Identification of Marine Biotechnology Value Chains with High Potential in the Northern Mediterranean Region.

Marine (blue) biotechnology is an emerging field enabling the valorization of new products and processes with massive potential for innovation and economic growth. In the Mediterranean region, this innovation potential is not exploited as well as in other European regions due to a lack of a clear identification of the different value chains and the high fragmentation of business innovation initiatives. As a result, several opportunities to create an innovative society are being missed. To address this problem, eight Northern Mediterranean countries (Croatia, France, Greece, Italy, Montenegro, Portugal, Slovenia and Spain) established five national blue biotechnology hubs to identify and address the bottlenecks that prevent the development of marine biotechnology in the region. Following a three-step approach (1. Analysis: setting the scene; 2. Transfer: identification of promising value chains; 3. Capitalization: community creation), we identified the three value chains that are most promising for the Northern Mediterranean region: algae production for added-value compounds, integrated multi-trophic aquaculture (IMTA) and valorization aquaculture/fisheries/processing by-products, unavoidable/unwanted catches and discards. The potential for the development and the technical and non-technical skills that are necessary to advance in this exciting field were identified through several stakeholder events which provided valuable insight and feedback that should be addressed for marine biotechnology in the Northern Mediterranean region to reach its full potential.

Prepared microplastics interaction with Artemia salina under low pH conditions representing ocean acidification; a simulated environmental exposure.

Ocean acidification and microplastic pollution are two of the major ecological concerns. The distribution of large quantities of plastic debris and microplastics all across the oceans emphasises the need to determine the influence of microplastics in ocean acidification and to evaluate its concomitant toxicological effects on aquatic life forms. Studies on the combined impact of both the stressors are very limited, but much needed in the current scenario. Where most of the present-day research use purchased microplastics of defined size and morphology (microspheres, fibres, rods, etc.), the present study employs prepared "true to life microplastics" that resemble the environmental microplastic pollutants in morphology and size heterogeneity. The present study focusses on evaluating the fate and impact of oceanic microplastics on the physiology and development of Artemia salina (Brine shrimp), one among the most ecologically significant zooplankton species. Natural sea water was acidified by controlled perturbation of carbon dioxide using a valve system. The hatching rate of A. salina cysts receded significantly (p < 0.05) upon singular exposures to microplastics and low pH (7.80), whereas combined effect was insignificant. The reactive oxygen species (ROS) elevated as a result of individual exposures to microplastics and low pH. However, only in 0.5 mg mL-1 PE treatments at pH 7.80, an additive impact was reported for ROS activity (p < 0.05). The SOD activities increased significantly but it can be attributed as the individual responses towards exposure to both the stressors. A significant additive impact was not observed for SOD activity (p > 0.05). But during the development, significant morphological anomalies were observed. Changes in the appendages of nauplii and juveniles as a result of combined exposure to microplastics and low pH treatments are significant findings. Our observations suggest that coupled exposure to microplastics and low pH could induce significant oxidative stress in the marine zooplanktons and also adversely affect their normal development. Findings from the current study emphasise the need for further research to understand the coupled toxicological impacts of ocean acidification and predominant pollutants such as microplastics to other marine animals as well.

Polyethylene, Polystyrene, and Polypropylene leachate impact upon marine microalgae Dunaliella tertiolecta.

In the aquatic environment, plastics may release several hazardous substances of severe ecotoxicological concern not covalently bound to the polymers. The aim of this study was to examine the adverse effects of leachates of different virgin polymers, polypropylene (PP), polyethylene (PE), and polystyrene (PS) on marine microalgae Dunaliella tertiolecta. The tests carried out on D. tertiolecta included: growth inhibition, oxidative stress (DCFH-DA), and DNA damage (COMET assay). Polypropylene and PS leachates produced growth inhibition at the lowest concentration (3.1% of leachate). In contrast, a hormesis phenomenon was observed with PE leachates. An algae inhibition growth ranking (PP>PS>PE) was noted, based upon EC50 values. Reactive oxygen species (ROS) generated were increased with leachates concentrations with PS exhibiting the highest ROS levels, while a marked genotoxic effect (30%) was found only with PP. All leachates were free from detectable quantities of organic compounds (GC/MS) but showed the presence of transition, post-transition and alkaline earth metals, metalloids, and nonmetals (

Early ecotoxic effects of ZnO nanoparticle chronic exposure in Mytilus galloprovincialis revealed by transcription of apoptosis and antioxidant-related genes.

