Polystyrene microplastic alters the redox state and arsenic metabolization in the freshwater bivalve Limnoperna fortunei.

Most organisms possess the capacity to metabolize arsenic (As) accumulating compounds to less toxic forms, thus minimizing the adverse effect induced by this metalloid. However, other contaminants may to interfere with As metabolism, contributing to the accumulation of more toxic compounds. Microplastics (MPs) are omnipresent in aquatic environment and may induce toxicological effects (alone or in combination with other contaminants) on living organisms. Therefore, the objective of the present study was to evaluate the effect of the exposure of the freshwater clam Limnoperna fortunei to a combination of MP (4 and 40 µg/L of polystyrene microbeads, 1.05 µm) and As (50 µg/L) for 48 h, evaluating the accumulation and metabolization of As and oxidative stress parameters, such as catalase (CAT), glutathione-S-transferase activities, total antioxidant competence, reduced glutathione (GSH), and lipid damage in the gills and digestive glands. Results revealed that low MP concentration disrupts the redox state of the digestive gland by a decrease in the antioxidant activity (CAT and total antioxidant capacity). GSH levels in the gills of animals exposed to MP (4 µg/L) alone and the combination of MP + As increased, concomitant with an increase in the percentage of toxic compounds, indicating the effect of MP on As metabolism. Although, few studies evaluated the effect of coexposure to MP + As by considering metabolization of metalloid in freshwater bivalve, our results revealed that exposure to MP reduced the metabolization capacity of As, favoring the accumulation of more toxic compounds besides the MP alone, which showed a pro-oxidant effect in L. fortunei.

Multi-benthic size approach to unveil different environmental conditions in a Mediterranean harbor area (Ancona, Adriatic Sea, Italy).

Harbors are hubs of human activity and are subject to the continuous discharge and release of industrial, agricultural, and municipal waste and contaminants. Benthic organisms are largely known to reflect environmental conditions they live in. Despite meio- and macrofauna interacting within the benthic system, they are ecologically distinct components of the benthos and as such may not necessarily respond to environmental conditions and/or disturbances in the same way. However, in a few field studies the spatial patterns of meio- and macrofauna have been simultaneously compared. In the present study, we assess the response and patterns in the abundance, diversity, and distribution of the two benthic size classes to the different environmental conditions they live in (i.e., sediment concentrations of selected trace metals and polycyclic aromatic hydrocarbons (PAHs); organic matter contents and grain size) characterizing the Ancona Harbor (Adriatic Sea). Meio- and macrofauna provided partially similar types of information depending on the indices used (univariate measures or community structure/species composition) and the different 'response-to-stress'. The community structure (i.e., taxa composition) of both benthic size components clearly showed differences among sampling stations located from inside to outside the harbor, reflecting the marked environmental heterogeneity and disturbance typically characterizing these systems. Notwithstanding, the univariate measures (i.e., meio- and macrofauna total abundance, diversity indices and equitability) didn't show similar spatial patterns. Meiofauna were likely to be more sensitive to the effects of environmental features and contaminants than macrofauna. Overall, trace metals and PAHs affected the community composition of the two benthic components, but only the meiofauna abundance and diversity were related to the environmental variables considered (i.e., quantity and quality of organic matter). Our results pinpoint the importance of studying both meio- and macrofauna communities, which could provide greater insight into the processes affecting the investigated area and reveal different aspects of the benthic ecosystems in response to harbor conditions.

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.

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.

Organochlorines and Polycyclic Aromatic Hydrocarbons as fingerprint of exposure pathways from marine sediments to biota.

