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Wastewater-based epidemiology (WBE) is a well-established approach that can provide objective and real-time data on the consumption of substances such as pharmaceuticals. However, most of the studies reported so far compares consumption data obtained using WBE with those derived from prescription data from public health systems, which is often incomplete and might represent a source of uncertainty. This study aims to compare the measured pharmaceutical consumption back calculated with the WBE approach with consumption derived from dispensed pharmaceuticals in two regions of Spain, managed by two different Health Systems. To do so, a group of 17 pharmaceuticals, including the most representative ones of every therapeutic family, were monitored in influent wastewater (IWW) samples collected over a week campaign in spring 2022 at four different wastewater treatment plants (WWTPs) in Spain: two WWTPs in Madrid city (center of Spain) and two WWTPs in Catalonia (Northeast of Spain). Population-normalized daily loads (PNDL) revealed that the patterns of pharmaceutical occurrence in the different WWTPs are very similar, being acetaminophen, 4-acetamidoantipyrine and valsartan the pharmaceuticals with the highest PNDL values: 17162 ± 1457 mg day -1 1000 inh -1 for acetaminophen, 2365 ± 696 and 2429 ± 263 mg day -1 1000 inh -1 for 4-acetamidoantipyrine, 2006 ± 541 and 2041 ± 352 mg day -1 1000 inh -1 for valsartan. Pharmaceutical PNLDs were then transformed into measured pharmaceutical consumption (MC) and compared with dispensed consumption (DC) data obtained from the pharmacies in the catchment area where the WWTPs are located. A ratio MC/DC within 0.8 to 1.2 was obtained for 11 out of the 17 studied pharmaceuticals. Highlighting a match in all the cardiovascular system pharmaceuticals, with the exception of losartan (1.29-1.39 ratio) and valsartan (1.35-1.43) in all WWTPs. In summary, the degree of correlation between MC/DC is higher than those previously reported comparing with the prescribed pharmaceutical consumption., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Drug consumption estimates are traditionally based on surveys or information from police seizures. Alternatively, residues of illicit drugs in untreated wastewater (influent) can be used to calculate mass loads and subsequently estimate drug consumption in the community throughout the week. For this purpose, wastewater is commonly sampled for seven consecutive days within the Sewage analysis CORe group Europe (SCORE), while other sampling schemes may be implemented in long-term studies outside this consortium. The current study demonstrates how sampling frequency of illicit drug residues in the influent of wastewater treatment plants (WWTPs) affects the derived weekly average. Thirty WWTPs were sampled over the course of 12 years and influents were analyzed for five drugs (metabolites): 3,4-methylenedioxymethamphetamine (MDMA), methamphetamine, amphetamine, benzoylecgonine (a metabolite of cocaine), and 11-nor-9-Carboxy tetrahydrocannabinol (THC-COOH). Subsequently, small and large WWTPs were grouped with a threshold of 100,000 inhabitants. After data curation, standardized loads were calculated (mg/d per 1000 inhabitants). Weekly averages of loads of the drug residues were calculated based on six scenarios (sampling one to six weekdays) and compared to the weekly average in the control situation (sampling seven weekdays) in a Monte Carlo simulation. Results indicate that drug residues with more dynamic loads over a week require more frequent sampling. The analysis illustrates that a decreased sampling frequency (4 or 5 days per week) still leads to a representative weekly average for all drugs tested when a deviation up to a factor of 1.25 is deemed acceptable. However, knowledge on typical levels is necessary to define outliers. We therefore recommend to study dynamics in drug residue loads for WWTPs before reducing sampling frequency in long term monitoring programs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Sewage surveillance is widely applied to track valid human excretion information and identify public health conditions during corona virus disease 2019 (COVID-19) pandemic. This approach can be applied to monitor the antibiotic resistance level in sewers and to assess the risk of spreading antibiotic resistance in municipal wastewater systems. However, there is still little information about human antibiotic resistance occurrence characteristics in sewer system. This study conducted a field trial for whole year to advance understanding on spatial and temporal occurrence of antibiotic resistance genes (ARGs) in gravity sewerage. The spatial distribution of ARGs along the drainage pipe line (from human settlements to wastewater treatement pant (WWTP)) was insignificant, which may be affected by irregular human emission alongside the pipeline. The correlation between ARGs and antibiotics in sewage was insignificant. The temporal distribution showed that the effect of temperature on ARGs abundance was evident, the ARGs abundance in sewage was generally higher during the cold season. Metagenomic analysis revealed that the detected ARGs were mainly distributed in Proteobacteria (47.51 %) and Antinobacteria (20.11 %). Potential hosts of ARGs in sewage were mainly identified as human gut microorganisms, including human pathogenic bacteria, such as Prevotella, Kocuria, and Propionibacterium, etc. This study provides a new insight into the sewerage surveillance tracking characteristics of human ARGs in sewer system, and suggesting that the sewage-carried ARGs surveillance is a promising method for assessment and management of antibiotic resistance level on population size., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In advanced wastewater treatment plants on pig farms, meticulous design aims to eliminate intrinsic pollutants such as organic matter, heavy metals, and biological contaminants. In our field survey across Southern China, a notable disparity in wastewater treatment procedures among various farming facilities lies in the utilization of terminal chemical oxidation post-sedimentation tank. However, recent focus in wastewater surveillance has predominantly centered on antibiotic resistance genes, leaving the efficacy of virus removal in different effluent systems largely unexplored. To profile virus composition at the effluent, assess the virus elimination efficiency of chemical oxidation at the effluent end, and the potential environmental driver of virus abundance, we deployed a meta-transcriptomics approach to first determine the total virome in effluent specimens of terminal clean water tank system (CWT) and terminal chemical oxidation system (TCO) in Southern China pig farms, respectively. From these data, 172 viruses were identified, with a median reads per million (RPM) of 27,789 in CWT and 19,982 in TCO. Through the integration of analyses encompassing the co-occurrence patterns within viral communities, the ecology of viral diversity, and a comparative assessment of average variation degrees, we have empirically demonstrated that the procedure of TCO may perturb viral communities and diminish their abundance, particularly impacting RNA viral communities. However, despite the diminished abundance, pathogenic viruses such as PEDV and PRRSV persisted in the effluent following chemical deoxidation at a moderate RPM value, indicating a substantial in situ presence at effluent. Our environmental driver modeling, employing GLM and mantel tests, substantiated the intricate nature of virus community variation within the effluent, influenced heterogeneously by diverse factors. Notably, pond temperature emerged as the foremost determinant, while fishing farming exhibited a positive correlation with virus diversity (p < 0.05). This revelation of the cryptic persistence of virus communities in wastewater effluent expands our understanding of the varied responses of different virus categories to oxidation. Such insights transcend mere virus characterization, offering valuable implications for enhancing biosafety measures in farming practices and informing wastewater-based epidemiological surveillance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Trace organic contaminants (TrOCs) are omnipresent in wastewater treatment plants (WWTPs), yet, their removal during wastewater treatment is oftentimes incomplete and underlying biotransformation mechanisms are not fully understood. In this study, we elucidate how different factors, including pre-exposure levels and duration, influence microbial adaptation towards catabolic TrOC biodegradation and its potential role in biological wastewater treatment. Four sequencing batch reactors (SBRs) were operated in parallel in three succeeding phases, adding and removing a selection of 26 TrOCs at different concentration levels. After each phase of SBR operation, a series of batch experiments was conducted to monitor biotransformation kinetics of those same TrOCs across various spike concentrations. For half of our test TrOCs, we detected increased biotransformation in sludge pre-exposed to TrOC concentrations ≥5 µg L -1 over a 30-day period, with most significant differences observed for the insect repellent DEET and the artificial sweetener saccharin. Accordingly, 16S rRNA amplicon sequencing revealed enrichment of taxa that have previously been linked to catabolic biodegradation of several test TrOCs, e.g., Bosea sp. and Shinella sp. for acesulfame degradation, and Pseudomonas sp. for caffeine, cyclamate, DEET, metformin, paracetamol, and isoproturon degradation. We further conducted shotgun metagenomics to query for gene products previously reported to be involved in the TrOCs' biodegradation pathways. In the future, directed microbial adaptation may be a solution to improve bioremediation of TrOCs in contaminated environments or in WWTPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Persistent and mobile (PM) chemicals are considered detrimental for drinking water resources as they may pass through all barriers protecting these resources against pollution. However, knowledge on the occurrence of PM chemicals in the water cycle, that make their way into drinking water resources, is still limited. The effluents of six municipal wastewater treatment plants (WWTPs, n = 38), surface water of two rivers (n = 32) and bank filtrate of one site (n = 15) were analyzed for 127 suspected PM chemicals. In the rivers, median concentrations of 92 detected analytes ranged from 0.3 ng/L to 2.6 µg/L (tetrafluoroborate, BF4). Lower than average dilution from WWTP effluent to surface water of 43 PM chemicals suggests significant discharge from other sources. Many of these compounds were industrial chemicals, including cyanoguanidine, trifluoromethanesulfonic acid and BF4. River bank filtration (RBF) reduced the total concentration of 40 quantified compounds by 60 % from 19.5 µg/L in surface water to 8.4 µg/L in bank filtrate, on average. Of these, 20 compounds showed good removal (> 80 %), 14 intermediate (80 - 20 %) and 6 no removal (≤ 20 %), among them carbamazepine, hexafluorophosphate, and 2-pyrrolidone. 