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Plastic pollution is a notable environmental issue, being plastic widespread and characterized by long lifetime. Serious environmental problems are caused by the improper management of plastic end-of-life. In fact, plastic litter is currently detected in any environment. Biodegradable Polymers (BPs) are promising materials if correctly applied and managed at their end of life, to minimize environmental problems. However, poor data on the fate and toxicity of BPs on marine organisms still limit their applicability. In this work we tested the effects of five biodegradable polymers (polybutylene succinate, PBS; polybutylene succinate-co-butylene adipate, PBSA; polycaprolactone, PCL; poly (3-hydroxybutyrates, PHB; polylactic acid, PLA) widely used for several purposes. Adult individuals of the isopod Idotea balthica basteri were fed on these polymers for twenty-seven days by adding biodegradable microplastic polymers (BMPs) to formulated feeds at two concentrations, viz. 0.84 and 8.4 g/kg feed. The plastic fragments affected the mortality rates of the isopods, as well as the expression levels of eighteen genes (tested by Real Time qPCR) involved in stress response and detoxification processes. Our findings confirmed that I. balthica basteri is a convenient model organism to study the response to environmental pollution and emerging contaminants in the aquatic environment, and highlighted the need for the correct use of BMPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Increasing plastic contamination poses a serious threat to marine organisms. Microplastics (MPs) ingestion can represent a risk for the organism itself and for the ultimate consumer. Through the analysis of the gastrointestinal tract, coupled with stable isotope analysis on the muscle tissue, this study provides insights into the relationship between MPs pollution and ecology in two commercial marine species caught in the Central Tyrrhenian Sea: Aristaeomorpha foliacea and Parapenaeus longirostris . Stable isotope analysis was conducted to determine the trophic position and the trophic niche width. The gastrointestinal tracts were processed, and the resultant MPs were analysed under FT-IR spectroscopy to estimate the occurrence, abundance, and typology of the ingested MPs. The trophic level of the species was similar ( P. longirostris TP = 3 ± 0.10 and A. foliacea TP = 3.1 ± 0.08), with an important trophic niche overlap, where 38% and 52% of P. longirostris and A. foliacea has ingested MPs, respectively. Though species-level differences may not be evident regarding MP's abundance per individual, a high degree of dissimilarity was noted in the typologies of ingested particles. This research provides valuable insights into how MPs enter marine trophic webs, stressing that isotopic niche analysis should be combined with other methods to explain in detail the differences in MPs ingestion.

