Although simulated studies have provided valuable knowledge regarding the communities of planktonic bacteria and biofilms, the lack of systematic field studies have hampered the understanding of microbiology in real-world service lines and premise plumbing. In this study, the bacterial communities of water and biofilm were explored, with a special focus on the lifetime development of biofilm communities and their key influencing factors. The 16S rRNA gene sequencing results showed that both the planktonic bacteria and biofilm were dominated by Proteobacteria. Among the 15,084 observed amplicon sequence variants (ASVs), the 33 core ASVs covered 72.8 %, while the 12 shared core ASVs accounted for 62.2 % of the total sequences. Remarkably, it was found that the species richness and diversity of biofilm communities correlated with pipe age. The relative abundance of ASV2 (f_Sphingomonadaceae) was lower for pipe ages 40–50 years (7.9 %) than for pipe ages 10–20 years (59.3 %), while the relative abundance of ASV10 (f_Hyphomonadaceae) was higher for pipe ages 40–50 years (19.5 %) than its presence at pipe ages 20–30 years (1.9 %). The community of the premise plumbing biofilm had significantly higher species richness and diversity than that of the service line, while the steel-plastics composite pipe interior lined with polyethylene (S-PE) harbored significantly more diverse biofilm than the galvanized steel pipes (S-Zn). Interestingly, S-PE was enriched with ASV27 (g_Mycobacterium), while S-Zn pipes were enriched with ASV13 (g_Pseudomonas). Moreover, the network analysis showed that five rare ASVs, not core ASVs, were keystone members in biofilm communities, indicating the importance of rare members in the function and stability of biofilm communities. This manuscript provides novel insights into real-world service lines and premise plumbing microbiology, regarding lifetime dynamics (pipe age 10–50 years), and the influences of pipe types (premise plumbing vs. service line) and pipe materials (S-Zn vs. S-PE).

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Shower systems provide unique environments that are conducive to biofilm formation and the proliferation of pathogens. The water heating temperature is a delicate decision that can impact microbial growth, balancing safety and energy consumption. This study investigated the impact of different heating temperatures (39 °C, 45 °C, 51 °C and 58 °C) on the shower hose biofilm (exposed to a final water temperature of 39 °C) using controlled full-scale shower setups. Whole metagenome sequencing and metaproteomics were employed to unveil the microbial composition and protein expression profiles. Overall, the genes and enzymes associated with disinfectant resistance and biofilm formation appeared largely unaffected. However, metagenomic analysis revealed a sharp decline in the number of total (86,371 to 34,550) and unique genes (32,279 to 137) with the increase in hot water temperature, indicating a significant reduction of overall microbial complexity. None of the unique proteins were detected in the proteomics experiments, suggesting smaller variation among biofilms on the proteome level compared to genomic data. Furthermore, out of 43 pathogens detected by metagenomics, only 5 could actually be detected by metaproteomics. Most interestingly, our study indicates that 45 °C heating temperature may represent an optimal balance. It minimizes active biomass (ATP) and reduces the presence of pathogens while saving heating energy. Our study offered new insights into the impact of heating temperature on shower hose biofilm formation and proposed optimal parameters that ensure biosafety while conserving energy.

