Predictors of deviation in neurovascular bundle preservation during robotic prostatectomy.

INTRODUCTION: Neurovascular bundle (NVB) preservation during robot-assisted radical prostatectomy (RARP) directly affects patient functional outcomes. Despite careful surgical planning, many NVB preservation techniques are changed intraoperatively from their preoperative plan. Our objective was to identify risk factors predicting intraoperative change in NVB preservation technique during RARP. MATERIALS AND METHODS: Prospective data from 578 RARPs performed by a single surgeon between 2010 and 2017 at a tertiary care center. Side-specific NVB preservation technique was planned preoperatively. Surgical techniques were either complete nerve sparing (CNS), or incomplete nerve sparing (INS). Variables included age, tumor grade, prostate volume, number of lifetime biopsies, history of post-biopsy sepsis, and laterality. Variables were modeled in multivariable logistic regressions as potential predictors of deviation in surgical technique. Functional and oncological outcomes were also assessed. RESULTS: A total of 46.9% of cases underwent some intraoperative change in NVB preservation from their preoperative plan. A total of 37.7% of 880 prostate sides planned for CNS underwent unplanned INS. Older age, Gleason ≥ 3+4, post-biopsy sepsis, prostate volume, and left-sided dissections were significantly associated with unplanned INS. Number of lifetime biopsies was not a predictor of unplanned INS. Patients with an intraoperative change to INS had poorer potency and continence. Study limitations included the retrospective nature of analysis and lack of pathological assessment of NVB preservation. CONCLUSIONS: Age, Gleason ≥ 3+4, post-biopsy sepsis, prostate volume, and laterality were significant predictors of unplanned INS during RARP, which should guide patient counseling when discussing risks and functional outcomes. The number of lifetime biopsies did not predict unplanned INS, a valuable finding for patients on active surveillance. Our findings highlight the importance of careful preoperative planning and novel adjuncts such as multiparametric MRI.

The Effect of Intramural Myomas Without an Intracavity Component on In Vitro Fertilization Outcomes in Single Fresh Blastocyst Transfer Cycles.

STUDY OBJECTIVE: To assess clinical pregnancy rate (CPR) and live birth rate (LBR) in the presence of non-cavity-deforming intramural myomas in single fresh blastocyst transfer cycles. DESIGN: Retrospective cohort study (Canadian Task Force classification II-2). SETTING: Academic fertility center. PATIENTS: A total of 929 fresh single blastocyst transfer cycles were included, 94 with only non-cavity-distorting intramural myomas and 764 without myomas. Cleavage embryo transfers were excluded to reduce bias based on embryo quality. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: CPR and LBR were assessed. There were no differences noted in gravidity, parity, or body mass index between patients with myomas and those without myomas. Women with myomas required higher doses of gonadotropins (mean, 2653 ± 404 IU vs 2350 ± 1368 IU; p = .04) than women without myomas. However, the total number of mature oocytes collected and the total number of blastocysts created were similar. CPR (47% vs 32%; p = .005) and LBR (37.8% vs 25.5%; p = .02) were lower in patients who had intramural myomas compared with those without myomas. CPR and LBR were significantly reduced in the presence of even 1 myoma (odds ratio [OR], 0.53; 95% confidence interval [CI], 0.33-0.83 and OR, 0.56; 95% CI, 0.35-0.92, respectively). In patients with myomas >1.5 cm, LBR was also significantly reduced, even after adjusting for age, smoking, quality of embryo transferred, antral follicle count, and dose of gonadotropins (OR, 0.53; 95% CI, 0.29-0.97). This LBR finding was not significant if all myomas were included (including those <1.5 cm in diameter), but CPR was still significantly reduced. CONCLUSION: Relatively small (>1.5 cm) non-cavity-distorting intramural myomas negatively affect CPR and LBR in in vitro fertilization cycles, even in the presence of only 1 myoma.

Do trained reproductive endocrinologists perform better than their trainees? Comparing clinical pregnancy rates and live birth rates after transfer of single fresh blastocysts.