To elucidate the dynamics of a suite of organochlorine contaminants (PCBs, HCB), PAHs and Hg and verify the potential of these pollutants as reliable fingerprints of sources, an ensemble of marine sediments and organisms (finfish, shellfish species and Mytilus galloprovincialis) were analysed from the contaminated Augusta Bay (Southern Italy). The Hg and HCB concentration in the sediments exceeded the EQS of the Directive 2000/60/EU. Similarly, ∑PCB and selected PAHs were above the threshold limit set by regulation. The marine organisms showed Hg concentrations above CE 1881/2006. Contaminants in transplanted mussel evidenced an increased accumulation overtime and different distribution patterns between sampling sites. Analysis of the homolog composition of PCB congeners revealed comparable patterns between sediments and marine organisms and offered the opportunity to define a robust fingerprint for tracing contaminants transfer from the abiotic to the biotic compartments. These results were confirmed by the Fluoranthene/Pyrene, Hg and HCB distribution modes.

Co-exposure to nTiO2 impairs arsenic metabolism and affects antioxidant capacity in the marine shrimp Litopenaeus vannamei.

Aquatic animals are vulnerable to arsenic (As) toxicity. However, rarely does a contaminant occur alone in the aquatic environment. For this reason, this study was conducted to evaluate whether titanium dioxide nanoparticles (nTiO2) can interfere with the effects induced by As in Litopenaeus vannamei. Arsenic accumulation and metabolic capacity; expression and enzymatic activity of GSTΩ (glutathione-S-transferase omega isoform); antioxidant responses such as GSH, GR, and GST (reduced glutathione levels, glutathione reductase, and glutathione-S-transferase activity, respectively); and lipid peroxidation in the gills and hepatopancreas of shrimp were evaluated. The results are summarized as follows: (1) higher accumulation of As occurred in both tissues after exposure to As alone; (2) co-exposure to nTiO2 affected the capacity to metabolize As; (3) GSTΩ gene expression was not modified, but its activity was decreased by co-exposure to both contaminants; (4) As alone increased the GSH levels in the hepatopancreas, and co-exposure to nTiO2 reduced these levels in both tissues; (5) a decrease in the GST activity in the gills occurred with all treatments; (6) in the gills, GR activity was increased by As, and nTiO2 reversed this increase, whereas in the hepatopancreas co-exposure inhibited enzyme activity; (7) only in the hepatopancreas lipid damage was observed when animals were exposed to As or nTiO2 but not in co-exposure. The results showed that the As induces toxic effects in both tissues of shrimp and that co-exposure to nTiO2 can potentiate these effects and decrease the capacity to metabolize As, favoring the accumulation of more toxic compounds.

Environmental pharmaceuticals and climate change: The case study of carbamazepine in M. galloprovincialis under ocean acidification scenario.

Contaminants of emerging concern and ocean changes are key environmental stressors for marine species with possibly synergistic, but still unexplored, deleterious effects. In the present study the influence of a simulated ocean acidification scenario (pH = 7.6) was investigated on metabolism and sub-lethal effects of carbamazepine, CBZ (1 µg/L), chosen as one of the most widely diffused pharmaceuticals in marine organisms. A multidisciplinary approach was applied on mussels, M. galloprovincialis, integrating measurement of drug bioaccumulation with changes in the whole transcriptome, responsiveness of various biochemical and cellular biomarkers including immunological parameters, lipid and oxidative metabolism, onset of genotoxic effects. Chemical analyses revealed a limited influence of hypercapnia on accumulation and excretion of CBZ, while a complex network of biological responses was observed in gene expression profile and functional changes at cellular level. The modulation of gamma-aminobutyric acid (GABA) pathway suggested similarities with the Mechanism of Action known for vertebrates: immune responses, cellular homeostasis and oxidative system represented the processes targeted by combined stressors. The overall elaboration of results through a quantitative Weight of Evidence model, revealed clearly increased cellular hazard due to interactions of CBZ with acidification compared to single stressors.

Human pharmaceuticals in marine mussels: Evidence of sneaky environmental hazard along Italian coasts.