In the last years biodegradable polymers (BPs) were largely used as real opportunity to solve plastic pollution. Otherwise, their wide use in commercial products, such as packaging sector, is causing a new pollution alarm, mainly because few data reported about their behaviour in the environment and toxicity on marine organisms. Our previous results showed that embryos of the sea urchin Paracentrotus lividus (Lmk) exposed to poly(ε-caprolactone) (PCL), poly(3-hydroxybutyrate) (PHB) and poly(lactic acid) (PLA) showed delay of their development and morphological malformations, also affecting at the molecular levels the expression of several genes involved in different functional responses. In the present work for the first time, we tested the effects of five microplastics (MPs) obtained from BPs such as PBS, poly(butylene succinate), PBSA, poly(butylene succinate-co-butylene adipate), PCL, PHB and PLA, upon grazing activity of the sea urchin revealed by: i. histological analysis seeing at the gonadic tissues; ii. morphological analysis of the deriving embryos; iii. molecular analyses on these embryos to detect variations of the gene expression of eighty-seven genes involved in stress response, detoxification, skeletogenesis, differentiation and development. All these results will help in understanding how MP accumulated inside various organs in the adult sea urchins, and more in general in marine invertebrates, could represent risks for the marine environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Marine plastic pollution is a well-recognised and debated issue affecting most marine ecosystems. Despite this, the threat of plastic pollution on seagrasses has not received significant scientific attention compared to other marine species and habitats. The present review aims to summarise the scientific data published in the last decade (January 2012-2023), concerning the evaluation of plastic pollution, of all sizes and types, including bio-based polymers, on several seagrass species worldwide. To achieve this goal, a comprehensive and critical review of 26 scientific papers has been carried out, taking into consideration the investigated areas, the seagrass species and the plant parts considered, the experimental design and the type of polymers analysed, both in field monitoring and in laboratory-controlled experiments. The outcomes of the present review clearly showed that the dynamics and effects of plastic pollution in seagrass are still under-explored. Most data emerged from Europe, with little or no data on plastic pollution in North and South America, Australia, Africa and Antarctica. Most of the studies were devoted to microplastics, with limited studies dedicated to macroplastics and only one to nanoplastics. The methodological approach (in terms of experimental design and polymer physico-chemical characterisation) should be carefully standardised, beside the use of a model species, such as Zostera marina, and further laboratory experiments. All these knowledge gaps must be urgently fulfilled, since valuable and reliable scientific knowledge is necessary to improve seagrass habitat protection measures against the current plastic pollution crisis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aquatic organisms are exposed to ever-changing complex mixtures of chemicals throughout their lifetime. Component-Based Mixture Risk Assessment (CBMRA) is a well-established methodology for water contaminant-mixture management, the use of which is growing due to improved access to reference ecotoxicity data and extensive monitoring datasets. It enables the translation of measured exposure concentrations of chemicals into biological effect values, and thus to quantitatively estimate the risk of the whole water sample (i.e., as a mixture). However, many factors can bias the final risk decision by impacting the risk metric components; thus, a careful design of the CBMRA is needed, taking into primary consideration the specific features of the dataset and mixture risk assessment assignments. This study systematically addressed the effects of the most common approaches used for handling the concentrations of chemicals below the limit of detection/quantification (LOD/LOQ) in CBMRA. The main results included: i) an informed CBMRA procedure that enables the tracking of the risk decisions triggered by substances below LOD/LOQ, ii) a conceptual map and guidance criteria to support the selection of the most suitable approach for specific scenarios and related interpretation; iii) a guided implementation of the informed CBMRA on dataset of pesticide concentrations in Italian rivers in 2020 (702,097 records)., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Plastic pollution is a remarkable environmental issue. In fact, plastic is widespread in the lifetime and serious environmental problems are caused by the improper management of plastic end of life, being plastic litter detected in any environment. Efforts are put to implement the development of sustainable and circular materials. In this scenario, biodegradable polymers, BPs, are promising materials if correctly applied and managed at the end of life to minimize environmental problems. However, a lack of data on BPs fate and toxicity on marine organisms, limits their applicability. In this research, the impact of microplastics obtained from BPs, BMPs, were analyzed on Paracentrotus lividus. Microplastics were produced from five biodegradable polyesters at laboratory scale by milling the pristine polymers, under cryogenic conditions. Morphological analysis of P. lividus embryos exposed to polycaprolactone (PCL), polyhydroxy butyrate (PHB) and polylactic acid (PLA) showed their delay and malformations, which at molecular level are due to variation in expression levels of eighty-seven genes involved in various cellular processes, such as skeletogenesis, differentiation and development, stress, and detoxification response. Exposure to poly(butylene succinate) (PBS) and poly(butylene succinate-co-adipate) (PBSA) microplastics showed no detectable effects on P. lividus embryos. These findings contribute with important data on the effect of BPs on the physiology of marine invertebrates., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Plastic pollution is a distinctive element of the globalized world. In fact, since the 1970s the expansion and use of plastics, particularly in the consumer and commercial sectors, has given this material a permanent place in our lives. The increasing use of plastic products and the wrong management of end-of-life plastic products have contributed to increasing environmental pollution, with negative impacts on our ecosystems and the ecological functions of natural habitats. Nowadays, plastic pollution is pervasive in all environmental compartments. As aquatic environments are the dumping points for poorly managed plastics, biofouling and biodegradation have been proposed as promising approaches for plastic bioremediation. Known for the high stability of plastics in the marine environment, this represents a very important issue to preserve marine biodiversity. In this review, we have summarized the main cases reported in the literature on the degradation of plastics by bacteria, fungi, and microalgae and the degradation mechanisms involved, to highlight the potential of bioremediation approaches to reduce macro and microplastic pollution.

