. Therefore, the use of DCOIT as a booster biocide in antifouling paints should be further assessed, as it may cause environmental hazards to marine organisms.

Fish have defense systems that are capable of repairing damages caused by xenobiotics like benzo[a]pyrene (BaP), so the aims of this study were to identify BaP toxicity in melanomacrophages (MMs) cytoskeleton, evaluate the melanin area in MMs, and analyze genotoxicity. Rainbow trout juveniles (n = 24) were split in 48h and 7d treatments that received 2 mg/kg of BaP. After the experiment, blood samples were collected and liver was removed, to proceed with the analysis: EROD activity, MMs melanin area quantification, melanosomes movements, and a genotoxicity test. The results revealed increased in EROD activity after 48-h and 7-day BaP exposure. The group 7d displayed a reduction in MMs pigmented area, melanosomes aggregation, in addition to an increased frequency of micronucleus. By means of the EROD assay, it was possible to confirm the activation of BaP biotransformation system. The impairment of the melanosomes' movements possibly by an inactivation of the protein responsible for the pigment dispersion consequently affects the melanin area and thus might negatively impact the MMs detoxification capacity. In addition to this cytotoxicity, the increased frequency of micronucleus might also indicate the genotoxicity of BaP in this important fish species.

The knowledge about the effects of pharmaceuticals on aquatic organisms has been increasing in the last decade. However, due to the variety of compounds presents in the aquatic medium, exposure scenarios and exposed organisms, there are still many gaps in the knowledge on how mixtures of such bioactive compounds affect exposed non target organisms. The crayfish Procambarus clarkii was used to analyze the toxicity effects of mixtures of ciprofloxacin, flumequine and ibuprofen at low and high concentrations (10 and 100 μg/L) over 21 days of exposure and to assess the recovery capacity of the organism after a depuration phase following exposure during additional 7 days in clean water. The crayfish accumulated the three compounds throughout the entire exposure in the hepatopancreas. The exposure to the mixture altered the abundance of proteins associated with different cells functions such as biotransformation and detoxification processes (i.e. catalase and glutathione transferase), carbohydrate metabolism and immune responses. Additionally changes in expression of genes encoding antioxidant enzymes and in activity of the corresponding enzymes (i.e. superoxide dismutase, glutathione peroxidase and glutathione transferase) were reported. Alterations at different levels of biological organization did not run in parallel under all circumstances and can be related to changes in the redox status of the target tissue. No differences were observed between control and exposed organisms for most of selected endpoints after a week of depuration, indicating that exposure to the drug mixture did not produce permanent damage in the hepatopancreas of P. clarkii.

Intrauterine growth restriction (IUGR) results in abnormal morphology and gastrointestinal function, such as reduced villi height and crypt depth, thinner mucosa and muscle layers, and reduced brush border enzyme activities, delayed gastric emptying, increased stress response. As a gastrointestinal growth factor, the manner by which the porcine glucagon-like peptide-2 (pGLP-2) microsphere administration restored the gastrointestinal function and growth performance of IUGR piglets was investigated. Fourteen newborn Duroc × (Yorkshire × Landrace) IUGR piglets (0.92 ± 0.113 kg) were assigned into the IUGR (negative control group) and pGLP-2 microsphere groups. The piglets in group pGLP-2 were intraperitoneally administered with 100 mg pGLP-2 microspheres on day 1 after birth. From days 15 to 26 of trial, the body weight of the pGLP-2 group was significantly higher than that of the control. IUGR piglets of group pGLP-2 showed a significantly increased pancreas weight, serum insulin content and activity of lipase and amylase. Injection of pGLP-2 microspheres restored the intestinal absorptive capacity by significantly increasing the mRNA expression of the sodium-glucose cotransporter 1 in the jejunum and the peptide transporter 1 in the jejunum. It also restored the redox balance by increasing the catalase mRNA expression and decreasing the heat shock protein 70 mRNA expression. In addition, this improvement was associated with the significant increase in gut diameter, length and weight. Therefore, it was concluded that the injection of pGLP-2 microspheres was a suitable therapeutic strategy for compensatory growth in low birth weight IUGR piglets.