PURPOSE: To compare clinical pregnancy rates and live birth rates of single blastocyst transfers performed by attending physicians or fellows in reproductive endocrinology and infertility program. METHODS: Retrospective study in an academic reproductive center. We evaluated 932 fresh single blastocyst transfer cycles performed by fellows in training (389 embryo transfers) and by attending physicians (543 embryo transfers). RESULTS: There were no differences in the baseline characteristics and IVF cycle parameters between patients who had transfers performed by fellows or attending physicians. Transfers performed by attending physicians or fellows resulted in similar CPR (46.5 vs. 42.9%, p = 0.28) and LBR (38.3 vs. 34.2%, p = 0.11). Multivariate logistic regression analysis showed that even after adjusting for possible confounders (age, gravity, parity, baseline FSH, antral follicle count, dose of gonadotropins, stimulation protocol, and quality of embryo transferred), CPR (OR 0.81, CI 0.62-1.07) and LBR (OR 0.79, CI 0.6-1.05) in the two groups were comparable. CONCLUSION: Clinical pregnancy rate and live birth rate after embryo transfer performed by attending staffs or fellows are comparable. This finding reassures fellowship programs that allowing fellows to perform embryo transfers does not compromise the outcome.

Evaluating the reliability of environmental concentration data to characterize exposure in environmental risk assessments.

Environmental risk assessments often rely on measured concentrations in environmental matrices to characterize exposure of the population of interest-typically, humans, aquatic biota, or other wildlife. Yet, there is limited guidance available on how to report and evaluate exposure datasets for reliability and relevance, despite their importance to regulatory decision-making. This paper is the second of a four-paper series detailing the outcomes of a Society of Environmental Toxicology and Chemistry Technical Workshop that has developed Criteria for Reporting and Evaluating Exposure Datasets (CREED). It presents specific criteria to systematically evaluate the reliability of environmental exposure datasets. These criteria can help risk assessors understand and characterize uncertainties when existing data are used in various types of assessments and can serve as guidance on best practice for the reporting of data for data generators (to maximize utility of their datasets). Although most reliability criteria are universal, some practices may need to be evaluated considering the purpose of the assessment. Reliability refers to the inherent quality of the dataset and evaluation criteria address the identification of analytes, study sites, environmental matrices, sampling dates, sample collection methods, analytical method performance, data handling or aggregation, treatment of censored data, and generation of summary statistics. Each criterion is evaluated as "fully met," "partly met," "not met or inappropriate," "not reported," or "not applicable" for the dataset being reviewed. The evaluation concludes with a scheme for scoring the dataset as reliable with or without restrictions, not reliable, or not assignable, and is demonstrated with three case studies representing both organic and inorganic constituents, and different study designs and assessment purposes. Reliability evaluation can be used in conjunction with relevance evaluation (assessed separately) to determine the extent to which environmental monitoring datasets are "fit for purpose," that is, suitable for use in various types of assessments. Integr Environ Assess Manag 2024;20:981-1003. © 2024 Society of Environmental Toxicology & Chemistry (SETAC). This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Determination of perfluorinated compounds in aquatic organisms: a review.

Bioaccumulation of PFAS in aquatic organisms is an environmental problem of growing concern around the world. This problem has been tackled by regulatory bodies by proposing EQS for biota in EU water bodies and tolerable daily intake for food. The introduction of regulatory limits requires the availability of harmonised and validated analytical methods of sufficient sensitivity. This paper reviews recent advances in analytical methods for analysis of PFAS in aquatic organisms. The methods available for biota analysis are mostly based on three extraction procedures: ion-pair extraction, solvent liquid extraction, and alkaline digestion. The resulting extracts are then subjected to different clean-up or enrichment steps on solid adsorbents, for example graphitized carbon black, C(18), and WAX phases. All methods reviewed in this work give reliable results but are partially validated only, because of the lack of certified reference materials and regular interlaboratory exercises. The few interlaboratory exercises performed on real unspiked samples did not afford satisfactory results for PFAS other than PFOS, especially for matrices with high lipid content, for example mussels. The reasons for those partially negative results have been identified, and can mainly be attributed to calibration procedures and availability and purity of standards. The urgent need for certified reference materials for this type of analysis is emphasized.

Evaluation of the Use of Anticoagulotherapy in Cancer Patients in Palliative Care Residence.

