Honey bees (Apis mellifera) are sophisticated bioindicators due to their capacity to integrate contamination signals from multiple environmental matrices. This study investigated pesticide residues and multi-element accumulation, including rare earth elements (REEs), in worker honey bees across four distinct ecological contexts in Calabria (Southern Italy): two protected montane forests (S1 and S2, Sila National Park), one suburban, and one urban site. A total of 778 agrochemicals and 46 trace elements were targeted using gas and liquid chromatography-tandem mass spectrometry (GC-MS/MS and LC-ESI-MS/MS), and inductively coupled plasma-mass spectrometry (ICP-MS), respectively. Chemometric analyses were applied to assess spatial variability and evaluate the discriminant power of the elemental fingerprint. Pesticide residues were predominantly below quantification limits, with the exception of the fungicide ametoctradin at site S2, likely reflecting localized agricultural proximity. Elemental analyses revealed pronounced, site-specific accumulation patterns driven by landscape context. Urban honey bees (S4) exhibited significant enrichments in Pb, Ni, Cr, and Se, consistent with traffic-related emissions, while suburban bees (S3) showed higher levels of Ba, Mn, As, and Zn. Surprisingly, bees from the protected Natura 2000 site (S2) displayed the highest diversity and concentrations of REEs, suggesting an interplay between lithogenic sources and agricultural inputs. Furthermore, we identified striking interindividual variability in accumulation-a critical, yet often overlooked, dimension likely driven by age, foraging range, and micro-scale environmental heterogeneity. These findings challenge current biomonitoring paradigms and underscore the role of honey bees as high-resolution tools for assessing both emerging contaminants (REEs) and anthropogenic pressures in terrestrial ecosystems.

A rapid and environmentally sustainable analytical method was developed for the simultaneous determination of nitrophenols, nitroguaiacols, and nitrocatechols in environmental water samples. The proposed strategy integrates aqueous-phase derivatization with ethyl chloroformate, dispersive pipette extraction (DPX), and gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) into a streamlined workflow that minimizes organic solvent consumption, sample handling, and total analysis time. Derivatization was performed directly in the aqueous matrix under mild conditions, enabling seamless coupling with the DPX procedure. Multivariate experimental design was applied to optimize both derivatization parameters and programmed temperature vaporization (PTV) injection conditions, efficiently identifying the most influential factors while minimizing the number of experiments required. Under optimized conditions, the method provided extraction recoveries of 55-82%, excellent linearity (R2 >= 0.9986), and limits of quantification ranging from 0.3 to 5 mu g/L. Accuracy ranged between 84 and 119% with relative standard deviations below 17.5%, while matrix effects were negligible for the investigated water matrices. Notably, the proposed method enables, for the first time, the simultaneous determination of nitrophenols, including dinitrophenols, nitroguaiacols, and nitrocatechols in environmental water samples, significantly expanding the analytical scope beyond previously reported approaches. Greenness was evaluated using the AGREEprep metric, confirming the low environmental impact of the protocol. The developed method represents a powerful, reliable, and analytically comprehensive tool for the large-scale monitoring of nitroaromatic phenols in environmental waters.