Since it has been demonstrated that urban effluents can have adverse effects on aquatic organisms, a multibiomarker study was used to evaluate the effects of wastewater treatment plant (WWTP) effluents discharged into the marine and freshwater environments on clams in Cádiz, Spain. One bioassay was performed in the Bay of Cádiz, exposing Ruditapes philippinarum (marine) to a reference site as well as two sites close to WWTP discharges for 14 days. A second bioassay was performed in the Guadalete River, exposing Corbicula fluminea (fresh water) to three sites for 21 days. The biomarkers analysed included defence mechanisms and various toxic effects. Results indicated that WWTP effluents activated defence mechanisms and induced toxic effects in clams exposed to both environments, thus indicating bioavailability of contaminants present in water. Elevated enzymatic activity was found in clams deployed in La Puntilla and El Trocadero compared to control clams and those exposed to the reference site, and 96% of clams deployed at G2 in the Guadalete River died before day 7. Clams exposed to G1 and G3 indicated significant differences in all biomarkers analysed with respect to control clams (p < 0.05). Both species were sensitive to contaminants present in studied sites. This is the first time that these species were used in cages to assess the environmental risk of wastewater effluent discharges in freshwater and marine column environments. The multibiomarker approach provided important ecotoxicological information and is useful for the assessment of the bioavailability and effect of contaminants from WWTP effluents on marine and fresh water invertebrates.

The presence of trace metals in the sediment, water, and biota of the Epe lagoon has been recently linked to oil exploration and municipal perturbations around the lagoon. The study was aimed at assessing the concentrations and associated health risks of Fe, Zn, Cu, Ni, Pb, Cd, Cr, Mn, Co and V in the water, sediment, and Gymnarchus niloticus of Epe lagoon and to evaluate the role of the enteric parasite Nilonema gymnarchi in bioaccumulation of the metals in the fish. The temperature, pH, redox potential, conductivity, turbidity, dissolved oxygen (DO), total dissolved solids (TDS), and salinity were determined in-situ using a handheld multi-parameter probe (Horiba Water Checker Model U-10). The concentrations of Fe, Zn, Cu, Ni, Pb, Cd, Cr, Mn, Co, and V were determined in the surface water, bottom sediment, Gymnarchus niloticus, and its enteric parasites, Nilonema gymnarchi in Epe lagoon using the Flame Atomic Absorption Spectrometer (Philips model PU 9100). The bioaccumulation factors and target hazard quotients of the trace metals in the infected and uninfected fish were estimated and compared. The intestinal tissue sections of the infected and uninfected fish were examined using a binocular dissecting microscope (American Optical Corporation, Model 570) using hematoxylin and eosin (H&E) stain. Biochemical markers such as reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation (MDA) were determined in the liver of the infected and uninfected fish. The SOD level was higher in the uninfected fish than the infected ones supports the indications deduced from the bioaccumulation analysis. Strong positive correlations between SOD and most of the metals- Fe (0.916), Zn (0.919), Cu (0.896), and Ni (0.917) suggests that the metals may have inflicted more toxicity in the uninfected. The histopathological comparisons made between the uninfected and infected fish showed consistency with the outcomes of other comparisons made in this study. These evidence were marked by tissue alterations in the infected fish ranging from no observed changes to mild alterations, while the uninfected exhibited more severe tissue injuries such as hemorrhagic lesions, severe vascular congestion, edema, the increased connective tissue of the submucosa, and vascular congestion. The condition factors of the infected (0.252) and uninfected (0.268) fish indicated slenderness and unfitness possibly due to environmental stressors such as trace metals. The parasitized fish showing better-coping potentials than the uninfected, coupled with the significant bioaccumulation interferences exhibited by the parasite Nilonema gymnarchi is an indication that the parasites may be a good metal sequestration agent for the fish and can be used to forestall the significant health hazard quotient posed by the current level of iron and the synergy of all metals analyzed in the lagoon.

