Alice Rotini

Phytotoxic antibiotic sulfadimethoxine elicits a complex hormetic response in the weed Lythrum salicaria L

In order to evaluate the hormetic response of the weed Lythrum salicaria to drug exposure we investigated the effects of the antibiotic Sulfadimethoxine by growing Lythrum plants for 28 days on culture media containing different drug concentrations (between 0.005 and 50 mg.L−1). The antibiotic was absorbed by plants and can be found in plant tissue. The plant response was organ-dependent: roots, cotyledons and cotyledon petioles, were always affected by a toxic effect, whilst internodes and leaves length, showed a variable dose-depending response, with an increased growth at the lower drug concentrations and toxic effects at the higher ones. This variable response was probably dependant on different levels of local contamination resulting from a balance between accumulation rate and drug dilution in the increasing plant biomass. As a consequence, drug toxicity or hormetic response varied according to concentration and were different in each of the examined plant organ/tissue. Thus, even if hormesis can be considered a general plant response, each plant organ/tissue responds differently, depending on the local drug concentration and exposure time.

LOW DOSES OF TETRACYCLINE TRIGGER THE E. COLI GROWTH: A CASE OF HORMETIC RESPONSE

Hormesis is a biphasic dose-response relationship, occurring when low concentrations of toxic agents elicit apparent improvements. In this work, the ability of sub-inhibitory concentrations of Tetracycline to induce hormetic response in a model organism was investigated. To this aim a reference strain of Escherichia coli, MG1655, was exposed to six decreasing doses of Tetracycline (between 0.12 and 0.00375 μg/ml), much lower than the Minimal Inhibitory Concentration (4 μg/ml). An hormetic increase was observed at the intermediate concentrations (0.015–0.03 μg/ml) of the tested range. The Colony Forming Unit number, indeed, rose up to 141% and 121% as compared to the control. At the highest (0.12 μg/ml) and lowest (0.00375 μg/ml) concentrations a slight decrease in CFU number was found. Results demonstrated that, in Escherichia coli, low concentrations of Tetracycline bias the bacterial numerical increase through a hormetic response; the dose-response curve describing this numerical increase is an U-inverted curve. Furthermore, these data confirm that hormesis is common to many - if not all - living systems, including bacteria; they underline the relevance of a deepened knowledge of both the effects and the possible consequences of exposure to low doses of contaminants.

Insights into the CuO nanoparticle ecotoxicity with suitable marine model species

Metal oxide nanoparticles, among them copper oxide nanoparticles (CuO NPs), are widely used in different applications (e.g. batteries, gas sensors, superconductors, plastics and metallic coatings), increasing their potential release in the environment. In aquatic matrix, the behavior of CuO NPs may strongly change, depending on their surface charge and some physical-chemical characteristics of the medium (e.g. ionic strength, salinity, pH and natural organic matter content). Ecotoxicity of CuO NPs to aquatic organisms was mainly studied on freshwater species, few tests being performed on marine biota. The aim of this study was to assess the toxicity of CuO NPs on suitable indicator species, belonging to the ecologically relevant level of consumers. The selected bioassays use reference protocols to identify Effect/Lethal Concentrations (E(L)C), by assessing lethal and sub-lethal endpoints. Mortality tests were performed on rotifer (Brachionus plicatilis), shrimp (Artemia franciscana) and copepod (Tigriopus fulvus). While moult release failure and fertilization rate were studied, as sub-lethal endpoints, on T. fulvus and sea urchin (Paracentrotus lividus), respectively. The size distribution and sedimentation rates of CuO NPs, together with the copper dissolution, were also analyzed in the exposure media. The CuO NP ecotoxicity assessment showed a concentration-dependent response for all species, indicating similar mortality for B. plicatilis (48hLC50 = 16.94 ± 2.68mg/l) and T. fulvus (96hLC50 = 12.35 ± 0.48mg/l), followed by A. franciscana (48hLC50 = 64.55 ± 3.54mg/l). Comparable EC50 values were also obtained for the sub-lethal endpoints in P. lividus (EC50 = 2.28 ± 0.06mg/l) and T. fulvus (EC50 = 2.38 ± 0.20mg/l). Copper salts showed higher toxicity than CuO NPs for all species, with common sensitivity trend as follows: P. lividus ≥ T. fulvus (sublethal endpoint) ≥ B. plicatilis >T. fulvus (lethal endpoint) >A. franciscana. CuO NP micrometric aggregates and high sedimentation rates were observed in the exposure media, with different particle size distributions depending on the medium. The copper dissolution was about 0.16% of the initial concentration, comparable to literature values. The integrated ecotoxicological-physicochemical approach was used to better describe CuO NP toxicity and behavior. In particular, the successful application of ecotoxicological reference protocols allowed to produce reliable L(E)C data useful to identify thresholds and assess potential environmental hazard due to NPs.