The late part of the 20th century the advancement of knowledge regarding nutrition and\ndisease prevention provided an opportunity for individuals to affect their own health.\nThis expanding body of information helped people to understand how the environment\nand their own behaviour affected their body. People now had powerful tools to help\nmaintaining and protecting their health. The understanding of how our diet affects our\nwell being has dramatically changed the lifestyles and attitudes of people, who began to\nmake menu and purchasing decisions based on how foods would affect their heath. A\nshift toward healthier lifestyles and healthier diets began. Food processors and marketers\nhad to refocus their efforts from promoting foods for pleasure to promoting foods that fit\nin to a healthy diet. Primarily, the focus was on reducing fat and cholesterol in the diet\nand supplementing vitamins and minerals. Research began to demonstrate the presence\nof various phytochemicals in wine and juice make it from fruits and vegetables( such as\nstilbenes, falvoniods, polyphenols…) and specially in olive oil as ( polyphenol in\ndialdehyed form: oleochanthal, hydroxyoleocanthal..) These compounds have come to\nbe known as nutraceuticals. The list of nutraceuticals present in wine, fruit drink and\nolive oil that are believed to have positive biological properties has been expanding.\nFood processors and developers have become very interested in exploiting these\nnutraceuticals for the production of foods that are not only part of a healthy diet but also\nimprove the consumer’s health in another specific way. These foods have become\nknown as “functional foods” means quality marker.\nThe aim of this thesis has been to develop a analytical methods to determine the\nconcentration of a group of these nutraceuticals in food , such as, quantitative\ndetermination of resveratrol in wine, pterostilbene in blueberry juice and dialdehyde\nform in olive oil using a sensitive high-performance liquid chromatographic separation\nmethod coupling with tandem-mass and isotope dilution to order to optimal the\nconditions for the analysis method, such as extraction procedure, matrices, column,\nquality controls, wavelength, mobile phases, run time, optimal separation (gradient,\nretention times), temperature, capillary voltage, cone voltages, vacuum and labelled\ninternal standards, resulting in the best sensitivity and selectivity,The goodness and satisfactory of the method was performed according to, containing\nlinear measuring range, quantification, lower limit of quantification (LLOQ), lower limit\nof detection (LLOD), quality controls, precision(RSD %), accuracy, recovery, stability\nand matrix effects.\nIn conclusion, the described high-performance liquid chromatographic separation\nmethod with tandem-mass spectrometry detection and isotope dilution showed a\nsatisfactory overall analytical performance well suited for applications in food quality\ncontrol.

Nitroaromatic compounds, encompassing nitrophenols (NPs), nitrocatechols (NCs), and nitroguaiacols (NGs), have garnered significant attention due to their potentially harmful effects on human health and the environment. However, to date, no studies have been conducted on the analysis of these compounds in indoor dust. In order to address this lacuna, an analytical procedure was developed for the determination of NPs, NCs, and NGs in indoor dust. The proposed method is based on the extraction of target analytes by microwave assisted extraction (MAE) with an eco-friendly water-ethanol (50:50, v/v) mixture, employing a MAE modified setup based on smaller containers that reduced both sample and solvent volumes and enabled up to 96 simultaneous extractions. The MAE extract was then derivatized using chloroformates, a strategy that yielded mass spectra with enhanced sensitivity and specificity compared to those obtained thorough conventional acetic anhydride derivatization. The resulting derivatives were subsequently isolated by solid-phase microextraction (SPME) and analyzed by gas chromatography-tandem mass spectrometry (GC-MS/MS). Key parameters for derivatization, MAE, and SPME were optimized by means of the multivariate approach of experimental design and factorial analysis. Method validation demonstrated satisfactory LLOQ values (0.5 ng/mL for mononitro compounds and 10 ng/mL for dinitro compounds), accuracy (ranging from 81 to 120 %), and precision (RSD% between 0.2 and 12.6 %). The application of the proposed method to real samples revealed, for the first time, the presence of 4-NP (0.059-0.116 µg/g), 4-NG (0.064-0.093 µg/g), 4-NC (0.014-0.156 µg/g), and 4-Me-5-NC (0.010-0.048 µg/g) in indoor dust, thereby underscoring the potential role of this matrix as a pathway for human exposure to these contaminants.

