Suspect screening of wastewaters to trace anti-COVID-19 drugs: Potential adverse effects on aquatic environment.

During the first period of the SARS-CoV-2 pandemic, the lack of specific therapeutic treatments led to the provisional use of a number of drugs, with a continuous review of health protocols when new scientific evidence emerged. The management of this emergency sanitary situation could not take care of the possible indirect adverse effects on the environment, such as the release of a large amount of pharmaceuticals from wastewater treatment plants. The massive use of drugs, which were never used so widely until then, implied new risks for the aquatic environment. In this study, a suspect screening approach using Liquid Chromatography-High Resolution Mass Spectrometry techniques, allowed us to survey the presence of pharmaceuticals used for COVID-19 treatment in three WWTPs of Lombardy region, where the first European cluster of SARS-CoV-2 cases was detected. Starting from a list of sixty-three suspect compounds used against COVID-19 (including some metabolites and transformation products), six compounds were fully identified and monitored together with other target analytes, mainly pharmaceuticals of common use. A monthly monitoring campaign was conducted in a WWTP from April to December 2020 and the temporal trends of some anti-COVID-19 drugs were positively correlated with those of COVID-19 cases and deaths. The comparison of the average emission loads among the three WWTPs evidenced that the highest loads of hydroxychloroquine, azithromycin and ciprofloxacin were measured in the WWTP which received the sewages from a hospital specializing in the treatment of COVID-19 patients. The monitoring of the receiving water bodies evidenced the presence of eight compounds of high ecological concern, whose risk was assessed in terms of toxicity and the possibility of inducing antibiotic and viral resistance. The results clearly showed that the enhanced, but not completely justified, use of ciprofloxacin and azithromycin represented a risk for antibiotic resistance in the aquatic ecosystems.

Identification and characterisation of thiolated polysulfides in must and wine using online SPE UHPLC-HRMS.

3-Mercaptohexanol and 4-methyl-4-mercaptopentanone are volatile thiols with a low perception threshold and are found in relatively low concentrations in several types of wines, e.g. Sauvignon blanc. They contribute positively to the flavour of a wine when in their free form, but they can oxidise, especially in the presence of Cu2+ and sulfur residues originating from pesticide treatments on the grapes. This condensation reaction results in the formation of polysulfides, which during storage can cause the release of H2S: a compound known to give rise to off-flavour in wine. The formation of these polysulfides has been proposed to originate from cysteinyl and glutathionyl S-conjugate precursors, but they have not yet been characterised. In this work, a method using ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry and online solid-phase extraction was designed and optimised for detection of these S-conjugates. The method allowed the detection of 21 suspected symmetrical and asymmetrical S-conjugates in thiol-supplemented aqueous solutions, of which 17 were also recovered in supplemented synthetic musts and wines. Moreover, the proposed method was used to investigate polysulfide formation upon addition of two different types of sulfur. Differences in formation of S-conjugates were evident in the synthetic samples: a higher relative abundance was observed upon addition of wettable sulfur compared to washed sulfur. For the commercial wines, differences in polysulfide formation were minor and merely related to the differences between the wines and not to the type of sulfur added.

Urinary metabolomics reveals kynurenine pathway perturbation in newborns with transposition of great arteries after surgical repair.

INTRODUCTION: Transposition of the great arteries (TGA) is a cyanotic congenital heart defect that requires surgical correction, with the use of cardiopulmonary-bypass (CPB), usually within 3 weeks of life. The use of CPB in open heart surgery results in brain hypoperfusion and in a powerful systemic inflammatory response and oxidative stress. OBJECTIVE: We aimed to develop a novel untargeted metabolomics approach to detect early postoperative changes in metabolic profile following neonatal cardiac surgery. METHODS: We studied 14 TGA newborns with intact ventricular septum undergoing arterial switch operation with the use of CPB. Urine samples were collected preoperatively and at the end of the surgery and were analyzed using an untargeted metabolomics approach based on UHPLC-high resolution mass spectrometry. RESULTS: Since post surgery metabolic spectra were heavily contaminated by metabolites derived from administered drugs, we constructed a list of drugs used during surgery and their related metabolites retrieved from urine samples. This library was applied to our samples and 1255 drugs and drug metabolites were excluded from the analysis. Afterward, we detected over 39,000 unique compounds and 371 putatively annotated metabolites were different between pre and post-surgery samples. Among these metabolites, 13 were correctly annotated or identified. Metabolites linked to kynurenine pathway of tryptophan degradation displayed the highest fold change. CONCLUSIONS: This is the first report on metabolic response to cardiac surgery in TGA newborns. We developed an experimental design that allowed the identification of perturbed metabolic pathways and potential biomarkers of brain damage, limiting drugs interference in the analysis.

