Processes involved in biochemical response to pesticides by lizard Podarcis siculus (Rafinesque-Schmaltz, 1810) - A field study.

Although reptiles are non-target organisms of pesticide applications, their ecological niche and trophic role suggest that the use of these compounds in agriculture can have toxicological effects on them. Our recent field study on Italian wall lizard Podarcis siculus in hazelnut orchards evidenced that the use of pesticides-mixtures, consisting of thiophanate-methyl (TM), tebuconazole (TEB), deltamethrin (DM), lambda-cyhalothrin (LCT), besides copper sulphate, induced an increase of the total antioxidant capacity toward hydroxyl radicals and caused DNA damage; however, it did not cause neurotoxicity, and did not induce the glutathione-S-transferases' activities. These results raised some questions which were answered in this study by carrying out analyses on 4 biomarkers and 5 chemical substances in the tissues of non-target organisms coming from treated fields: cytochrome P450, catalase, total glutathione, and malondialdehyde, TM, TEB, DM, LCT and Cu. Our results highlighted a partial accumulation of different chemicals, the involvement of two important mechanisms of defence, and some cellular damages after exposure to the considered pesticides. In details, 1) LCT and DM were not accumulated in lizard muscle, copper remained at basal levels, whereas TM and TEB were uptaken with a partial metabolization of TM; 2) the cytochrome P450 and the catalase were involved in lizard biochemical responses to pesticides-mixtures used for "conventional" farming treatment; 3) "conventional" treatment with pesticides caused damage to lipids, besides DNA, probably related to the excess of hydroxyl radicals.

Corrigendum: 10-year time course of Hg and organic compounds in Augusta Bay: Bioavailability and biological effects in marine organisms.

[This corrects the article DOI: 10.3389/fpubh.2022.968296.].

10-year time course of Hg and organic compounds in Augusta Bay: Bioavailability and biological effects in marine organisms.

In the last century, many Mediterranean coastal areas have been subjected to anthropogenic disturbances from industrial activities, uncontrolled landfills, shipyards, and high maritime traffic. The Augusta Bay (eastern Sicily, Italy) represents an example of a strongly impacted coastal environment with an elevated level of sediments contamination due to the presence of one of the largest European petrochemical plants, combined with an extensive commercial and military harbor. The most significant contaminants were represented by mercury (Hg) and hexachlorobenzene (HCB), derived from a former chlor-alkali plant, and other organic compounds like polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs). Since the 1970s, Augusta Bay has become internationally recognized as a contaminated marine environment, although very little information is available regarding the temporal trend of contaminants bioavailability and biological impacts on aquatic organisms. In this study, the Hg and HCB concentrations were investigated over 10 years (from 2003 to 2013) in sediments and invertebrate and vertebrate organisms; these two contaminants' ecotoxicity was further evaluated at a biochemical and cellular level by analyzing the induction of organic biotransformation processes and DNA damages. The results showed high concentrations of Hg and HCB in sediments and their strong bioaccumulation in different species with significantly higher values than those measured in reference sites. This trend was paralleled by increased micronuclei frequency (DNA damage biomarker) and activity of the biotransformation system. While levels of chemicals in sediments remained elevated during the time course, their bioavailability and biological effects showed a gradual decrease after 2003, when the chlor-alkali plant was closed. Environmental persistence of Hg and HCB availability facilitates their bioaccumulation and affects the health status of marine organisms, with possible implications for environmental risk, pollutants transfer, and human health.

Supporting SMEs during COVID-19: The case for targeted equity injections.

We analyze the potential role of equity injections in addressing solvency risks among small and medium-sized enterprises (SMEs) after the COVID-19 crisis. Building on firm-level balance sheet projections for a sample of European economies, we simulate selected policy interventions and find that equity injections are quite effective at dampening the rise in insolvencies. Cost effectiveness requires careful targeting, however; under an illustrative scenario, leaving aside any costs arising from imperfect information and implementation, the cost of a program targeting only those SMEs worth saving is just a tenth of the cost of an untargeted approach directed to all insolvent firms. Overall, our paper provides a case for governments to rely more on targeted equity injections in responding to major shocks that trigger mass solvency risks.

Chronic sublethal effects of ZnO nanoparticles on Tigriopus fulvus (Copepoda, Harpacticoida).

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) Zn2+ from ZnO NP suspension after centrifugation (supernatant) and (c) ZnSO4 H2O. 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 Zn2+ and ZnSO4 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.

Biological responses of the polychaete Hediste diversicolor (O.F.Müller, 1776) to inorganic mercury exposure: A multimarker approach.