Recently, China became one of the largest nanomaterial markets in the world. The wide use of ZnO nanoparticles in a number of products implies an increasing release in marine environment and consequently the evaluation of the potential effects upon marine organisms largely cultured in China for commercial purposes, such as invertebrate bivalves is a current need. To this aim, survival, bioaccumulation, and transcription pattern of key genes, p53, PDRP, SOD, CAT, and GST, involved in DNA damage/repair and antioxidation, in Mytilus galloprovincialis digestive gland, exposed to ZnO NPs (<100 nm) and ZnO bulk (150-200 nm) for 4 weeks, were evaluated. ZnSO4 was also assessed to appraise the role of zinc ions. Starting from 72 h, increasing mortality values along the exposure time were observed for all ZnO compounds. The highest difference was evident after 28 d when NPs resulted three times more toxic than bulk, (LC50) = 0.78 mg Zn/L (confidence limits: 0.64, 1.00) and 2.62 mg Zn/L (confidence limits: 1.00, 4.00), respectively. For ZnSO4 the (LC50) was always the lowest reaching the minimum value at 28 d 0.25 mg Zn/L (confidence limits: 0.10-0.40). Digestive gland showed higher uptake rate of ionic Zn respect to ZnO NPs and bulk during the first three days of exposure. In particular at the end of the exposure time (28 d) at 1 mg Zn/L the rank of Zn uptake rate was Zinc ion > ZnO NPs > ZnO bulk. The relative expression of investigated genes evidenced that distinct actions of apoptosis and antioxidation occurred in M. galloprovincialis exposed to ZnO NPs with a peculiar pattern dependent on exposure time and concentration. Application of the qRT-PCR technique revealed evidence of sensitivity to the nanomaterial since the first time of exposure.

Risk assessment of selected priority pollutants coming from boating activities.

In this work, we evaluated the risk posed to aquatic organisms in the coastal waters of Albania and Apulia (Italy) by two priority pollutants (PPs), Irgarol 1051 and Diuron, used as biocides in antifouling paints on boat hulls. With this aim, we carried out an extensive 3-year monitoring in ports and marinas along the coasts of both countries, which showed a widespread occurrence of both PPs, with Irgarol 1051 concentrations usually being lower than the Diuron ones. The measured concentrations were compared with regulatory Environmental Quality Standards (EQS) (Directive 2008/105/EC) and used to perform a probabilistic Ecological Risk Assessment (ERA), for a thorough evaluation of the potential adverse effects upon marine ecosystem. Irgarol 1051 amounts above the Annual Average Concentration (AA-EQS, 2.5 ng/L) were often detected in Apulia and, less frequently, in Albania. Moreover, in Apulia, sometimes the Maximum Allowable Concentrations (MAC-EQS, 16 ng/L) was exceeded. In Apulia, where levels exceeded MAC/AA-EQS, ERA found not negligible probabilities of exceeding the toxicity level (6-18 %). A less critical situation was observed for Diuron whose levels were always below the MAC-EQS (1800 ng/L) in both countries and, in Albania, also below the AA-EQS (200 ng/L). On the other hand, in Apulia, this limit was exceeded in some locations. Correspondingly, ERA determined a not negligible risk in these sites (probability of exceedance 4-7 %).

Application of a toxicity test battery integrated index for a first screening of the ecotoxicological threat posed by ports and harbors in the southern Adriatic Sea (Italy).