Dispersants are approved for use in many countries (UK, South Korea, Australia, Egypt, France, Greece, Indonesia, Italy, Japan, Malaysia, Norway, Singapore, Spain, Thailand, and several coastal African, South American, and Middle Eastern countries). Here, the protocols of the most advanced (France, Norway, UK, Spain, Greece, Italy, USA, and Australia) are compared for identifying possible harmonization of approval procedures. Pre-toxicity testing, recognized oil datasets, common thresholds, standardized protocols, zoning, and monitoring are some of the aspects that can be discussed between countries., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Plastic is a ubiquitous material in our life, and its durability represents a great problem for the environment. Several studies reported the occurrence of plastic litter in different environmental compartments and, consequently, numerous efforts are currently focused on how improving its recycling process and produce environmentally friendly solutions. In recent years, biodegradable polymers/plastics (BPs) have been proposed to reduce environmental impacts in specific applications (e.g., when conventional plastics are difficult or expensive to remove from the environment). Their wide use in commercial products, especially in the packaging sector, is causing new pollution alarm. Research studies are ongoing to improve BPs manufacturing and characteristics, but few data are reported about their behavior and toxicity into the marine environment. This paper reviewed the current state of the art highlighting that, even though the degradation of BPs in simulated or real marine environments is quite investigated, only eleven papers reported their effects on marine organisms (e.g., behavioral and oxidative stress and potential cascading effects on marine ecosystems). Presently, the main benefits of BPs are linked to waste management (including collection and recycling of organic waste). Due to the existing knowledge gaps, BPs cannot be deemed yet as a solution to marine plastic pollution., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Due to the continuous development, production and consumption of nanoparticles (NPs), their release, fate and effects in marine coastal environment can represent a major concern. The aim of this study was to evaluate the toxicity of ZnO nanoparticles (ZnO NPs) and compare it to bulk ZnSO 4 on three macroinvertebrates: the isopod Cymodoce truncata (i.e. used for the first time in ecotoxicology), the amphipod Gammarus aequicauda and the sea urchin Paracentrotus lividus. This study showed concentration- and time-dependent relationships for all biological models for both ZnO NPs and ZnSO 4 . Both Zn forms elicited high toxicity to G. aequicauda and C. truncata juveniles, but ZnO NPs induced comparable responses to both species (96h-LC 50 = 0.30 and 0.37 mg/L for G. aequicauda and C. truncata, respectively; p > 0.05), while differences were found after ZnSO 4 exposure (96h-LC 50 = 0.28 and 0.63 mg/L, respectively; p < 0.05). ZnO NPs generated sub-lethal effects on P. lividus embryos (72h-EC 50 = 0.04 (0.03, 0.05) mg/L), not significantly different from ZnSO 4 ones (72h-EC 50 = 0.06 (0.05, 0.07) mg/L). Effects of ZnO NPs were similar to existing literature data for other testing species. C. truncata can be considered as a promising new biological model in (nano)ecotoxicology., (© 2021. The Author(s).)

Nanoplastics are considered contaminants of emerging concern at the global scale. The recent evidence of their occurrence in seawater from the Mediterranean Sea calls for a thorough evaluation of their impact on marine life and in particular on vulnerable life stages such as planktonic embryos. Here, we investigated the impact of increasing nominal concentrations of 50 nm amino-modified (PS-NH 2 ) and 60 nm carboxy-modified (PS-COOH) polystyrene nanoparticles (PS NPs) on the embryonic development of the ascidian Ciona robusta (phylum Chordata), a common benthic invertebrate living in Mediterranean coastal areas with the peculiarity of being an early chordate developmental model. A strong agglomeration of PS-COOH (approx. 1 µm) was observed in natural sea water (NSW) already at time 0, while PS-NH 2 resulted still monodispersed (approx. 130 nm) but largely aggregated after 22 h with a microscale dimension similar to those negatively charged. However, their effect on C. robusta embryos development largely differed at 22 h: PS-COOH did not affect larvae phenotypes nor their development, while PS-NH 2 caused a dose-dependent effect (EC 50 (22 h) of 7.52 μg mL -1 ) with various degrees of phenotype malformations (from mild to severe) and impairment of larval swimming. Embryos (up to 30%) exposed to 15 µg mL -1  PS-NH 2 resulted not developed and the majority was unable to hatch. Calculated PS-NH 2 EC 50 resulted higher than those available for other marine invertebrate species, suggesting a protective role of the egg envelopes surrounding C. robusta embryos toward nanoplastics exposure.

