Subcellular effects and lipid metabolism alterations in the gilthead seabream Sparus aurata fed on ovatoxins-contaminated mussels.

The marine microalgae Ostreopsis cf. ovata are a well-known producer of palytoxin (PlTXs) analogues, i.e. ovatoxins (OVTXs) among others, which arouse concern for animal and human health. Both in field and laboratory studies, presence of OVTXs, detected in species directly feeding on O. cf. ovata, was frequently correlated with impairment on organisms' physiology, development and behaviour, while similar knowledge is still lacking for animals feeding on contaminated preys. In this study, transfer and toxicity of OVTXs were evaluated in an exposure experiment, in which gilthead seabream Sparus aurata was fed with bivalve mussel Mytilus galloprovincialis, contaminated by a toxic strain of O. cf. ovata. Mussels exposed to O. cf. ovata for 21 days accumulated meanly 188 ± 13 µg/kg OVTXs in the whole tissues. Seabreams fed with OVTX-contaminated mussels started to reject the food after 6 days of contaminated diet. Although no detectable levels of OVTXs were measured in muscle, liver, gills and gastro-intestinal tracts, the OVTX-enriched diet induced alterations of lipid metabolism in seabreams livers, displaying a decreased content of total lipid and fatty acid, together with overexpression of fatty acid biosynthetic genes, downregulation of ß-oxidation genes and modulation of several genes related to lipid transport and regulation. Results from this study would suggest the hypothesis that OVTXs produced by O. cf. ovata may not be subject to bioaccumulation in fish fed on contaminated preys, being however responsible of significant biological effects, with important implications for human consumption of seafood products.

Cellular effects of microplastics are influenced by their dimension: Mechanistic relationships and integrated criteria for particles definition.

The definition of microplastics (MPs) is nowadays too generic from a biological perspective, since different characteristics of these particles might influence their effects. To provide experimental evidence that size is an important factor to be considered, Mediterranean mussels Mytilus galloprovincialis were exposed to five size classes of polyethylene fragments (PE-MPs, 20-50 µm, 50-100 µm, 100-250 µm, 250-500 µm, 500-1000 µm). After 10 days of exposure, MPs ingestion and mechanistic relationships between particles size and cellular effects were analysed through a wide panel of biological alterations, including immune system responses, cholinergic function, antioxidant system, lipid metabolism and peroxidation. Results were further elaborated through a Weight of Evidence approach, summarizing the overall biological significance of obtained results in a hazard index based on the number and magnitude of variations and their toxicological relevance. PE-MPs 500-1000 µm were identified as the less biologically reactive size class due to the limited ingestion of particles coupled with the lack of biological effects, followed by PE-MPs 250-500 µm, which slightly altered the cholinergic function and lysosomal membranes. Conversely, PE-MPs smaller than 250 µm provoked a more consistent onset of biological alterations in terms of immune system composition and functioning, redox homeostasis, and lipid metabolism. The overall findings of this study highlight the importance of considering the size of particles for monitoring and risk assessment of MPs, introducing a more integrated evaluation of plastic pollution that, beside particles concentration, should adequately weigh those characteristics triggering the onset of biological effects.

Acute and Sublethal Effects of Deltamethrin Discharges from the Aquaculture Industry on Northern Shrimp (Pandalus borealis Krøyer, 1838): Dispersal Modeling and Field Investigations.

Pharmaceutical deltamethrin (Alpha Max), used as delousing treatments in aquaculture, has raised concerns due to possible negative impacts on the marine environment. A novel approach combining different scientific disciplines has addressed this topic. Acute (mortality) and sublethal effects (i.e., fitness, neurological, immunological, and oxidative responses) of exposure of northern shrimp (Pandalus borealis) were studied in laboratory experiments. Passive water sampling combined with sediment analyses revealed environmental concentrations. Finally, dispersal modeling was performed to predict environmental concentrations. Ecotoxicological analyses showed mortality in shrimp after 1 h of exposure to 2 ng L-1 (1000-fold dilution of treatment dose), revealing a high sensitivity to deltamethrin. Sublethal effects included induction of acetylcholinesterase and acyl CoA oxidase activities and oxidative impairment, which may be linked to neurotoxic responses. Field concentrations of 10-200 ng L-1 in water (100 m from the pens) and

Toxicological effects of cigarette butts for marine organisms.