Ports and harbors may represent a threat for coastal ecosystems due to pollutant inputs, especially those derived from maritime activities. In this study, we report a first assessment of the ecotoxicological threat posed by six ports and harbors of opposite coastal regions, Apulia and Albania, in the southern Adriatic Sea (Italy). A bioassay battery consisting of four different species representing different trophic levels, algae Dunaliella tertiolecta, bacteria Vibrio fischeri, crustacean Artemia salina, and echinoids Paracentrotus lividus, has been used to assess sediment elutriates, pore waters, and sediment suspensions. Two different approaches of toxicity data integration, worst case and integrated index, have been used to determine the most appropriate procedure for the investigated sites. All sites with the worst case approach showed high toxicity levels. The chronic test with algae was the most sensitive identifying the highest effects in the battery. This effect can be attributable to contaminants derived from antifouling paints. The sediments, evaluated with V. fischeri test, often showed toxicity not found in the aqueous matrices of the same sites and that can be mainly linked to organic compounds. The test battery used in this study allowed us to perform a preliminary screening of the ecotoxicological risk of the studied area. In fact, the species utilized for toxicity tests responded differently to the investigated samples, showing different sensitivity. The test battery integrated index did not allow highlighting the differences among the sites and showed a general high ecotoxicological risk. A larger number of tests with higher sensitivity together with a tailored attribution of weights to endpoints and matrices will improve the final site evaluation.

Assessing genotoxic effects of plastic leachates in Drosophila melanogaster.

Plastic polymers were largely added with chemical substances to be utilized in the items and product manufacturing. The leachability of these substances is a matter of concern given the wide amount of plastic waste, particularly in terrestrial environments, where soil represents a sink for these novel contaminants and a possible pathway of human health risk. In this study, we integrated genetic, molecular, and behavioral approaches to comparatively evaluate toxicological effects of plastic leachates, virgin and oxodegradable polypropylene (PP) and polyethylene (PE), in Drosophila melanogaster, a novel in vivo model organism for environmental monitoring studies and (eco)toxicological research. The results of this study revealed that while conventional toxicological endpoints such as developmental times and longevity remain largely unaffected, exposure to plastic leachates induces chromosomal abnormalities and transposable element (TE) activation in neural tissues. The combined effects of DNA damage and TE mobilization contribute to genome instability and increase the likelihood of LOH events, thus potentiating tumor growth and metastatic behavior ofRasV12 clones. Collectively, these findings indicate that plastic leachates exert genotoxic effects in Drosophila thus highlighting potential risks associated with leachate-related plastic pollution and their implications for ecosystems and human health.

Do new cement-based mortars pose a significant threat to the aquatic environment?

Nowadays, the use of multi-functional mortars has increased significantly, with interesting applications in the sustainable construction. In the environment, the cement-based materials are subjected to leaching, so the assessment of potential adverse effects upon aquatic ecosystem is necessary. This study focuses on the evaluation of the ecotoxicological threat and of a new type of cement-based mortar (CPM-D) and its raw materials leachates. A screening risk assessment were performed by Hazard Quotient methods. The ecotoxicological effects were investigated by a test battery with bacteria, crustacean, and algae. Two different procedures, Toxicity test Battery Index (TBI) and Toxicity Classification System (TCS), to obtain a single value for toxicity rank were used. Raw materials showed the highest metal mobility and in particular, for Cu, Cd and V potential hazard was evidenced. Leachate toxicity assessment evidenced the highest effects linked to cement and glass while the mortar showed the lowest ecotoxicological risk. TBI procedure allows a finer classification of effect linked to materials with respect to TCS which is based on worst case approach. A safe by design approach taking into account the potential and the effective hazard of the raw materials and of their combinations could allow to achieve sustainable formulations for building materials.

Toxic effects of pentachlorophenol, azinphos-methyl and chlorpyrifos on the development of Paracentrotus lividus embryos.

