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.

Microplastics in seawater and marine organisms: Site-specific variations over two-year study in Giglio Island (North Tyrrhenian Sea).

Geographical and temporal differences of microplastic occurrence were documented in water and fish collected in 2017 and 2019 from the Giglio Island (North Tyrrhenian Sea) close to the area where the Costa Concordia sank in January 2012. Results on water samples showed a site-dependent difference, suggesting the role of surface current dynamics in the microplastic local distribution, while tested Neuston nets (200 µm and 330 µm mesh size) did not influence microplastic retention efficiency. Fish exhibited in 2019 a higher frequency of specimens positive to microplastic ingestion with respect to 2017, with an occurrence higher than those typically observed in other Mediterranean areas. Both in water and fish, fragments were the dominating shape, polypropylene and polyethylene were the prevalent polymers, without particular difference between sites and years. This study highlights the importance of applying microplastic investigation in biotic and abiotic matrices for an effective monitoring of this pollution in the marine environment.

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.

Microplastics pollution after the removal of the Costa Concordia wreck: First evidences from a biomonitoring case study.

Microplastics (MPs) represent a matter of growing concern for the marine environment. Their ingestion has been documented in several species worldwide, but the impact of specific anthropogenic activities remains largely unexplored. In this study, MPs were characterized in different benthic fish sampled after 2.5 years of huge engineering operations for the parbuckling project on the Costa Concordia wreck at Giglio Island. Fish collected in proximity of the wreck showed a high ingestion of microplastics compared to both fish from a control area and values reported worldwide. Also the elevated percentage of nylon, polypropylene lines and the presence of polystyrene are quite unusual for marine organisms sampled in natural field conditions, thus supporting the possible relationship of ingested microplastics with maritime operations during wreck removal. On the other hand, the use of transplanted mussels revealed a lower frequency of ingested MPs, and did not discriminate differences between the wreck and the control area. Some variations were observed in terms of typology and size of particles between surface- and bottom-caged mussels highlighting the influence of a different distribution of MPs along the water column. In conclusion, this study demonstrated that MPs pollution in the area of Costa Concordia was more evident on benthonic environment than on seawater column, providing novel insights on the possibility of using appropriate sentinel organisms for monitoring specific anthropogenic sources of MPs pollution in the marine environment.

Effects of exposure to halogenated organic compounds combined with dietary restrictions on the antioxidant defense system in herring gull chicks.

The Herring gull (HG) (Larus argentatus) is naturally exposed to halogenated organic compounds such as polychlorinated biphenyls (PCBs) through its diet. During periods of food scarcity, arctic seabirds experience lipid mobilization, allowing stored lipid soluble contaminants to re-enter the body circulation. In the present study, we investigated the effects of PCB exposure and fasting on the antioxidant defense system in HG chicks. Forty newly hatched chicks were exposed to contaminated cod liver oil for 6weeks and then fasted for 1week. We assessed the hepatic total oxyradical scavenging capacity (TOSC) against peroxynitrite, hydroxyl and peroxyl radicals, and measured glutathione (reduced: GSH, and oxidized: GSSG) levels and the enzymatic activities of catalase, glutathione peroxidase, glutathione reductase and superoxide dismutase. The results show that fasting significantly increased the HOC levels in the HG chick livers. Limited effects were observed on antioxidant responses; significant effects were only found for catalase (CAT) activity, Se-dependent GPX activity and the GSH/GSSG ratio in the exposed and fasted group. CAT and Se-dependent GPX activities correlated negatively with the PCB concentrations within this group, and a nonlinear relationship between glutathione and contaminant levels was also found. These effects were generally not observed after exposure or fasting alone and were likely related to the high PCB levels induced by the combination of exposure and fasting.

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, for monitoring the impact of an offshore platform in the Adriatic Sea.