Metals in Mediterranean aquatic species.

Metals such as arsenic (As), cadmium (Cd), copper (Cu), chromium (Cr), nickel (Ni), lead (Pb), vanadium (V), have been determined in species of Mediterranean marine organisms collected from areas supposed to be at background contamination levels. The Inductively Coupled Plasma Mass Spectrometry (ICP-MS) approach was adopted for the determination of all the metals. Arsenic, Cd and Pb determined in the 42 samples, do not exceed the pertinent maximum level except a sample of hake. In wild fish, the concentration range for Cr, Ni, V and Cu was, respectively: 0.07-0.09, 87.6-124, 0.022-0.075 and 0.79-1.74 µg/g fresh weight (fw). The farmed fish samples show concentration levels below the wild fish ones, except for Cr which range at the same levels. Cadmium and Pb show a high sample number under the quantification limit. The elements do not bio-magnify among the species considered and appear to show low variations in relation to organisms' position in the food chain and at sampling sites.

Persistent toxic substances in Mediterranean aquatic species.

Fish and fishery products may represent one of the main sources of dietary exposure to persistent toxic substances (PTSs) such as polychlorinated dibenzodioxins, dibenzofurans, and biphenyls; polybromodiphenyl ethers; organochlorine pesticides; perfluorooctanoic acid and perfluorooctane sulfonate; and inorganic mercury and methyl mercury. In this study, PTS contamination of Mediterranean fish and crustaceans caught in Italian coastal waters was investigated in order to increase the representativeness of the occurrence database for wild species. The objectives were to verify the suitability of regulatory limits for PTSs, identify background concentrations values, if any, and examine the possible sources of variability when assessing the chemical body burdens of aquatic species. Twelve wild species of commercial interest and two farmed fish species were chosen. Excluding methyl mercury, chemical concentrations found in wild species fell generally towards the low ends of the concentration ranges found in Europe according to EFSA database and were quite lower than the tolerable maximum levels established in the European Union; farmed fish always showed contamination levels quite lower than those detected in wild species. The data obtained for wild species seemed to confirm the absence of local sources of contamination in the chosen sampling areas; however, species contamination could exceed regulatory levels even in the absence of specific local sources of contamination as a result of the position in the food web and natural variability in species' lifestyle. A species-specific approach to the management of contamination in aquatic organisms is therefore suggested as an alternative to a general approach based only on contaminant body burden. A chemical-specific analysis performed according to organism position in the food chain strengthened the need to develop this approach.

Artificial-turf playing fields: contents of metals, PAHs, PCBs, PCDDs and PCDFs, inhalation exposure to PAHs and related preliminary risk assessment.

The artificial-turf granulates made from recycled rubber waste are of health concern due the possible exposure of users to dangerous substances present in the rubber, and especially to PAHs. In this work, we determined the contents of PAHs, metals, non-dioxin-like PCBs (NDL-PCBs), PCDDs and PCDFs in granulates, and PAH concentrations in air during the use of the field. The purposes were to identify some potential chemical risks and to roughly assess the risk associated with inhalation exposure to PAHs. Rubber granulates were collected from 13 Italian fields and analysed for 25 metals and nine PAHs. One further granulate was analysed for NDL-PCBs, PCDDs, PCDFs and 13 PAHs. Air samples were collected on filter at two fields, using respectively a high volume static sampler close to the athletes and personal samplers worn by the athletes, and at background locations outside the fields. In the absence of specific quality standards, we evaluated the measured contents with respect to the Italian standards for soils to be reclaimed as green areas. Zn concentrations (1 to 19 g/kg) and BaP concentrations (0.02 to 11 mg/kg) in granulates largely exceeded the pertinent standards, up to two orders of magnitude. No association between the origin of the recycled rubber and the contents of PAHs and metals was observed. The sums of NDL-PCBs and WHO-TE PCDDs+PCDFs were, respectively, 0.18 and 0.67×10(-5) mg/kg. The increased BaP concentrations in air, due to the use of the field, varied approximately from <0.01 to 0.4 ng/m(3), the latter referring to worst-case conditions as to the release of particle-bound PAHs. Based on the 0.4 ng/m(3) concentration, an excess lifetime cancer risk of 1×10(-6) was calculated for an intense 30-year activity.