Polycyclic musk compounds have been identified in environmental matrices (water, sediment and air) and in biological tissues in the last decade, yet only minimal attention has been paid to their chronic toxicity in the marine environment. In the present research, the clams Ruditapes philippinarum were exposed to 0.005, 0.05, 0.5, 5 and 50 μg/L of the fragrances Galaxolide® (HHCB) and Tonalide® (AHTN) for 21 days. A battery of biomarkers related with xenobiotics biotransformation (EROD and GST), oxidative stress (GPx, GR and LPO) and genotoxicity (DNA damage) were measured in digestive gland tissues. HHCB and AHTN significantly (p < 0.05) induced EROD and GST enzymatic activities at environmental concentrations. Both fragrances also induced GPx activity. All concentrations of both compounds induced an increase of LPO and DNA damage on day 21. Although these substances have been reported as not acutely toxic, this study shows that they might induce oxidative stress and genotoxicity in marine organisms.

This study assesses the sex-specific effects induced by CdTe QDs, on the marine mussel Mytilus galloprovincialis in comparison to its dissolved counterpart. A 14 days exposure to CdTe QDs and dissolved Cd was conducted (10 μg Cd L^-1), analysing Cd accumulation, oxidative stress, biotransformation, metallothionein and oxidative damage in the gonads. Both Cd forms caused significant antioxidant alterations, whereby QDs were more pro-oxidant, leading to oxidative damage, being females more affected. Overall, biochemical impairments on gonads of M. galloprovincialis demonstrate that the reproductive toxicity induced by CdTe QDs in mussels are sex-dependent and mediated by oxidative stress and lipid peroxidation. It is crucial to acknowledge how gametes are affected by metal-based nanoparticles, such as Cd-based QDs. As well as understanding the potential changes they may undergo at the cellular level during gametogenesis, embryogenesis and larval development potentially leading to serious impacts on population sustainability and ecosystem health.

Light is an essential factor for organisms and affects the endocrine and stress regulation of fish in nature. However, sudden changes in light and dark conditions in artificial environments can negatively impact fish. In the present study, to evaluate the physiological and oxidative stress responses of goldfish (Carassius auratus) exposed to two different light conditions, sudden light changes and slowly dimming light changes for 24 h, we analyzed the mRNA expression and activity of stress indicators [corticotropin-releasing hormone (CRH) and pro-opiomelanocortin (POMC)], levels of plasma cortisol and glucose, mRNA expression of glucocorticoid receptor (GR), and activity of plasma oxidative stress indicators (superoxide dismutase and catalase). Consequently, the mRNA expressions and activities of CRH and POMC, plasma levels of cortisol and glucose, and mRNA expression of GR were found to be significantly increased during the light changes, particularly in the control group. Additionally, plasma levels of cortisol and glucose in the control group were significantly higher than those in the dimming group during the light changes. However, no significant differences in mRNA expression levels and activities of antioxidant enzymes both in the control and dimming groups were observed. These results indicate that dimming light induces less stress than sudden changes in light.

Silver is a ubiquitous metal in the marine environment which can be accumulated by marine organisms. In order to assess the effect of dissolved silver (AgD) and AgNPs in R. philippinarum, the organisms were exposed to 20 μg L^-1 of AgD and AgNPs (15 nm) over 7 days. Bioaccumulation of the metal and oxidative and detoxification biomarkers were studied in control and exposed clams. Ag was accumulated in gills and digestive glands. Results for biochemical biomarkers (superoxide dismutase, catalase and glutathione reductase activity, lipid peroxidation and metallothionein provoked a general increase in the integrated biomarker response index (IBR) values) indicating the induction of oxidative stress in the clams exposed to both Ag treatments. Therefore, the presence of Ag forms at the tested concentration in the aquatic medium represent a risk for R. philippinarum.