An ecotoxicological protocol with caged mussels, Mytilus galloprovincialis, was developed to evaluate the potential impact of an offshore gas platform in the central Adriatic Sea. Reference organisms were collected on a seasonal basis from an unpolluted site and transplanted for four weeks in both the sampling area and to the investigated platform. Chemical analyses of trace metals in mussel tissues were integrated with a multi-biomarker approach for the early detection of biological responses at several cellular targets. Induction of metallothioneins, peroxisomal proliferation and activity of acetylcholinesterase were measured as markers for specific classes of chemicals. Special attention was given to oxyradical metabolism and appearance of oxidative-mediated toxicity to reveal a more general onset of cellular disturbance. In addition to individual antioxidants (superoxide dismutase, catalase, glutathione S-transferases, glutathione reductase, Se-dependent and Se-independent glutathione peroxidases, and levels of total glutathione), the total oxyradical scavenging capacity (TOSC) allowed a quantification of the overall capability to neutralize specific forms of intracellular reactive oxygen species (ROS; i.e. peroxyl and hydroxyl radicals). Cellular damages were evaluated as lysosomal destabilization (membrane stability, accumulation of lipofuscin and neutral lipids), lipid peroxidation products (malondialdehyde) and DNA integrity (strand breaks and micronuclei); the air survival test was finally applied to evaluate the overall physiological condition of mussels. Concentration of trace metals (As, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Zn) revealed only limited variations in transplanted mussels during various experimental periods and such changes appeared partly related to natural fluctuations. Among biological responses, variations of antioxidants and lysosomal stability were confirmed as sensitive early warning signals for biological disturbance of both natural and anthropogenic origin. The presented protocol with caged mussels allowed marked biological effects caused by the investigated platform to be excluded, and represented a useful approach that is easy to extend for monitoring the impact of offshore activities in the Adriatic sea.

Vitellogenin gene expression in males of the Antarctic fish Trematomus bernacchii from Terra Nova Bay (Ross Sea): a role for environmental cadmium?

Synthesis of vitellogenin (VTG) in male fish is a widely recognized effect for estrogenic pollutants in temperate environments, while similar investigations are still lacking for Antarctic organisms. In this study, a preliminary characterization of vitellogenin gene expression was performed by RT-PCR in the key species Trematomus bernacchii sampled in different phases of reproductive cycle and food availability. Females exhibited the highest gene expression during the spawning period, but VTG mRNA was always detected also in males; a significant increase of gene expression was observed both in males and females at the end of the feeding season. These results were not fully supported by a differential exposure to phyto- or anthropogenic estrogens during the planctonic cycle; on the other side, the endocrine properties of cadmium, naturally elevated in Terra Nova Bay and increasing during algal bloom, would explain both the presence of VTG mRNA in males and the seasonal changes of gene induction. Laboratory exposures did not reveal short-term estrogenic effects of cadmium while an elevated responsiveness of T. bernacchii was observed toward a classical estrogenic receptor agonist (17beta-estradiol). Different hypotheses were considered to suggest delayed endocrine effects of cadmium, including the early interaction with other cellular detoxification systems or alterations at multiple levels of the hypothalamus-pituitary-gonad-liver axis. Although molecular mechanisms of VTG gene expression in males of T. bernacchii remain unclear, obtained results provide interesting insights on this species which should stimulate future research activities.

Pro-oxidant effects of extremely low frequency electromagnetic fields in the land snail Helix aspersa.

Pro-oxidant effects of extremely low frequency (ELF) 50-Hz magnetic fields were investigated in the land snail Helix aspersa exposed both in short-term laboratory treatments and under field conditions by maintaining the organisms in the proximity of a power line for up to 2 months. Oxidative perturbations were investigated as individual antioxidants (catalase, glutathione reductase, glutathione S-transferases, and total glutathione) and total scavenging capacity toward peroxyl radicals and hydroxyl radicals. Accumulation of lipid peroxidation products, destabilization of lysosomal membranes, and loss of DNA integrity were also evaluated as markers of cell damage. The overall results indicated an oxidative challenge caused by ELF magnetic fields with particularly prompt and sensitive responses for catalase, glutathione reductase, and the overall capability to neutralize peroxyl radicals. Cell injuries occurred to different extents according to duration and intensity of electromagnetic exposure and confirmed complex cause-effect relationships between pro-oxidant factors, efficiency of antioxidant defenses, and the onset of oxidative toxicity. This study highlights the importance of a multimarker approach for detecting a wide panel of biological responses, the necessity of investigating the long-term effects of early oxidative responses, and the role of ELF in enhancing susceptibility to other forms of pathologies or diseases.