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Microplastics (MPs), miniscule plastic particles measuring less than 5 mm in size, have become a concern in terrestrial ecosystems, with primarily agricultural and wetland soils being the soils with highest plastic loadings. The adverse effect of MPs might lead to changes in physicochemical and biological characteristics of soil including soil properties, microbial communities, plants, as well as the potential or affirmed correlations among them. Therefore, understanding the risks and effects of MPs, particularly within the soil-plant-microbe context is challenging and have become a subject of substantial scientific inquiry. This comprehensive review is focused on the effects of MPs on the rhizosphere and plant-microbe symbiotic relationships, with implications for plant growth and ecosystem-level nutrient fluxes. MPs alter soil physicochemical properties, microbial community composition, and enzymatic activities in the rhizosphere, influencing nutrient availability and uptake by plants. These changes in the rhizosphere can disrupt plant-microbe symbiotic interactions, such as mycorrhizal associations and nitrogen-fixing symbioses, ultimately impacting plant growth and the cycling of nutrients within ecosystems. Furthermore, we elaborate on the effects of MPs on the rhizosphere and plant-microbe symbiotic relationships carrying implications for plant growth and ecosystem-level nutrient fluxes. Future research directions and solutions to the microplastics menace acknowledging combined effects of MPs and other contaminants, advanced technologies for MPs identification and quantification, and microbial engineering for MPs remediation. This knowledge of MPs-induced impacts on soil-plant-microbe interactions is essential to generate mitigating actions in soil environmental management and conservation.

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Photodegradation of microplastics (MPs) induced by sunlight plays a crucial role in determining their transport, fate, and impacts in aquatic environments. Dissolved black carbon (DBC), originating from pyrolyzed carbon, can potentially mediate the photodegradation of MPs owing to its potent photosensitization capacity. This study examined the impact of pyrolyzed wood derived DBC (5 mg C/L) on the photodegradation of polystyrene (PS) MPs in aquatic solutions under UV radiation. It revealed that the photodegradation of PS MPs primarily occurred at the benzene ring rather than the aliphatic segments due to the fast attack of hydroxyl radical (•OH) and singlet oxygen (¹O₂) on the benzene ring. The photosensitivity of DBC accelerated the degradation of PS MPs, primarily attributed to the increased production of •OH, ¹O₂, and triplet-excited state DBC (³DBC*). Notably, DBC-mediated photodegradation was related to its molecular weight (MW) and chemical properties. Low MW DBC (<3 kDa) containing more carbonyl groups generated more •OH and ¹O₂, accelerating the photodegradation of MPs. Nevertheless, higher aromatic phenols in high MW DBC (>30 kDa) scavenged •OH and generated more O₂•^-, inhibiting the photodegradation of MPs. Overall, this study offered valuable insights into UV-induced photodegradation of MPs and highlighted potential impacts of DBC on the transformation of MPs.

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Constructed wetlands (CWs) have been proven to effectively immobilize plastic particles. However, little is known about the differences in the impact of varying sized plastic particles on nitrous oxide (N₂O) release, as well as the intervention mechanisms in CWs. Here, we built a lab-scale wetland model and introduced plastic particles of macro-, micro-, and nano-size at 100 μg/L for 370 days. The results showed that plastic particles of all sizes reduced N₂O release in CWs, with the degrees being the strongest for the Nano group, followed by Micro and Macro groups. Meanwhile, ¹⁵N- and ¹⁸O-tracing experiment revealed that the ammoxidation process contributed the most N₂O production, followed by denitrification. While for every N₂O-releasing process, the contributing proportion of N₂O in nitrification-coupled denitrification were most significantly cut down under exposing to macro-sized plastics and had an obvious increase in nitrifier denitrification in all groups, respectively. Finally, we revealed the three mechanism pathways of N₂O release reduction with macro-, micro-, and nano-sized plastics by impacting carbon assimilation (RubisCO activity), ammonia oxidation (gene amo abundance and HAO activity), and N-ion transmembrane and reductase activities, respectively. Our findings thus provided novel insights into the potential effects of plastic particles in CWs as an eco-technology.

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Drinking water biosafety has become an increasing concern for public health. Chlorination is widely used as the main disinfection strategy worldwide but has clear and well-known byproduct issues. The Netherlands has successfully demonstrated unchlorinated approach for almost 20 years but has not been widely adopted by other countries. To chlorine or not chlorine is becoming a critical question in front of all the water utilities. This review aims to provide a good overview of current biosafety management strategies, their disadvantages, as well as the latest developments and future trends. Firstly, the advantages and deficiencies of conventional disinfection and non-disinfection were discussed. Secondly, the commonly used and promising methods for biostability assessment are described. Finally, critical views on the strategy selection for ensuring drinking water biosafety were discussed. It is recommended to achieve both biological and chemical balance by removing pathogens while minimizing the organic matter and dosing a minimum level of disinfectants, which would represent the compromise choice between the current chlorine-based disinfection and chlorine-free strategy. It's worth noting that the complexity of ensuring biosafety lies in the variations among different regions, the selection of suitable methods should be tailored to specific situations on a case-by-case basis.

