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Integrated compensatory responses of physiological systems towards homeostasis are generally overlooked when it comes to analysing alterations in biochemical parameters indicative of such processes. Here an hypothesis-driven multivariate analysis accounting for interactive multibiomarker responses was used to investigate effects of long-term exposure of Carcinus maenas to Hazardous and Noxious Substances (HNS). Adult male crabs were exposed to low and high post-spill levels of acrylonitrile (ACN) or aniline (ANL) for 21d. Bioaccumulation, feeding behaviour, and biomarkers related to mode-of-action (MoA) (detoxification, neurotransmission and energy production) were evaluated over time. Distinct temporal patterns of response to low and high exposure concentrations were depicted, with a main set of interactive multibiomarker predictors identified for each HNS (five for ACN and three for ANL), useful to follow coupled evolvement of biomarker responses. ACN caused peripheral neurotoxic effects coupled with enhanced biotransformation and significant oxidative damage particularly relevant in gills. ANL elicited alterations in central neurotransmission affecting ventilation coupled with very low levels of oxidative damage in gills. Results indicate chronic toxicity data are determinant to improve HNS hazard assessment if the aim is to obtain reliable risk calculations, and develop effective predictive models avoiding overestimation but sufficiently protective. Accounting for multibiomarker interactions brought otherwise overlooked information about C. maenas responses and MoA of ACN and ANL., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Shipping industry and seaborne trade have rapidly increased over the last fifty years, mainly due to the continuous increasing demand for chemicals and fuels. Consequently, despite current regulations, the occurrence of accidental spills poses an important risk. Hazardous and noxious substances (HNSs) have been raising major concern among environmental managers and scientific community for their heterogeneity, hazardous potential towards aquatic organisms and associated social-economic impacts. A literature review on ecotoxicological hazards to aquatic organisms was conducted for seven HNSs: acrylonitrile, n-butyl acrylate, cyclohexylbenzene, hexane, isononanol, trichloroethylene and xylene. Information on the mechanisms of action of the selected HNS was also reviewed. The main purpose was to identify: i) knowledge gaps in need of being addressed in future research; and ii) a set of possible biomarkers suitable for ecotoxicological assessment and monitoring in both estuarine and marine systems. Main gaps found concern the scarcity of information available on ecotoxicological effects of HNS towards marine species and their poorly understood mode of action in wildlife. Differences were found between the sensitivity of freshwater and seawater organisms, so endpoints produced in the former may not be straightforwardly employed in evaluations for the marine environment. The relationship between sub-individual effects and higher level detrimental alterations (e.g. behavioural, morphological, reproductive effects and mortality) are not fully understood. In this context, a set of biomarkers associated to neurotoxicity, detoxification and anti-oxidant defences is suggested as potential indicators of toxic exposure/effects of HNS in marine organisms. Overall, to support the development of contingency plans and the establishment of environmental safety thresholds, it will be necessary to undertake targeted research on HNS ecotoxicity in the marine environment. Research should address these issues under more realistic exposure scenarios reflecting the prevailing spatial and temporal variability in ecological and environmental conditions., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Among organic pollutants existing in coastal areas, polycyclic aromatic hydrocarbons (PAHs) are of great concern due to their ubiquity and carcinogenic potential. The aim of this study was to evaluate the seasonal patterns of PAHs in the digestive gland and arm of the common octopus (Octopus vulgaris) from the Northwest Atlantic Portuguese coast. In the different seasons, 18 PAHs were determined