Despite the increasing interest for pharmaceuticals in the marine environment, their accumulation in wild organisms and consequent environmental hazards are still poorly known. The Mediterranean Sea is highly challenged by the density of coastal populations, large consumption of pharmaceuticals and their often limited removal by Wastewater Treatment Plants (WWTPs). In this respect, the present study aims to provide the first large-scale survey on the distribution of such contaminants of emerging concern in native mussels, Mytilus galloprovincialis from Italian coasts. Organisms were collected from 14 sites representative of relatively unpolluted marine waters along the Adriatic and Tyrrhenian Sea and analysed for 9 common pharmaceuticals including Non-Steroidal Anti-Inflammatory Drugs (NSAIDs: Diclofenac DIC, Ibuprofen IBU, Ketoprofen KET and Nimesulide NIM), the analgesic Acetaminophen AMP, the antiepileptic Carbamazepine CBZ, the antihypertensive Valsartan VAL, the anxiolytic Lormetazepam LOR and the antidepressant Paroxetine PAR. Results indicated the widespread occurrence of the majority of pharmaceuticals in mussel tissues: CBZ was measured in >90% of analysed samples, followed by VAL (>50%), PAR (>40%), and DIC (>30%), while only AMP and KET were never detected. Heterogeneous tissue concentrations ranged from a few units up to hundreds of ng/g (d.w.), while seasonal and interannual variability, investigated over 4 years, did not highlight any clear temporal trend. Limited differences obtained between the Adriatic and Tyrrhenian Sea, as well as coastal versus off-shore sampling sites, suggest that analysed levels of pharmaceuticals in mussels tissues should be considered as baseline concentrations for organisms collected in unpolluted areas of the Mediterranean. This study provided the first unambiguous evidence of the widespread occurrence of pharmaceuticals in marine mussels from Italian coasts, giving novel insights on the potential ecotoxicological hazard from such compounds in marine species.

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.

Impact of different crystalline forms of nTiO2 on metabolism and arsenic toxicity in Limnoperna fortunei.

Arsenic (As) is a ubiquitous contaminant in the environment and it is known to induce oxidative stress in aquatic organisms. In an attempt to remove As from water, some studies have suggested the titanium dioxide nanomaterial (nTiO2) as a promising alternative. However, it has been observed that nTiO2 can induce toxicity alone or in combination with metals, and this toxicity is dependent on its crystalline form of nanomaterial (mainly rutile as nTiO2R and anatase as nTiO2A, respectively). Considering that both (nTiO2 and As) can occur together, the objective of this study was to evaluate if co-exposure to rutile and anatase may influence accumulation, metabolisation, and toxicity of arsenite (As+3) in the golden mussel Limnoperna fortunei after 48 h of co-exposure to nTiO2 (1 mg/L) and As (50 µg/L). Accumulation and chemical speciation of As in organisms were determined. Also, biochemical analyses, such as the activity of the enzymes glutathione S-transferase omega (GSTΩ), catalase (CAT) and glutathione S-transferase (GST), as well as lipid peroxidation (LPO) were investigated. Results showed that co-exposure to nTiO2A + As changed accumulation pattern of metalloid in gills and digestive gland. Both crystalline forms of nTiO2 affected the metabolisation capacity favoring the accumulation of more toxic As compounds and nTiO2A alone or in combination with As showed induce oxidative stress in gills of L. fortunei. In this way, it has a high potential risk of the co-exposure of these contaminants to aquatic organisms, and it also needs to consider the nanomaterial (nTiO2) properties and their application in the environmental remediation, carefully and judiciously.

Role of the chronic air pollution levels in the Covid-19 outbreak risk in Italy.