13 substances occurred at concentrations ≥ 0.1 µg/L in bank filtrate; for six of them toxicological data were insufficient for a health-based risk assessment. The regulatory definition of P and M chemicals, if used together with existing data on environmental half-lives (P) and K oc (M), showed little power to discriminate between chemicals well removed in RBF and those that were hardly removed. This comprehensive field study shows that RBF is a useful but incomplete barrier to retain PM chemicals from surface water. Thus, PM chemicals are, indeed, a challenge for a sustainable water supply., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Pharmaceutical wastewater often contains significant levels of antibiotic residues, which continuously induce and promote antibiotic resistance during the sewage treatment process. However, the specific impact of antibiotics on the emergence of antibiotic resistance genes (ARGs), microbiomes, and mobile genetic elements (MGEs), as well as the dose-response relationship remain unclear. Herein, through metagenomic sequencing and analysis, we investigated the fate, transmission, and associated risk of ARGs over a ten-year period of exposure to a gradient of sulfonamide antibiotics at a pharmaceutical wastewater treatment plant (PWWTP), an associated wastewater treatment plant (WWTP), and the receiving river. Through abundance comparison and principal co-ordinates analysis (PCoA), our results revealed distinct ARG, microbiome, and MGE profiles across different antibiotic concentrations. Notably, there was a decreasing trend in the abundance of ARGs and MGEs as the antibiotic concentrations were attenuated (p < 0.05). Further partial least squares path modeling analysis, Procrustes analysis and network analysis indicated that variation in MGEs and microbiomes were the driving forces behind the distribution of ARGs. Based on these findings, we proposed an antibiotic-microbiome-MGE-ARG dissemination paradigm, in which integrons as key drivers were closely associated with prevalent ARGs such as sul1, sul2, and aadA. With a focus on human pathogenic bacteria and the associated health risks of ARGs, we conducted pathogen source analysis and calculated the antibiotic resistome risk index (ARRI). Our findings highlighted potential risks associated with the transition from PWWTP to WWTP, raising concerns regarding risk amplification due to the mixed treatment of antibiotic-laden industrial wastewater and domestic sewage. Overall, the results of our study provide valuable information for optimizing wastewater treatment practices to better manage antibiotic resistance., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Na Wang reports financial support was provided by Ministry of Science and Technology of the People's Republic of China. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024. Published by Elsevier Ltd.)

The concept of incorporating foam fractionation in aerated bioreactors at wastewater treatment plants (WWTPs) for the removal of per- and polyfluoroalkyl substances (PFAS) has recently been proposed. The extent of PFAS enrichment in aerated bioreactors' foams, as indicated by enrichment factors (EFs), has been observed to vary widely. Laboratory evidence has shown that factors affecting PFAS enrichment in foams include conductivity, surfactant concentrations and initial PFAS concentrations. However, real wastewaters are complex heterogenous matrices with physical, chemical and biological characteristics potentially contributing to the phenomenon of PFAS partitioning into foams. In this study, we characterised mixed liquor suspensions, including conductivity, filament content, aqueous PFAS concentrations, surface tension and total suspended solids concentrations (TSS) as well as foams, including bubble size and half-life. We used statistical tools - linear mixed-effects model - to establish relationships between PFAS enrichment in aerated bioreactor foams and the examined characteristics. We found that some of the examined characteristics, specifically filament content, surface tension and TSS concentrations measured in mixed liquor suspension and foam half-life, are negatively and significantly associated with the enrichment of longer chain PFAS (with perfluorinated carbon number ≥ 6). Of these, filament content is the important determinant of PFAS enrichment, potentially leading to an increase in, for example, perfluorooctanoic acid (PFOA) EF from 3 to 100 between typical filamentous and non-filamentous suspended biomass. However, enrichment of shorter chain PFAS (with perfluorinated carbon number ≤ 5) is negligible and is not affected by the characteristics that were measured. The findings of our study may serve as valuable information for the implementation of foam fractionation at WWTPs by elucidating the drivers that contribute to the enrichment of longer chain PFAS, under conditions typically found at WWTPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Fate and transport of per- and polyfluoroalkyl substances (PFAS) in wastewater treatment plant (WWTP) effluent discharged to rapid infiltration basins (RIBs) is investigated using data from 26 WWTPs in Michigan, USA. PFAS were found to accumulate in groundwater downgradient from RIBs with median groundwater-effluent enrichment factors for ten commonly detected, terminal-form perfluoroalkyl acids (PFAAs) ranging from 1.3 to 5.2. Maximum contaminant levels for drinking water were exceeded in groundwater at all WWTPs with available PFAS data. Numerical models of unsaturated fluid flow and PFAS transport honoring RIB site properties, such as median vertical separation distance to the water table and a realistic range of area-normalized effluent fluxes, show long-chain PFAS undergo significant delays from air-water interface (AWI) adsorption, requiring up to 15 times longer to reach maximum mass flux to the saturated zone under low-flux conditions, where AWI area is 2.5 times greater. Short-chain PFAS commonly detected in effluent are only minimally affected by AWI adsorption and show little to no attenuation under high-flux conditions. The nonlinear inverse relationship between water content and AWI area highlights the important role of AWI adsorption in modulating unsaturated transport of long-chain PFAS to underlying groundwater due to the broad range of flux rates applied to RIB systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The wide utilization of poly dimethyl diallyl ammonium chloride (polyDADMAC) in industrial conditions leads to its accumulation in waste activated sludge (WAS), thereby affecting subsequent WAS treatment processes. This work investigated the interaction between polyDADMAC and WAS components from the perspective of anaerobic digestion (AD) performance and anaerobes adaptability variation. The results showed that polyDADMAC decreased the content of biodegradable organic substrates (i.e., soluble protein and carbohydrate) by binding with the functional groups and then settling to the solid phase, thus impeding the subsequent utilization. Higher concentrations of polyDADMAC prompted an initial protective response of excreting organic substrates into extracellular environment, but its toxicity to archaea was irreversible. Consequently, polyDADMAC inhibited the processes of AD and induced a 30 % reduction in methane production with 0.05 g polyDADMAC/g total suspended solid (TSS) addition. Changes in microbial community structure indicated that archaea involved in methane production (e.g., Anaerolineaceae sp. and Methanosaeta sp.) were inhibited when exposed to polyDADMAC. However, several adaptive bacteria with the ability of utilizing complex organics and participating in nitrogen cycle (e.g., Aminicenantales sp. and Ellin6067 sp.) were enriched with the above dosage. Specifically, the decreased abundance of genes relevant to methane metabolism pathway (i.e., mer and cdh) and increased abundance of genes involved in metabolism of cofactors and vitamins (e.g., nad and thi) indicated the toxicity of polyDADMAC and the irritant response of microflora. Moreover, polyDADMAC underwent degradation in AD system, resulting in a 12 % reduction in 15 days, accompanied by an increase in the -NO 2 functional group. In general, this study provided a thorough understanding of the interaction between polyDADMAC and WAS components, raising concerns regarding the elimination of endogenous pollutants during AD., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Diclofenac (DCF) is the first drug on the EU Watch List of Priority Substances due to its extensive uses, incomplete removal in wastewater treatment plants (WWTPs), and toxic effects. Once in the environment, DCF undergoes processes that yield various transformation products (TPs) or metabolites, whichcan be more toxic than DCF. While these TPs or metabolites often dominate the majority of the drug load in municipal wastewater, they have been largely ignored. This review critically examines recent data on the formation, occurrence, fate, and toxicology of DCF TPs in the aquatic environment. This review reveals some important findings. First, DCF TPs may constitute >57 % of DCF in wastewater influent, ∼60 % in effluent, and ∼30 % in surface waters. Second, TPs persistently retain the core structure of DCF and may pose greater environmental risks than the parent drug. Third, some metabolites may revert back to the parent drug. Fourth, WWTPs serve as a consistent source that release DCF and its by-products into the environment. Fifth, mixtures of DCF and its metabolites, along with other contaminants, may pose elevated and synergistic environmental risks than individual compounds. These findings suggest that current risk assessment practices, which ignore the impacts of the metabolites and the chemical interactions/synergies, may seriously underestimate the overall toxicity of DCF and likely other pharmaceuticals. Further studies are needed to monitor the long-term fate and toxicity of the metabolites, as well as new analytical methods and standards to unveil the hidden metabolites and the associated environmental risks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Nitrification, a process carried out by aerobic microorganisms that oxidizes ammonia to nitrate via nitrite, is an indispensable step in wastewater nitrogen removal. To facilitate energy and carbon savings, applying low dissolved oxygen (DO) is suggested to shortcut the conventional biological nitrogen removal pathway, however, the impact of low DO on nitrifying communities within activated sludge is not fully understood. This study used genome-resolved metagenomics to compare nitrifying communities under extremely low- and high-DO. Two bioreactors were parallelly operated to perform nitrification and DO was respectively provided by limited gas-liquid mass transfer from the atmosphere (AN reactor, DO < 0.1 mg/L) and by sufficient aeration (AE reactor, DO > 5.0 mg/L). Low DO was thought to limit nitrifiers growth; however, we demonstrated that complete nitrification could still be achieved under the extremely low-DO conditions, but with no nitrite accumulation observed. Kinetic analysis showed that after long-term exposure to low DO, nitrifiers had a higher oxygen affinity constant and could maintain a relatively high nitrification rate, particularly at low levels of DO (<0.2 mg/L). Community-level gene analysis indicated that low DO promoted enrichment of nitrifiers (the genera Nitrosomonas and Nitrospira, increased by 2.3- to 4.3-fold), and also harbored with 2.3 to 5.3 times higher of nitrification functional genes. Moreover, 46 high-quality (>90 % completeness and <5 % contamination) with 3 most abundant medium-quality metagenome-assembled genomes (MAGs) were retrieved using binning methods. Genome-level phylogenetic analysis revealed the species succession within nitrifying populations. Surprisingly, compared to DO-rich