In this review, a 5-year overview on environmentally friendly approaches for the extraction of the most relevant organic pollutants in soil, sediment, particulate matter, and sewage sludge coupled with chromatographic analysis is reported. Organic contaminants encompass various compounds derived from personal care products, industrial chemicals, microplastics, organic matter combustion, agricultural practices, and plasticizer material. The principles of green analytical chemistry (GAC) and green sample preparation (GSP) serve as a guideline for the development of more environmentally sustainable analytical protocols. This study focuses attention on microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), matrix solid-phase dispersion (MSPD), and microextraction techniques, such as solid-phase microextraction (SPME), stir bar sorptive extraction (SBSE), hollow-fiber liquid-phase microextraction (HF-LPME), spray-assisted droplet formation-based liquid-phase microextraction (SADF-LPME), and dispersive liquid–liquid extraction (DLLME). These approaches represent the most relevant eco-friendly sample preparation for the advanced extraction of target analytes from environmental solid samples.

The recovery of lithium from extracts obtained from a black mass of spent lithium-ion batteries treated with a ternary solvent system at acidic pH was investigated using flat-sheet nanofiltration (NF) membranes operated according to a dead-end configuration. Specifically, four samples obtained at different pH values (2.5 and 5) and extraction times (48, 96 and 168 h) were treated in selected operating conditions by using two commercial polymeric membranes (denoted DK and HL, with an approximate molecular weight cut-off of 150–300 Da) up to a volume reduction factor (VRF) of 4. Membrane performance was assessed in terms of productivity and selectivity towards specific ions, including lithium. For most treated samples, the HL membrane exhibited higher permeate fluxes in comparison to the DK membrane. However, the DK membrane performed better in terms of lithium rejection than the HL membrane, with a negative rejection at VRF 4 observed for all treated samples. More than 90% of multivalent ions were rejected by both membranes independently of the VRF. The membrane ability to retain multivalent ions led to their progressive concentration in the retentate as the VRF increased. The extraction time did not impact the NF performance of both membranes in terms of ion rejection. For the DK membrane conditions of extraction of 96 h and pH 5 represented the best trade-off between flux, ion rejection, and total lithium recovery.

Tyre and road wear particles (TRWPs) are a significant source of micro- and nanoparticles contamination of aquatic environments. However, their occurrence and biological effects in terrestrial ecosystems remain poorly investigated, including their role as carriers of trace elements. This study, therefore, examines the accumulation of trace elements in Tenebrio molitor following exposure to TRWPs. Adults were fed organic wheat flour contaminated with 5 % and 10 % (w/w) TRWP powder, simulating concentration in soils near high-traffic areas. Chemical analyses of TRWPs and treated and control beetles at 10- and 20-days post-eclosion were performed using inductively coupled plasma-mass spectrometry (ICP-MS), supported by unsupervised pattern recognition techniques. A total of 36 trace elements, including heavy metals and rare earth elements (REEs), were identified as being associated with TRWPs. Element transfer from contaminated food to beetle tissues was observed, with accumulation of essential (Na, Mg, Zn, Ca, Ni), and non-essential (Ba, Al, V, Cr, Fe, As, Ga, Pb, Cd) elements and REEs (Gd, La, Nd, Pr, Sm, Y). Our findings suggest that TWRPs may pose a risk of contaminant transfer in the environment, potentially impacting food webs in ecosystems near high-traffic areas.

Lithium-ion batteries (LIBs) are central to sustainable energy technologies, and while lithium (Li) is the most recognized component, its rapid growth has also intensified demand for other critical metals, particularly manganese (Mn-(II)), nickel (Ni-(II)), and cobalt (Co-(II)). Recovering these valuable metal cations from spent LIBs and associated wastewater is essential to reduce production costs, conserve resources, and mitigate environmental risks. Here, we present mixed matrix membranes (MOF-PES MMMs) as versatile adsorbents for the simultaneous recovery of Ni-(II), Co-(II), and Mn-(II) from multicomponent solutions. Five metal-organic frameworks (MOFs)ZIF-8, MIL-53-(Al), UiO-66, and their amino-functionalized analogues (NH2-MIL-53-(Al) and NH2-UiO-66)were synthesized and incorporated into poly-(ether sulfone) (PES) membranes. The choice of MOF filler significantly influenced metal affinity, with amino functionalization enhancing adsorption relative to the parent structures. Among all formulations, ZIF-8-PES exhibited broad-spectrum performance, achieving >90% removal efficiency for all three metal ions simultaneously as well as excellent reusability for at least three cycles. These resultsachieved using oligomineral water with common interferons present in solutionhighlight MOF-PES MMMs as efficient, scalable platforms for multimetal recovery, offering a sustainable pathway for LIB recycling.