Cigarette butts (CBs), one of the most common litter items found on beaches, represent a still unexplored environmental hazard. This study aimed at a multidisciplinary characterization of their toxicological risks on marine organisms integrating chemical analyses of released compounds with a wide panel of biological responses, such as ecotoxicological bioassays on species of different trophic positions, molecular responses in an ex vivo model (Precision-Cut Tissue Slices, PCTS of mussels digestive glands), bioavailability and cellular biomarkers in mussels exposed to CBs in laboratory experiments. Trace metals, aliphatic and polycyclic aromatic hydrocarbons, nicotine and cotinine were released in artificial seawater after 24 h which determined a significant inhibition of bacterial bioluminescence, oyster embryo development and growth in different algal species. Modulation of peroxisomal proliferation and antioxidant gene expression was observed in mussels PCTS, while the in vivo exposure determined accumulation of chemicals and significant alterations of immune system, antioxidant and neurotoxic responses, peroxisomal proliferation and genotoxic damage. Using a quantitative Weight of Evidence model, the risks of CBs to the marine environment were summarized, highlighting the importance of integrating chemical analyses, batteries of ecotoxicological bioassays, molecular and cellular biomarkers to assess the impact of these hazardous materials on marine environment.

Corrigendum: 10-year time course of Hg and organic compounds in Augusta Bay: Bioavailability and biological effects in marine organisms.

[This corrects the article DOI: 10.3389/fpubh.2022.968296.].

10-year time course of Hg and organic compounds in Augusta Bay: Bioavailability and biological effects in marine organisms.

In the last century, many Mediterranean coastal areas have been subjected to anthropogenic disturbances from industrial activities, uncontrolled landfills, shipyards, and high maritime traffic. The Augusta Bay (eastern Sicily, Italy) represents an example of a strongly impacted coastal environment with an elevated level of sediments contamination due to the presence of one of the largest European petrochemical plants, combined with an extensive commercial and military harbor. The most significant contaminants were represented by mercury (Hg) and hexachlorobenzene (HCB), derived from a former chlor-alkali plant, and other organic compounds like polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs). Since the 1970s, Augusta Bay has become internationally recognized as a contaminated marine environment, although very little information is available regarding the temporal trend of contaminants bioavailability and biological impacts on aquatic organisms. In this study, the Hg and HCB concentrations were investigated over 10 years (from 2003 to 2013) in sediments and invertebrate and vertebrate organisms; these two contaminants' ecotoxicity was further evaluated at a biochemical and cellular level by analyzing the induction of organic biotransformation processes and DNA damages. The results showed high concentrations of Hg and HCB in sediments and their strong bioaccumulation in different species with significantly higher values than those measured in reference sites. This trend was paralleled by increased micronuclei frequency (DNA damage biomarker) and activity of the biotransformation system. While levels of chemicals in sediments remained elevated during the time course, their bioavailability and biological effects showed a gradual decrease after 2003, when the chlor-alkali plant was closed. Environmental persistence of Hg and HCB availability facilitates their bioaccumulation and affects the health status of marine organisms, with possible implications for environmental risk, pollutants transfer, and human health.

Marine heatwaves hamper neuro-immune and oxidative tolerance toward carbamazepine in Mytilus galloprovincialis.

The increased frequency and intensity of short-term extreme warming phenomena have been associated to harsh biological and ecosystem outcomes (i.e., mass mortalities in marine organisms). Marine heatwaves (MHWs), occurring when seasonal temperature threshold is exceeded for at least 5 consecutive days, may reduce the tolerance of coastal species toward additional pressures, but interactions between such multiple stressors are virtually unexplored. The present study aimed to characterize in Mytilus galloprovincialis the influence of a simulated MHW scenario on the toxicological effects of the pharmaceutical carbamazepine (CBZ), ubiquitously detected in the marine environment and chosen as model compound for this relevant class of emerging contaminants. The bioaccumulation of CBZ and responsiveness of various biological parameters, including immune system, antioxidant status, lipid metabolism and cellular integrity, were analyzed in exposed mussels both during and after the end of the heatwave. MHW appeared to strongly modulate accumulation of CBZ, paralleled by weakened immunocompetence and onset of oxidative disturbance that finally evolved to cellular damages and lipid metabolism disorders. Elaboration of the overall results through a quantitative Weight of Evidence model, revealed the highest hazard in organisms exposed to both the stressors 10 days after the end of the heatwave, suggesting that MHWs could leave a footprint on the capability of mussels to counteract CBZ toxicity, thus affecting their vulnerability and predisposition to adverse effects toward multiple stressors.

Community profiling focus group: An empowering tool for immigrant community groups.