After the initial outbreak in China, the diffusion in Italy of SARS-CoV-2 is exhibiting a clear regional trend with more elevated frequency and severity of cases in Northern areas. Among multiple factors possibly involved in such geographical differences, a role has been hypothesized for atmospheric pollution. We provide additional evidence on the possible influence of air quality, particularly in terms of chronicity of exposure on the spread viral infection in Italian regions. Actual data on Covid-19 outbreak in Italian provinces and corresponding long-term air quality evaluations, were obtained from Italian and European agencies, elaborated and tested for possible interactions. Our elaborations reveal that, beside concentrations, the chronicity of exposure may influence the anomalous variability of SARS-CoV-2 in Italy. Data on distribution of atmospheric pollutants (NO2, O3, PM2.5 and PM10) in Italian regions during the last 4 years, days exceeding regulatory limits, and years of the last decade (2010-2019) in which the limits have been exceeded for at least 35 days, highlight that Northern Italy has been constantly exposed to chronic air pollution. Long-term air-quality data significantly correlated with cases of Covid-19 in up to 71 Italian provinces (updated April 27, 2020) providing further evidence that chronic exposure to atmospheric contamination may represent a favourable context for the spread of the virus. Pro-inflammatory responses and high incidence of respiratory and cardiac affections are well known, while the capability of this coronavirus to bind particulate matters remains to be established. Atmospheric and environmental pollution should be considered as part of an integrated approach for sustainable development, human health protection and prevention of epidemic spreads but in a long-term and chronic perspective, since adoption of mitigation actions during a viral outbreak could be of limited utility.

Graphene oxide and GST-omega enzyme: An interaction that affects arsenic metabolism in the shrimp Litopenaeus vannamei.

Arsenic (As) is one of the most widespread contaminants; it is found in almost every environment. Its toxic effects on living organisms have been studied for decades, but the interaction of this metalloid with other contaminants is still relatively unknown, mainly whether this interaction occurs with emerging contaminants such as nanomaterials. To examine this relationship, the marine shrimp Litopenaeus vannamei was exposed for 48 h to As, graphene oxide (GO; two different concentrations) or a combination of both, and gills, hepatopancreas and muscle tissues were sampled. Glutathione S-transferase (GST)-omega gene expression and activity were assessed. As accumulation and speciation (metabolisation capacity) were also examined. Finally, a molecular docking simulation was performed to verify the possible interaction between the nanomaterial and GST-omega. The main finding was that GO modulated the As toxic effect: it decreased GST-omega activity, a consequence related to altered As accumulation and metabolism. Besides, the molecular docking simulation confirmed the capacity of GO to interact with the enzyme structure, which also can be related to the decreased GST-omega activity and subsequently to the altered As accumulation and metabolisation pattern.

Helium, inorganic and organic carbon isotopes of fluids and gases across the Costa Rica convergent margin.

In 2017, fluid and gas samples were collected across the Costa Rican Arc. He and Ne isotopes, C isotopes as well as total organic and inorganic carbon concentrations were measured. The samples (n = 24) from 2017 are accompanied by (n = 17) samples collected in 2008, 2010 and 2012. He-isotopes ranged from arc-like (6.8 RA) to crustal (0.5 RA). Measured dissolved inorganic carbon (DIC) δ13CVPDB values varied from 3.55 to -21.57‰, with dissolved organic carbon (DOC) following the trends of DIC. Gas phase CO2 only occurs within ~20 km of the arc; δ13CVPDB values varied from -0.84 to -5.23‰. Onsite, pH, conductivity, temperature and dissolved oxygen (DO) were measured; pH ranged from 0.9-10.0, conductivity from 200-91,900 µS/cm, temperatures from 23-89 °C and DO from 2-84%. Data were used to develop a model which suggests that ~91 ± 4.0% of carbon released from the slab/mantle beneath the Costa Rican forearc is sequestered within the crust by calcite deposition with an additional 3.3 ± 1.3% incorporated into autotrophic biomass.

Genotoxic effect of dimethylarsinic acid and the influence of co-exposure to titanium nanodioxide (nTiO2) in Laeonereis culveri (Annelida, Polychaeta).