Ecophysiological Plasticity and Bacteriome Shift in the Seagrass Halophila stipulacea along a Depth Gradient in the Northern Red Sea

Halophila stipulacea is a small tropical seagrass species. It is the dominant seagrass species in the Gulf of Aqaba (GoA; northern Red Sea), where it grows in both shallow and deep environments (1–50 m depth). Native to the Red Sea, Persian Gulf, and Indian Ocean, this species has invaded the Mediterranean and has recently established itself in the Caribbean Sea. Due to its invasive nature, there is growing interest to understand this species’ capacity to adapt to new conditions, which might be attributed to its ability to thrive in a broad range of ecological niches. In this study, a multidisciplinary approach was used to depict variations in morphology, biochemistry (pigment and phenol content) and epiphytic bacterial communities along a depth gradient (4–28 m) in the GoA. Along this gradient, H. stipulacea increased leaf area and pigment contents (Chlorophyll a and b, total Carotenoids), while total phenol contents were mostly uniform. H. stipulacea displayed a well conserved core bacteriome, as assessed by 454-pyrosequencing of 16S rRNA gene reads amplified from metagenomic DNA. The core bacteriome aboveground (leaves) and belowground (roots and rhizomes), was composed of more than 100 Operational Taxonomic Units (OTUs) representing 63 and 52% of the total community in each plant compartment, respectively, with a high incidence of the classes Alphaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria across all depths. Above and belowground communities were different and showed higher within-depth variability at the intermediate depths (9 and 18 m) than at the edges. Plant parts showed a clear influence in shaping the communities while depth showed a greater influence on the belowground communities. Overall, results highlighted a different ecological status of H. stipulacea at the edges of the gradient (4–28 m), where plants showed not only marked differences in morphology and biochemistry, but also the most distinct associated bacterial consortium. We demonstrated the pivotal role of morphology, biochemistry (pigment and phenol content), and epiphytic bacterial communities in helping plants to cope with environmental and ecological variations. The plant/holobiont capability to persist and adapt to environmental changes probably has an important role in its ecological resilience and invasiveness.

Salinity-Based Toxicity of CuO Nanoparticles, CuO-Bulk and Cu Ion to Vibrio anguillarum

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 EC50 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. CuSO4 5H2O 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; Cu2+ 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: CuSO4 5H2O > 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 Cu2+ 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.

Cytotoxicity and genotoxicity of CuO nanoparticles in sea urchin spermatozoa through oxidative stress

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.

Ecotoxicological effects of polystyrene microbeads in a battery of marine organisms belonging to different trophic levels

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.

The trade-off between digestibility and phenol content influences the food choice of the obligate seagrass-feeding neritid snail Smaragdia souverbiana

Grazing influences the structure and function of seagrass meadows. Like terrestrial plants, seagrasses have evolved nutritional, structural and chemical mechanisms to either tolerate or resist grazing. Phenols are common secondary metabolites that have multiple roles, including grazing deterrence, and their content in seagrass tissues can vary both among species and within a single species in response to biotic and abiotic processes. Such variability influences the grazing behaviour of species that consume seagrass, but few studies have addressed mesograzers that consume or damage seagrass, such as the nerite Smaragdia souverbiana. We assessed the effect of environmentally- mediated differences in phenol concentration on these seagrass mesograzers by testing: ( 1) if phenol content is different between sites and seagrass species ( Zostera muelleri vs Halophila ovalis) and ( 2) if S. souverbiana inflicts different amounts of damage on different seagrasses from different sites, according to their phenol content. Phenol content was consistently lower in Z. muelleri than H. ovalis, and concentrations were about 60% lower in both species at one of two sites in eastern Moreton Bay, Queensland. These differences corresponded to different foraging patterns of S. souverbiana, with the snails generally choosing the seagrass species with the lowest phenol content. These results suggest that S. souverbiana chooses among locally available seagrasses based on a trade- off between phenol content and digestibility ( related to cell size). This work demonstrates how subtle local processes can create complex changes in seagrass- grazer interactions with potential ecological consequences at local and regional scales.

Dispersant approval procedures in France and Italy: A comparative ecotoxicity study

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 (LC50 ≥ 10 times reference toxicant), while only one dispersant met the Italian approval criterion (EC50 > 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.