and the detoxification capacity of the species was evaluated. Ethoxyresorufin O-deethylase (EROD) and ethoxycoumarin O-deethylase (ECOD) activities were measured to assess phase I biotransformation capacity. Individual PAH ratios were used for major source (pyrolytic/petrogenic) analysis. Risks for human consumption were determined by the total toxicity equivalence approach. Generally, low levels of PAHs were detected in the digestive gland and in the arm of octopus, with a predominance of low molecular over high molecular weight compounds. PAHs exhibited seasonality in the concentrations detected and in their main emission sources. In the digestive gland, the highest total PAH levels were observed in autumn possibly related to fat availability in the ecosystem and food intake. The lack of PAH elimination observed in the digestive gland after captivity could be possibly associated to a low biotransformation capacity, consistent with the negligible/undetected levels of EROD and ECOD activity in the different seasons. The emission sources of PAHs found in the digestive gland varied from a petrogenic profile observed in winter to a pyrolytic pattern in spring. In the arm, the highest PAH contents were observed in June; nevertheless, levels were always below the regulatory limits established for food consumption. The carcinogenic potential calculated for all the sampling periods in the arm were markedly lower than the ones found in various aquatic species from different marine environments. The results presented in this study give relevant baseline data for environmental monitoring of organic pollution in coastal areas., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In order to further demonstrate that TBT-induced imposex through RXR signaling is not species-specific, Hexaplex trunculus was selected as an experimental model species. We first isolated RXR in H. trunculus, and determined gene transcription through quantitative real-time PCR in key tissues (e.g., penis/penis-forming area and central nervous system:- CNS), upon exposure to tributyltin (TBT) (5 and 50 ng TBTL(-1)). Two months of exposure to TBT induced imposex and led to a significant increase in the severity of the phenomenon in females and an increase in male penis lengths. Exposure to TBT altered RXR gene transcription in a tissue and sex-specific manner. In the CNS, there were no significant changes in RXR gene transcription between control and TBT-exposed females. A similar trend was observed in male CNS. On the contrary, in the penis-forming area/penis of females exposed to TBT, a significant increase in RXR gene transcription was observed in the 50 ng TBTL(-1) group. Furthermore, a positive correlation was observed between overall female penis lengths and RXR gene transcription. In males, although a trend towards an increase in RXR gene transcription in penis was observed, differences did not reach significance. Overall, the results of the present study give further support to a local role of RXR in the penis-forming area during the development of imposex by TBT, thus suggesting a conserved function of RXR in penis formation at least within prosobranch gastropods., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The protein levels and tissue distribution patterns of P-glycoprotein (Pgp) and cytochrome P450 (CYP1A) were investigated in Nile tilapia (Oreochromis niloticus) after waterborne exposure to different benzo(a)pyrene (BaP) concentrations, using immunochemical approaches. The Pgp mammalian monoclonal antibody (mAb) C219 cross reacted with a ∼170kDa protein, almost exclusively localized to the bile canaliculi, while probing with the Pgp mammalian mAb C494, resulted in a positive reaction in liver, gills and intestine of Nile tilapia and covered a wider set of cell types. Levels of Pgp expression were not altered after in vivo exposure to BaP. CYP1A, detected with the mAb C10-7, reacted positively in liver, gills and intestine and followed a BaP dose-dependent fold induction. Taken together, these results indicate that CYP1A is involved in BaP metabolism in liver, gills and intestine, however, further studies are needed to elucidate the possible interaction of the efflux protein Pgp with BaP and/or its metabolites., (Copyright © 2013 Elsevier B.V. All rights reserved.)