The application of many current-use pesticides has increased after the disuse of persistent, bioaccumulative or toxic ones as DDT or chlordane. Many of the used pesticides are considered less dangerous towards the environment for their physico-chemical properties. This study investigated the toxic effects of three current-use pesticides, pentachlorophenol (PCP), azinphos-methyl (AZM), and chlorpyrifos, on Mediterranean sea urchin Paracentrotus lividus early development and offspring quality. The experimental results showed that the most toxic pesticides were PCP and AZM at EC50 level. Nevertheless at low concentration PCP resulted the less toxic compound and showed EC1 value more protective than NOEC. PCP at high concentration seemed to modify cytoskeleton assembly, while at low concentrations, it could alter the deposition of the larval skeleton. OPs at low concentrations until 300 µg/l showed a similar toxicological behaviour with a trend corresponding to the pesticide concentrations. At high concentration (500 µg/l) the effect mainly observed was the embryos pre-larval arrest. This investigation highlighted the relevance to evaluate, in coastal seawaters, the levels of the used pesticides to understand the real impact on benthic populations mainly in sites characterized by intensive agriculture or floriculture activities, such as the coastal areas of the Mediterranean Sea.

Physical and chemical threats posed by micro(nano)plastic to sea urchins.

The awareness of the plastic issue is rising in recent years. Our seas and coastal seawaters are investigated with the aim to evaluate the possible fate, behavior and the impact of these novel contaminants upon marine biota. In particular, benthic organisms are exposed to micro(nano)plastics that sink and accumulated on the seabed. Sea urchins can be prone to the plastic impact for all their lifespan with effect that can be extended upon the trophic cascade since their key role as grazer organisms. Moreover, they are largely used in the assessment of contaminant impact both as adult individuals and as early larval stages. This review analyzes the recent literature about the chemical and physical hazards posed by diverse polymers to sea urchins, in relation to their peculiar characteristics and to their size. The search was based on a query of the keyword terms: microplastic _ OR nanoplastic_AND Sea urchins in Web of Science and Google Scholar. The effects provoked by exposure of different sea urchin biological form are highlighted, considering both laboratory exposure and collection in real world. Additional focus has also been given upon the exposure methods utilized in laboratory test and in the existing limitations in the testing procedures. In conclusion, the micro(nano)plastics major impact seemed to be attributable to leaching compounds, however variability and lacking of realisms in the procedures do not allow a full understanding of the hazard posed by micro(nano)plastics for sea urchins. Finally, the work provides insights into the future research strategies to better characterize the actual risk for sea urchins.

Gathering new knowledge from existing monitoring dataset of Campania marine coastal area (Southern Italy).

In this study we propose, a multi-step strategy of selection and characterization of long-term dataset of contaminant concentrations in different environmental matrices (i.e., water and sediment). Starting from a high quality and homogeneous dataset of chemical parameters, a selection of a usable refined dataset followed by statistical characterization and hazard assessment was performed. The database of chemical contamination data from monitoring activities in the coastal marine water area of the Campania Region (Italy) produced by the Regional Agency for Environmental Protection of Campania (ARPAC) between 2013 and 2019 was utilized. Descriptive and multivariate statistics were applied to the extracted data subset to describe spatial variability and to investigate the relationships between matrices and contaminants. In addition, the impact on the sediment matrix was considered using the contamination factor (Cf), the sediment quality guidelines (SQG), and the probability of exceedance (PoE) of given thresholds for metals and organic compounds. The results highlighted the main anthropogenic pressures between the Gulf of Pozzuoli and the Gulf of Napoli, and the potential hazard posed in particular by metals (i.e., Pb, C, and Hg), TBT, and PAHs. A wide range of As concentration along the Campania coastline and Ni occurrence in the southern part of the region, mainly attributable to geogenic origin, was also evidenced. This approach allows extracting new knowledge from large dataset systematically collected by ARPAC monitoring activity, to support possible actions of contamination control and mitigation.

Suitability of lysosomal membrane stability in Eisenia fetida as biomarker of soil copper contamination.