Community psychology profiling, a complex and lengthy intervention methodology, can play an important role in enhancing social cohesion and integrating immigrants into local communities. In this pilot research intervention, we wanted to investigate if the Community Profiling Focus Group, which is flexible and a less time-consuming methodology could be employed to initiate empowering processes in marginalized community groups such as immigrants. Five focus groups were carried out with different immigrants. Overall our research shows that, though immigrants had deep concerns mostly on work issues and low acceptance of foreigners, participating in the Community Profiling Focus Group enabled them to be more proactive and to identify objectives they could reach jointly to increase their empowerment and integration in their communities.

Long-lasting effects of chronic exposure to chemical pollution on the hologenome of the Manila clam.

Chronic exposure to pollutants affects natural populations, creating specific molecular and biochemical signatures. In the present study, we tested the hypothesis that chronic exposure to pollutants might have substantial effects on the Manila clam hologenome long after removal from contaminated sites. To reach this goal, a highly integrative approach was implemented, combining transcriptome, genetic and microbiota analyses with the evaluation of biochemical and histological profiles of the edible Manila clam Ruditapes philippinarum, as it was transplanted for 6 months from the polluted area of Porto Marghera (PM) to the clean area of Chioggia (Venice lagoon, Italy). One month post-transplantation, PM clams showed several modifications to its resident microbiota, including an overrepresentation of the opportunistic pathogen Arcobacter spp. This may be related to the upregulation of several immune genes in the PM clams, potentially representing a host response to the increased abundance of deleterious bacteria. Six months after transplantation, PM clams demonstrated a lower ability to respond to environmental/physiological stressors related to the summer season, and the hepatopancreas-associated microbiota still showed different compositions among PM and CH clams. This study confirms that different stressors have predictable effects in clams at different biological levels and demonstrates that chronic exposure to pollutants leads to long-lasting effects on the animal hologenome. In addition, no genetic differentiation between samples from the two areas was detected, confirming that PM and CH clams belong to a single population. Overall, the obtained responses were largely reversible and potentially related to phenotypic plasticity rather than genetic adaptation. The results here presented will be functional for the assessment of the environmental risk imposed by chemicals on an economically important bivalve species.

Susceptibility of polar cod (Boreogadus saida) to a model carcinogen.

Studies that aim to characterise the susceptibility of the ecologically relevant and non-model fish polar cod (Boreogadus saida) to model carcinogens are required. Polar cod were exposed under laboratory conditions for six months to control, 0.03 µg BaP/g fish/week and 0.3 µg BaP/g fish/week dietary benzo(a)pyrene (BaP), a reference carcinogen. The concentrations of the 3-OH-BaP bile metabolite and transcriptional responses of genes involved in DNA adduct recognition (xpc), helicase activity (xpd), DNA repair (xpf, rad51) and tumour suppression (tp53) were assessed after 0, 1, 3 and 6 months of exposure, alongside body condition indexes (gonadosomatic index, hepatosomatic index and condition factor). Micronuclei and nuclear abnormalities in blood and spleen, and liver histopathological endpoints were assessed at the end of the experiment. Fish grew steadily over the whole experiment and no mortality was recorded. The concentrations of 3-OH-BaP increased significantly after 1 month of exposure to the highest BaP concentration and after 6 months of exposure to all BaP concentrations showing the biotransformation of the mother compound. Nevertheless, no significant induction of gene transcripts involved in DNA damage repair or tumour suppression were observed at the selected sampling times. These results together with the absence of chromosomal damage in blood and spleen cells, the subtle increase in nuclear abnormalities observed in spleen cells and the low occurrence of foci of cellular alteration suggested that the exposure was below the threshold of observable effects. Taken together, the results showed that polar cod was not susceptible to carcinogenesis using the BaP exposure regime employed herein.

Transcriptional and Catalytic Responsiveness of the Antarctic Fish Trematomus bernacchii Antioxidant System toward Multiple Stressors.