Background: Several patients admitted to palliative care residences are on anticoagulotherapy (AC). Given the risks of venous thromboembolism (VTE) and bleeding, the decision to continue or stop AC on admission remains clinically challenging. Objectives: To determine the prevalence of AC use and incidence of suspected VTE and bleeding events in palliative care patients. Methods: Retrospective cohort study including all deceased patients at a Canadian palliative care residence over two years. Results: Among the 453 patients' charts reviewed (369 with cancer), 183 (40%) were on AC at admission or <30 days earlier. Only 64 (35%) continued AC, with 78% discontinuing it during their stay. Demographic parameters were similar in the AC and non-AC groups. The incidence of suspected VTE was lower in patients pursuing AC post-admission than in those who stopped: (4.6% vs. 6.7%) and, conversely, the incidence of bleeding was higher in patients on AC: (10.8% vs. 7.6%), though these differences were not statistically significant. The risk of death in cancer patients within 72 hours of suspected VTE or bleeding event was 80% and 30%, respectively. Patients on AC had a 33% reduced risk of VTE but a 44% increased risk of bleeding. Conclusion: This study provides information on the AC use in palliative care patients. In term of survivorship, it suggests a possible advantage to continue AC to prevent a symptomatic or distressing death. Given the low incidence of events, larger powered studies will be necessary to further characterize the risks/benefits of pursuing AC in patients in palliative care residences.

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.

Integrated Exposure and Algal Ecotoxicological Assessments of Effluents from Secondary and Advanced-Tertiary Wastewater-Treatment Plants.

The great concern over the environmental impact of wastewaters has led to the designing of advanced treatment processes to upgrade conventional treatment plants and achieve a significant reduction of contaminants in receiving waters. In the present study we combined chemical and ecotoxicological analyses, aiming to evaluate the reduction of toxicity effects associated with the removal of micropollutants and to define the contribution of the detected compounds to the overall toxicity of the mixtures in a series of wastewater effluents collected from a secondary treatment (OUT 2) and from a tertiary activated carbon treatment (OUT 3) plant. The target compounds were selected after a screening procedure among pharmaceuticals, musk fragrances, and trace metals. The classical algal growth inhibition test was conducted on the original effluent samples and on different fractions obtained by solid-phase extraction (SPE) treatment. A good accordance was found between the removal of toxicity (30%-80%) and organic compounds (70%-80%) after the tertiary treatment, suggesting its high efficiency to improve the wastewater quality. The discrepancy between the contribution to the overall toxicity of the nonadsorbable compounds (i.e., inorganic or very polar organic compounds) as experimentally measured by the SPE bioassays (18%-76%) and calculated by the concentration addition approach (>97%) could be mitigated by including the bioavailability correction in metal-toxicity modeling of wastewater mixtures. For the organic compounds, the toxic equivalency method enabled us to quantify the portion of toxicity explained by the detected chemicals in both OUT 2 (82%-104%) and OUT 3 (5%-57%), validating the selection of the target molecules. The applied integrating approach could be implemented by the inclusion of both additional target chemicals and toxicity endpoints. Environ Toxicol Chem 2022;41:2404-2419. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Determination of perfluoroalkyl acids in different tissues of graminaceous plants.

A method for the determination of 12 perfluoroalkyl acids (PFAA) in vegetal samples was proposed. The analytical procedure was developed to optimize the detection of short-chain PFAA (C < 8) due to their higher potential to be translocated and bioaccumulated in plants than long-chain congeners. The method, based on ultrasonic extraction, clean-up and HPLC-MS/MS analysis, determined PFAA in different plant tissues allowing the PFAA distribution and partition in vegetal compartments to be studied. The performance of this analytical procedure was validated by analysing samples (root, stem and leaf) of reed grass. The validated method was then applied to graminaceous plants from an agricultural area impacted by a fluorochemical plant discharge (Northern Italy). The PFAA congeners were detected in most of the samples with ΣPFAA concentrations in the whole plant ranging from

Exposure assessment of PFAS-contaminated sites using avian eggs as a biomonitoring tool: A frame of reference and a case study in the Po River valley (Northern Italy).