Accumulated metals in soils negatively affect dwelling organisms. Earthworms, which are widespread and perform various essential functions, are able to accumulate metals that can damage the coelomic cells. The aim of this work was to evaluate the effect on Eisenia fetida lysosomal membrane stability both during and after copper exposure, and finally to link this to internal concentrations. E. fetida specimens were exposed to a reference soil and two Cu-spiked soils (35 and 350 mg kg(-1) d.w.) for 14 days (uptake period) and then transferred into the reference soil for other 18 days (elimination period). After 3 days of uptake, internal Cu concentrations increased and were higher in the specimens exposed to soils spiked with 350 mg Cu kg(-1) d.w. After 2 days of elimination, a strong decrease of internal Cu concentrations was always observed. The lysosomal membrane stability, measured as neutral red retention-times, was approximately 50 min for the earthworms exposed to the reference soils, whereas it decreased, at the end of the uptake period, to 21 and 13 min, respectively, for the organisms exposed to soils spiked with 35 and 350 mg Cu kg(-1) d.w. A full recovery of the lysosomal membrane stability was reached after 14 and 18 days of the elimination period, respectively, for the organisms exposed to soils spiked with 35 and 350 mg Cu kg(-1) d.w. The neutral-red assay would seem a good biomarker since the lysosomal membrane stability of E. fetida appeared to respond rapidly and strongly to soil copper contamination.

Assessment of the effects of Cr, Cu, Ni and Pb soil contamination by ecotoxicological tests.

This study aimed to assess soil quality by chemical and ecotoxicological investigations and to check the correspondence between soil metal concentrations and ecotoxicity. For these purposes, surface soils collected at four adjacent roadside urban parks and at a former industrial area were characterized for C/N, organic matter content, texture, and pH. Cr, Cu, Ni and Pb, chosen among the most representative soil metal contaminants, were measured as total content and as available and water soluble fractions. In addition, the total concentrations of the investigated metals were used to calculate two chemical indices: the contamination and the potential ecological risk factors. The toxicity of the investigated soils was evaluated by an ecotoxicity test battery carried out on both soil samples (Vibrio fischeri, Heterocypris incongruens and Sinapis alba) and elutriates (Vibrio fischeri, Daphnia magna and Selenastrum capricornutum). The findings, both by the chemical and ecotoxicological approaches, would suggest that the soils with high metal contamination pose ecological risks. On the other hand, moderately metal contaminated soils did not exclude soil ecotoxicity. In fact, toxic effects were also highlighted in soils with low metal content, toxicity being affected by metal availability and soil characteristics. Moreover, the results suggest the importance of using a battery of tests to assess soil ecotoxicity.

Cadmium, lead and their mixtures with copper: Paracentrotus lividus embryotoxicity assessment, prediction, and offspring quality evaluation.

The aim of this research was to assess the combined effects of three heavy metals (copper, lead, cadmium) on the fertilization and offspring quality of the sea urchin Paracentrotus lividus at EC50, NOEL, and EC1 concentrations. The observed data were compared with the predictions derived from approaches of Concentration Addition (CA) and Independent Action (IA) in order to evaluate the proper prediction of the observed mixture toxic effect. The P. lividus embryotoxicity of trace metals decreases as follows: Cu > Pb > Cd at all toxicity concentration tested. EC50 mixture revealed less toxic only than Cu; EC50 was 0.80 (± 0.07) mg/l, the offspring malformations were mainly P1 type (skeletal alterations) up to 20% mixture concentration, and P2 type from 70% concentration. The NOEL and EC1 mixtures evidenced that all compounds contribute to the overall toxicity, even if present at low concentrations: the former EC50 was 0.532 (± 0.058) mg/l and the latter was 1.081 (± 0.240) mg/l. The developmental defects observed were mainly P1 type in both mixtures. Both CA and IA models did not accurately predict mixture toxicity for EC50 and NOEL mixtures. Instead, EC1 mixture effects seemed well represented by the IA model. The protective action of the CA model, although quite accurate when applied to simple biological systems like algae and bacteria, but failed to represent the worst-case in this study with more complex organisms. It would be useful to introduce in the models one or more factors that take into account the complexity of these biological systems.