Few data are available about the effect of dimethylated forms (DMA) on aquatic organisms. As rarely a contaminant occurs alone, studies evaluating the combined effect of different contaminants in aquatic organisms are needed. In fact, the presence of nanomaterials, such as titanium dioxide nanoparticles (nTiO2), in the aquatic environment is now a reality due to its intensive production and use. So, this study evaluated the toxicological effects of DMA in an acute exposure condition and considered the potential influence of nTiO2 on the effects induced by DMA in the polychaete, Laeonereis culveri. The animals were exposed over 48 h to DMA (50 and 500 µg/l) alone or in combination with nTiO2 (1 mg/l). Biochemical parameters such as concentration of reactive oxygen species (ROS), glutathione-S-transferase (GST) activity, levels of reduced glutathione levels (GSH) and macromolecular (lipid and DNA) damage were evaluated, as well the DNA repair system. In addition, the accumulation of total As and the chemical speciation of the metalloid in the organisms was determined. The results showed that: (1) only the group exposed to 500 µg of DMA/l accumulated As and when co-exposed to nTiO2, this accumulation was not observed. (2) The levels of ROS increased in the group exposed to 50 µg/l of DMA alone and the effect was reversed when this group was co-exposed to nTiO2 (3) None of the treatments showed altered GST activity or GSH levels. (4) All groups that received nTiO2 (alone or in combination with DMA) showed lipid peroxidation. (5) The exposure to DMA (both concentrations) alone or in combination with nTiO2 induced DNA damage in L. culveri. These results showed that DMA exhibits a genotoxic effect and that co-exposure to nTiO2 had an influence on its toxicity. So the occurrence of both contaminants simultaneously can represent a threat to aquatic biota.

Different crystalline forms of titanium dioxide nanomaterial (rutile and anatase) can influence the toxicity of copper in golden mussel Limnoperna fortunei?

Although some studies have showed the effects of different crystalline structures of nTiO2 (anatase and rutile) and their applicability in several fields, few studies has analyzed the effect of coexposure with other environmental contaminants such as copper. Thus, the objective of this study was to evaluate if the coexposure to nTiO2 (nominal concentration of 1 mg/L; anatase or rutile) can increase the incorporation and toxic effect induced by Cu (nominal concentration of 56 µg/L) in different tissues of Linmoperna fortunei after 120 h of exposure. Our results showed that the coexposure increased the accumulation of Cu in the gills and adductor muscle independently of the crystalline form and can positively or negatively modulate the antioxidant system, depending on the tissue analyzed. However, exposure only to rutile nTiO2 induced damage in the adductor muscle evidenced by the infiltration of hemocytes in this tissue. Additionally, histomorphometric changes based on fractal dimension analysis showed that coexposure to both forms of nTiO2 induced damage in the same tissue. These results suggest that both crystalline forms exhibited toxicity depending on the analyzed tissue and that coexposure of nTiO2 with Cu may be harmful in L. fortunei, indicating that increased attention to the use and release of nTiO2 in the environment is needed to avoid deleterious effects in aquatic biota.

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.

Antibiotic and heavy metal resistance in enterococci from coastal marine sediment.

Sediment samples from three coastal sites - two beach resorts (Beach 1 and Beach 2 sites) and an area lying between an oil refinery and a river estuary (Estuarine site) - were analyzed for antibiotic- and heavy metal (HM)-resistant enterococci. A total of 123 enterococci, 36 E. faecium, 34 E. casseliflavus, 33 E. hirae, 5 E. faecalis, 3 E. durans, 3 E. gallinarum, and 9 Enterococcus spp, were recovered. Strains resistant to erythromycin, tetracycline and quinupristin/dalfopristin (Q/D) were recovered from all sites, whereas multidrug-resistant isolates were recovered only from "Beach 2" (14%) and "Estuarine" (3.7%). As regards HM resistance, the strains showed a high frequency (68%) of cadmium and/or copper resistance and uniform susceptibility to mercury. The prevalence of cadmium-resistant strains was significantly higher among erythromycin-resistant than among erythromycin-susceptible strains. A significant association between cadmium or copper resistance and Q/D resistance was also observed at "Estuarine" site. The levels of the two HMs in sediment from all sites were fairly low, ranging from 0.070 to 0.126 µg/g, for cadmium and from 1.00 to 7.64 µg/g for copper. Mercury was always undetectable. These findings are consistent with reports that low HM concentrations may contribute to co-selection of antibiotic-resistant bacterial strains, including enterococci.

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.