Pentachlorophenol, polychlorodibenzodioxin and polychlorodibenzofuran in eggs from hens exposed to contaminated wood shavings.

Laying hens may be exposed to pentachlorophenol (PCP) present in bedding materials derived from treated timber. As a result, this chemical and its contaminants or degradation products, such as polychlorodibenzodioxins (PCDDs) and polychlorodibenzofurans (PCDFs), may be present in eggs. The litter-to-eggs transfer and depletion of these compounds were studied in a flock of laying hens reared on contaminated wood shavings. PCP determination was carried out via high resolution gas chromatography coupled to low resolution mass spectrometry/mass spectrometry (HRGC-LRMS/MS); PCDDs and PCDFs were quantified by HRGC-HRMS (SIM). After substitution of the litter contaminated with PCDDs and PCDFs at an average of 50 pg WHO-TE g(-1) and with PCP at 15 microg g(-1), pooled eggs from six selected hens were sampled twice a month for the depletion study. At steady state, PCDDs and PCDFs showed a transfer ratio of 0.9; for PCP it was 0.03. PCP concentration in eggs (500 ng g(-1) whole weight) fell sharply the second week after exposure withdrawal; for PCDDs and PCDFs (47 pg WHO-TE g(-1) fat, at day 0), the overall TEQ half-life was estimated at an average of 3.8 weeks. Due to differences in toxicokinetics (a faster depletion), PCP does not appear to be a suitable marker of PCDD and PCDF violative levels. However, the prominent analytical contribution of H(7)CCD and O(8)CCD in the contamination profile may help to trace the source of contamination. Among congeners, 2,3,7,8 T(4)CDF exhibited a different depletion pattern, indicating a possible mechanism of active transport.

Levels and profiles of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and polychlorinated biphenyls in feedstuffs and milk from farms in the vicinity of incineration plants in Tuscany, Italy.

Levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorobiphenyls (PCBs) were determined in samples of bovine and ovine milk collected in farms located in the province of Grosseto, Tuscany, in the vicinity of two incineration plants as well as in farms located in areas with presumable background levels of contamination. Samples of feedstuffs of local origin used in the investigated farms were also collected and analysed. The cumulative levels of PCDDs, PCDFs, and dioxin-like PCBs (DL-PCBs) in feedstuffs ranged from 0.25 to 0.61 pg WHO-TE/g fresh weight (fw) in the farms under impact from incinerator emissions and from 0.21 to 0.34 pg WHO-TE/g fw in the control area farms. The sums of the six non-dioxin-like indicator PCB (NDL-PCB) congeners were 0.13 to 9.3 ng/g fw and 1.2 to 1.9 ng/g fw, respectively. In milk samples, the levels detected were 0.71 to 2.9 pg WHO-TE/g fat and 0.52 to 0.59 pg WHO-TE/g fat in farms under impact from the incinerators and in the control area farms, respectively. The corresponding sums of the six indicator PCB congeners were 1.4 to 8.2 ng/g fat and 0.90 to 1.6 ng/g fat. In all samples, contamination levels were below the limits set by the European Community (EC; Commission Directive 2006/13/EC and Commission Regulation 1881/2006/EC.) No relevant differences were found between samples collected in potentially exposed areas and control areas for total toxic equivalents and cumulative analytic levels of PCDDs, PCDFs, DL-PCBs, and NDL-PCBs (p > 0.05, Mann-Whitney U test). Observed levels were in agreement with those found in other countries in areas with background levels of exposure. Congener levels and profiles of PCDDs, PCDFs, and PCBs in feedstuffs and milk samples that were compared describe congener-specific transfer behavior.

PCDD and PCDF depletion in milk from dairy cows according to the herd metabolic scenario.