Ocean-warming and acidification jeopardize Antarctic marine species, adapted to cold and constant conditions and naturally exposed to high pro-oxidant pressures and cadmium (Cd) bioavailability. The aim of this study was to investigate if projected temperature increase and pH reduction may affect the accumulation and the effects of Cd in the rockcod Trematomus bernacchii. Organisms were exposed for 14 days to six scenarios, combining environmental or increased temperature (-1 °C, +1 °C) and control or reduced pH (8.05, 7.60), either with or without Cd (40 µg/L). Responses in liver and gills were analyzed at different levels, including mRNA and functional measurements of metallothioneins and of a wide battery of antioxidants, integrated with the evaluation of the total antioxidant capacity and onset of oxidative damages. In the gills, metallothioneins and mRNA of antioxidant genes (nrf2, keap1, cat, gpx1) increased after Cd exposure, but such effects were softened by warming and acidification. Antioxidants showed slighter variations at the enzymatic level, while Cd caused glutathione increase under warming and acidified scenarios. In the liver, due to higher basal antioxidant protection, limited effects were observed. Genotoxic damage increased under the combined stressors scenario. Overall results highlighted the modulation of the oxidative stress response to Cd by multiple stressors, suggesting the vulnerability of T. bernacchii under predicted ocean change scenarios.

Microplastics in real wastewater treatment schemes: Comparative assessment and relevant inhibition effects on anaerobic processes.

The occurrence, fate and removal of microplastics (MPs) in a wastewater treatment plant (WWTP) in Central Italy were investigated together with their potential adverse effects on anaerobic processes. In the influent of the WWTP, 3.6 MPs.L-1 were detected that mostly comprised polyester fibers and particles in the shape of films, ranging 0.1-0.5 mm and made of polyethylene and polypropylene (PP). The full-scale conventional activated sludge scheme removed 86% of MPs, with the main reduction in the primary and secondary settling. MPs particles bigger than 1 mm were not detected in the final effluent and some loss of polymers types were observed. In comparison, the pilot-scale upflow granular anaerobic sludge blanket (UASB) + anaerobic membrane bioreactor (AnMBR) configuration achieved 94% MPs removal with the abatement of 87% of fibers and 100% of particles. The results highlighted an accumulation phenomenon of MPs in the sludge and suggested the need to further investigate the effects of MPs on anaerobic processes. Accordingly, PP-MPs at concentrations from 5 PP-MPs.gTS-1 to 50 PP-MPs.gTS-1 were spiked in the pilot-scale UASB reactor that was fed with real municipal wastewater, where up to 58% decrease in methanogenic activity was observed at the exposure of 50 PP-MPs.gTS-1. To the best of our knowledge, the presented results are the first to report of PP-MPs inhibition on anaerobic processes.

Integrated characterization and risk management of marine sediments: The case study of the industrialized Bagnoli area (Naples, Italy).

The aim of the present work is to demonstrate the practical importance of a multidisciplinary approach and weighted criteria to synthesize and integrate different typologies of data (or lines of evidence, LOEs), including chemical levels in marine sediments, their bioavailability to specific indicator species, ecotoxicological effects measured through subcellular biomarkers and batteries of bioassays, and potential impacts of pollution on local benthic communities. The area of Bagnoli (Gulf of Naples, Southern Italy) was selected as a model case-study, as it is a coastal area chronically impacted by massive industrial contamination (trace metals and hydrocarbons), and dismissed decades ago without any subsequent remediation or habitat restoration. The results of each LOE were elaborated to provide specific hazard indices before their overall integration in a weight of evidence (WOE) evaluation. Levels of some trace metals and PAHs revealed a severe contamination in the entire study area. Bioavailability of hydrocarbons was evident particularly for high molecular weight PAHs, which also caused significant variations of cellular biomarkers, such as cytochrome P450 metabolization in fish, lysosomal membrane destabilization in mussels, genotoxic effects both in fish and molluscs. The results of a battery of bioassays indicated less marked responses compared to those obtained from chemical and biomarkers analyses, with acute toxicity still present in sediments close to the source of contamination. The analysis of benthic assemblages showed limited evidence of impact in the whole area, indicating a good functioning of local ecosystems at chronic contamination. Overall, the results of this study confirm the need of combining chemical and biological data, the quantitative characterization of various typologies of hazard and the importance of assessing an integrated environmental WOE risk, to orientate specific and scientifically-supported management options in industrialized areas.

Effects of ocean warming and acidification on accumulation and cellular responsiveness to cadmium in mussels Mytilus galloprovincialis: Importance of the seasonal status.