conditions, low-DO conditions were found to efficiently suppressed the ordinary heterotrophic microorganisms (e.g., the families Anaerolineales, Phycisphaerales, and Chitinophagales), but selected for the specific candidate denitrifiers (within phylum Bacteroidota). This study provides new microbial insights to demonstrate that low-DO favors the enrichment of autotrophic nitrifiers over heterotrophs with species-level successions, which would facilitate the optimization of energy and carbon management in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Dewatering is an indispensable link in sludge treatment, but its effect on the microplastics (MPs) remains inadequately understood. This study investigated the physicochemical changes and leaching behavior of MPs during the mechanical dewatering of sludge, as well as the impact of MP leachates on activated sludge (AS). After sludge dewatering, MPs exhibit rougher surfaces, decreased sizes and altered functional groups due to the addition of dewatering agents and the application of mechanical force. Meanwhile, plastic additives, depolymerization products, and derivatives of their interactions are leached from MPs during sludge dewatering process. The concentration of MP-based leachates in sludge is 2-25 times higher than that in water. The enhancement of pH and ionic strength caused by dewatering agents induces the release of MP leachates enriched with protein-like, fulvic acid-like, and soluble microbial by-product-like substances. The reflux of MP leachates in sludge dewatering liquor to the wastewater treatment system negatively impacts AS, leading to a decrease in COD removal rate and inhibition of the extracellular polymeric substances secretion. More importantly, MP leachates cause oxidative stress to microbial cells and alter the microbial community structure of AS at the phylum and genus levels. These findings confirm that MPs undergo aging and leaching during sludge dewatering process, and MP leachates may negatively affect the wastewater treatment system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Rivers play a pivotal role in global carbon (C) and nitrogen (N) biogeochemical cycles. Urbanized rivers are significant hotspots of greenhouse gases (GHGs, N 2 O, CO 2 and CH 4 ) emissions. This study examined the GHGs distributions in the Guanxun River, an effluents-receiving subtropical urbanized river, as well as the key environmental factors and processes affecting the pattern and emission characteristics of GHGs. Dissolved N 2 O, CO 2 , and CH 4 concentrations reached 228.0 nmol L -1 , 0.44 mmol L -1 , and 5.2 μmol L -1 during the wet period, and 929.8 nmol L -1 , 0.7 mmol L -1 , and 4.6 μmol L -1 during the dry period, respectively. Effluents inputs increased C and N loadings, reduced C/N ratios, and promoted further methanogenesis and N 2 O production dominated by incomplete denitrification after the outfall. Increased urbanization in the far downstream, high hydraulic residence time, low DO and high organic C environment promoted methanogenesis. The strong CH 4 oxidation and methanogenic reactions inhibited by the effluents combined to suppress CH 4 emissions in downstream near the outfall, and the process also contributed to CO 2 production. The carbon fixation downstream from the outfall were inhibited by effluents. Ultimately, it promoted CO 2 emissions downstream from the outfall. The continuous C, N, and chlorine inputs maintained the high saturation and production potential of GHGs, and altered microbial community structure and functional genes abundance. Ultimately, the global warming potential downstream increased by 186 % and 84 % during wet and dry periods on the 20-year scale, and increased by 91 % and 49 % during wet and dry periods on the 100-year scale, respectively, compared with upstream from the outfall. In urbanized rivers with sufficient C and N source supply from WWTP effluents, the large effluent equivalently transformed the natural water within the channel into a subsequent "reactor". Furthermore, the IPCC recommended EF 5r values appear to underestimate the N 2 O emission potential of urbanized rivers with high pollution loading that receiving WWTP effluents. The findings of this study might aid the development of effective strategies for mitigating global climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Research has demonstrated that depending on the type and concentration, microplastics affect anaerobic digestion (AD). Owing to the high abundance of polyamide-6 (PA6) in wastewater treatment plants and limited understanding of its behavior, this study investigates PA6 microplastics' effect in AD. Biochemical methane potential experiments were performed under mesophilic (35 °C) and thermophilic (55 °C) conditions using PA6 at concentrations from 0 to 200 particles/g total solids (TS). Under both conditions, methane production increased in the highest (200 particles/g TS) PA6-dosed reactors, with thermophilic conditions having a statistically significant effect. Methane yield increased from 403.1 ± 5.3 mL/g VS to 436.6 ± 9.2 mL/g VS under thermophilic and from 332.1 ± 1.5 to 340.6 ± 6.6 mL/g VS under mesophilic conditions for the 200 particles/g TS dose, corresponding to increases of 8.3% and 2.6% respectively. PA6 crystallinity decreased from 32.8% to 27.1% and 26.8%, corresponding to decreases of 17.4% in mesophilic and 18.2% in thermophilic reactors compared to pristine control. Similarly, crystallinity decreased in PA6 microplastics collected from abiotic reactors, with thermophilic conditions showing a greater effect. The carbonyl index (CI) values were similar between biotic and abiotic reactors, but PA6 from all reactors had significantly higher CI than pristine PA6, suggesting abiotic factors also affect carbonyl bonds. Additionally, an increase in average PA6 mass was observed for mesophilic and thermophilic conditions by 22.0 % and 23.0 %, respectively. The study shows that temperature and other abiotic factors, like sludge chemistry, significantly influence the fate and effect of PA6 microplastics in digesters. Including abiotic reactors seems crucial for a full understanding of the impact of microbial and non-microbial factors in microplastic studies in the AD process. Studying the effects of microplastics on AD is only one part of the picture, whereas simultaneously examining their fate in digestion is necessary for a complete understanding., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Optimizing nitrogen removal is crucial for ensuring the efficient operation of wastewater treatment plants (WWTPs), but it is susceptible to variations in influent conditions and operational parameter constraints, and conflicts with the energy-saving and carbon emission reduction goals. To address these issues, this study proposes a hybrid framework integrating process simulation, machine learning, and multi-objective genetic algorithms for nitrogen removal diagnosis and optimization, aiming to predict the total nitrogen in effluent, diagnose nitrogen over-limit risks, and optimize the control strategies. Taking a full-scale WWTP as a case study, a process time-lag simulation-enhanced machine learning model (PTLS-ML) was developed, achieving R 2 values of 0.94 and 0.79 for the training and testing sets, respectively. The proposed model successfully identified the potential reasons of nitrogen over-limit risks under different influent conditions and operational parameters, and accordingly provided optimization suggestions. In addition, the multi-objective optimization (MOO) algorithms analysis further demonstrated that maintaining 4-6 mg/L total nitrogen concentration in effluent by adjusting process operational parameters can effectively balance multiple objectives (i.e., effluent water quality, operating costs, and greenhouse gas emissions), achieving coordinated optimization. This framework can serve as a reference for stable operation, energy-saving, and emission reduction in the nitrogen removal of WWTPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Hospital wastewater is a major hotspot for the spread of antimicrobial resistance (AMR) in aquatic ecosystems. This study aimed to investigate the prevalence of antibiotic resistance genes (ARGs) and their correlation with crAssphage in a hospital wastewater treatment plant (HWWTP) and a receiving canal. Water samples were analyzed for 94 ARGs and crAssphage relative to the 16S rRNA using high-throughput quantitative polymerase chain reaction (HT-qPCR). Subsequently, 7 ARGs and crAssphage were selected and quantified using qPCR. The results showed that the detected genes ranged from 79 to 93 out of 95 genes. The raw wastewater (WW) samples had the highest gene diversity compared to the upstream canal, which had less diversity than downstream samples, as determined by HT-qPCR. The bla GES was the most abundant in WW samples, while qacEΔ1, merA, IS6100, tnpA, and IS26 showed high prevalence throughout the treatment processes. The concentrations of intI1, sul1, bla TEM, bla NDM, bla VIM, tetQ, mcr-1, crAssphage, and 16S rRNA, measured using qPCR, were the highest in WW and significantly reduced in treated water samples. Although some water quality parameters, such as total suspended solids and dissolved oxygen, did not significantly differ before and after treatment, removal efficiency ranged from 0.60 to 3.23 log reduction values (LRV). The highest LRV was observed for the tetQ, whereas the mcr-1 had the lowest LRV. Strong positive correlations among the absolute concentrations of ARGs and crAssphage were observed (Spearman's rho = 0.6-1.0), and biochemical oxygen demand correlated with bla TEM and bla VIM (Spearman's rho = 0.6). These results indicate that crAssphage and water quality could reflect the distribution of other ARGs throughout the HWWTP. Further studies are needed to underscore the importance of monitoring ARGs and genetic markers such as crAssphage in HWWTPs and their receiving waters to enhance our understanding of ARG distribution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cyclic methylsiloxanes (CMSs), widely found in wastewater treatment plants (WWTPs), are potentially hazardous to the environment and human health. In this study, the environmental behavior and human exposure risks of three CMSs (D4-D6) were evaluated in WWTPs located in Beijing and Kunming, Yunnan province. D5 had the highest concentrations in air, water, and sludge, with seasonal variation that consisted of a high concentration in summer and low concentration in winter. The CMS concentrations in air were 3-4-fold higher in the A 2 /O (Anaerobic-Anoxic-Oxic) treatment units than in the other units. CMS emissions to air, soil, and water from the Beijing WWTP were in the ranges of 3.4 × 10 4 -5.0 × 10 4  kg·a -1 , 4.5 × 10 2 -7.5 × 10 2  kg·a -1 , and 2.5 × 10 2 -2.9 × 10 2  kg·a -1 , constituting 98 %, 1.3 %, and 0.7 % of the total emissions, respectively. Total daily inhalation exposure doses of CMSs (ADD inh,CMSs ) associated with four different jobs in WWTPs showed that wastewater treatment technicians had the highest ADD inh,CMSs (51 μg/kg/day), indicating that these people had the highest occupational exposure risk in WWTPs. Therefore, this study identified that atmospheric emission was the main environmental fate of CMSs in WWTPs, and provide a basis for the improvement of WWTP process and risk management decisions. ENVIRONMENTAL IMPLICATION: Assessing the environmental fate and occupational exposure risk of cyclic methylsiloxanes (CMSs) found in wastewater treatment