For many years, eggs of diverse bird species have been used as monitoring tools in studies investigating perfluoroalkyl substances (PFAS) contamination, especially in marine and remote areas. Avian eggs are a suitable monitoring matrix because they are relatively easy to collect and their yolks store diverse maternally transferred PFAS. Moreover, the concentrations of PFAS detected in the eggs are a good proxy for maternal exposure and allow the assessment of the potential risk for birds. These features support the use of avian eggs as a key monitoring tool in exposure assessment of PFAS-contaminated sites. We first review the recent application of avian eggs in PFAS monitoring in environmental risk assessment schemes, highlighting strengths and limitations and suggesting which criteria should be considered when selecting a proper study species and structuring the sampling and analytical protocol. Eventually, we report findings from a field study realized in 2020 near a perfluoropolymer factory site in the upper Po plain (Northern Italy), revealing an unprecedented contamination level of PFOA and C6O4 in three species of wild passerines. In future, long-term monitoring of PFAS contamination using avian eggs should be maintained, to provide crucial information on the temporal trend of fluorochemical production and waste disposal, while facilitating early identification of emerging PFAS as well as the quantification of their biomagnification across the trophic web. Integr Environ Assess Manag 2021;17:733-745. © 2021 SETAC.

Sediment quality assessment framework for per- and polyfluoroalkyl substances: Results from a preparatory study and regulatory implications.

A preparatory study was performed to develop a sediment quality and risk assessment strategy for Switzerland, addressing the following questions: the sediment fraction to be analyzed chemically (< 63 µm, or alternatively < 2 mm); the suitability of using perfluorooctanesulfonic acid (PFOS) as an indicator of per- and polyfluoroalkyl substances (PFAS) contamination in sediments; the availability of data for the derivation of sediment quality guidelines; and the suitability of normalization to total organic carbon (TOC). The results confirmed PFOS as a suitable indicator of PFAS contamination in sediments from small streams, being the most detected and on average with the highest concentrations among the analyzed PFAS. The fine fraction (< 63 µm) was more appropriate to screening for possible sources and studying the compound profiles at the study sites, but the analysis of the < 2 mm fraction and the normalization to a sample consisting of 100% of the < 63 µm fraction was, in principle, feasible for PFOS. Sediment quality guidelines for PFOS aiming to protect benthic invertebrates from generic adverse effects could be derived, but the available toxicity database is still too sparse. It was only possible to derive preliminary values. Sediment quality guidelines to protect wildlife and human health from secondary poisoning, using the equilibrium partitioning approach and simple trophic web models, were also derived. The use of food web models to derive sediment quality guidelines has not been validated, and the available database of Biota-Sediment Accumulation Factors and Trophic Magnification Factors for PFOS remains limited. However, it is still larger than the effect concentrations database based on sediment studies using benthic invertebrates. Normalization to TOC content in the field (0.2%-12.9%) was decisive in the quality assessment outcome. This is a widely accepted practice for hydrophobic organic contaminants, but its endorsement for PFOS would also benefit from validation. Integr Environ Assess Manag 2021;17:716-725. © 2021 SETAC.

Occurrence, distribution and pollution pattern of legacy and emerging organic pollutants in surface water of the Kongsfjorden (Svalbard, Norway): Environmental contamination, seasonal trend and climate change.

This work aimed to investigate the contamination pattern in Kongsfjorden marine environment (Svalbard, 79°N 12°E) and to disentangle primary and secondary emissions. Surface seawater, sampled in two seasons, was analysed by GC-MS and LC-MS/MS to detect polychlorobiphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), nonylphenols (NPs), bisphenol A (BPA) and perfluoroalkyl and polyfluoroalkyl substances (PFASs). In summer, average ΣPAHs, BPA, ΣNPs, ΣPFASs and ΣPCBs concentrations were 17.3 ± 11.1 ng/L, 0.9 ± 0.3 ng/L, 10.0 ± 6.9 ng/L, 0.4 ± 0.7 ng/L and 1.8 ± 1.3 pg/L, respectively; while in winter, they were 13.6 ± 10.1 ng/L, 0.5 ± 0.2 ng/L, 6.8 ± 3.3 ng/L,

Within- and Among-Clutch Variation of Yolk Perfluoroalkyl Acids in a Seabird from the Northern Adriatic Sea.