High level of PCDD+PCDF contamination in bulk milk (9.7 pg WHO-TE g(-1) fat) from 1604 Holstein Fresian lactacting cows was observed just four weeks after the beginning of their exposure to a feed supplement contaminated at 10.4 ng WHO-TE kg(-1) dry matter. In-farm produced hay and silage showed levels not exceeding 0.2 ng WHO-TE kg(-1) dry matter. After the supplement withdrawal, it was possible to monitor the depletion phase for a following 75-day period in milk, until the levels dropped well below 3.0 pg WHO-TE g(-1) fat, the EU regulatory Maximum Residue Level for PCDD+PCDF. During this phase, the half-life was calculated as 17+/-3 days, on WHO-TEQ basis. The full availability of farm data on both cow nutrition and milk production allowed the calculation of the carry-over rate (COR) (PCDD+PCDF milk excretion vs. feed), which was 46% at the end of the exposure. This COR value is justified from the main TE contribution of Penta-CDD and -CDF congeners (63%), and the half-life is among the shortest of all those described in the literature both for experimental and naturally-exposed dairy cows. A fugacity-based model predicts a bulk milk contamination of 5 pg WHO-TE g(-1) fat, compared to the 10 pg WHO-TE g(-1) fat level observed. Such findings are discussed in light of the lactation and metabolic status of the herd for which the transition period, characterised by a negative metabolic energy balance and a consequent adipose tissue mobilization, could play a relevant role.

Modeling of DR CALUX bioassay response to screen PCDDs, PCDFs, and dioxin-like PCBs in farm milk from dairy herds.

A recent issue in the EU legislation is the evaluation of the toxicologically-equivalent contribution of dioxin-like polychlorobiphenyls (DL-PCBs) in addition to that coming from polychlorodibenzodioxins (PCDDs) and polychlorodibenzofurans (PCDFs) as contaminants in foods for a total of 29 congeners. This fact is determining the need to revise analytical criteria both for confirmatory and screening analysis. In this work, a modeling was developed to check the reliability of the outcomes of the DR CALUX bioassay when applied to farm milk samples characterized by large differences in congener patterns. To reproduce some field conditions where DL-PCB contributions up to 90% of total WHO-TEQs (HRGC-HRMS assessment) were recorded in dairy products, goat milk samples from a common bulk were fortified at different TEQ levels with mixtures containing either PCDDs and PCDFs or non-ortho substituted DL-PCBs. Fortification ranged approximately 4.5-15 pgWHO-TEQ/g fat. Based on the results, DR CALUX relative potency value (REP) of DL-PCB 126 was estimated 0.061 against the canonical WHO-TEF of 0.1. The value of 0.061 together with the other DR CALUX REPs from the literature for the remaining 28 congeners were used to model DR CALUX response (C-TEQs) in milk samples with different congener patterns. The theoretical underestimation of DR CALUX data could be mitigated by correcting the latter with the linear correlation experimentally obtained between C-TEQs and the WHO-TEQs. Under these conditions, the use as calibrants of reference samples with different analytical patterns could help those laboratories involved in a high throughput routine to set the most appropriate decision limits to optimize screening output.

Synthesis and characterization of a dehydrogenation product arising from the oxidation of aminoethylcysteine ketimine decarboxylated dimer.

While investigating the antioxidant properties of aminoethylcysteine ketimine decarboxylated dimer (1) (a natural substance occurring in biological fluids such as human urine and plasma and in bovine cerebellum), a previously unreported oxidation product was obtained. This compound was identified and characterized through comparison with an authentic sample prepared via Pd-catalyzed dehydrogenation of 1. This molecule is an example of an alternative oxidation pathway involving 1.

Depletion of selected polychlorinated biphenyl, dibenzodioxin, and dibenzofuran congeners in farmed rainbow trout (Oncorhynchus mykiss): a hint for safer fish farming.