Polymetallic seafloor massive sulphide deposits are potential targets for deep-sea mining, but high concentrations of metals (including copper - Cu) may be released during exploitation activities, potentially inducing harmful impact. To determine whether shallow-water shrimp are suitable ecotoxicological proxies for deep-sea hydrothermal vent shrimp the effects of waterborne Cu exposure (3 and 10 days at 0.4 and 4 μM concentrations) in Palaemon elegans, Palaemon serratus, and Palaemon varians were compared with Mirocaris fortunata. Accumulation of Cu and a set of biomarkers were analysed. Results show different responses among congeneric species indicating that it is not appropriate to use shallow-water shrimps as ecotoxicological proxies for deep-water shrimps. During the evolutionary history of these species they were likely subject to different chemical environments which may have induced different molecular/biochemical adaptations/tolerances. Results highlight the importance of analysing effects of deep-sea mining in situ and in local species to adequately assess ecotoxicological effects under natural environmental conditions.

In this study, the performance of two native bivalves in responding to sediment resuspension was investigated during dredging operations of a semi-arid bay (Mucuripe, NE Brazil). The clam Anomalocardia flexuosa and the oyster Crassostrea rhizophorae were selected and caged in two sites influenced by the dredging plume. We assessed the bioaccumulation of metals and hydrocarbons in both species as biomarkers of exposure and then, biomarkers' responses were assessed in gills and digestive glands over a 28 days period, at 7 days intervals: the activities of phase I and II, and antioxidant enzymes, levels of lipid peroxidation and DNA strand breaks. Both transplanted bivalves accumulated Cu, polycyclic aromatic hydrocarbons (PAHs), and linear alkylbenzenes (LABs) in their whole-body tissues, whereas Ni, Pb and Zn accumulation was species-dependent. The exposure time set at day 28 was considered appropriate. Biomarkers exhibited time-related responses in both species, but gills exhibited a more sensitive response, indicating a function of barrier against the uptake of chemicals and also a relevant tissue to be targeted. In clams, Phase I and II enzymes (ethoxyresorufin O-deethylase and glutathione S-transferase) were induced during the period of intense dredging, while in oysters they were activated at the end of operations. Induction of antioxidant enzymes (glutathione peroxidase and glutathione reductase) and elevated levels DNA damage were observed in both exposure surveys. Clams and oysters were sensitive and responded to the exposure of resuspended sediments and the biomarkers of effects were associated with bioaccumulation of contaminants in the integrated analysis. These results indicate that The clam A. flexuosa and the oyster C. rhizophorae are suitable models to be used in monitoring programs or field exposure experiments in tropical regions.

The increasing production of Ag nanoparticle (AgNP) containing products has inevitably led to a growing concern about their release into the aquatic environment, along with their potential behaviour, toxicity, and bioaccumulation in marine organisms exposed to NPs released from these products. Hence, this study is focused on the effects of AgNPs in Saccostrea glomerata (rock oyster) in artificial seawater (ASW); evaluating the NP's stability, dissolution, and bioaccumulation rate. AgNPs NM300K (20 ± 5 nm) in concentrations of 12.5 μgL^-1 and 125 μgL^-1 were used to conduct the experiments, and were compared to a blank and a positive control of 12.5 μgL^-1 AgNO₃. Dissolution in ASW was measured by ICP-OES and stability was assessed by TEM after 1 h and 3, 5, and 7 days of exposure. Bioaccumulation in gills and digestive glands was measured after 7 days of exposure. The higher concentration of AgNPs induced more aggregation, underwent less dissolution, and showed less bioaccumulation, while the lower concentration showed less aggregation, more dissolution and higher bioaccumulation. Five biomarkers (EROD: ethoxyresorufin-o-deethylase, DNA strand breaks, LPO: lipid peroxidation, GST: glutathione S-transferase and GR: glutathione reductase) were analysed at 0, 3, 5 and 7 days. Significant differences compared to the initial day of exposure (day 0) were reported in DNA strand breaks after 5 and 7 days of exposure, GST, from the third day of exposure, in all the Ag samples, and in some samples for LPO and GR biomarkers, while no significant induction of EROD was observed. A combined effect for each type of treatment and time of exposure was also reported for DNA strand breaks and GST biomarkers measured at the digestive glands. In general, the significant inductions measured showed the following trend: 125 μgL^-1 AgNPs >12.5 μgL^-1 AgNPs ∼12.5 μgL^-1 AgNO₃ even though bioaccumulation followed the opposite trend.