Mercury (Hg) is a global priority pollutant given its relevance in terms of environmental damage and threat to human health. Its ecotoxicity was tested using the benthic keystone species Hediste diversicolor as target species. After 10 days of exposure to different levels of inorganic Hg (10 and 50 µg L-1), bioaccumulation and a wide range of biological responses were evaluated at different biological levels, including biomarkers of exposure, neurotoxicity, oxidative stress, genotoxicity and cytochemistry. In controlled laboratory conditions, Hg was taken up by H. diversicolor in a dose-response manner and caused a range of biological responses, including oxidative stress (GSTs, GPx, GSH-2GSSG, and TOSCA), neurotoxicity (AChE), and cellular damages at the membrane level (LFs, NLs, Ca2+-ATPase); however, it did not cause significant DNA damage or mortality. This study confirms the capability of H. diversicolor to tolerate high levels of metals and clarifies the mechanisms underlying the damage caused by waterborne Hg and the defense mechanisms, activated in this species. In particular, detoxification of the inorganic form of Hg in this species was found to be strongly related to glutathione expression and several antioxidant enzymes of the antioxidant system. This process also efficiently minimized negative effects on DNA and prevented death, but was not sufficient to avoid neurotoxicity and some cellular damages, mainly at the intestinal level.

PAH (Polycyclic Aromatic Hydrocarbon) bioaccumulation and PAHs/shell weight index in Ruditapes philippinarum (Adams & Reeve, 1850) from the Vallona lagoon (northern Adriatic Sea, NE Italy).

The Vallona lagoon is a transitional area located in the Po River delta (NE, ITALY) traditionally exploited for Manila clam (Ruditapes philippinarum) farming. During 2007-2008, a pipeline was buried in the middle of the lagoon to connect an off-shore structure to facilities on land. PAH levels were monitored in Manila clams and sediments before, during and after the pipeline construction to assess the impact of the activities through the pattern of distribution of the PAH compounds. PAH bioaccumulation in clams displayed seasonal fluctuations with higher levels in autumnal and wintry surveys than in spring-summer. Principal component analysis applied to PAHs in clams highlighted a petrogenic input during ante operam period and a pyrolytic origin during the burying activities. On the contrary, sediment PAH concentrations resulted quite similar both among sites and periods. Biota-Sediment-Accumulation-Factor values also confirmed that sediments were not the major source of PAH pollution for clams in this study. The welfare of clams was examined through two physiological indices (condition index and survival in air) to check the effects of the activities on a commercial resource. Both physiological indices exhibited seasonal variations connected to natural endogenous and exogenous factors; however survival in air was the most sensitive index in highlighting the effects of the pipeline burying activities. Finally, to ensure that PAH bioavailability assessment was not affected by seasonal variation of soft tissues of molluscs, PAHs/shell weight index was applied. Higher levels of this index were observed before and during the burying activities, whilst, after that, values significantly lowered. Moreover, the normalization enabled us to highlight the PAH uptake from clams in some particular periods and to compare different populations in a long-term biomonitoring program with data obtained from different periods of the year.

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.

Temporal evolution of the environmental quality of the Vallona Lagoon (Northern Mediterranean, Adriatic Sea).

Guidance Document 25/2010, suggests sediment and biota are the most suitable matrices for the trend monitoring purpose, because they integrate the pollution over time and space. So, from 2005 to 2014, the sediment and biota concentrations of heavy metals (As, Cd, Cr, Hg, Ni, Pb) were analysed in the Vallona Lagoon (northern Adriatic Sea, Italy), widely used for intensive and extensive bivalve farming. The contamination levels in sediment and biota were compared with Environmental Quality Standard (EQS) and threshold levels (TL) for human health. The results identified critical issues related to Cd in sediment samples as well as to Hg and Pb in biota which were not only ascribable to the physiological and seasonal variability of organisms. The Cr and Ni levels in sediment were higher than the EQS. However, the concentration increases at biota stations close to sites where EQS excesses were observed in sediment were not verified.

The marine sedimentary record of natural and anthropogenic contribution from the Sulcis-Iglesiente mining district (Sardinia, Italy).

Intensive exploitation of base metal deposits in the Sulcis-Iglesiente district (Sardinia, Italy), lasted from the 1850s to the 1990s, determined a high environmental impact on the coastal area, but the effects on marine environment have never been investigated. A marine sediment core, dated with 14C, was characterized for grain size, chemical and mineralogical composition, in order to reconstruct the sedimentary history of the area and to assess the environmental impact of mining. The comparison of chemical and mineralogical characteristics of recent sediments with those of pre-industrial age allowed discriminating the real anthropogenic impact from the natural metal enrichment. The correspondence, in the upper core, of anthropogenic trace metal enrichment with the presence of mine waste minerals is attributed to the exploiting over industrial scale; the still high metal enrichment in sediment surface levels suggests a still existing impact due to mine dumps and tailings weathering.

Establishing geochemical background levels of selected trace elements in areas having geochemical anomalies: The case study of the Orbetello lagoon (Tuscany, Italy).