Perfluoroalkyl substances (PFAS) are surface-active agents used in diverse industrial and commercial applications. They contaminate both freshwater and marine ecosystems, are highly persistent, and accumulate through trophic transfer. Seabirds are exposed to environmental contaminants due to their high trophic position in food webs and relatively long lifespan. We measured levels of 10 perfluoroalkyl acids (PFAAs) in egg yolks of yellow-legged gulls (Larus michahellis) breeding in the northern Adriatic Sea (Northeast Italy). We examined variations in PFAAs within clutches (between eggs of different laying order) and among clutches. Perfluorooctane sulfonate (PFOS) was the most abundant yolk PFAA (mean = 42.0 ng/g wet wt), followed by perfluorooctanoic acid (PFOA; 3.8 ng/g wet wt) and perfluorododecanoic acid (PFDoDa; 2.8 ng/g wet wt). The ∑PFAAs averaged 57.4 ng/g wet weight, ranging between 26.5 and 115.0 ng/g wet weight. The PFAA levels varied substantially among clutches (0.29-0.79 of the total variation), whereas the effects of laying order were considerably weaker (0.01-0.13). Egg-laying order effects were detected for ∑PFAAs, PFOS, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnA), and PFDoDa, whereby the last-laid eggs exhibited lower PFAA concentrations than early-laid eggs. Our results indicate that seagulls from the northern Adriatic basin deposit measurable amounts of PFAAs in their eggs. The large among-clutches differences in PFAAs suggest that exposure of yellow-legged gull females to these compounds is highly variable. Environ Toxicol Chem 2021;40:744-753. © 2020 SETAC.

Per- and Polyfluoroalkyl Substances (PFAS) in Fish from European Lakes: Current Contamination Status, Sources, and Perspectives for Monitoring.

Concentrations in fish of per- and polyfluoroalkyl substances (PFAS) were reported for 7 deep lakes in the European subalpine area: Lakes Geneva, Lugano, Maggiore, Iseo, Como, Garda, and Mergozzo; one shallow lowland lake (Varese); and 2 high-altitude alpine lakes (>2000 m a.s.l). Fillets and, in selected cases, other body fractions (viscera, liver, and residual carcass) from 8 fish species were analyzed. The possibility of harmonizing the monitoring protocols was tested. Results suggest that the sampling season is not critical for PFASs and the total protein content cannot be used for normalization of tissue concentrations because PFASs bind to specific proteins. Moreover, the polar lipid content could be used to reduce the variability of PFAS concentrations in phospholipid rich fractions of fish such as viscera and carcass. The data comparison and analysis show that the PFAS contamination in lake fish is generally correlated with the degree of urbanization of the lake catchment; however, it is sometimes difficult to compare absolute concentrations in lake fish because the lake hydro-morphological characteristics play a substantial role in determining the chemical concentrations of persistent and mobile contaminants. Environ Toxicol Chem 2021;40:658-676. © 2020 SETAC.

New compounds, old problems. The case of C6O4 - a substitute of PFOA - and its effects to the clam Ruditapes philippinarum.

C6O4 (difluoro{[2,2,4,5-tetrafluoro-5-(trifluoromethoxy)-1,3-dioxolan-4-yl]oxy}acetic acid) is a new surfactant and emulsifier used as a substitute of perfluorooctanoic acid (PFOA). Recently, C6O4 has been detected in aquatic environments, but, at present, no information concerning the effects of C6O4 on aquatic species, such as bivalves, are available in the literature. Therefore, in this study we evaluated for the first time the effects of C6O4 (0.1 and 1 µg/L) and PFOA (1 µg/L) to the clam Ruditapes philippinarum. Short-term (7 days) and long-term (21 days) exposures of clams to the two compounds were carried out and numerous biomarkers were measured in haemocytes/haemolymph, as well as in gills and digestive gland. The MANOVA analysis demonstrated statistically significant effects of the independent variables "treatment", "time" and "treatment-time interaction" on the whole dataset of biomarker responses. The two-way ANOVA analysis performed for each biomarker response indicated that the two compounds affected most of the cellular and tissue parameters measured. Despite preliminary, the results obtained suggested that C6O4 - similarly to PFOA - can affect both cellular and biochemical parameters of clams.

What's in the water? - Target and suspect screening of contaminants of emerging concern in raw water and drinking water from Europe and Asia.