Interactions between metabolism of trace metals and xenobiotic agonists of the aryl hydrocarbon receptor in the antarctic fish Trematomus bernacchii: environmental perspectives.

Although Antarctica is a pristine environment, organisms are challenged with contaminants either released locally or transported from industrialized regions through atmospheric circulation and marine food webs. Organisms from Terra Nova Bay also are exposed to a natural enrichment of cadmium, but to our knowledge, whether such environmental conditions influence biological responses to anthropogenic pollutants has never been considered. In the present study, the Antarctic rock cod (Trematomus bernacchii) was exposed to model chemicals, including polycyclic aromatic hydrocarbons (benzo[a]pyrene), persistent organic pollutants (2,3,7,8-tetrachlorodibenzo-p-dioxin [TCDD]), cadmium, and a combination of cadmium and TCDD. Analyzed parameters included chemical bioaccumulation, activity, and levels of biotransformation enzymes (cytochrome P4501A); metallothioneins and the efficiency of the antioxidant system measured as individual defenses (catalase, glutathione, glutathione reductase, glutathione S-transferases, and glutathione peroxidases); and total scavenging capacity toward peroxyl and hydroxyl radicals. Reciprocal interactions between metabolism of inorganic and organic pollutants were demonstrated. Dioxin enhanced the accumulation of cadmium, probably stored within proliferating endoplasmic reticulum, and cadmium suppressed the inducibility of cytochrome P4501A, allowing us to hypothesize a posttranscriptional mechanism as the depletion of heme group availability. Clear evidence of oxidative perturbation was provided by the inhibition of antioxidants and enhanced sensitivity to oxyradical toxicity in fish exposed to organic chemicals. Exposure to cadmium revealed counteracting responses of glutathione metabolism; however, these responses did not prevent a certain loss of antioxidant capacity toward peroxyl radicals. The pattern of antioxidant responses exhibited by fish coexposed to cadmium and TCDD was more similar to that observed for cadmium than to that observed for TCDD. The overall results suggest that elevated natural levels of cadmium in Antarctic organisms from Terra Nova Bay can limit biotransformation capability of polycyclic (halogenated) hydrocarbons, thus influencing the bioaccumulation and biological effects of these chemicals in key sentinel species.

Antioxidant efficiency in early life stages of the Antarctic silverfish, Pleuragramma antarcticum: responsiveness to pro-oxidant conditions of platelet ice and chemical exposure.

The Antarctic silverfish Pleuragramma antarcticum is a key organism in the ecology of Southern Ocean. Eggs with fully developed yolk-sac embryos and newly hatched larvae have been recently observed to occur in the platelet ice accumulating below the sea-ice layer. This environment has strong pro-oxidant characteristics at the beginning of austral spring, when the rapid growth of algal ice communities, the massive release of nutrients and the photoactivation of dissolved organic carbon and nitrates, all represent important sources for oxyradical formation. Such processes are concentrated in a short period of a few weeks which overlaps with the final development of P. antarcticum in platelet ice. The aim of this work was to characterize the antioxidant system in embryos of P. antarcticum and the responsiveness toward the natural increase of pro-oxidant conditions in early spring. Considering the lack of ecotoxicological data on this species and its pivotal importance in the ecosystem of Southern Ocean, the sensitivity of its early life stages was also evaluated after laboratory exposures to environmentally relevant doses of benzo(a)pyrene, as a model chemical potentially released from anthropogenic activities. Obtained results revealed a marked temporal increase of antioxidants in embryos of P. antarcticum as adaptive counteracting responses to oxidative conditions of platelet ice. Particularly prompt responses were observed for glutathione metabolism which, however, did not prevent formation of increasing levels of lipid peroxidation products; from the analysis of total oxyradical scavenging capacity (TOSC), the overall efficiency to neutralize peroxyl radicals remained almost constant while slightly lower TOSC values were obtained toward hydroxyl radicals at the end of sampling period. Laboratory exposures to 0.5-5 microg/l BaP caused a significant accumulation of this PAH but no significant effects on the activity of cytochrome P450. Antioxidants of exposed embryos showed less marked variations than embryos in field conditions suggesting that the elevated pro-oxidant challenge, to which these organisms are naturally adapted, might be responsible for the moderate responsiveness to pro-oxidant chemicals.