This study investigated for the first time the effects of ZnO nanoparticle (NP) chronic exposure (28 days) on Tigriopus fulvus. Acute toxicity (48 h) of three Zn chemical forms was assessed as well including the following: (a) ZnO nanoparticles (NPs), (b) Zn 2+ from ZnO NP suspension after centrifugation (supernatant) and (c) ZnSO 4 H 2 O. Physical-chemical and electronic microscopies were used to characterize spiked exposure media. Results showed that the dissolution of ZnO NPs was significant, with a complete dissolution at lowest test concentrations, but nano- and micro-aggregates were always present. Acute test evidenced a significant higher toxicity of Zn 2+ and ZnSO 4 compared to ZnO NPs. The chronic exposure to ZnO NPs caused negative effects on the reproductive traits, i.e. brood duration, brood size and brood number at much lower concentrations (≥ 100 μg/L). The appearance of ovigerous females was delayed at higher concentrations of ZnO NPs, while the time required for offspring release and the percentage of non-viable eggs per female were significantly increased. ZnO NP subchronic exposure evidenced its ability to reduce T. fulvus individual reproductive fitness, suggesting that ZnO NPs use and release must be carefully monitored. Graphical abstract Graphical Abstract.

After the accidental release of crude oil in marine environment, dispersants are applied on sea surface transferring oil into the water column where it can be broken down by biodegradation, thereby reducing potential pollution to coastal areas. Before they can be used in the wild, the ecotoxicity of such dispersants is usually evaluated with toxicity assays using algae, crustacean and fishes. Nowadays, there is a need to find alternative species to reduce the use of vertebrates both for ethical considerations and for reducing the cost of bioassays. Ciona robusta is a solitary ascidian that inhabits shallow waters and marine coastal areas. This species has been recently adopted as valuable biological model for ecotoxicity studies, thanks to its rapid embryonic and larval development, resemblance to vertebrates, and low risk of ethical issues. On this ground, the lethal and sublethal toxicity of two dispersants has been evaluated on Ciona juveniles. At this stage, the organisms become filter-feeders and the morphological alterations of the organs can be easily observed. The median lethal concentrations at 96 h (96hLC 50 ) for Dispersant 1 (non-ionic surfactant) and for Dispersant 2 (mixture of non-ionic surfactants and anionic surfactants) are 41.6 mg/L (38.6-44.9) and 92.5 mg/L (87.7-97.5), respectively. The Ciona juvenile model was compared to Dicentrarchus labrax fish juveniles test, and it showed increased sensitivity for Ciona to these compounds. These results suggest that 96 h mortality test bioassay could be a good alternative method to 96 h mortality assay with D. labrax, limiting the use of vertebrates for dispersant toxicity.

In recent years, EU countries have recognized national policies to authorize dispersant use to mitigate the petroleum hydrocarbon contamination in case of unintentional oil spills at sea. A harmonization of dispersant approval procedures is needed because the application of different methodologies agrees on dissimilar toxic responses for the same dispersant in different European countries. Actually, different dispersant approval procedures are applied in France and Italy with one French mandatory toxicity test and three Italian bioassays accompanied with different criteria of toxicity classification. In this paper, a harmonized tiered approach is proposed to address the dispersant ecotoxicological assessment in these two nations. Our approach, applicable at the European level, introduces two mandatory tests (algal growth inhibition test and mortality test with crustaceans) and one discretionary test (fish mortality test), by reducing use of vertebrates as much as possible in accordance with humane principles and animal welfare.