Ocean warming and acidification could represent an additional threat to marine organisms already coping with other anthropogenic impacts, such as chemical contamination in coastal areas. In this study, interactions between such multiple stressors and their synergistic effects in terms of accumulation, detoxification and biological effects of metals were investigated in the Mediterranean mussel Mytilus galloprovincialis. Organisms sampled during the winter period were exposed for 28 days to different combinations of two temperatures (10 °C and 15 °C), two pH/pCO2 (8.20/∼400µatm and 7.4/∼3000µatm) and two cadmium concentrations (0 and 20 µg/L). Cadmium concentrations increased in digestive glands and gills of metal-exposed mussels and were further enhanced by co-exposure at higher temperature. Interactive effects of temperature and/or pH were observed on Cd-mediated metallothionein induction, responsiveness of antioxidant system and onset of oxidative damages in lipids, with tissue-specific effects. Immunological effects showed a generalized sensitivity of lysosomal membrane stability toward the investigated stressors with major effects in co-exposed organisms. Cadmium and temperature affected phagocytosis efficiency and composition of haemocyte populations probably influencing the micronucleus frequency through varied mitotic rate. Several differences were highlighted between these results and those previously obtained from mussels exposed in summer, supporting the importance of season when addressing the tolerance of temperate organisms to variations of environmental factors. The elaboration of the whole biomarker results through weighted criteria allowed to summarize specific hazard indices, highlighting tissue-specific sensitivity toward multiple stressors and the need of improving the knowledge on interactions between multiple stressors.

Oxidative and interactive challenge of cadmium and ocean acidification on the smooth scallop Flexopecten glaber.

Ocean acidification (OA) may affect sensitivity of marine organisms to metal pollution modulating chemical bioavailability, bioaccumulation and biological responsiveness of several cellular pathways. In this study, the smooth scallop Flexopecten glaber was exposed to various combinations of reduced pH (pH/pCO2 7.4/∼3000 µatm) and Cd (20 µg/L). The analyses on cadmium uptake were integrated with those of a wide battery of biomarkers including metallothioneins, single antioxidant defenses and total oxyradical scavenging capacity in digestive gland and gills, lysosomal membrane stability and onset of genotoxic damage in haemocytes. Reduced pH slightly increased concentration of Cd in scallop tissues, but no effects were measured in terms of metallothioneins. Induction of some antioxidants by Cd and/or low pH in the digestive gland was not reflected in variations of the total oxyradical scavenging capacity, while the investigated stressors caused a certain inhibition of antioxidants and reduction of the scavenging capacity toward peroxyl radical in the gills. Lysosomal membrane stability and onset of genotoxic damages showed high sensitivity with possible synergistic effects of the investigated factors. The overall results suggest that indirect effects of ocean acidification on metal accumulation and toxicity are tissue-specific and modulate oxidative balance through different mechanisms.

Antioxidant defenses in polar cod (Boreogadus saida) and responsiveness toward dietary crude oil exposure.

Increasing anthropogenic activities in the Arctic pose the risk for accidental oil spills but our knowledge of ecotoxicological effects is still limited for Arctic organisms. This study aimed to provide insights in baseline levels of antioxidants in polar cod (Boreogadus saida) from different Arctic locations and investigate the susceptibility of this species to oxidative stress during a 32 day dietary crude oil exposure. Baseline levels of individual antioxidants and total oxyradical scavenging capacity (TOSC) varied among different Arctic fjords. Upon crude oil exposure, dose- and time-dependent polycyclic aromatic hydrocarbons (PAHs) biotransformation was evident through hepatic ethoxyresorufine-O-deethylase activity, glutathione S-transferase mRNA expression and biliary PAH metabolites. Changes in the activity of individual antioxidants and TOSC was, however, slight and transient upon exposure. In conclusion, biotransformation was clearly induced by the sub-lethal levels of crude oil in polar cod, while the antioxidant defense responded with high variability.

The Impact Of Dispositional Variables Of Elders, Relatives, And Paid Caregivers On Elders' Empowerment And Life Satisfaction.

Background/Study Context: The impact of dispositional characteristic of elders' closest network members on elders' life satisfaction and empowerment has remained largely unexplored. This innovative study aimed to assess the impact of dispositional variables of elders, relatives, and paid caregivers on life satisfaction and empowerment of elders. METHODS: One hundred forty-three triads (one elder, one paid caregiver, and one relative) completed six scales measuring modifiable personality characteristics (loneliness, optimism, regulation of positive and negative emotions), life satisfaction, and sense of empowerment. RESULTS: Two dispositional variables of relatives (positivity and ability to set goals) and one dispositional variable of care workers (self-satisfaction) were significantly related to life satisfaction of elders. Positivity of elders and the capacity to set objectives of the family members affected empowerment of elders. CONCLUSION: Future interventions aiming to improve life satisfaction and empowerment of elders should focus on these modifiable dispositional variables.

Nrf2 and regulation of the antioxidant system in the Antarctic silverfish, Pleuragramma antarctica: Adaptation to environmental changes of pro-oxidant pressure.