Bio-active ethylcellulose (EC) polymeric films have been obtained by incorporating curcumin (curc) and Ag(I)-based compounds, known for their antioxidant and antimicrobial activity, respectively, within the polymeric matrix. The recently reported Ag(I) coordination polymer, in both its structural forms (alpha-[(bpy)Ag(OTf)]infinity and beta-{[(bpy)Ag][OTf]}infinity), and the [(bpy)Ag(OTf)]infinity-curc polymeric co-crystal (bpy=2,2 '-bipyridine; OTf=trifluoromethanesulfonate) have been selected as Ag(I) species. The hybrid composite films have been prepared through the simple solvent casting method and characterized through Powder X-Ray Diffraction (PXRD), Differential Scanning Calorimetry (DSC) and Fourier Transform Infrared (FTIR) spectroscopy, Scanning Electron Microscope (SEM), UV-vis spectroscopy. The deep investigation of the film samples highlighted the non-inert behaviour of EC towards these specific active ingredients. Antimicrobial tests showed that EC films embedding the Ag(I)-based compounds present good antimicrobial performance, in particular against Staphylococcus aureus, used as a model of Gram-positive bacteria. In addition, Silver migration tests, performed on the Ag(I)-incorporating EC films, evidenced low values of silver release particularly in the case of the EC films incorporating [(bpy)Ag(OTf)]infinity-curc. Ethylcellulose films embedding the [(bpy)Ag(OTf)]infinity polymer and the binary co-crystal [(bpy)Ag(OTf)]infinity-curc, as active ingredients, have been prepared through solvent casting. The not innocent role of Ethylcellulose along the film forming process was highlighted by the deep investigation performed on the prepared samples. High antibacterial activity with low Silver release was found for the films incorporating the Ag(I) species. image

This work presents a novel method for the analysis of polycyclic aromatic hydrocarbons (PAHs) in saliva samples using solid phase microextraction (SPME) coupled with gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS). The protocol utilizes the latest commercially available overcoated fiber (PDMS/DVB/PDMS) for direct immersion extraction of the target analytes, enabling the determination of thirteen PAHs, including low-volatile compounds. The SPME extraction method was optimized using a central composite design (CCD). The evaluation of the fiber coating's robustness over time demonstrated excellent extraction performance with no significant degradation. The validation procedure confirmed good performance for all parameters, with LOQ values (100 ng/L for ten analytes and 500 ng/L for three analytes) comparable to other chromatographic methods. The environmental impact of the protocol was objectively assessed using two recently proposed metrics: the Green Analytical Procedure Index (GAPI) and the Analytical Greenness metric for sample preparation (AGREEprep). Both metrics indicated good overall environmental friendliness, with AGREEprep providing a satisfactory comprehensive score despite the use of highly impactful instrumentation. These characteristics make the developed method suitable for routine analysis in environmental and epidemiological monitoring.