Olive oil production generates large volumes of wastewaters mostly in peri-Mediterranean countries with adverse impacts on the biota of the receiving aquatic systems. Few studies have however documented its toxicity on aquatic species, with an almost total lack of relative studies on fish. We assessed the acute and sub-chronic OMW toxicity, as well as the acute and sub-chronic behavioural, morphological and biochemical effects of OMW exposure on the mosquitofish Gambusia holbrooki. LC₅₀ values of the acute bioassays ranged from 7.31% (24 h) to 6.38% (96 h). Behavioural symptoms of toxicity included hypoactivity and a shift away from the water surface, coupled with a range of morphological alterations, such as skin damage, excessive mucus secretion, hemorrhages, fin rot and exophhalmia, with indications also of gill swelling and anemia. Biochemical assays showed that OMW toxicity resulted in induction of catalase (CAT) and inhibition of acetylcholinesterase (AChE) activities. The implications of our results at the level of environmental policy for the sustainable management of the olive mill industry, i.e. the effective restriction of untreated OMW disposal of in adjacent waterways, as well as the implementation of new technologies that reduce their impact (detoxification and/or revalorization of its residues) are discussed.

The red-eared slider (Trachemys scripta elegans), a freshwater turtle, is an invasive species in many parts of the world where it survives in both freshwater and coastal saline habitats. High salinity can induce reactive oxygen species (ROS) production and lead to oxidative damage. In this study, we investigate the antioxidant defense mechanisms of T. s. elegans in response to salinity stress. The results showed that the mRNA expression levels of superoxide dismutase (SODs), catalase (CAT) and glutathione peroxidase (GSH-PXs) were significantly increased in both 5 psu and 15 psu groups at the early stages of salinity exposure (generally 6-48 h), but typically returned to control levels after the longest 30 d exposure. In addition, hepatic and cardiac mRNA levels of the NF-E2-related factor 2 (Nrf2), showed a similar upregulation as an early response to stress, but decreased at 30 d in the 5 psu and 15 psu groups. The mRNA levels of the negative regulator of Nrf2, kelch-like ECH associating protein 1 (Keap1), exhibited the opposite pattern. Moreover, mRNA expression levels of target of rapamycin (TOR) and ribosomal protein S6 kinase 1 (S6K1) in liver and heart showed roughly similar patterns to those for Nrf2. Furthermore, the content of malondialdehyde (MDA) was significantly increased in liver, especially in the 15 psu group by ~2.5-fold. Taken together, these results indicate that T. s. elegans may activate the TOR-Nrf2 pathway to modulate antioxidant genes transcription in order to promote enhanced antioxidant defense in response to salinity stress.

Concerns are growing about the presence of fluoxetine (FLX) in environmental matrices, as well as its harmful effects on non-target organisms. FLX in aquatic ecosystems has been detected in a range varying from pg/L to ng/L, while adverse effects have been reported in several organisms inhabiting freshwater and marine environments. The present study quantifies FLX concentrations in seawater samples from Santos Bay, Brazil and assesses metabolic responses and sublethal effects on the tropical brown mussel Perna perna. Levels of ethoxyresorufin‑O‑deethylase, dibenzylfluorescein dealkylase, glutathione S-transferase, glutathione peroxidase, cholinesterase, lipoperoxidation, and DNA damage were assessed in the gills and digestive gland of these animals, and lysosomal membrane stability was also assessed in hemocytes. FLX altered phase I and II enzyme activities, caused cytogenotoxic effects, and negatively impacted the overall health of mussels exposed to environmentally relevant concentrations. These findings contribute to characterize the risks of introducing this drug into the marine environment.