Oxylipins are diatom-derived secondary metabolites, deriving from the oxidation of polyunsatured fatty acids that are released from cell membranes after cell damage or senescence of these single-celled algae. Previous results revealed harmful toxic effects of polyunsaturated aldehydes (PUAs) and hydroxyacids (HEPEs) on sea urchin Paracentrotus lividus embryonic development by testing individual compounds and mixtures of the same chemical group. Here, we investigated the combined effects of these compounds on sea urchin development at the morphological and molecular level for the first time. Our results demonstrated that oxylipin mixtures had stronger effects on sea urchin embryos compared with individual compounds, confirming that PUAs induce malformations and HEPEs cause developmental delay. This harmful effect was also confirmed by molecular analysis. Twelve new genes, involved in stress response and embryonic developmental processes, were isolated from the sea urchin P. lividus ; these genes were found to be functionally interconnected with 11 genes already identified as a stress response of P. lividus embryos to single oxylipins. The expression levels of most of the analyzed genes targeted by oxylipin mixtures were involved in stress, skeletogenesis, development/differentiation, and detoxification processes. This work has important ecological implications, considering that PUAs and HEPEs represent the most abundant oxylipins in bloom-forming diatoms, opening new perspectives in understanding the molecular pathways activated by sea urchins exposed to diatom oxylipins., Competing Interests: The authors declare no conflict of interest.

Oxylipins such as polyunsaturated aldehydes (PUAs) and hydroxyacids (HEPEs) are signaling molecules derived from the oxidation of polyunsaturated fatty acids. They are common in diatoms that constitute a major group of microalgae in freshwater and oceanic ecosystems. Although HEPEs represent the most common oxylipins produced by diatoms, little information is available on their effects on marine invertebrates, and most of the information has been obtained by testing individual HEPEs. Our previous studies reported that four hydroxyacids, i.e., 5-, 9-, 11-, and 15-HEPE, were able to induce malformations and a marked developmental delay in sea urchin Paracentrotus lividus embryos, which had not been reported for other oxylipins. Here, we tested a mixture of 5-, 9-, 11-, and 15-HEPE at different concentrations for the first time. The results showed that mixtures of HEPEs have synergistic effects that are much more severe compared to those of individual HEPEs: The HEPE mixtures induced malformations in sea urchin embryos at lower concentrations. Increasing HEPE mixture concentrations induced a marked increase in the number of delayed embryos, until all embryos were delayed at the highest concentration tested. At the molecular level, the HEPE mixtures induced variations in the expression of 50 genes involved in different functional processes, mainly down-regulating these genes at the earliest stages of embryonic development. These findings are ecologically significant, considering that during diatom blooms, sea urchins could accumulate HEPEs in concentrations comparable to those tested in the present study.

Diatom-derived polyunsaturated aldehydes (PUAs), decadienal, heptadienal and octadienal, derive from the oxidation of fatty acids and have cytotoxic and anticancer effects. PUAs, tested separately, induce malformations in sea urchin Paracentrotus lividus embryos. Decadienal induces the worst malformations and lowest survival rates. Interestingly, decadienal, heptadienal and octadienal place in motion several genes to counteract their negative effects. To date, no studies are available reporting on the effects of PUA mixtures on marine invertebrates. Here we test binary and ternary mixtures on embryonic development of P. lividus. Our findings demonstrate that mixtures of PUAs act (i) at morphological level in synergistic way, being much more severe compared to individual PUAs; (ii) at molecular level also reveal an additive effect, affecting almost all fifty genes, previously tested using individual PUAs. This study is relevant from an ecological point of view since diatoms are a major food source for both pelagic and benthic organisms. This work opens new perspectives for understanding the molecular mechanisms that marine organisms use in reacting to environmental natural toxin mixtures such as diatom PUAs.