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Increasing wildfire frequency, a consequence of global climate change, releases incomplete combustion byproducts such as aquatic pyrogenic dissolved organic matter (DOM) and black carbon (DBC) into waters, posing a threat to water security. In August 2022, a series of severe wildfires occurred in Chongqing, China. Samples from seven locations along the Yangtze and Jialing Rivers revealed DBC, quantified by the benzene poly(carboxylic acid) (BPCA) method, comprising 9.5-19.2% of dissolved organic carbon (DOC). High concentrations of BPCA-DBC with significant polycondensation were detected near wildfire areas, likely due to atmospheric deposition driven by wind. Furthermore, Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) revealed that wildfires were associated with an increase in condensed aromatics, proteins, and unsaturated hydrocarbons, along with a decrease in lignins. The condensed aromatics primarily consisted of dissolved black nitrogen (DBN), contributing to abundant high-nitrogen-containing compounds in locations highly affected by wildfires. Meanwhile, wildfires potentially induced the input of recalcitrant sulfur-containing protein-like compounds, characterized by high oxidation, aliphatic nature, saturation, and low aromaticity. Overall, this study revealed the appearance of recalcitrant DBC and dissolved organic sulfur in river waters following wildfire events, offering novel insights into the potential impacts of wildfires on water quality and environmental biogeochemistry.

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This study assessed the evolution of wastewater systems during the rapid urbanization of Beijing, with special focuses on the carbon footprints and growing underground WWTPs (u-WWTPs). Specifically, the Bishui plant (in situ constructed u-WWTP) was assessed in detail regarding eco-environmental benefits. Our results showed that, the direct emission intensity of 65 WWTPs decreased from 0.47 to 0.24 kg CO_2eq/m³, when the electricity intensity increased from 0.22 to 0.39 kWh/m³ from 2010 to 2020. Bishui u-WWTP emitted 36.6 kt CO_2eq/year (0.09 kg CO_2eq/m³), with electricity intensity of 0.43 kg CO_2eq/m³. Additionally, compare to the hypothetical relocating scenario, it saved 6.67 × 10⁴ m² land and 33.0 kt CO_2eq/year, and the created urban river carries 6.5 × 10¹³ J/year heat outside town. The evaluation and balance of choice for conventional or underground WWTP should be made case by case. However, this study demonstrated that u-WWTP is not only a construction manner, but a sustainable management model with positive eco-environment effects, algin with future city expansion, and circular economy visions.

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The excessive use and accumulation of water-soluble polymers (WSPs, known as “liquid plastics”) in the environment can pose potential risks to both ecosystems and human health, but the environmental fate of WSPs remains unclear. Here, the adsorption behavior of WSPs with different molecular weight on kaolinite (Kaol) and montmorillonite (Mt) were examined. The results showed that the adsorption of PEG and PVP on minerals were controlled by hydrogen bond and van der Waals force. The Fourier transform infrared (FTIR) spectra and two-dimensional correlation spectroscopy (2D-COS) analysis revealed that there were interactions between the Al-O and Si-O groups of the minerals and the polar O- or N-containing functional groups as well as the alkyl groups of PEG and PVP. The adsorption characteristics of WSPs were closely related to their molecular weight and the pore size of minerals. Due to the relatively large mesopore size of Kaol, both PEG and PVP were absorbed into inner spaces, for which the adsorption capacity increased with molecular weight of the polymers. For Mt, all types of PEG could enter its micropores, while PVP with larger molecular weights appeared to be confined externally, leading to a decrease in the adsorption capacity of PVP with increasing molecular weight. The findings of this study provide a theoretical basis for scientific evaluation of environmental processes of WSPs.