Despite the key importance of Nrf2-Keap1 in regulating antioxidant system in vertebrates, this system is still poorly investigated in marine species. The present study focused on the Antarctic silverfish Pleuragramma antarctica which, during the final phases of embryo development in platelet ice, is challenged by a sudden enhancement of environmental oxidative conditions associated to ice melting. Partial coding sequences were identified for Nrf2, its repressor Keap1 and for typical Nrf2-target antioxidant genes, like catalase, glutathione peroxidase isoform 1 and Cu/Zn-dependent superoxide dismutase. Compared to temperate homologues, the protein sequences showed an elevated conservation of amino acids essential for catalytic functions, while a few specific substitutions in non-essential regions may represent a molecular adaptation to improve flexibility and accessibility to active site at cold temperatures. The role of the Nrf2-Keap1 pathway in modulating the activation of antioxidant defences was demonstrated at both transcriptional and functional levels with a clear temporal increase of antioxidant protection in embryos before the hatching. Such findings confirm the importance of Nrf2 and highlight regulation of antioxidants as an adaptive strategy in P. antarctica to protect the early life stages toward the environmental changes of pro-oxidant pressure.

Dispositional characteristics, relational well-being and perceived life satisfaction and empowerment of elders.

OBJECTIVES: The broad purpose of this research is to identify the key modifiable variables most related to elders' life satisfaction and empowerment in order to improve the efficacy of interventions projects. Our study aims to integrate the theoretical perspectives of personality and community psychology focusing both on dispositional characteristics and relational well-being of elders, investigating triads, composed by an elder, a paid caregiver and the most involved relative. This study explores the impact of (1) some socio-demographic characteristics of elders, (2) some modifiable dispositional variables of elders and (3) elders' relational well-being on elders' empowerment and life satisfaction. METHOD: The study involved 429 people in 143 triads. Semi-structured interviews with elders, paid caregiver and close relatives were used to construct a new pilot measure of elders' relational well-being. Life Satisfaction, Empowerment, Loneliness, Positivity, Humor and Emotions self-efficacy scales were also administered. Hierarchical multiple regressions were performed. RESULTS: Elders' positivity, relational well-being of elders and living alone were significantly related to empowerment. Elders' relational well-being and positivity significantly contributed to life satisfaction. DISCUSSION: Interventions to increase empowerment and life satisfaction should focus primarily on augmenting positivity and relational well-being integrating the theoretical premises of both personality and community psychology.

Indirect effects of climate changes on cadmium bioavailability and biological effects in the Mediterranean mussel Mytilus galloprovincialis.

Despite the great interest in the consequences of climate change on the physiological functioning of marine organisms, indirect and interactive effects of rising temperature and pCO2 on bioaccumulation and responsiveness to environmental pollutants are still poorly explored, particularly in terms of cellular mechanisms. According to future projections of temperature and pH/pCO2, this study investigated the main cellular pathways involved in metal detoxification and oxidative homeostasis in Mediterranean mussels, Mytilus galloprovincialis, exposed for 4 weeks to various combinations of two levels of pH/pCO2 (8.2/∼400 µatm and 7.4/∼3000 µatm), temperature (20 and 25 °C), and cadmium addition (0 and 20 µg/L). Bioaccumulation was increased in metal exposed organisms but it was not further modulated by different temperature and pH/pCO2 combinations. However, interactions between temperature, pH and cadmium had significant effects on induction of metallothioneins, responses of the antioxidant system and the onset of oxidative damages, which was tissue dependent. Multiple stressors increased metallothioneins concentrations in the digestive gland revealing different oxidative effects: while temperature and cadmium enhanced glutathione-dependent antioxidant protection and capability to neutralize peroxyl radicals, the metal increased the accumulation of lipid peroxidation products under acidified conditions. Gills did not reveal specific effects for different combinations of factors, but a general stress condition was observed in this tissue after various treatments. Significant variations of immune system were mainly caused by increased temperature and low pH, while co-exposure to acidification and cadmium enhanced metal genotoxicity and the onset of permanent DNA damage in haemocytes. Elaboration of the whole biomarker data in a cellular hazard index, corroborated the synergistic effects of temperature and acidification which increased the toxicological effects of cadmium. The overall results confirmed that climate change could influence ecotoxicological effects of environmental contaminants, highlighting the importance of a better knowledge of cellular mechanisms to understand and predict responsiveness of marine organisms to such multiple stressors.