BaP is one of the most studied PAH, due to its ubiquitous presence in aquatic environments and toxicity to aquatic organisms. The main goal of this study was to assess BaP effects in Nile Tilapia after waterborne and dietary exposures, through the evaluation of EROD and GST activities in liver, gills and intestine, and BaP metabolites in bile; and also to evaluate the usefulness of these commonly used biomarkers after two different routes of exposure. Waterborne exposure to BaP led to a significant induction of EROD in all tissues analyzed (644%, 1640% and 2880% in relation to solvent in liver, gill and intestine respectively) while in dietary exposures EROD was induced only in intestine (3143%) after exposure to high BaP concentrations. GST activities with CDNB were slightly induced in liver (40%) and in gill (66%) after water exposure to BaP, and in intestine after dietary exposure to low BaP concentrations (182%). BaP metabolites in bile increased after both exposure routes, and were highly correlated with EROD activity after water exposure. In summary, this work has shown that the effects of BaP on biotransformation pathways depend on the route of exposure. Moreover, barrier tissues like gills and intestine also have an important role in the first-pass metabolism of BaP, reducing the amount of parent compound that reaches the liver to be metabolized. For that reason, EROD activity as a biomarker of exposure should also be applied in extrahepatic organs, like gills and intestine, in monitoring studies. Biliary BaP type metabolites are good reflectors of contamination levels under both exposure routes, while GST activity with CDNB as substrate, as a phase II enzyme, does not seem a reliable biomarker of exposure to BaP regardless the route of exposure., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

This study aimed to investigate the relationships between levels of organochlorine compounds (PCBs and DDTs), polycyclic aromatic hydrocarbons (PAHs), metals and biomarkers of exposure (EROD and GST) and oxidative stress biomarkers, in wild and cultured European seabass (Dicentrarchus labrax). Additionally, the fatty acid profile was determined in muscle to assess differences between cultivated and wild specimens of marketable size. Although results pointed to relatively low concentration of contaminants, wild specimens presented enhanced accumulation of metals and cultivated specimens higher levels of organic compounds. Biomarkers were correlated with contaminants in tissues, showing that higher levels can cause effects at the biochemical level. Wild specimens have presented higher content on omega-3 fatty acids in muscle. Besides the useful application of biomarkers in wild species, this study has shown that they can also be applied to monitor biological effects in the fish farming process., (Copyright 2010 Elsevier Inc. All rights reserved.)

Metals are environmentally ubiquitous and can be found at high concentrations in seawater and subsequently in marine organisms. Metals with high redox potential can trigger oxidative stress mechanisms with damaging effects in biological tissues. In aquatic species, oxidative stress has been evaluated by assessing antioxidant enzymes activities and oxidative damages in tissues. The purpose of this study was to evaluate oxidative stress biomarkers and metal residues in white seabream (Diplodus sargus), a species entering aquaculture production in Portugal. Metal residues (Cu, Cd, As and Pb), in liver and muscle, as well as oxidative stress biomarkers were assessed at different stages in the life cycle of white seabream under culture conditions and in wild specimens, of a marketable size. Metal concentrations in tissues were low, and below the established limits. However, wild white seabream showed higher accumulation than cultured ones. Antioxidant enzymes, namely catalase (CAT) and superoxide dismutase (SOD), were correlated with metal accumulation. Oxidative damages to tissues were low, with wild white seabream showing lower levels than cultured fish. This study showed that white seabream has a good antioxidant defense system, capable of reducing oxidative damages in tissues resulting from the presence of metals.

Thirty three PCB congeners were analyzed in three size classes of seabass (Dicentrarchus labrax), from a semi-intensive fish farm, in food and in abiotic compartment. Water and suspended particulated matter (SPM) showed higher concentrations of lower chlorinated PCBs, whereas fish tissues and food showed higher concentrations of CB153 and CB138. In order to examine the influence of different inputs to seabass contamination, bioaccumulation factors (BAF), biota-suspended particulate matter bioaccumulation factors (BSMAF) and biomagnification factors (BMF) were calculated. In general, the log BAF and log BSMAF showed good correlations with log K(ow), for congeners with log K(ow)>6.1. In three fish size classes, PCB congeners with log K(ow)<6.1 were found at concentrations higher than what was expected based on the BAF calculated for the more hydrophobic congeners, suggesting a different contribution of sources. The application of a mass balance model for accumulation of some congeners in seabass estimated concentrations in fish 3- to 38-fold higher than the measured values, probably by overestimating gill absorption. The model was applied with other scenarios in order to maximize uptake from food, which may have a significant influence in accumulation, and not only from diet pellets. Its contribution to fish contamination for highly chlorinated PCBs may reach a maximum of 54-64%, and for lower chlorinated PCBs its contribution is lower than 20%. In this work it was proved that to reduce fish contamination the quality of the water should be controlled.