Green energy transition has supposed to give a huge boost to the electric vehicle rechargeable battery market. This has generated a compelling demand for raw materials, such as cobalt and nickel, which are key common constituents in lithium-ion batteries (LIBs). However, their existing mining protocols and the concentrated localization of such ores have made cobalt and nickel mineral conundrums, and their supplies experience shortages, which threaten to slow the progress of the renewable energy transition. Aiming to contribute to the sustainable recycling of these valuable metals from LIBs and wastewater, in this work, we explore the use of four mixed matrix membranes (MMMs) embedding different metal-organic frameworks (MOFs), i.e., MIL-53(Al), MIL-53(Fe), MIL-101(Fe), and {(SrCu6II)-Cu-II[(S,S)-serimox](3)(OH)(2)(H2O)}39H(2)O (SrCu(6)Ser) in polyether sulfone (PES), for the recovery of cobalt(II) and nickel(II) metal cations from mixed cobalt-nickel aqueous solutions containing common interfering ions. Whereas the neat PES membrane slightly contributes to the adsorption of metal ions, showing reduced removal efficiency values of 10.2 and 9.5% for Ni(II) and Co(II), respectively, the inclusion of MOFs in the polymeric matrix substantially improves the adsorption performances. The four MOF@PES MMMs efficiently remove these metals from water, with MIL-53(Al)@PES being the one that presents better performance, with a removal efficiency up to 95% of Ni(II) and Co(II). Remarkably, SrCu(6)Ser@PES exhibits outstanding selectivity toward cobalt(II) cations compared to of nickel(II) ones, with removal efficiencies of 63.7 and 15.1% for Co(II) and Ni(II), respectively. Overall, the remarkable efficiencies, versatility, high environmental robustness, and cost-effective synthesis shown by this family of MOF@PES MMMs situate them among the best adsorbents for the extraction of this kind of contaminants.

Heavy metal ions are a common source of water pollution. In this study, two novel membranes with biobased metal-organic frameworks (BioMOFs) embedded in a polyacrylonitrile matrix with tailored porosity were prepared via nonsolvent induced phase separation methods and designed to efficiently adsorb heavy metal ions from oligomineral water. Under optimized preparation conditions, stable membranes with high MOF loading up to 50 wt % and a cocontinuous sponge-like morphology and a high water permeability of 50-60 L m-2 h-1 bar-1 were obtained. The tortuous flow path in combination with a low water flow rate guarantees maximum contact time between the fluid and the MOFs, and thus a high heavy metal capture efficiency in a single pass. The performances of these BioMOF@PAN membranes were investigated in the dynamic regime for the simultaneous removal of Pb2+, Cd2+, and Hg2+ heavy metals from aqueous environments in the presence of common interfering ions. The new composite adsorbing membranes are capable of reducing the concentration of heavy metal pollutants in a single pass and at much higher efficiency than previously reported membranes. The enhanced performance of the mixed matrix membranes is attributed to the presence of multiple recognition sites which densely decorate the BioMOF channels: (i) the thioether groups, deriving from the S-methyl-l-cysteine and (S)-methionine amino acid residues, able to recognize and capture Pb2+ and Hg2+ ions and (ii) the oxygen atoms of the oxamate moieties, which preferentially interact with Cd2+ ions, as revealed by single crystal X-ray diffraction. The flexibility of the pore environments allows these sites to work synergically for the simultaneous capture of different metal ions. The stability of the membranes for a potential regeneration process, a key-factor for the effective feasibility of the process in real life applications, was also evaluated and confirmed less than 1% capacity loss in each cycle.

A new structural form of the already reported silver based 1D coordination polymer (CP) [(bpy)Ag (OTf)](8) (bpy = 2,2'-bipyridine; OTf = trifluoromethanesulfonate) has been synthesized and fully characterized through single-crystal and powder XRD, DSC, and FTIR analysis. The reaction of both structural forms of the Ag(I) CP with curcumin (curc) afforded the formation of a new solid form, specifically an inorganic polymeric co-crystal between curc and [(bpy)Ag (OTf)] in a 1: 4 molar ratio, respectively. The co-crystal has been characterized both in solution and in the solid state through H-1 NMR, PXRD, DSC, IR, and UV-vis spectroscopies. In addition, a thorough computational study has been carried out to reveal the possible interaction modes between the co-crystal components. The Ag(I) compounds here presented have been preliminarily tested for antimicrobial activity against Staphylococcus aureus and Escherichia coli strains, all showing promising results.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