Farmed fish can be exposed to persistent organic contaminants--such as polychlorinated biphenyls (PCBs), dibenzodioxins (PCDDs), and dibenzofurans (PCDFs)--via feed, this eventually resulting in accumulation levels of health concern. To study the correlation between feed contamination, chemical accumulation in fish muscle (fillet), and chemical depletion, an all-vegetal base (or blank) feed was prepared and fortified with a commercial PCB mixture (Aroclor 1254) and six PCDD and PCDF congeners (namely, 2,3,7,8-T(4)CDD, 2,3,7,8-T(4)CDF, 1,2,3,7,8-P(5)CDD, 1,2,3,7,8-P(5)CDF, O(8)CDD, and O(8)CDF) to reproduce realistic low, medium, and high contamination levels. After a 1-month exposure, trout (Oncorhynchus mykiss) were fed with the blank feed and sacrificed every 0.5 months over a 3-month period from exposure end; fillet specimens were sampled at each time. In all groups, the average fish weight increased linearly through the observation period. The chemical diminishing patterns observed were due to the combined effect of clearance and growth dilution: for 10 PCB and four PCDD and PCDF congeners, patterns were described with an empirical one-compartment (fish muscle) model. The canonical pseudo-first-order kinetic equation used was also modified into the form C=[C(0)exp(-k(C)t)] (m(W)t+1)(-1) to distinguish between the contributions to depletion from clearance, exp(-k(C)t), and growth dilution, (m(W)t+1)(-1). Most mean clearance half-life (HL(C)) estimates appear to be greater than 4 months, in a number of cases reaching magnitudes well over 10 months or even negative, thus clearly indicating a non-negligible contribution from a second compartment. Based on means and their 95% confidence intervals, the depletion HL(D) estimates of the 14 selected congeners seem to be comprised between 1.2-3.4 and 1.0-5.0 months, respectively: these values, accounting for both clearance and growth dilution, provide an indication of the relevance of a blank feed as a management option to reduce the overall PCB, PCDD, and PCDF content in farmed trout. Due to a lack of bioaccumulation, O(8)CDD and O(8)CDF yielded no results for evaluation, whereas for many PCB congeners results were insufficient for empirical modelling.

Persistent and toxic substances in the Venice lagoon biota: an approach for quantitative data analysis for risk management.

An approach based on hypothesis testing for the management of persistent inorganic and organic toxic chemicals (PTS/POPs) detected in clams and mussels from the Venice lagoon is presented. The chemicals of interest for this evaluation were the polychlorinated biphenyls (PCBs), dibenzodioxins (PCDDs), and dibenzofurans (PCDFs), hexachlorobenzene (HCB), and the heavy metals cadmium (Cd), mercury (Hg), and lead (Pb). Two statistically different populations of data for PCDDs+PCDFs (TEQs), HCB, Cd, and Pb, associated with biota samples collected respectively in the lagoon central district and in the southern and northern districts were identified. The central district is under the impact of the Porto Marghera industrial settlement and the city of Venice, whereas the southern and northern districts are subject to a general impact. Of the aforementioned chemicals, those with more discriminating power were found to be HCB and PCDDs+PCDFs.

Determination of surfactants and some of their metabolites in untreated and anaerobically digested sewage sludge by subcritical water extraction followed by liquid chromatography-mass spectrometry.

Enormous amounts of sewage sludge are worldwide generated and released into the environment. Analysis of the most common and/or toxic chemicals in sludge should be mandatory before deciding its destination. Surfactants and some of their breakdown products are invariably the most common organic contaminants in domestic sewage sludge. For determining these compounds, we have developed a method based on extraction with subcritical water followed by liquid chromatography-mass spectrometry. On extracting surfactants and their metabolites from 50 mg of sludge, the efficiency of the water extraction device was evaluated in terms of pH of the extractant, temperature, and time of the static extraction. The best extraction conditions were obtained by using carbonate buffer (pH 9.4) at 200 degrees C as extractant, 10 min of static extraction at the pressure of 100 bar followed by 17 min of dynamic extraction. Analyte collection was performed by inserting a solid-phase extraction cartridge downstream the extraction cell. Compared to 16-h Soxhlet extraction with methanol, this procedure was remarkably more efficient in extracting anionic surfactants and acidic metabolites of nonylphenol ethoxylates (NPECs). A short survey was conducted to estimate concentration changes of target compounds after 14-d sludge anaerobic digestion. Results showed that 54-74% of both neutral and weakly acidic ethoxylate species were removed after residence of the sludge in the digester. On the contrary, little, if any, removal of anionic surfactants was observed after the digestion treatment. As expected, the level of nonylphenol increased under anaerobic conditions.