plants (WWTPs) is crucial. This is the first study to identify that atmospheric emission was the main environmental fate of CMSs in WWTPs, especially D5; the inhalation exposure doses of CMSs were all significantly higher in the occupational population working in WWTPs. The results described in our study will help enhance the understanding of current knowledge base of environmental fate and exposure risk of CMSs in WWTPs, and provide a basis for the improvement of WWTP process and risk management decisions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Emerging contaminants are pervasive in aquatic environments globally, encompassing pharmaceuticals, personal care products, steroid hormones, phenols, biocides, disinfectants and various other compounds. Concentrations of these contaminants are detected ranging from ng/L to μg/L. Even at trace levels, these contaminants can pose significant risks to ecosystems and human health. This article systematically summarises and categorizes data on the concentrations of 54 common emerging contaminants found in the influent and effluent of wastewater treatment plants across various geographical regions: North America, Europe, Oceania, Africa, and Asia. It reviews the occurrence and distribution of these contaminants, providing spatial and causal analyses based on data from these regions. Notably, the maximum concentrations of the pollutants observed vary significantly across different regions. The data from Africa, in particular, show more frequent detection of pharmaceutical maxima in wastewater treatment plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The increasing levels of industrialization and urbanization have led to the generation of significant amounts of wastewater and waste products, often containing chemicals like per- and poly-fluoroalkyl substances (PFASs) commonly found in consumer products. PFASs are known for their persistence, ubiquity, and ecotoxicological impacts, raising concerns about potential harm to ecosystems. This paper reports the occurrence and evaluates the ecological risks of PFASs in aquatic ecosystems and wastewater treatment works (WWTWs) across Africa. We reviewed 32 papers published in the period 2009-2024 and identified a total of 35 PFAS compounds in surface waters, wastewater, sediments, fish, crocodiles, and invertebrates. Much of the reported studies came from South Africa, followed by Kenya and Nigeria. PFAS concentrations in Africa were <0.7-390.0 ng L -1 in surface waters, 0.05-772 ng g -1 dw in sediments, and <0.2-832 ng L -1 in wastewater, while the highest levels in fish and invertebrates were 460.7 and 35.5 ng g -1 ww, respectively. The PFAS levels were in the same range of data as those reported globally. However, the high concentrations of PFASs in sediments and wastewater suggest areas of point contamination and a growing risk to aquatic ecosystems from effluent discharges. Calculated risk quotients suggested that, in Africa, organisms in river systems face greater risks due to exposure to PFASs compared to those in lakes, while marine organisms might face higher risks compared to freshwater organisms. Future studies should focus on PFAS contamination sources, especially WWTWs, as emerging sources of PFASs in aquatic systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study is the first of its kind in terms of focusing on the seasonal monitoring of bromine species (bromide - and bromate) and some of the main physicochemical parameters in the surface water of stations inside and in front of the El Noubareya and El-Umum drains that flow directly or indirectly to the Egyptian Mediterranean coast at A (El Noubareya Drain) and B (El-Mex Bay) sites. Among the bromine species, bromate (BrO 3 - ) is a disinfection byproduct considered by many international agencies to have a potential carcinogenic effect in humans and is also known to be ecologically toxic to aquatic organisms. Drain water samples collected from the studied sites A and B had a bromide/chlorinity ratio (3.85E-03 - 6.25E-03 and 3.27E-03 - 6.97E-03, respectively) significantly higher than the typical value for open seawater (3.50E-03), showing significant dilution with wastewater at drain stations in the investigated sites. The source and origin of bromine species and the major ions studied associated with the wastewater units were identified and tracked by calculating the ion/chlorinity ratio and multivariate analysis. The total hazard quotient (THQ) for bromate intake and dermal exposure in children, females, and males demonstrates negligible harm to human health. The toxic unit (TU) and the sum of toxic units (STU) values of the three trophic levels in the surface water for the two sites under investigation yielded approximately comparable values for risk quotient (RQ) and mixture risk characterization ratios (RCR mix(MEC/PNEC) ), indicating that invertebrates are more sensitive to bromate dangers than fish and algae. The study highlights the importance of conducting large-scale laboratory tests on the effluents resulting from wastewater treatment units, including bromide levels, to prevent the formation of dangerous side compounds such as bromate, which may have negative effects on populations and may lead to the toxicity of trophic levels in ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The disposal of sewage water in Gaza City has emerged as a significant issue with extensive environmental repercussions. This study seeks to investigate the pollution plume resulting from the discharge of untreated or partially treated sewage water through Gaza City's main outlet into the Mediterranean Sea. Additionally, it aims to predict scenarios for various design configurations of submerged outfalls with either single-port or multi-port diffusers and compare these scenarios against the Environmental Protection Agency (EPA) recreational water quality criteria. According to the EPA, the concentration of Enterococci bacteria at the edge of the mixing zone should not exceed 35 CFU/100 ml to minimize the adverse environmental impact on the marine ecosystem. CORMIX software was utilized as a modeling tool to simulate the dispersion and attenuation behavior of pollutants resulting from this process and to conduct a sensitivity analysis to optimize the design configuration of the sewage disposal system. The simulation considered the influence of ambient conditions (ambient velocity, wind, and seawater density), effluent characteristics (density, flow rate, pollutant concentration, and pollutant decay rate), outfall configuration, and sea bathymetry. Simulation results indicate that a single-port diffuser is unsuitable according to EPA recreation standards. Multi-port unidirectional diffusers, extending 490 m from the shore into the water, meet the required standards. To a lesser extent, the multi-port staged distributor also meets the standards and is more recommended for counter-current situations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Ship-generated greywater contains a variety of pollutants which, through various pathways, usually are discharged into the sea. To understand the seasonal variation in greywater volumes, the contaminant concentrations in, and the potential hazard of, ship-generated greywater streams, a four-phase strategy for sampling, characterization and hazard assessment of greywater was developed and implemented. Eight greywater streams, sampled from five ships, were characterized for selected pollutants. The metals Zn, Cu, Mn and the metalloid, As, collectively contributed 98 % to the Hazard Index. Laundry greywater had the highest average concentration of phosphorus (42 mg/l) while galley greywater had the highest average concentration of nitrogen (30 mg/l). The geometric means of COD-Cr, BOD 5 , TSS and P exceeded the IMO resolution MEPC 227(64) guideline values for sewage effluent from Advanced Wastewater Treatment Plants. The results establish the basis for and contribute to discussions on, the optimization of ship-generated greywater management and the establishment of potential regulatory strategies in the Baltic Region., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kent Salo reports financial support was provided by Swedish Agency for Marine and Water Management. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Alkaline anaerobic fermentation is an effective approach for resource utilization and reduction of waste activated sludge (WAS). Perfluorooctanoic acid (PFOA) is widespread in WAS, however, its potential impact on alkaline anaerobic fermentation of WAS remains largely unknown. Hence, this study focused on investigating the influence of PFOA on volatile fatty acids (VFAs) production and sludge reduction during alkaline anaerobic fermentation (pH = 10 ± 0.1), as well as the critical mechanisms. Results demonstrated that low PFOA concentration (5 mg/kg-TS) raised VFAs yield to 109.37%, while high levels of PFOA (25 and 50 mg/kg-TS) remarkably decreased VFAs production to 89.55% and 80.44% of the control. Mechanism exploration revealed that PFOA facilitated the solubilization process, and low PFOA level enhanced the accumulation of VFAs via increased bioavailable substrates and the activities of enzymes and microorganisms. On the contrary, high levels of PFOA were substantial biotoxicity, inducing excessive ROS production, causing oxidative damage, and reducing enzyme activity and functional microbial abundance, thereby decreasing VFAs production. Additionally, further analysis of sludge physicochemical properties confirmed that the effect of PFOA on WAS reduction exhibited the same trend as VFAs production. This work provides a basis for PFOA environmental risk assessment and WAS resource utilization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The land application of biosolids as a management practice is considered a beneficial use for improving crop yield and reducing the need for other fertilizers. PFAS enter wastewater treatment plants through collection networks, including industrial discharges, the use of PFAS-containing products, and runoff. Therefore, PFAS may be present in biosolids derived from sewage sludge. The objectives of this study were to evaluate PFAS levels in biosolids samples collected at two wastewater treatment plants operated by the Miami Dade Water and Sewer Department (MDWASD): (1) the South District Wastewater Treatment Plant (SDWWTP) which received landfill leachate and (2) the Central District Wastewater Treatment Plant (CDWWTP). Sludge samples were collected after thickening, anaerobic digestion, and dewatering processes. The samples were subjected to batch leaching tests for 30 days. After the leaching tests, the PFAS levels in the liquid and solid fractions were analyzed for 40 PFAS. The findings show that during the aeration process (i.e., activated sludge process), PFAS are removed from the wastewater and accumulate on the solids. When the thickened sludge is digested, some PFAS are released to the liquid phase as the volatile solids decompose. During the dewatering process by centrifugation, PFAS that are partitioned to the liquid phase are removed, reducing PFAS content in the dewatered biosolids. Of the 40 PFAS analyzed, 24 were detected in leachate or solid residue samples. Samples from the SDWWTP had higher levels of PFAS due to the contribution from landfill leachate discharged to this facility. The partitioning of PFAS between the liquid phase and solid residue after 30 days of mixing indicates that the majority of PFAS in the biosolids are highly soluble and