The herein presented work aims to the development of an easy method for the quantitative determination of parabens and bisphenols in human salivabased on the use of methyl chloroformate as a derivatizing agent, followed by solid-phase microextraction (SPME) and gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) analysis with selected reaction monitoring (SRM). Using multivariate analysis, two derivatization strategies were compared and optimized, demonstrating that the use of methyl chloroformate led to better sensitivity than the classical derivatization by acetic anhydride. Good performance in the sorption process of the derivatized target analytes was obtained using the most recent commercialized overcoated fiber (PDMS/DVB/ PDMS). The validation procedure of the final protocol led to satisfactory results in terms of linearity, limit of quantitation, accuracy, and precision. All parabens were quantified from 10 ng/L using the developed method, except for methylparaben, which was quantified from 100 ng/L along with all bisphenols. Intra-and inter-day accuracy and intra-and inter-day precision can be considered satisfactory for all analytes (values between 73% and 118%), except for the inter-day accuracy of BPF. Quite good results also in terms of matrix effect were obtained for the target compounds (range 71% to 118%, RSD% less than 13.6%), except for BPA at the middle concentration and MeP at the lowest concentration. The greenness of the method was evaluated and the results indicated that our approach is more eco-friendly than previously published methods. Based on its characteristics, the presented method can be considered a suitable approach to determine parabens and bisphenols in routine analysis for biomonitoring purposes.

The increasing use of agrochemicals, including fertilizers and herbicides, has led to worrying metal contamination of soils and waters and raises serious questions about the effects of their transfer to different levels of the trophic web. Accumulation and biomagnification of essential (K, Na, Mg, Zn, Ca), nonessential (Sr, Hg, Rb, Ba, Se, Cd, Cr, Pb, As), and rare earth elements (REEs) were investigated in newly emerged adults of Tenebrio molitor exposed to field-admitted concentrations of a metribuzin-based herbicide and an NPK blend fertilizer. Chemical analyses were performed using inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) supported by unsupervised pattern recognition techniques. Physiological parameters such as cuticle melanization, cellular (circulating hemocytes), and humoral (phenoloxidase enzyme activity) immune responses and mass loss were tested as exposure markers in both sexes. The results showed that NPK fertilizer application is the main cause of REE accumulation in beetles over time, besides toxic elements (Sr, Hg, Cr, Rb, Ba, Ni, Al, V, U) also present in the herbicide-treated beetles. The biomagnification of Cu and Zn suggested a high potential for food web transfer in agroecosystems. Gender differences in element concentrations suggested that males and females differ in element uptake and excretion. Differences in phenotypic traits show that exposure affects metabolic pathways involving sequestration and detoxification during the transition phase from immature-to-mature beetles, triggering a redistribution of resources between sexual maturation and immune responses. Our findings highlight the importance of setting limits for metals and REEs in herbicides and fertilizers to avoid adverse effects on species that provide ecosystem services and contribute to soil health in agroecosystems.

This work proposes a new method for the quantification of benzothiazoles (BTs), benzotriazoles (BTRs), and benzenesulfonamides (BSAs) in tap water, river water, and wastewater. The protocol involved the use of microextraction by packed sorbent (MEPS), applied for the first time for the extraction of the target analytes, combined with programmed temperature vaporization-gas chromatography-triple quadrupole mass spectrometry (PTV-GC-QqQ-MS). Considering the synergism between MEPS extraction and PTV injection, the experimental variables affecting their performance were simultaneously optimized by "experimental design", while principal component analysis (PCA) was used to find the overall optimal working conditions. Response surface methodology was used to gain a comprehensive understanding of the effects of working variables on method performance. The developed method achieved very good linearities and satisfactory intra- and inter-day accuracies and precisions. The protocol permitted the detection of the target molecules with limit of detection (LODs) values between 0.005 and 0.85 μg/L. The green character of the procedure was evaluated using three metrics: "Analytical Eco-Scale", "Green Analytical Procedure Index" (GAPI), and "Analytical Greenness metric for sample preparation (AGREEprep). The satisfactory results obtained with real water samples demonstrate the applicability of the method for monitoring campaigns and exposome studies.