The determination of background concentration values (BGVs) in areas, characterised by the presence of natural geochemical anomalies and anthropogenic impact, appears essential for a correct pollution assessment. For this purpose, it is necessary to establish a reliable method for determination of local BGVs. The case of the Orbetello lagoon, a geologically complex area characterized by Tertiary volcanism, is illustrated. The vertical concentration profiles of As, Cd, Cr, Cu, Hg, Ni, Pb and Zn were studied in four sediment cores. Local BGVs were determined considering exclusively samples not affected by anthropogenic influence, recognized by means of multivariate statistics and radiochronological dating ((137)Cs and (210)Pb). Results showed BGVs well-comparable with mean crustal or shale values for most of the considered elements except for Hg (0.87 mg/kg d.w.) and As (16.87 mg/kg d.w.), due to mineralization present in the catchment basin draining into the lagoon.

Environmental quality of Italian marine water by means of marine strategy framework directive (MSFD) descriptor 9.

ISPRA, on behalf of the Italian Ministry of Environment, carried out the initial assessment of environmental quality status of the 3 Italian subregions (Mediterranean Sea Region) on Descriptor 9. The approach adopted to define the GES started to verify that contaminants in fish and other seafood for human consumption did not exceed levels established by Community legislation (Reg. 1881/2006 and further updates). As the Marine Strategy Framework Directive (MSFD) requires to use health tools to assess the environment, Italy decided to adopt a statistical range of acceptance of thresholds identified by national (D.Lgs. 152/2006 concerning water quality required for mussel farms) and international legislation (Reg. 1881/2006 and further updates), which allowed to use the health results and to employ them for the assessment of environmental quality. Italy proposed that Good Environmental Status (GES) is achieved when concentrations are lower than statistical range of acceptance, estimated on samples of fish and fishery products coming from only national waters. GIS-based approach a to perform different integration levels for station, cell's grid and years, was used; the elaborations allowed to judge the environmental quality good.

The impact of the Bagnoli industrial site (Naples, Italy) on sea-bottom environment. Chemical and textural features of sediments and the related response of benthic foraminifera.

The coastal zone of the disused industrial site of Bagnoli, has been studied since 1999 in order to highlight chemical and ecological features of pollution, mainly due to a steel plant. This further study was performed in order to check the foraminiferal response to changes in sediment grain-size and contaminant concentrations and to recognise the actual effects of the environmental stress determined by industrial pollution on the foraminiferal assemblages. This was attained by considering a wider area than in the previous studies so as to recognise the possible reference conditions. Very high contamination, mainly due to Cu, Fe, Hg, Mn, Ni, Pb, Zn and PAHs, was recorded in the marine sediments close to the steel plant. Contaminant concentrations and sediment composition were recognised as important factors influencing the foraminiferal response by means of statistical analysis. The foraminiferal abnormality index (FAI) is positively correlated with heavy metals concentration, exceeding the natural threshold in front of the plant. Furthermore, increasing pollution levels correspond to the increase of pollution-tolerant species in the assemblage.

Methylmercury determination in marine sediment and organisms by Direct Mercury Analyser.

An analytical method for simple and rapid determination of methylmercury in sediment and organism samples is described. The proposed method employs the oxygen combustion-gold amalgamation using Direct Mercury Analyser (DMA-80) after complete removal of MeHg by organic extraction and back extraction to an aqueous medium. DMA-80 instrument is equally suitable for the analysis of solid and liquid materials and has a good detection limit. The analytical performance of this method was evaluated by analysis of certified reference materials (CRM-580, IAEA-405, DORM-2, DOLT-3, SRM-2976 and SRM-2977) assessing its quality in terms of accuracy, repeatability and quantification limit. Furthermore total mercury and methylmercury have been analysed in sediment and organism samples collected during the XXI Italian Antarctic Expedition in Terra Nova Bay (Ross Sea, Northern Victoria Land). The results obtained show the validity of the proposed method as ready-to-use analytical method to analyse real samples.

Seasonal variations of exposure biomarkers, oxidative stress responses and cell damage in the clams, Tapes philippinarum, and mussels, Mytilus galloprovincialis, from Adriatic sea.

This work investigated the natural variability of several biomarkers in Tapes philippinarum and Mytilus galloprovincialis, sampled from Northern Adriatic where these organisms are important sentinel species for future environmental impact assessment. Levels of metallothioneins, peroxisomal enzymes and acetylcholinesterase, showed a significant seasonality and marked differences between clams and mussels. Among antioxidant enzymes, catalase and GST decreased during the warmer period, the latter enzyme activity resulting particularly high in clams. The total oxyradical scavenging capacity toward peroxyl radicals decreased in mussels from winter to summer, indicating a prooxidant challenge due to higher seawater temperature and intensity of light irradiance. Lysosomal membrane stability did not exhibit significant seasonal variations, while some variations were observed for DNA damages. Overall results indicated a significant influence of seasonal variability on several biomarkers and species-specific differences which should be considered to discriminate the appearance of anthropogenic disturbance.

Application of nano-FIA-Direct-EI-MS to determine diethylene glycol in produced formation water discharges and seawater samples.

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.