The presence of pharmaceuticals in the aquatic ecosystem has become a topic of growing interest in recent years. In this study, the marine clam Ruditapes philippinarum was exposed during 14 days to concentrations close to those found in the environment: (15 μg L^-1) of carbamazepine (CBZ), diclofenac (DCF) and ibuprofen (IBU), three pharmaceuticals widely used worldwide and commonly found within the aquatic environment. Additionally, exposure was followed by a depuration phase (7 days). A battery of biomarkers (superoxide dismutase SOD, catalase CAT, glutathione reductase GR, total glutathione peroxidase T-GPx, glutathione transferase GST, lipid peroxidation LPO, acetylcholinesterase AChE and metallothionein MT) was evaluated throughout the exposure and depuration. The Integrated Biomarker Response index was calculated with all selected biomarkers and used as a complementary tool in the evaluation of the organisms' health status. Exposure induced changes in the clams' biochemical responses that led to the hypothesis of the harmful role of the pharmaceuticals resulting in negative effects (changes in enzyme activities, LPO and MT levels, related to ROS production) particularly after short-term exposure. However, the clams showed the ability to cope with these imbalances by recovering their general oxidative status by the end of exposure.

A passive treatment plant, located in the Iberian Pyrite Belt (Huelva, Southwest Spain), was designed for acid mine drainage remediation. Since its installation, the improvement of water quality in terms of hydrochemical composition has been demonstrated successfully. However, according to the Water Framework Directive, the treated effluent must have ecological values for potential living. The freshwater clam Corbicula fluminea was chosen to carry out bioassessments (survival, biomarker responses, and metal bioaccumulation in soft tissue) with effluents from the mining site, as well as, products from the passive treatment plant in order to determine the level of quality of that water from the biological point of view in toxicity tests. Results discarded mortality as endpoint for biomonitoring purposes. Only the lipid peroxidation of the cell membrane evidenced significant responses, even in correlation with the pollution degree of each effluent. Regarding bioaccumulation, some elements displayed a strong relationship (Fe, Cu, Co, and Zn) between concentrations in the environment and in the tissue. As final conclusion, the usage of the Asian clam was validated as biomonitor tool in short term exposure to acid mine drainage, and, as early warning responses (72 h), the chosen parameters would be lipid peroxidation and bioaccumulation of a specific set of elements (Fe, Cu, Co, Zn). Ecological water quality levels reached by the passive treatment plant were in agreement with the efficiency of hydrochemical improvements.

This study assesses the sediment quality of the Guadalquivir River watercourse between the Alcalá del Río dam and the city of Seville. The main objective of this work is to address sediment quality in the area using an integrative approach that links sediment contamination and toxicity using the Asiatic clam (Corbicula fluminea) under field conditions. This is the first study conducted in the area that use of a battery of biomarkers from exposure (GST, GPx) to adverse biological effect (DNA and histopathological damage) to identify the contamination adverse effects in a river area affected by a cocktail of different anthropogenic activities (urban, industrial, agricultural, etc.). The sediment quality characterized in the area shows a significant biological stress related to metal(loid)s at station located in Alcalá del Río in the river upper part of the studied area, being this stress toxic when approaching the city of Seville. The sediments located nearby this city showed toxicity by means of positive values in the biomarkers of effects measured in the caged clams and related to contaminants with an industrial and urban discharge origins. These results have shown the useful and strength of the biomarker approach used in this study that combines biomarker responses from exposure to effects and allows identifying the contamination adverse effects by means of using caging individuals of the Asian clam. It has been proved in the different experiments how once the exposure biomarkers reach a maximum value of their system the detoxification ability of the organisms is collapsed and then the biomarkers of effect are measured significantly in the different tissues. The use of field surveys using tolerant specie such as the Asian clam is recommendable to determine sediment quality under an integrative point of view as here reported.

The gastrointestinal tract (GIT) of fish is a target of contaminants since it can absorb these substances. We evaluated the morphophysiological alterations in the GIT of Sphoeroides testudineus collected in two estuaries presenting differences in their environmental quality (NIA and IA). The intestine was analyzed for histological and neuronal changes; liver and gills for biochemical markers; muscle tissues for neurotoxicity and peripheral blood for genotoxic damage. The results showed alterations in the GIT of the animals collected in the IA, such as muscle tunica and goblet cell density reduction, increased intraepithelial lymphocytes density and changes in neuronal density. Furthermore, changes were observed in MTs and LPO in the gills. Thus, we suggest that TGI is functioning as a barrier that responds to ingested contaminants, in order to reduce their absorption and translocation. Thus, alterations in morphophysiological and enteric neurons in S. testudineus can be used as biomarkers of environmental contamination.