Adverse effects of oxo-degradable plastic leachates in freshwater environment.

The production of biodegradable plastics is considered to be a way to reduce plastic waste issue. Among others, oxo-degradant additives enable a faster degradation of plastics in the environment. However, the introduction of these new materials could provoke the release of substances potentially toxic in the environment. This work determined and compared the toxicity of leachates from various additivated polymers (polyethylene, PE; polypropylene, PP; polystyrene, PS) upon different test organisms: plants (Sorghum saccharatum, Lepidium sativum, Sinapis alba, and Vicia faba), crustacean (Daphnia magna), and luminescent bacteria (Vibrio fischeri). Daphnia magna survival was mainly affected by PS and PP leachates (72% and 61% effect, respectively) while PS notably reduced the reproduction rate. On plants, only PP exerted a negative effect (S. saccharatum IG% 32.4), while V. fischeri always showed values around 50%. The data integration, through the Toxicity Test Battery Integrated Index (TBI) approach, allowed to rank the leachates toxicity as PE > PS > PP. This result could be mainly ascribable to the highest metals content in PE since no difference with organic compounds analysis was evidenced. In conclusion, since the polymers exerted dissimilar toxicity, the additive could not be considered the sole responsible of the measured toxicity, but its role in the enhancement of the virgin polymers leachates effects can be solidly hypothesized.

Bioremediation of diesel oil in a co-contaminated soil by bioaugmentation with a microbial formula tailored with native strains selected for heavy metals resistance.

The aim of the work is to assess the feasibility of bioremediation of a soil, containing heavy metals and spiked with diesel oil (DO), through a bioaugmentation strategy based on the use of a microbial formula tailored with selected native strains. The soil originated from the metallurgic area of Bagnoli (Naples, Italy). The formula, named ENEA-LAM, combines ten bacterial strains selected for multiple resistance to heavy metals among the native microbial community. The biodegradation process of diesel oil was assessed in biometer flasks by monitoring the following parameters: DO composition by GC-MS, CO2 evolution rate, microbial load and composition of the community by T-RFLP, physiological profile in Biolog ECOplates and ecotoxicity of the system. The application of this microbial formula allowed to obtain, in the presence of heavy metals, the complete degradation of n-C(12-20), the total disappearance of phenantrene, a 60% reduction of isoprenoids and an overall reduction of about 75% of the total diesel hydrocarbons in 42 days. Concurrently with the increase of metabolic activity at community level and the microbial load, the gradual abatement of the ecotoxicity was observed. The T-RFLP analysis highlighted that most of the ENEA-LAM strains survived and some minor native strains, undetectable in the soil at the beginning of the experiment, developed. Such a bioaugmentation approach allows the newly established microbial community to strike a balance between the introduced and the naturally present microorganisms. The results indicate that the use of a tailored microbial formula may efficiently facilitate and speed up the bioremediation of matrices co-contaminated with hydrocarbons and heavy metals. The study represents the first step for the scale up of the system and should be verified at a larger scale. In this view, this bioaugmentation strategy may contribute to overcome a critical bottleneck of the bioremediation technology.

DNA damages and offspring quality in sea urchin Paracentrotus lividus sperms exposed to ZnO nanoparticles.

The recent advances in nanotechnology lead to a potential increase of the release of nanoparticles (NPs) into marine environment through different routes, with possible toxic effects upon the living part of this ecosystem. One of the ways of NPs marine contamination gaining today increasing concern stems from the widespread use cosmetics containing ZnO NPs as UV-filter. Although the possible adverse effects on marine organisms have been already ascertained, the information about the possible genotoxicity of ZnO NPs is still scant. In this work the spermiotoxicity of ZnO particles of different sizes (ZnO Bulk > 200 nm, ZnO NPs 100 nm and ZnO NPs 14 nm) was assessed, using Paracentrotus lividus spermatozoa, by evaluating the DNA damage of the exposed sperm, fertilization capability and DNA damage transmission to progeny. Our results showed that ZnO NPs induced DNA damages in spermatozoa after 30 min of exposure. While the sperm fertilization capability was not affected, morphological alterations (skeletal alterations) in offspring were observed and a positive correlation between sperm DNA damage and offspring quality was reported. This study underlines that a possible spermiotoxic action of ZnO NPs at concentration close to those reported in marine coastal water could occur.

