Simona Schiavo

Genotoxic and cytotoxic effects of ZnO nanoparticles for Dunaliella tertiolecta and comparison with SiO2 and TiO2 effects at population growth inhibition levels

The increasing use of oxide nanoparticles (NPs) in commercial products has intensified the potential release into the aquatic environment where algae represent the basis of the trophic chain. NP effects upon algae population growth were indeed already reported in literature, but the concurrent effects at cellular and genomic levels are still largely unexplored. Our work investigates the genotoxic (by COMET assay) and cytotoxic effects (by qualitative ROS production and cell viability) of ZnO nanoparticles toward marine microalgae Dunaliella tertiolecta. A comparison at defined population growth inhibition levels (i.e. 50% Effect Concentration, EC50, and No Observed Effect Concentration, NOEC) with SiO2 and TiO2 genotoxic effects and previously investigated cytotoxic effects (Manzo et al., 2015) was performed in order to elucidate the possible diverse mechanisms leading to algae growth inhibition. After 72h exposure, ZnO particles act firstly at the level of cell division inhibition (EC50: 2mg Zn/L) while the genotoxic action is evident only starting from 5mg Zn/L. This outcome could be ascribable mainly to the release of toxic ions from the aggregate of ZnO particle in the proximity of cell membrane. In the main, at EC50 and NOEC values for ZnO NPs showed the lowest cytotoxic and genotoxic effect with respect to TiO2 and SiO2. Based on Mutagenic Index (MI) the rank of toxicity is actually: TiO2>SiO2>ZnO with TiO2 and SiO2 that showed similar MI values at both NOEC and EC50 concentrations. The results presented herein suggest that up to TiO2 NOEC (7.5mg/L), the algae DNA repair mechanism is efficient and the DNA damage does not result in an evident algae population growth inhibition. A similar trend for SiO2, although at lower effect level with respect to TiO2, is observable. The comparison among all the tested nanomaterial toxicity patterns highlighted that the algae population growth inhibition occurred through pathways specific for each NP also related to their different physicochemical behaviors in seawater.

Effects of PVP/PEI coated and uncoated silver NPs and PVP/PEI coating agent on three species of marine microalgae

In the last years, applications for silver nanoparticles (Ag NPs) continue to increase together with the concerns about their potential input and hazards in aquatic ecosystems, where microalgae are key organisms. The aim of the present study was to assess the relative sensitivity of three marine microalgae species with differences in cell wall composition/structure exposed to Poly N-vinyl-2-pirrolidone/Polyethyleneimine (PVP/PEI) coated 5nm Ag NPs and uncoated 47nm Ag NP. As limited attention has been paid to the role of coating agents in NP toxicity, the effect of PVP/PEI alone was also evaluated. After 72h in artificial seawater, 47nm Ag NPs formed around 1400nm size aggregates while PVP/PEI coated 5nm Ag NPs reached around 90nm. Ag+ release in seawater was around 3% for 47nm Ag NPs and 30% for PVP/PEI coated 5nm Ag NPs. PVP/PEI coated 5nm Ag NP aggregates entrapped the algal cells in a network of heteroaggregates, while uncoated 47nm Ag NPs interacted to a lesser extent with algae. The concentration of PVP/PEI coated 5nm Ag NPs that exerted the median effect (EC50) on algae growth pointed out differences in algae sensitivity: T. suecica was about 10 times more sensitive than I. galbana and P. tricornutum. Further, the coating agent alone was as toxic to algae as PVP/PEI coated 5nm Ag NPs, suggesting that presence of the coating agent was the main driver of toxicity of coated NPs. Uncoated 47nm Ag NPs instead, showed similar toxicity towards algae although P. tricornutum was slightly less sensitive than T. suecica and I. galbana, which agrees with the presence of a resistant silicified cell wall in the diatom. The present work demonstrates differences in sensitivity of three marine microalgae, possibly related to their cell surface and size characteristics.

Experimental investigation to evaluate the potential environmental hazards of photovoltaic panels.

Recently the potential environmental hazard of photovoltaic modules together with their management as waste has attracted the attention of scientists. Particular concern is aroused by the several metals contained in photovoltaic panels whose potential release in the environment were scarcely investigated. Here, for the first time, the potential environmental hazard of panels produced in the last 30 years was investigated through the assessment of up to 18 releasable metals. Besides, the corresponding ecotoxicological effects were also evaluated. Experimental data were compared with the current European and Italian law limits for drinking water, discharge on soil and landfill inert disposal in order to understand the actual pollution load. Results showed that less than 3% of the samples respected all law limits and around 21% was not ecotoxic. By considering the technological evolutions in manufacturing, we have shown that during the years crystalline silicon panels have lower tendency to release hazardous metals with respect to thin film panels. In addition, a prediction of the amounts of lead, chromium, cadmium and nickel releasable from next photovoltaic waste was performed. The prevision up to 2050 showed high amounts of lead (30t) and cadmium (2.9t) releasable from crystalline and thin film panels respectively.

Immune and reproductive system impairment in adult sea urchin exposed to nanosized ZnO via food

In marine environment the release and the consequent sedimentation of ZnO NPs, mainly used in sunscreens, could provoke toxic effects in particular in grazer organisms, like sea urchins. In this work, a first evaluation of DNA and cellular effects on adult sea urchins Paracentrotus lividus exposed through the diet to different sizes (100 and 14nm) ZnO NPs, was performed. Moreover, the consequent impact upon offspring quality was evaluated. Preliminarily results showed that the assumption of food containing ZnO NPs 100nm provoked in adult echinoids damages to immune cells (33% of damaged nucleus) and transmissible effects to offspring (75.5% of malformed larvae). Instead food with ZnO NPs 14nm provoked 64% of damaged nucleus in immune cells and 84.7% of malformed larvae.

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.

Different sizes of ZnO diversely affected the cytogenesis of the sea urchin Paracentrotus lividus

Today nanoparticles (NPs) have many applications in commercial products due to their small size and peculiar properties that, conversely, make them potentially toxic for humans and the environment. ZnO NPs are largely used in many personal care products, such as sunscreens and cosmetics. In this study the cytotoxic effects of ZnO particles with different sizes (ZnO Bulk, >100nm; ZnO NPs, 100nm and ZnO NPs, 14nm) upon the first developmental stages of the sea urchin Paracentrotus lividus, are evaluated. Morphological alterations are also assessed by embryotoxicity tests. The cytogenetic analysis highlighted that ZnO NPs interfere with cell cycle inducing a dose-dependent decrease of mitotic activity and chromosomal aberrations at higher concentrations (30μM). Moreover, the larval development was affected by ZnO NPs 100nm (EC50=0.46 [0.30-0.63] μM [Zn]) in a dose-dependent way. Size-dependent toxicity was instead not obtained for ZnO NPs. From our results could be highlighted that the presence of embryos, blocked in pre-larval stage, could be due to the induction of chromosome aberrations by ZnO particles, confirming that cytogenetic analyses allow evaluating possible NPs action mechanisms.

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.