There is growing worry that drinking water can be affected by contaminants of emerging concern (CECs), potentially threatening human health. In this study, a wide range of CECs (n = 177), including pharmaceuticals, pesticides, perfluoroalkyl substances (PFASs) and other compounds, were analysed in raw water and in drinking water collected from drinking water treatment plants (DWTPs) in Europe and Asia (n = 13). The impact of human activities was reflected in large numbers of compounds detected (n = 115) and high variation in concentrations in the raw water (range 15-7995 ng L-1 for ∑177CECs). The variation was less pronounced in drinking water, with total concentration ranging from 35 to 919 ng L-1. Treatment efficiency was on average 65 ± 28%, with wide variation between different DWTPs. The DWTP with the highest ∑CEC concentrations in raw water had the most efficient treatment procedure (average treatment efficiency 89%), whereas the DWTP with the lowest ∑177CEC concentration in the raw water had the lowest average treatment efficiency (2.3%). Suspect screening was performed for 500 compounds ranked high as chemicals of concern for drinking water, using a prioritisation tool (SusTool). Overall, 208 features of interest were discovered and three were confirmed with reference standards. There was co-variation between removal efficiency in DWTPs for the target compounds and the suspected features detected using suspect screening, implying that removal of known contaminants can be used to predict overall removal of potential CECs for drinking water production. Our results can be of high value for DWTPs around the globe in their planning for future treatment strategies to meet the increasing concern about human exposure to unknown CECs present in their drinking water.

The new PFAS C6O4 and its effects on marine invertebrates: First evidence of transcriptional and microbiota changes in the Manila clam Ruditapes philippinarum.

There is growing concern for the wide use ofperfluorooctanoic acid (PFOA) because of its toxic effects on the environment and on human health. A new compound - the so called C6O4 (perfluoro ([5-methoxy-1,3-dioxolan-4-yl]oxy) acetic acid) - was recently introduced as one of the alternative to traditional PFOA, however this was done without any scientific evidence of the effects of C6O4 when dispersed into the environment. Recently, the Regional Agency for the Protection of the Environment of Veneto (Italy) detected high levels of C6O4 in groundwater and in the Po river, increasing the alarm for the potential effects of this chemical into the natural environment. The present study investigates for the first time the effects of C6O4 on the Manila clam Ruditapes philippinarum exposed to environmental realistic concentrations of C6O4 (0.1 µg/L and 1 µg/L) for 7 and 21 days. Furthermore, in order to better understand if C6O4 is a valid and less hazardous alternative to its substitute, microbial and transcriptomic alterations were also investigated in clams exposed to 1 µg/L ofPFOA. Results indicate that C6O4 may cause significant perturbations to the digestive gland microbiota, likely determining the impairment of host physiological homeostasis. Despite chemical analyses suggest a 5 times lower accumulation potential of C604 as compared to PFOA in clam soft tissues, transcriptional analyses reveal several alterations of gene expression profile. A large part of the altered pathways, including immune response, apoptosis regulation, nervous system development, lipid metabolism and cell membrane is the same in C6O4 and PFOA exposed clams. In addition, clams exposed to C6O4 showed dose-dependent responses as well as possible narcotic or neurotoxic effects and reduced activation of genes involved in xenobiotic metabolism. Overall, the present study suggests that the potential risks for marine organism following environmental contamination are not reduced by replacing PFOA with C6O4. In addition, the detection of both C6O4 and PFOA into tissues of clams inhabiting the Lagoon of Venice - where there are no point sources of either compounds - recommends a similar capacity to spread throughout the environment. These results prompt the urgent need to re-evaluate the use of C6O4 as it may represent not only an environmental hazard but also a potential risk for human health.

Uptake and translocation of perfluoroalkyl acids (PFAAs) in hydroponically grown red chicory (Cichorium intybus L.): Growth and developmental toxicity, comparison with growth in soil and bioavailability implications.