have a high tendency to be mobilized (by runoff, irrigation, precipitation) after land application. The fate profiles of PFAS biosolids were evaluated in terms of their solubility and retardation characteristics., Competing Interests: Declaration of competing interest No conflict of interest exists. I wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aquatic pollution from pharmaceuticals is a growing environmental concern globally, particularly in Catalonia's primary water bodies, the Llobregat and Besòs rivers. This study investigates pharmaceutical residues in reclaimed water effluents from the Llobregat River and a wastewater treatment plant (WWTP) in the Besòs River, critical contributors to the region's water resources. Employing LC-MS/MS, 85 pharmaceutical residues were monitored, revealing elevated concentrations of tramadol, losartan, and gemfibrozil, commonly prescribed drugs in Catalonia. Surprisingly, downstream concentrations exceeded upstream levels significantly, indicating the adverse impact of reclaimed water on water quality. Furthermore, evaluation of WWTP efficiency displayed varying removal rates, from 10 % to 99.8 %, highlighting treatment inadequacies for certain compounds. Predictive environmental concentrations (PECs) aligned closely with measured values, affirming the utility of predictive models in early-stage research. Risk assessment via the risk quotient (RQ) method identified atorvastatin and chlorpromazine as surpassing toxicity thresholds. This study underscores the urgent need to address pharmaceutical contamination in urban rivers and reclaimed waters in Catalonia. By highlighting treatment inefficacies and potential ecological risks, it contributes to the development of sustainable water management strategies and environmental conservation efforts in the region. Efforts should focus on continuously monitoring specific compounds, evaluating their individual toxicity, and implementing appropriate remediation techniques in WWTPs to safeguard water quality and aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Cristian Gomez-Canela reports financial support was provided by Spain Ministry of Science and Innovation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Didecyldimethylammonium chloride (DDAC), a toxic quaternary ammonium compound (QAC) linked to multidrug resistance, is used widely in households and hospitals and on swine farms to prevent disease transmission. However, little is known about its occurrence in watersheds receiving livestock wastewaters or manure. We monitored DDAC and tracers (veterinary and human drugs) once a season over a year at 14 sites in a Japanese watershed where swine outnumbered humans 1.2 to 1 and where both swine and human wastewaters were largely treated on site. DDAC concentrations in sewage-treatment-plant effluent (33-52 ng/L) were close to, whereas those in river waters (3.6-16,672 ng/L) far exceeded, those reported worldwide. DDAC mass flows at the catchment outlet (1692-3816 μg/s) were higher than those of any of the drugs. DDAC concentrations were significantly correlated with total concentrations of veterinary drugs (Spearman's correlation coefficient, 0.95, P < 0.01), indicating that the major pathway of DDAC entry to surface waters was via effluent discharge from swine farms. Comparison of observed and predicted mass flows implied that a substantial percentage of DDAC was washed from the barn floor into swine excrement. To our knowledge, this is the first study to demonstrate QAC hotspots attributable to animal husbandry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Viruses are the most abundant yet understudied members that may influence microbial metabolism in activated sludge treating antibiotic production wastewater. This study comprehensively investigated virome community characteristics under the selection pressure of nine types and different concentrations of antibiotics using a metagenomics approach. Of the 15,514 total viral operational taxonomic units (tOTUs) recovered, only 37.5 % were annotated. Antibiotics altered the original viral community structure in activated sludge. The proportion of some pathogenic viral families, including Herpesviridae&#95;like, increased significantly in reactors treating erythromycin production wastewater. In total, 16.5 % of the tOTUs were associated with two or more hosts. tOTUs rarely carried antibiotic resistance genes (ARGs), and the ARG types in the tOTUs did not match the ARGs carried by the bacterial hosts. This suggests that transduction contributes little to the horizontal ARG transfer. Auxiliary metabolic genes (AMGs) were prevalent in tOTUs, and those involved in folate biosynthesis were particularly abundant, indicating their potential to mitigate antibiotic-induced host damage. This study provides comprehensive insights into the virome community in activated sludge treating antibiotic production wastewater and sheds light on the potential role of viral AMGs in mitigating antibiotic-induced stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In recent years especially during COVID-19, the increased usage of antiviral drugs has led to increased interest in monitoring their presence in wastewater worldwide. In this study, it was examined the occurrence, fate and environmental risks of favipiravir which is used for COVID-19 treatment in two wastewater treatment plants (WWTPs) with different treatment processes in Istanbul, Turkey. Favipiravir was measured in WWTPs influent samples, effluent samples and sludge samples with maximum concentrations of 97 μg/L, 64.11 μg/L and 182.47 μg/g, respectively. Favipiravir had removal efficiency below 55 % for both WWTPs. Mass balance analysis showed that favipiravir removal in WWTPs mainly attributed to biodegradation/biotransformation. Statistical analysis revealed a significant correlation between favipiravir concentration and COVID-19 incidence in Istanbul. The microbial distribution analysis indicated that comparison of collected COVID-19 pandemic sludge and post-pandemic period sludge samples, a noteworthy reduction in the Chloroflexi and Actinobacteriota phyla at the phylum level was observed. Environmental risk assessment using risk quotients ranged from 168 to 704, indicating that the presence of this antiviral drug posed significant ecological risks to aquatic organisms. The study concluded that WWTPs were releasing antiviral drugs into the environment, thereby posing risks to both the aquatic ecosystem and public health. The results of this study demonstrate the persistence of favipiravir in WWTPs and offer crucial supporting data for further research into the advancement of wastewater treatment technology. Also, this study shows wastewater based monitoring is supplementary and early warning system for determining the occurrence of antiviral drugs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bacterial assembly and key taxa during long-term acclimatization in refractory wastewater treatment systems is of paramount importance for optimizing system performance and improving management strategies. Therefore, this study employed high-throughput sequencing, coupled with machine learning models and statistical analysis approaches, to comprehensively elucidate key features of bacterial communities and assembly processes in pesticide wastewater treatment systems. A nine-month monitoring showed substantial variation in diversity and composition of bacterial community between two interconnected biological treatment units (designated as BA and PA). Dynamics of bacterial communities in both units were similar. Moreover, water quality played crucial roles in regulating the bacterial community structure of activated sludge, which were primarily driven by deterministic patterns. Homogeneous selection contributed to 62.85 % and 64.43 % of the variations in BA and PA samples, respectively. Additionally, network analysis revealed significant modularity in bacterial compositions in both groups. Linear regression analysis identified major bacterial modules associated with metabolism and degradation functions. Notably, Module 2 in PA samples has significant positive correlations with functions related to metabolism of nucleotide, amino acid, and xenobiotics, as well as benzoate degradation. Furthermore, key taxa in ecological modules identified by Random Forest model, such as Pseudomonas, Sphingobium, and PHOS-HE28, were dominant populations with metabolism and degradation functions. Particularly, Sphingobium, appeared to be a potential multifunctional degrading bacterium, related to amino acid and xenobiotics metabolism, as well as fatty acid, valine, leucine, isoleucine, fluorobenzoate, and aminobenzoate degradation. These findings are important for developing operating strategies to maintain stable system performance during refractory wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Per- and polyfluoroalkyl substances (PFAS) are a class of chemicals present in a wide range of commercial and consumer products due to their water-repellency, nonstick, or surfactant properties, resulting from their chemical and thermal stability. This stability, however, often leads to persistence in the environment when they are inevitability released. We utilized microbial microcosms from wastewater treatment plant (WWTP) sludge to determine how employing different functional groups such as heteroatom linkages, varying chain lengths, and hydrofluoroethers (HFEs) will impact the ultimate fate of these novel PFAS structures. A suite of five novel fluorosurfactant building blocks (F 7 C 3 OCHFCF 2 SCH 2 CH 2 OH (FESOH), F 3 COCHFCF 2 SCH 2 CH 2 OH (MeFESOH), F 7 C 3 OCHFCF 2 OCH 2 CH 2 OH (ProFdiEOH), F 7 C 3 OCHFCF 2 CH 2 OH (ProFEOH), and F 3 COCHFCF 2 OCH 2 CH 2 OH (MeFdiEOH)) and their select transformation products, were incubated in WWTP aerobic microcosms to determine structure-activity relationships. The HFE alcohol congeners with a thioether (FESOH and MeFESOH) were observed to transform faster than the ether congeners, while also producing second-generation HFE acid products (F 7 C 3 OCHFC(O)OH (2H-3:2 polyfluoroalkyl ether carboxylic acid [PFECA]) and F 3 COCHFC(O)OH (2H-1:2 PFECA). Subsequent biodegradation experiments with 2H-1:2 PFESA and 2H-1:2 PFECA displayed no further transformation over 74 days. Surface water Photofate experiments compared 2H-1:2 PFECA, and 2H-1:2 polyfluorinated ether sulfonate (PFESA) with their fully fluorinated ether acid counterparts, and demonstrated the potential for both HFE acid species to completely mineralize over extended periods of time, a fate that highlights the value of studying novel PFAS functionalization. Environ Toxicol Chem 2024;43:2297-2305. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Wastewater treatment facilities can filter out some plastics before they reach the open environment, yet microplastics often persist throughout these systems. As they age, microplastics in wastewater may both leach and sorb pollutants and fragment to provide an increased surface area for bacterial attachment and conjugation, possibly impacting antimicrobial resistance (AMR) traits. Despite this, little is known about the effects of persistent plastic pollution on microbial functioning. To address this knowledge gap, we deployed five different artificially weathered plastic types and a glass control into the final maturation pond of a municipal wastewater treatment plant in Ōtautahi-Christchurch, Aotearoa/New Zealand. We sampled the plastic-associated biofilms (plastisphere) at 2, 6, 26, and 52 weeks, along with the ambient pond water, at three different depths (20, 40, and 60 cm from the pond water surface). We investigated the changes in plastisphere microbial diversity and functional potential through metagenomic sequencing. Bacterial 16S ribosomal RNA genes composition did not vary among plastic types and glass controls ( P = 0.997) but varied among sampling times [permutational multivariate analysis of variance (PERMANOVA), P = 0.001] and depths (PERMANOVA, P = 0.011). Overall, there was no polymer-substrate specificity evident in the total composition of genes (PERMANOVA, P = 0.67), but sampling time (PERMANOVA, P = 0.002) and depth were significant factors (PERMANOVA, P = 0.001). The plastisphere housed diverse AMR gene families, potentially influenced by biofilm-meditated conjugation. The plastisphere also harbored an increased abundance of genes associated with the biodegradation of nylon, or nylon-associated substances, including nylon oligomer-degrading enzymes and hydrolases.IMPORTANCEPlastic pollution is pervasive and ubiquitous. Occurrences of plastics causing entanglement or ingestion, the leaching of toxic additives and persistent organic pollutants from environmental plastics, and their consequences for marine macrofauna are widely reported. However, little is known about the effects of persistent plastic pollution on microbial functioning. Shotgun metagenomics sequencing provides us with the necessary tools to examine broad-scale community functioning to further investigate how plastics influence microbial communities. This study provides insight into the functional consequence of continued exposure to waste plastic by comparing the prokaryotic functional potential of biofilms on five types of plastic [linear low-density polyethylene (LLDPE), nylon-6, polyethylene terephthalate, polylactic acid, and oxygen-degradable LLDPE], glass, and ambient pond water over 12 months and at different depths (20, 40, and 60 cm) within a tertiary maturation pond of a municipal wastewater treatment plant., Competing Interests: The authors declare no conflict of interest.

Water resources play a crucial role in sustaining life on earth yet chemicals of emerging concern (CECs) arising from extensive human applications are an increasing threat towards their existence. In this study, we examined the occurrence, removal and potential risk of CECs found in rivers and wastewater treatment plants (WWTPs) in western Kenya. Samples were prepared by solid-phase extraction and analysed using high performance liquid chromatography-mass spectrometry with a target list of 785 compounds. Out of these, 333 and 352 (influent 322, effluent 265) compounds were quantified in rivers and wastewater respectively, with pharmaceuticals, industrial compounds, and pesticides being frequently detected in both rivers and WWTPs. Compounds with highest concentrations included saccharin (9.9 μg/L), metformin (7.5 μg/L), and oxypurinol (6.5 μg/L) in rivers whereas caffeine (280 μg/L), deoxycholic acid (179 μg/L), 2-oxindole (10.9 μg/L) and ibuprofen (8.1 μg/L) were found at high concentrations in WWTPs. Based on the types of crops grown, samples from maize growing regions recorded the highest number of pesticides (75) which coincided with the spraying season. The WWTP showed the capacity to eliminate some compounds although the removal efficiencies varied greatly with 204 compounds exhibiting an average removal efficiency exceeding 50 %. Based on the risk assessment, crustaceans had the highest potential risk for toxicity with toxic unit (TU) values up to 5.4 driven primarily by diazinon and dichlorvos followed by algae (TU up to 0.07) and fish (TU up to 0.01) in rivers. A similar trend was observed in WWTP with diazinon (TU up to 5.5), diuron (TU up to 0.07) and carbendazim (TU up to 0.006) driving the risk for crustaceans, algae and fish respectively. These findings highlight the significance of surface water and WWTPs as sources and sinks of CECs in the environment translating to potential risks on aquatic organisms and humans., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Faith Kandie reports financial support was provided by UNESCO-TWAS. Faith Kandie reports financial support was provided by Federal Ministry of Education and Research (BMBF). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hospital wastewater is known to contain various pathogenic microorganisms and harmful substances. During the hospital wastewater treatment process, the bioaerosols released may encapsulate these pathogens, leading to human infection. This study undertook an investigation to compare the dispersion characteristics and seasonal variations of bioaerosols from hospital and municipal sewage. The results indicated that the airborne bacterial concentration from hospital sewage (119 ± 118 CFU/m 3 ) was higher than municipal sewage (46 ± 19 CFU/m 3 ), with the highest concentration observed in summer. The dominant bacterial genera present in bioaerosols from both sewages were alike, with the proportions varied by sewage types and the structure mainly influenced by seasonal factors. Bacteroides, Escherichia-Shigella and Streptococcus were identified as the most prevalent pathogenic genera in spring, summer and winter bioaerosols, respectively, while Pseudomonas and Acinetobacter were abundant in autumn. Although the non-carcinogenic risk associated with bioaerosols was low (<1), the presence of pathogenic species and their potential synergistic interactions elevated the overall exposure risk. The diffusion modeling results demonstrated that bioaerosol emissions from the surface of hospital sewage can reach up to 10570 CFU/m 3 in summer and can spread more than 300 m downwind. The potential pathogenicity of bioaerosols was also highest in summer, which may pose a health hazard to populations located downwind. Therefore, the management and control of bioaerosols from sewage should be strengthened, especially in summer., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Coastal cities, as hubs of social and economic activity, have witnessed rapid urbanization and population growth. This study explores the transformative changes in urban municipal wastewater treatment practices and their profound implications for greenhouse gas (GHG) emissions in Chinese coastal provinces. The approach employed in this study integrates comprehensive data analysis with statistical modeling to elucidate the complex interplay between urbanization, wastewater treatment practices, and GHG emissions. Results reveal a substantial surge in GHG emissions from coastal wastewater treatment, rising from 3367.1 Gg CO 2 e/yr in 1990-23644.8 Gg CO 2 e/yr in 2019. Spatially, the top 20 cities contribute 56.0% of emissions, with hotspots in the Bohai Sea Region, Yangtze River Delta, and Pearl River Delta. Initially dominated by emissions from untreated wastewater, post-2004, GHG emissions from treatment processes became the primary source, tied to electricity use. Growing population and urbanization rates escalated wastewater discharge, intensifying GHG emissions. From 1990 to 2019, average GHG intensity ranged between 320.5 and 676.6 g CO 2 e/m 3 wastewater, with an annual increase of 12.3 g CO 2 e/m 3 . GHG intensity variations relate to the wastewater treatment rate, impacting CH 4 , N 2 O, and CO 2 emissions, underscoring the need for targeted strategies to mitigate environmental impact., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Hospital wastewaters (HWWs) serve as critical reservoirs for disseminating antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB). However, the dynamics and noteworthy shifts of ARGs and their associated pathogenicity, mobility, and resistome risks during HWWs treatment processes remain poorly understood. Utilizing metagenomic sequencing and assembly, we identified 817 ARG subtypes conferring resistance to 20 classes of antibiotics across 18 HWW samples from influent to effluent. Genes encoding resistance to multidrug, aminoglycoside and beta&#95;lactam were the most prevalent ARG types, reflecting patterns observed in clinical settings. On-site treatment efforts decreased the relative abundance of ARGs by 77.4% from influent to secondary sedimentation, whereas chlorine disinfection significantly increased their abundance in the final effluent. Deterministic processes primarily drove the taxonomic assembly, with Proteobacteria being the most abundant phylum and serving as the primary host for 15 ARG types. Contig-based analysis further revealed 114 pathogenic ARB, with Escherichia coli, Pseudomonas alcaligenes, and Pseudomonas aeruginosa exhibiting multidrug-resistant. The contributions of host bacteria and pathogenic ARB varied throughout wastewater treatment. In addition, 7.10%-31.0 % ARGs were flanked by mobile genetic elements (MGEs), predominantly mediated by transposase (74.1%). Notably, tnpA exhibited the highest potential for ARG dissemination, frequently co-occurring with beta-lactam resistance genes (35.2%). Considering ARG profiles, pathogenic hosts, and transferability, raw influent exhibited the highest antibiotic resistome risk index (ARRI), followed by the final effluent. Chlorine disinfection exacerbated resistome risks by inducing potential pathogenic ARB and mobile ARGs, posing threats to the receiving environment. This study delineates ARG occurrence patterns, highlights mechanisms of ARG carriage and horizontal gene transfer, and provides insights for assessing resistance risks and prioritizing interventions in clinical settings., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Efforts to regulate and monitor emerging contaminants are insufficient because new chemicals are continually brought to market, and many are unregulated and potentially harmful. Domestic wastewater treatment plants are not designed to remove micropollutants and are important sources of emerging contaminants in the aquatic environment. In this study, non-target screening, an unbiased method for analyzing compounds without prior information, was used to identify compounds that may be emitted in wastewater treatment plant effluent and should be monitored. Nine wastewater treatment plants using different treatment methods were studied, and a non-target screening data-processing method was used. The frequencies at which the contaminants were detected and contaminant persistence through the treatment processes were considered, and then the contaminants were prioritized. The predicted no-effect concentration of each prioritized contaminant was used to determine whether further analysis and monitoring of the contaminant was necessary. Quantitative analyses of five compounds (amantadine, atenolol, benzotriazole, diphenhydramine, and sulpiride) were performed using reference standards. Probable molecular formulae and structures were proposed for 17 contaminants, and the risks posed by the contaminants were estimated using predicted no-effect concentrations. The results provide valuable insights into how unregulated micropollutants can be identified and prioritized for monitoring in future studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Globally, pharmaceuticals and personal care products (PPCPs) are detected in surface waters receiving wastewater, yet their presence in biota, remain largely understudied. To address this, we conducted a study that measured 46 PPCPs in spot water samples and fish caught up- and downstream from wastewater treatment plants (WWTPs) in Victoria, Australia. We sampled 15 sites located along four waterways following a 3-site