The environmental quality of an acid mine drainage polluted river (Odiel River) in the Iberian Pyrite Belt (SW Spain) was assessed by combining analyses of biomarkers (DNA strand breaks, LPO, EROD, GST, GR, GPx) in freshwater clams (Corbicula fluminea) exposed during 14 days and correlated with metal(loid) environmental concentrations. Results pointed that enzymatic systems are activated to combat oxidative stress in just 24 h. Along exposure, there were homeostatic regulations with the glutathione activity that influenced in lipid peroxidation oscillations, provoking significant DNA strand damage after 14 exposure days. EROD activity showed no changes throughout the exposure period. The Asian clam displayed balance biomarkers of exposure-antioxidant activity under non-stressfully environments; meanwhile, when was introduced into acid polymetallic environments, such as the acid mine drainage, its enzymatic activity was displaced towards biomarkers of effect and the corresponding antioxidant activity.

The antihypertensive losartan (LOS) has been detected in wastewater and environmental matrices, however further studies focused on assessing the ecotoxicological effects on aquatic ecosystems are necessary. Considering the intensive use of this pharmaceutical and its discharges into coastal zones, our study aimed to determine the environmental concentrations of LOS in seawater, as well as to assess the biological effects of LOS on the marine bivalve Perna perna. For this purpose, fertilization rate and embryolarval development were evaluated through standardized assays. Phase I (ethoxyresorufin O‑deethylase EROD and dibenzylfluorescein dealkylase DBF) and II (glutathione S-transferase GST) enzymes, glutathione peroxidase (GPx), Cholinesterase (ChE), lipoperoxidation (LPO) and DNA damage were used to analyze sublethal responses in gills and digestive gland of adult individuals. Lysosomal membrane stability was also assessed in hemocytes. Our results showed the occurrence of LOS in 100% of the analyzed water samples located in Santos Bay, Sao Paulo, Brazil, in a range of 0.2 ng/L-8.7 ng/L. Effects on reproductive endpoints were observed after short-term exposure to concentrations up to 75 mg/L. Biomarker responses demonstrated the induction of CYP450 like activity and GST in mussel gills exposed to 300 and 3000 ng/L of LOS, respectively. GPx activity was also increased in concentration of exposure to 3000 ng/L of LOS. Cyto-genotoxic effects were found in gills and hemocytes exposed in concentrations up to 300 ng/L. These results highlighted the concern of introducing this class of contaminants into marine environments, and pointed out the need to include antihypertensive compounds in environmental monitoring programs.

Lead (Pb) is a non-essential metal. Its occurrence in the environment is related principally to anthropogenic contamination. Pb is toxic to aquatic organisms and can provoke damage to membranes and inhibit the activity of essential enzymes. The filter-feeding, Manila clam Ruditapes philippinarum is widely used as a biomonitor organism to assess metal toxicity. Among biomarkers related to the Pb toxicity, the enzymatic activity of δ-aminolevulinic acid dehydratase (δ-ALAD) has been adopted as a specific tool. Metallothionein (MT), lipid peroxidation (LPO) and antioxidant enzymes activities, such as catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST) and superoxidase dismutase (SOD) have also been employed to assess the toxic effect of metals. Two target tissues, the gills and the digestive gland, were selected to examine biomarker responses. In order to assess the effects of Pb accumulation and the mechanisms involved in the recovery from it, clams were exposed at two Pb levels (10 and 100 µg/L) for 7 days and were later maintained in clean water for 7 days as a depuration period. Pb accumulation was dependent on the exposure concentration and higher Pb levels were observed in the gills compared to the digestive gland. Inhibition of δ-ALAD, GST and SOD and the induction of MT and LPO over the exposure period were observed in the gills and the digestive gland of R. philippinarum. The depuration period showed a continuous inhibition of the δ-ALAD activity and induction of MT and LPO in both tissues. These results demonstrate that lead induced an exposure effect and the 7 days of depuration were not sufficient to recover the basal health status of the clams.