Heavy metal contamination is recognized worldwide as a serious threat to human health and wildlife, and reducing their emissions is a priority of international and EU actions. Due to its persistence, high bioaccumulation tendency, and toxicity properties, lead (Pb) is one of the heavy metals of greatest concern. Even at low concentrations, lead induces various clinical and subclinical conditions in both humans and animals, and it has been included in the priority list of hazardous substances. In the present study, we used zebrafish's early stages as a model, given their well-acknowledged predictive value in the risk assessment of chemicals. This study was designed to investigate the morphological and morphometric alterations induced by Pb during zebrafish's early development and disclose the putative effects stage- and/or dose-dependent. We examined injuries induced by two environmentally relevant and extremely low concentrations of Pb (2.5 μg/L and 5 μg/L) during two exposure windows: early (between 1 and 7 dpf) and late (between 2 and 8 dpf). We clearly demonstrated that the incidence and severity of morphological abnormalities increased with increasing Pb dose and exposure time in both early and late-exposed groups. Furthermore, we revealed that malformation severity was significantly higher in the early exposed group than in the late exposure group at all exposure times and for both tested doses, thus highlighting the high sensitivity of zebrafish during the initial stages of development. The information presented in this paper emphasizes the effectiveness of morphological biomarkers in unveiling threatening situations and supports the role of zebrafish embryos and larvae in risk assessment and environmental monitoring.

Lead (Pb), due to its high toxicity and bioaccumulation tendency, is one of the top three pollutants of concern for both humans and wildlife and occupies second place in the Priority List of Hazardous Substances. In freshwater fish, Pb is mainly absorbed through the gills, where the greatest accumulation occurs. Despite the crucial role of gills in several physiological functions such as gas exchange, water balance, and osmoregulation, no studies evaluated the effects of environmentally relevant concentrations of Pb on this organ, and existing literature only refers to high levels of exposure. Herein we investigated for the first time the molecular and morphological effects induced by two low and environmentally relevant concentrations of Pb (2.5 and 5 μg/L) on the gills of Danio rerio, a model species with a high translational value for human toxicity. It was demonstrated that Pb administration at even low doses induces osmoregulatory dysfunctions by affecting Na+/K+-ATPase and AQP3 expression. It was also shown that Pb upregulates MTs as a protective response to prevent cell damage. Modulation of SOD confirms that the production of reactive oxygen species is an important toxicity mechanism of Pb. Histological and morphometric analysis revealed conspicuous pathological changes, both dose- and time-dependent.

Pendimethalin-based herbicides are used worldwide for pre-emergence selective control of annual grasses and weeds in croplands. The endurance of herbicides residues in the environment has an impact on the soil biodiversity and fertility, also affecting non-target species, including terrestrial invertebrates. Carabid beetles are known as natural pest control agents in the soil food web of agroecosystems, and feed on invertebrates and weed seeds. Here, a mass spectrometry untargeted profiling of haemolymph is used to investigate Pterostichus melas metabolic response after to pendimethalin-based herbicide exposure. Mass spectrometric data are examined with statistical approaches, such as principal component analysis, for possible correlation with biological effects. Those signals with high correlation are submitted to tandem mass spectrometry to identify the associated biomarker. The time course exposure showed many interesting findings, including a significant downregulation of related to immune and defense peptides (M-lycotoxin-Ls4a, Peptide hormone 1, Paralytic peptide 2, and Serine protease inhibitor 2). Overall, the observed peptide deregulations concur with the general mechanism of uptake and elimination of toxicants reported for Arthropods.