design: WWTP-discharge('hotspot'), 'upstream'(∼2 km) and 'downstream'(∼2 km). Spot water and fish were also sampled at reference sites >100 km from WWTP discharge (n = 3). Additionally, spot water samples were taken from WWTP effluent outflows (n = 3). From each locality, we analysed 3-12 fish (n = 131 total). In waterways, passive samplers (POCIS; ∼28d, n = 19 PPCPs) were also deployed. Individual fish (axial muscle) and water were analysed with LC-MS-MS. We found that PPCP concentrations in environmental surface water ranged from<0.02-0.97 μg/L. In WWTP effluent, the range was <0.02-1.4 μg/L. Of the 46 PPCPs analysed, 12 were detected in spot water samples and five in fish. In water, the highest concentration detected was for antidepressant venlafaxine (3 μg/L). The most frequently detected PPCPs: venlafaxine (54.9%), metoprolol (41.2%), propranolol (29.4%), carbamazepine (29.4%), caffeine (17.6%) and sulfamethoxazole (17.6%). Out of 131 fish analysed, 35 fish had detectable levels of PPCPs in the muscle tissue. The highest muscle concentrations were: venlafaxine (150 μg/kg, redfin perch), and sertraline (100 μg/kg, eel). Bioaccumulation factors ranged from 104 to 341L/kg for venlafaxine in redfins, 21-1,260L/kg for carbamazepine in redfins and eels, and 367-3,333L/kg for sertraline in eels. Based on our human health risk calculations for venlafaxine, carbamazepine, sertraline, triclosan, and caffeine, consumption of fish does not pose a significant risk to human health. Despite this, most of the detected PPCPs in surface waters exceeded 10 ng/L trigger value, which has led to further investigations by EPA. Our study highlights the need for using multiple lines of evidence for estimating risks of PPCPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2024. Published by Elsevier Ltd. All rights reserved.)

A diversity of contaminants of emerging concern (CECs) are present in wastewater effluent, posing potential threats to receiving waters. It is urgent for a holistic assessment of the occurrence and risk of CECs related to wastewater treatment plants (WWTP) on national and regional scales. A data mining-based risk prioritization method was developed to collect the reported contaminants and their respective concentrations in municipal and industrial WWTPs and their receiving waters across China over the past 20 years. A total of 10,781 chemicals were reported in 8336 publications, of which 1037 contaminants were reported with environmental concentrations. While contaminant categories varied across WWTP types (municipal vs. industrial) and regions, pharmaceuticals and cyclic hydrocarbons were the most studied CECs. Contaminant composition in receiving water was closer to that in municipal than industrial WWTPs. Publications on legacy pesticides and polycyclic aromatic hydrocarbons in WWTP decreased recently compared to the past, while pharmaceuticals and perfluorochemicals have received increasing attention, showing a changing concern over time. Detection frequency, concentration, removal efficiency, and toxicity data were integrated for assessing potential risks and prioritizing CECs on national and regional scales using an environmental health prioritization index (EHPi) approach. Among 666 contaminants in municipal WWTP effluent, trichlorfon and perfluorooctanesulfonic acid were with the highest EHPi scores, while 17ɑ-ethinylestradiol and bisphenol A had the highest EHPi scores among 304 contaminants in industrial WWTPs. The prioritized contaminants varied across regions, suggesting a need for tailoring regional measures of wastewater treatment and control., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Huizhen Li, Jing You reports financial support was provided by Department of Science and Technology of Guangdong Province. Jing You, Huizhen Li reports financial support was provided by National Natural Science Foundation of China. Jing You reports financial support was provided by Innovative Research Team of Department of Education of Guangdong Province. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Wastewater treatment plants are hotspots for the release of antimicrobial resistant pathogenic bacteria into aquatic ecosystems, significantly contributing to the cycle of antimicrobial resistance. Special attention should be paid to antimicrobial resistant ESKAPE bacteria, which have been identified as high-priority targets for control measures. Among them, Klebsiella pneumoniae is particularly noteworthy. In this study, we collected wastewater samples from the inlet, sedimentation tank, and effluent water of a wastewater treatment plant in June, July, October, and November of 2018. We detected and characterized 42 K. pneumoniae strains using whole genome sequencing (15 from the inlet, 8 from the sedimentation tank, and 19 from the effluent). Additionally, the strains were tested for their antimicrobial resistance phenotype. Using whole genome sequencing no distinct patterns were observed in terms of their genetic profiles. All strains were resistant to tetracycline, meanwhile 60%, 47%, and 37.5% of strains isolated from the inlet, sedimentation tank, and effluent, respectively, were multidrug resistant. Some of the multidrug resistant isolates were also resistant to colistin, and nearly all tested positive for the eptB and arnT genes, which are associated with polymyxin resistance. Various antimicrobial resistance genes were linked to mobile genetic elements, and they did not correlate with detected virulence groups or defense systems. Overall, our results, although not quantitative, highlight that multidrug resistant K. pneumoniae strains, including those resistant to colistin and genetically unrelated, being discharged into aquatic ecosystems from wastewater treatment plants. This suggests the necessity of monitoring aimed at genetically characterizing these pathogenic bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The concern on the fate and distribution of contaminants of emerging concern (CECs) is a burning topic due to their widespread occurrence and potential harmful effects. Particularly, antibiotics have received great attention due to their implications in antimicrobial resistance occurrence. The impact of wastewater treatment plants (WWTP) is remarkable, being one of the main pathways for the introduction of CECs into aquatic systems. The combination of novel analytical methodologies and risk assessment strategies is a promising tool to find out environmentally relevant compounds posing major concerns in freshwater ecosystems impacted by those wastewater effluents. Within this context, a multi-target approach was applied in three Spanish river basins affected by different WWTP treated effluents for spatio-temporal monitoring of their chemical status. Solid phase extraction followed by ultra-high-performance liquid chromatography were used for the quantification of a large panel of compounds (n = 270), including pharmaceuticals and other consumer products, pesticides and industrial chemicals. To this end, water samples were collected in four sampling campaigns at three locations in each basin: (i) upstream from the WWTPs; (ii) WWTP effluent discharge points (effluent outfall); and (iii) downstream from the WWTPs (500 m downriver from the effluent outfall). Likewise, 24-h composite effluent samples from each of the WWTPs were provided in all sampling periods. First the occurrence and distribution of these compounds were assessed. Diverse seasonal trends were observed depending on the group of emerging compounds, though COVID-19 outbreak affected variations of certain pharmaceuticals. Detection frequencies and concentrations in effluents generally exceeded those in river samples and concentrations measured upstream WWTPs were generally low or non-quantifiable. Finally, risks associated with maximum contamination levels were evaluated using two different approaches to account for antibiotic resistance selection as well. From all studied compounds, 89 evidenced environmental risk on at least one occasion in this study., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Pesticides are among the main drivers posing risks to aquatic environments, with effluents from wastewater treatment plants (WWTPs) serving as a major source. This study aimed to identify the primary pesticides for which there was a risk of release into aquatic environments through WWTP effluents, thereby enabling more effective contamination management in public water bodies. In this study, monitoring, risk assessment, and risk-based prioritization of 87 pesticides in effluents from three WWTPs in the Yeongsan River Basin, Korea, were conducted. A total of 59 pesticides were detected at concentrations from 0.852 ng/L to 82.044 μg/L and exhibited variable patterns across different WWTP locations. An environmental risk assessment based on the risk quotient (RQ) of individual pesticides identified 13 substances implicated in significant ecotoxicological risks, as they exceeded RQ values of 1 at least once. An optimized risk (RQ f )-based prioritization, considering the frequency of the measured environmental concentration (MEC) exceeding the predicted environmental concentration (PNEC), was conducted to identify pesticides that potentially posed risks and thus should be managed as a priority. Four pesticides had an RQ f value >1; metribuzin exhibited the highest RQ f value of 4.951, followed by 3-phenoxybenzoic acid, atrazin-2-hydroxy, and atrazine. Additionally, five pesticides (terbuthylazine, methabenzthiazuron, diuron, thiacloprid, and fipronil) and another four pesticides (propazine, imidacloprid, hexaconazole, and hexazione) had RQ f values >0.1 and > 0.01, respectively. By calculating the contributions of individual pesticides to the RQ f of these mixtures (RQ f, mix ) based on the concentration addition model, it was determined that >95 % of the sum of RQ f, mix was driven by the top seven pesticides. These findings highlight the importance of prioritizing pesticides for effective management of contamination sources., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Sang Don Kim reports financial support was provided by National Research Foundation of Korea. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In response to the rapid emergence and dissemination of antimicrobial resistant bacteria (ARB) and genes (ARGs), integrated surveillance systems are needed to address antimicrobial resistance (AMR) within the One Health Era. Wastewater analyses enable biomarker monitoring at the sewershed level, offering timely insights into pathogen circulation and ARB/ARGs trends originating from different compartments. During two consecutive epidemic waves of the COVID-19 pandemic in Portugal, taxonomic and functional composition of raw urban wastewater from two wastewater treatment plants (WWTPs) representing one million in equivalent population, located in the main urban areas of the country, were profiled by shotgun metagenomics. Hospital wastewater from two central hospitals located in the WWTPs catchment areas were also sequenced. The resistome and virulome were profiled using metagenomic assemblies without taxonomic constraint, and then specifically characterized for ESKAPE pathogens. Urban and hospital wastewater exhibited specific microbiota signatures, Pseudomonadota dominated in the first and Bacteroidota in the latter. Correlation network analyses highlighted 85 (out of top 100) genera co-occurring across samples. The most frequent ARGs were classified in the multidrug, tetracyclines, and Macrolides, Lincosamides, Streptogramins (MLS) classes. Links established between AMR determinants and bacterial hosts evidenced that the diversity and abundance of ARGs is not restricted to ESKAPE, being also highly predominant among emergent enteropathogens, like Aeromonas and Aliarcobacter, or in the iron (II) oxidizer Acidovorax. The Aliarcobacter genus accumulated high abundance of sulphonamides and polymyxins ARGs, while Acinetobacter and Aeromonas hosted the highest abundance of ARGs against beta-lactams. Other bacteria (e.g. Clostridioides, Francisella, Vibrio cholerae) and genes (e.g. vanA-type vancomycin resistance) of public health interest were detected, with targeted monitoring efforts being needed to establish informative baseline data. Altogether, results highlight that wastewater monitoring is a valuable component of pathogen and AMR surveillance in healthy populations, providing a community-representative snapshot of public health trends beyond priority pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The abundance and fate of microplastics (MPs) in wastewater treatment plants (WTPs) has been reported extensively. However, in the wastewater, the extent to which hazardous chemicals such as persistent organic pollutants (POPs) accumulated by MPs not been clearly explored. In this study, MPs was sampled from influents and effluents in WTPs to characterize POPs in sorption within MPs. The highest concentrations of PCDD/Fs, PBDD/Fs, PBDEs, and PCBs in sorption within MPs from untreated influents were 5310, 2310, 5,220,000, and 22,700 pg/g, respectively. The most toxic congener, 1,2,3,7,8-PeCDD, accounts for up to 32.3 % of the contribution to PCDD/Fs within MPs. Furthermore, the concentration of PCDD/Fs within MPs from untreated influents could be up to 27.7 times higher than that in microplastic pellets on the coastal beach. This study highlights the quantitative evidence of the POPs within MPs present in untreated influents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Antimicrobial resistance (AMR) is a significant threat that demands surveillance to identify and analyze trends of the emerging antibiotic resistance genes (ARGs) and potential microbial carriers. The influent of the wastewater treatment plants (WWTPs) reflects the microbes derived from the population and effluent being the source of dissemination of potential pathogenic microbes and AMR. The present study aimed to monitor microbial communities and antibiotic resistance genes in WWTPs employing a whole metagenome shotgun sequencing approach. The samples were collected from a sewage treatment plant (STP) and a common effluent treatment plant (CETP) in Delhi, India. The results showed the influent of STP to be rich in Bifidobacterium, Bacteroides, Escherichia, Arcobacter, and Pseudomonas residents of gut microbiota and known to cause diseases in humans and animals; whereas the CETP sample was abundant in Aeromonas, Escherichia, and Shewanella known to be involved in the degradation of different compounds. Interestingly, the effluent samples from both STPs and CETP were rich in microbial diversity, comprising organic and xenobiotic compound degrading and disease-causing bacteria, indicating the effluent being the source of dissemination of concerning bacteria to the environment. The functional profile at both sites displayed similarity with an abundance of housekeeping function genes as analyzed by Clusters of Orthologous Genes (COG), KEGG Orthology (KO), and subsystem databases. Resistome profiling by MEGARes showed the dominance of ARGs corresponding to beta-lactams having relative abundance ranging from 16% to 34% in all the metagenome datasets, followed by tetracycline (8%-16%), aminoglycosides (7%-9%), multi-drug (5%-9%), and rifampin (3%-9%). Also, AMR genes oxa, ant3-DPRIME, and rpoB, which are of clinical importance were predominantly and most prevalently present in all the samples. The presence of AMR in effluents from both types of treatment plants indicates that wastewater from both sources contributes to the spread of pathogenic bacteria and resistance genes, increasing the environmental AMR burden and therefore requires tertiary treatment before discharge. This work will facilitate further research towards the identification of suitable biomarkers for monitoring antibiotic resistance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Water occurrence states in sewage sludge, influenced by sludge physicochemical properties, are crucial for sludge dewaterability and have recently been regarded as a research hotspot. Here, the multifold characteristics of sludge flocs during hydrothermal treatment, including rheological properties, solid-water interfacial interactions, and the polarity distribution and molecular structure of extracellular polymeric substances (EPS), were systematically investigated, and the impact of these characteristics on sludge dewaterability was explored in depth. Hydrothermal treatment at 80 °C and 100 °C induced the conversion of free water into bound water, while an increase in temperature to 180 °C resulted in a significant decrease in bound water content, approximately 4-fold lower than at 100 °C. In addition to the conventional view of decreased sludge surface hydrophilicity at high temperatures, the decline in bound water was associated with the reduction in sludge apparent viscosity. XAD resin fractionation identified the hydrophobic/hydrophilic EPS (HPO-/HPI) ratio as an important factor determining water occurrence states. Especially, hydrolysis of HPI-related hydrophilic proteins and subsequent increase in HPO-related tryptophan-like substances played a dominant role in reducing sludge viscosity and facilitating the release of bound water. Protein conformational analysis revealed that the disruption of α-helix structures and disulfide bonds significantly reduced EPS water-holding capacity, providing strong evidence for the potential of targeting these dense structure units to enhance sludge dewaterability. These findings provide a holistic understanding of multidimensional drivers of water occurrence states in sludge, and guide directions for optimizing sludge treatment efficiency through EPS modification., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Wastewater treatment plants (WWTPs) are considered a significant microplastic discharge source. To evaluate the amount and characteristics of microplastics discharged from WWTPs in South Korea, we selected 22 municipal WWTPs nationally and investigated microplastics at each treatment stage. The mean microplastic removal efficiency by WWTPs was >99%, and most of the microplastics were removed by sedimentation with the second clarifier during wastewater treatment. Consequently, the microplastic removal efficiency of WWTPs did not significantly differ from that of the adopted wastewater treatment technology because a second clarifier was applied in most WWTPs. However, for WWTPs operating a tertiary treatment process, the removal efficiency was enhanced compared with that of WWTPs discharging after a second clarifier. Although the microplastic removal efficiency was high by WWTP, the discharge contribution to the water environment could not be ignored because of the amount of treated wastewater, resulting in an increase of 5.8-270.9 items/m 3 of microplastics in the receiving water. The characteristics of microplastics in WWTPs, including their components, shape, and size, were also evaluated. The most detected components included polytetrafluoroethylene and polyester. Most microplastics detected were categorized as fragments and fibers, while other types were hardly detected. The size of more than 70% of the microplastics detected in WWTPs was under 300 μm, implying that the size of microplastics required to control in WWTPs was much smaller than the defined size of microplastics. An evaluation of the correlation between other pollution factors and microplastic abundance did not reveal positive correlations, and microplastic occurrence was not affected by changing seasons, which may need to be evaluated with further studies. Research should also be performed on the effect of influent sources on the level of microplastic abundance and fate of ultrafine plastics in WWTPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Anammox granular sludge (AnGS) has received considerable attention due to its low carbon footprint (less aeration energy and carbon source consumption) and high biomass density, but growth rate and stability are still the bottlenecks of AnGS process. Calcium ion (Ca 2+ ) is essential for the growth of anaerobic ammonium oxidation bacteria (AnAOB) and plays an important role in the formation and stability of AnGS. Response of AnGS to Ca 2+ under different concentrations was comprehensively investigated by multi-spectral and metagenomics analysis in four aspects: nitrogen removal performance, surface morphology, extracellular polymeric substance (EPS) composition and characterization, and microbial community. The nitrogen removal efficiency was significantly enhanced at appropriate Ca 2+ concentration (2 mmol/L), owning to the more favorable morphology and functional microbial composition of AnGS. However, the nitrogen removal performance of AnGS declined with the Ca 2+ concentration increased from 2 to 8 mmol/L, due to the negative effects of excess Ca 2+ on EPS, mass transfer efficiency, and functional microorganisms. Meanwhile, an unexpected slight "rebound" of nitrogen removal efficiency was observed at Ca 2+ = 6 mmol/L and attributed to the defense mode transformation of AnGS (from "ion stabilization" to "precipitate shield" modes) against excess Ca 2+ stress. Based on the findings, the response mechanism of AnGS to Ca 2+ with different concentrations was established. Our results enhanced the understanding of the interaction between AnGS and Ca 2+ , which may be valuable for filling the theoretical gap in enhancing the granulation and stability of AnGS and providing a reference for the practical operation of the AnGS process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Identifying priority pollutants in wastewater treatment plant (WWTP) effluents is crucial for optimizing monitoring efforts, improving regulations, and developing targeted mitigation strategies. Despite the presence of numerous trace organic pollutants in WWTP effluents, a comprehensive prioritization scheme is lacking, hindering effective control. This study screened 216 micropollutants, including pharmaceuticals, pesticides, and industrial chemicals, which had been detected in effluents from 46 WWTPs across China. A multi-criteria prioritization method was developed, considering exposure potential based on median concentrations and detection frequencies, as well as hazard potential determined by persistence, bioaccumulation, in vitro toxicity, and in vivo toxicity. Pollutants with low exposure or hazard potential were filtered out, and a priority index was calculated to rank the remaining 59 substances. The top 15 priority pollutants included regulated persistent organic pollutants like perfluorooctanoic acid and their alternatives such as perfluorobutane sulfonate, pesticide transformation products, and emerging contaminants such as bisphenol A, which are not currently regulated in WWTP effluents. This study provides a systematic approach to identify priority pollutants and generates a guiding framework for monitoring, regulation, and control of both well-recognized and overlooked contaminants in WWTP effluents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)