Short-chain perfluoroalkyl acids (PFAAs) have shown a high potential for plant (crop) uptake, making them possibly significant contributors to the total dietary exposure to PFAAs. The plant uptake of PFAAs is a complex process that needs better characterization, as it does not only depend on perfluoroalkyl chain length, but also on their polar terminal group, on the plant species and the exposure media. Here, a plant uptake study with nine perfluoroalkyl acids (PFAAs) was carried out under the hydroponic (soilless) exposure conditions. Red chicory was grown in a nutrient solution, spiked with PFAAs mixture at three different concentrations (i.e. 62.5, 125 and 250 µg/L), in order to extend the range of levels tested and reported in the literature so far. Bioaccumulation metrics and transpiration stream concentration factors (TSCFs) were employed for the plant uptake characterization and consequent comparison with the results of soil uptake experiment we previously performed with the same crop. The results showed that calculated root concentration factors (RCFs) increase with PFAA chain length, while the opposite chain length dependence was present for shoots. Plants from two treatments with the highest PFAAs concentrations manifested physiological changes (discoloration, inhibited roots and leaves growth), despite of the used exposure concentrations being much lower than previously published phytotoxicity thresholds. A comparison among RCFs and TSCFs derived from hydroponic and from the soil experiment has emphasized their different magnitudes and PFAAs chain length dependence patterns. They could not be ascribed only to soil sorption as a process decreasing PFAAs bioavailability for plants, but also to developmental differences between the root systems formed in soil and in nutrient solution and to the potential competitive PFAAs sorption to roots in hydroponics. The interchangeable use of bioaccumulation and translocation parameters derived in hydroponic and soil systems would lead to erroneous conclusions and plant uptake predictions.

Uptake and translocation of perfluoroalkyl acids (PFAA) in red chicory (Cichorium intybus L.) under various treatments with pre-contaminated soil and irrigation water.

Perfluoroalkyl acids (PFAAs), particularly short-chained ones, have high potential for crop uptake, posing a threat to human health in contaminated areas. There is a scarcity of studies using contaminated water as the medium for PFAAs delivery to crops, and a lack of data on the partitioning of PFAA mixtures in growing media. In this context, a controlled experimental study was carried out in a greenhouse to investigate the uptake of a PFAA mixture into red chicory, a typical crop from a major PFAA contamination hot-spot in northern Italy, under treatments with environmentally relevant concentrations in spiked irrigation water and soil, separately and simultaneously. To our knowledge, this is the first study involving multiple exposure media and laboratory adsorption/desorption batch tests as a way of assessing the decrease in the bioavailability of PFAAs from soil. Exposure concentrations for each of the 9 utilized PFAAs were 0, 1, 10 and 80 µg/L in irrigation water and 0, 100 and 200 ng/gdw in soil, combined into 12 treatments. The highest bioaccumulation was measured for PFBA in roots (maximum of 43 µg/gdw), followed by leaves and heads of the chicory plants in all treatments, with the concentrations exponentially decreasing with an increasing PFAA chain length in all plant compartments. The use of irrigation water as the delivery medium increased the transport of PFAAs to the aerial chicory parts, long-chain substances in particular. Additionally, the distribution of PFAAs in the soil was assessed by depth and compared with laboratory measured soil-water equilibrium partition coefficients, revealing only partial dependency of PFAAs bioavailability on the adsorption in soil.

Presence and infectivity of SARS-CoV-2 virus in wastewaters and rivers.

The presence of SARS-CoV-2 in raw wastewaters has been demonstrated in many countries affected by this pandemic. Nevertheless, virus presence and infectivity in treated wastewaters, but also in the receiving water bodies are still poorly investigated. In this study, raw and treated samples from three wastewater treatment plants, and three river samples within the Milano Metropolitan Area, Italy, were surveyed for SARS-CoV-2 RNA detection by means of real time RT-PCR and infectivity test on culture cells. SARS-CoV-2 RNA was detected in raw, but not in treated wastewaters (four and two samples, respectively, sampled in two dates). The isolated virus genome was sequenced, and belonged to the strain most spread in Europe and similar to another found in the same region. RNA presence in raw wastewater samples decreased after eight days, probably following the epidemiological trend estimated for the area. Virus infectivity was always null, indicating the natural decay of viral pathogenicity in time from emission. Samples from receiving rivers (three sites, sampled in the same dates as wastewaters) showed in some cases a positivity to real time RT-PCR, probably due to non-treated, or inefficiently treated discharges, or to the combined sewage overflows. Nevertheless, also for rivers infectivity was null. Risks for public health should be limited, although a precautionary approach to risk assessment is here advocated, giving the preliminary nature of the presented data.