The aim of this study was to detect ecotoxicological effects of 0.1 μm polystyrene microbeads in marine organisms belonging to different trophic levels. MP build up, lethal and sub-lethal responses were investigated in the bacterium Vibrio anguillarum (culturability), in the green microalga Dunaliella tertiolecta (growth inhibition), in the rotifer Brachionus plicatilis (mortality and swimming speed alteration) and in the sea urchin Paracentrotus lividus (immobility and swimming speed alteration) exposed to a wide range of microplastic (MP) concentrations (from 0.001 to 10 mg L -1 ). Survival was not affected in all organisms up to 10 mg L -1 , while algal growth inhibition, rotifer and sea urchin larvae swimming behaviour alterations were observed after exposure to MPs. Ingestion was only observed in rotifers and it was directly correlated with sub-lethal effects. These results account for the ecotoxicological risk associated to the polystyrene microbeads, which are able to affect different endpoints in primary producers and consumers (rotifers and sea urchins) since no effects were observed in decomposers. This study points out the importance of using a battery of marine organisms belonging to different trophic levels by studying acute toxicity of MPs at low and high contamination levels, and investigating sub-lethal responses. Further investigations aimed at studying the transfer of these materials through the web are particularly recommended., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Copper oxide nanoparticles (CuO NPs) are extensively used in various industrial and commercial applications. Despite their wide application may lead to the contamination of marine ecosystem, their potential environmental effects remain to be determined. Toxicity assessment studies have primarily focused on investigating the effects of CuO NPs on fertilization success and embryo development of different sea urchin species while the impact on sperm quality have never been assessed. In this line, this study aims to assess the effects of CuO NPs on the spermatozoa of the sea urchin Paracentrotus lividus. After sperm exposure to CuO NPs, biomarkers of sperm viability, cytotoxicity, oxidative stress, and genotoxicity as well as morphology were evaluated. Results showed that CuO NPs exposure decreased sperm viability, impaired mitochondrial activity and increased the production of reactive oxygen species (ROS) and lipid peroxidation. Furthermore, CuO NPs exposure caused DNA damage and morphological alterations. Together with the antioxidant rescue experiments, these results suggest that oxidative stress is the main driver of CuO NP spermiotoxic effects. The mechanism of toxicity is here proposed: the spontaneous generation of ROS induced by CuO NPs and the disruption of the mitochondrial respiratory chain lead to production of ROS that, in turn, induce lipid peroxidation and DNA damage, and result in defective spermatozoa up to induce sperm cytotoxicity. Investigating the effects of CuO NPs on sea urchin spermatozoa, this study provides valuable insights into the mechanism of reproductive toxicity induced by CuO NPs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Several different chemical products are used on oil platforms to aid oil-water separation during the production process. These chemicals may enter into the sea by means of production water (PW), the main discharge derived from oil and gas offshore platforms. Consequently, toxic effects may occur in the marine environment, causing reductions in wildlife numbers, degrading ecosystem functions and threatening human health. For most of these chemicals, environmental toxicity and safety thresholds in marine ecosystems have not been fully investigated as yet. In this work, a numerical-ecotoxicological approach is proposed to assess the potential environmental risk associated with the discharge of five oilfield production chemicals (deoiler, scale inhibitor, corrosion inhibitor, catalyst, dehydrating agent) from a platform in the southern Adriatic Sea (Mediterranean Sea). Their concentrations in the seawater are numerically predicted, under different seasonal conditions, starting from the real concentrations used during the production process. The predicted concentrations are then evaluated in terms of possible toxic effects in order to assess the potential risk of oilfield production chemicals discharged into the sea.

Bacteria are used in ecotoxicology for their important role in marine ecosystems and their quick, reproducible responses. Here we applied a recently proposed method to assess the ecotoxicity of nanomaterials on the ubiquitous marine bacterium Vibrio anguillarum , as representative of brackish and marine ecosystems. The test allows the determination of 6-h EC 50 in a wide range of salinity, by assessing the reduction of bacteria actively replicating and forming colonies. The toxicity of copper oxide nanoparticles (CuO NPs) at different salinities (5-20-35 ‰) was evaluated. CuSO 4 5H 2 O and CuO bulk were used as reference toxicants (solubility and size control, respectively). Aggregation and stability of CuO NP in final testing dispersions were characterized; Cu 2+ dissolution and the physical interactions between Vibrio and CuO NPs were also investigated. All the chemical forms of copper showed a clear dose-response relationship, although their toxicity was different. The order of decreasing toxicity was: CuSO 4 5H 2 O > CuO NP > CuO bulk. As expected, the size of CuO NP aggregates increased with salinity and, concurrently, their toxicity decreased. Results confirmed the intrinsic toxicity of CuO NPs, showing modest Cu 2+ dissolution and no evidence of CuO NP internalization or induction of bacterial morphological alterations. This study showed the V. anguillarum bioassay as an effective tool for the risk assessment of nanomaterials in marine and brackish environments.