Leachates of micronized plastic toys provoke embryotoxic effects upon sea urchin Paracentrotus lividus.

Microplastics are defined as plastic fragments <5 mm, and they are found in the ocean where they can impact on the ecosystem. Once released in seawater, microplastics can be internalized by organisms due to their small size, moreover they can also leach out several additives used in plastic manufacturing, such as plasticizers, flame retardants, etc., resulting toxic for biota. The aim of this study was to test the toxicity of micronized PVC products with three different colors, upon Paracentrotus lividus embryos. In particular, we assessed the effects of micronized plastics and microplastic leachates. Results showed a decrease of larval length in plutei exposed to low concentrations of micronized plastics, and a block of larval development in sea urchin embryos exposed to the highest dose. Virgin PVC polymer did not result toxic on P. lividus embryos, while an evident toxic effect due to leached substances in the medium was observed. In particular, the exposure to leachates induced a development arrest immediately after fertilization or morphological alterations in plutei. Finally, PVC products with different colors showed different toxicity, probably due to a different content and/or combination of heavy metals present in coloring agents.

Assessment of the effects of soil PAH accumulation by a battery of ecotoxicological tests.

Surface soils were collected at remote, urban and industrial sites in the Southern of Italy in order to evaluate PAH concentrations and assess the toxic effects by a battery of ecotoxicological tests. The tests were performed on whole soils and on both organic and aqueous extracts. Further goal of this study was to integrate the results coming from each test and matrix in a synthetic toxicity index. The highest summation sigmaPAH concentrations were measured at the industrial soil, although this one did not show an high ecotoxicological risk. Among the performed tests, the phytotoxicity tests showed the highest sensitivity. For whole soil, the worst case always has been represented by test through bacteria. Our results could represent the first step toward the selection of a proper battery to characterize the soil ecotoxicological risk.

Testing ZnO nanoparticle ecotoxicity: linking time variable exposure to effects on different marine model organisms.

Zinc oxide nanoparticles (ZnO NPs) are increasingly used in several personal care products, with high potential to be released directly into marine environment with consequent adverse impact on marine biota. This paper aimed to compare the ecotoxicological effect of ZnO NPs (< 100 nm) towards three marine organisms widely used in toxicity assessment: an algal species (Dunaliella tertiolecta), a bioluminescent bacterium (Vibrio fischeri), and a crustacean (Artemia salina). Bulk ZnO (ZnO bulk, 200 nm) and ionic zinc were also investigated for understanding the role of size and of ionic release in the ZnO toxic action. To this aim, different ecotoxicological tests were used: the inhibition of bioluminescence with V. fischeri at three exposure times (5, 15, and 30 min); the D. tertiolecta growth inhibition at 24, 48, and 72 h; the A. salina mortality at 24-96 h, and A. salina mortality and body growth each 3 days along chronic exposure (14 days). For all selected species, ZnO NPs toxicity was strictly dependent on the exposure time and different sensitivities were recorded: ZnO NPs were more toxic towards algae (EC50 2.2 mg Zn/L) but relatively less toxic towards bacteria (EC50 17 mg Zn/L) and crustaceans (EC50 96 h 58 mg Zn/L). During the 14-day chronic exposure of A. salina, ZnO NPs had a significant inhibition of vitality and body length (EC5014d 0.02 mg Zn/L), while the effect of ZnSO4 was not statistically different from the control. ZnO NP toxicity was related to zinc ions and to interactions of particle/aggregates with target organisms and therefore to NP behavior in the testing matrix and to the different testing time exposures.