A research project has been performed to the request of the RAMOGE Executive Secretariat to identify differences between dispersant approval procedures in France and Italy and propose ways to harmonize them. A collaborative study has been conducted by CEDRE (Centre of Documentation, Research and Experimentation on Accidental Water Pollution) and ISPRA (Italian Institute for Environmental Protection and Research) to: a) compare current approval procedures in Italy and France with identification of differences and commonalities; b) carry out toxicity tests using both procedures on two selected dispersants; c) propose a common approach between Italy and France. The results showed that, because of the differences in ecotoxicological tests and in the evaluation criteria used, the outcomes on the same products could be different in Italy and in France. Both tested dispersants met the French requirements for approval (LC 50 ≥ 10 times reference toxicant), while only one dispersant met the Italian approval criterion (EC 50 > 10mg/L). A possible way of harmonizing the approval procedures could be to increase the number of test organisms in the French procedure, which currently only uses one crustacean species. Furthermore, a common criterion for toxicity assessment should be discussed and agreed., (Copyright © 2017. Published by Elsevier Inc.)

Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food webs. Toxicity testing based on microorganisms are widely used because they are relatively quick, reproducible, cheap, and are not associated with ethical issues. Here, we describe an ecotoxicological method to evaluate the biological response of the marine bacterium Vibrio anguillarum. This method assesses the acute toxicity of chemical compounds, including new contaminants such as nanoparticles, as well as environmental samples. The endpoint is the reduction of bacterial culturability (i.e., the capability to replicate and form colonies) due to exposure to a toxicant. This reduction can be generally referred to as mortality. The test allows for the determination of the LC50, the concentration that causes a 50% decrease of bacteria actively replicating and forming colonies, after a 6 h exposure. The culturable bacteria are counted in terms of colony forming units (CFU), and the "mortality" is evaluated and compared to the control. In this work, the toxicity of copper sulphate (CuSO4) was evaluated. A clear dose-response relationship was observed, with a mean LC50 of 1.13 mg/L, after three independent tests. This protocol, compared to existing methods with microorganisms, is applicable in a wider range of salinity and has no limitations for colored/turbid samples. It uses saline solution as the exposure medium, avoiding any possible interferences of growth medium with the investigated contaminants. The LC50 calculation facilitates comparisons with other bioassays commonly applied to ecotoxicological assessments of the marine environment.

Swimming speed alteration and mortality assays with the marine crustacean Artemia franciscana were carried out. EC50 and LC50 values after 24-48h exposures were calculated for two reference toxicants, copper sulphate pentahydrate (CuSO4·5H2O) and Sodium Dodecyl Sulphate (SDS), and an ecotoxicological concern organic compound, Diethylene Glycol (DEG). Different end-points have been evaluated, in order to point out their sensitivity levels. The swimming speed alteration (SSA) was compared to mortality values and also to the hatching rate inhibition (literature data). SSA resulted to be more sensitive than the mortality and with a sensitivity comparable to (or even higher than) the hatching rate endpoint., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Acute heavy metal toxicity was compared in nauplii of an autochthonous Artemia parthenogenetica (Branchiopoda, Anostraca) from Saline di Cervia (Ravenna, Italy) and of a non native commercially available A. franciscana. No significant difference in sensitivity was detected between two species.

Diethylene glycol (DEG) is commonly used to dehydrate natural gas in off-shore extraction plants and to prevent formation of gas hydrates. It may be released into the sea accidentally or in discharged produced waters (PWs). PWs samples from off-shore gas platforms in the Adriatic Sea (Italy) have been used in this study. The objectives of the study were: a) to evaluate the toxicity of DEG for marine organisms; b) to evaluate if a high DEG content in PWs may alter their toxicity; c) to verify whether the DEG threshold concentration established by the Italian legislation (3.5 g/l) for PWs discharged at sea is safe for marine environment. Ten different species (Vibrio fischeri, Phaeodactylum tricornutum, Dunaliella tertiolecta, Brachionus plicatilis, Artemia franciscana, Tigropus fulvus, Mytilus galloprovincialis, Crassostrea gigas, Tapes philippinarum and Dicentrarchus labrax) have been exposed to DEG; four of these species were also exposed to PWs in combination with DEG. The results showed that: a) DEG is not toxic at levels normally detected in Adriatic PWs; b) DEG in combination with PW showed mainly additive or synergistic effects; c) short-term bioassays showed that the DEG limit of 3.5 g/l could be acceptable., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded. . Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

This paper reports the concentrations of eight trace metals (Cr, Zn, Cu, Pb, Ni, As, Cd, and Hg) in the surficial sediments of the Berre lagoon. This is a typical Mediterranean lagoon, seriously affected by human activities since the 1930s, when it became one of the most important industrial sites in France. The overall range of concentrations is 38-428 mg kg(-1) for chromium, 50-151 mg kg(-1) for zinc, 11-48 mg kg(-1) for copper, 18-82 mg kg(-1) for lead, 18-56 mg kg(-1) for nickel, 4-10 mg kg(-1) for arsenic, 0.2-1.6 mg kg(-1) for cadmium, and 0.15-0.40 mg kg(-1) for mercury. The spatial distribution of concentrations largely mirrors the localization of major impact sources, with the most affected area restricted to a radius of a few kilometers from point sources, due to the environmental conditions of the outfall zone (fine sediments, low hydrodynamic regime, confinement), which favor the in situ accumulation of pollutants. The extent of contamination from trace metals in the lagoon sediments is evaluated through a three-pronged approach: (i) by comparison with other areas (see Table 3 for an up-to-date review of trace metal concentrations in lagoon systems and coastal regions of the world ocean); (ii) by evaluating the metal enrichment in the sediments through the calculation of concentration factors (CF) for each trace metal and by combining the CFs to estimate a Metal Pollution Index (MPI) taking into account the differences in toxicity of trace metals; and (iii) by defining a potential level of biological risk by the use of quality criteria such as the Threshold Effect Level (TEL) and Effects Range-Low (ER-L) benchmarks. On the basis of the calculated CFs, sediments appear as particularly enriched with Pb and Cd throughout the lagoon and with Cr and Zn at some sites. Although no site can be classified as "unpolluted" when looking at the calculated MPIs, two regions--the Vaïne pond and the southwestern area--have been identified as critical points of contamination. Trace metal inputs to the lagoon need to be kept under strict control in the future, with particular regard to Ni, Cr, Pb, and Cu that, on the basis of TEL and ER-L benchmarks, already appear associated with a potential biological risk.

Diethylene glycol (DEG) is extensively used on offshore gas platforms to prevent the hydrate formation during the gas-water separation process and to inhibit corrosion events. This chemical might enter in the marine environment via the produced formation water (PFW) discharge. In this study, a new approach was applied to the investigation of the DEG content in PFW discharges and seawater samples from four gas installation platforms in the Adriatic Sea (Italy). The method includes an off-line solid-phase extraction/pre-concentration technique, followed by a nanoscale flow injection/direct-electron ionization (EI) mass spectrometric analysis. Direct-EI is a novel and miniaturized interface for directly coupling a liquid chromatograph with an electron ionization mass spectrometer. The capability to acquire EI spectra, and to operate in selected ion monitoring mode during actual sample analyses, allowed a precise quantification of DEG with a method limit of detection of 31microg/l. In addition, a careful evaluation of the matrix effect showed that, as opposed to electrospray ionization, the response of the Direct-EI interface was not affected by sample interferences.