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Treated drinking water is delivered to customers through drinking water distribution systems (DWDSs). Although studies have focused on exploring the microbial ecology of DWDSs, knowledge about the effects of different water treatments on the bacterial community of biofilm and loose deposits in DWDS is limited. This study assessed the effects of additional treatments on the bacterial communities developed in 10 months’ old pilot DWDSs. The results showed a similar bacterial community in the pipe-wall biofilm, which was dominated by Novosphingobium spp. (20–82 %) and Sphingomonas spp. (11–53 %), regardless of the treatment applied. The bacterial communities that were retained in the distribution systems (including pipe-wall biofilm and loose deposits) were similar to the particle-associated bacteria (PAB) in the corresponding supply water. The additional treatments showed clear effects of the removal and/or introduction of particles. The genera Aeromonas spp., Clostridium spp., Legionella spp., and Pseudomonas spp., which contain opportunistic pathogenic species, were only detected among the PAB in ion exchange system. Our study demonstrated that the biofilm community is consistent across treatments, and the contribution from bacteria in loose deposits is important but can be controlled by removing particles. These findings offer more insight into the origin and development of microbial ecology in DWDSs and suggest paths for further research on the possibility of managing the microbial ecology in distribution systems.

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Drinking water distribution systems are increasingly vulnerable to sewage intrusion due to aging water infrastructure and intensifying water stress. While the health risks associated with sewage intrusion have been extensively studied, little is known about the impacts of intruded bacteria and dissolved organic matter (DOM) on microbiology in drinking water. In this dynamic study, we demonstrate that the intrusion of 1 % sewage into tap water resulted in immediate contamination, including an 8-fold increase in biomass (TCC), a 48.9 % increase in bacterial species (ASVs), a 12.5 % increase in organic carbon content (DOC), and a 13.5 % increase in unique DOM molecular formulae. Over time, sewage intrusion altered tap water microbiology by accelerating bacterial growth rates (5-fold faster), selectively promoting ASVs in community succession, and producing 998 more unique DOM formulae. More significantly, statistical analysis revealed that the intrusion of 1 % sewage shifted the driving force of bacterial and DOM composition covariance from a DOM-dependent process in tap water to a bacterial-governed process post-intrusion. Our results clearly demonstrate the disruptive effects of sewage intrusion into tap water, emphasizing the urgent need to consider the long-lasting impacts of sewage intrusion in drinking water distribution systems, in addition to its immediate health risks.

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Micro- and nanoplastics (MNPs) are attracting increasing attention due to their persistence and potential ecological risks. This review critically summarizes the effects of photo-oxidation on the physical, chemical, and biological behaviors of MNPs in aquatic and terrestrial environments. The core of this paper explores how photo-oxidation-induced surface property changes in MNPs affect their adsorption toward contaminants, the stability and mobility of MNPs in water and porous media, as well as the transport of pollutants such as organic pollutants (OPs) and heavy metals (HMs). It then reviews the photochemical processes of MNPs with coexisting constituents, highlighting critical factors affecting the photo-oxidation of MNPs, and the contribution of MNPs to the phototransformation of other contaminants. The distinct biological effects and mechanism of aged MNPs are pointed out, in terms of the toxicity to aquatic organisms, biofilm formation, planktonic microbial growth, and soil and sediment microbial community and function. Furthermore, the research gaps and perspectives are put forward, regarding the underlying interaction mechanisms of MNPs with coexisting natural constituents and pollutants under photo-oxidation conditions, the combined effects of photo-oxidation and natural constituents on the fate of MNPs, and the microbiological effect of photoaged MNPs, especially the biotransformation of pollutants.@en

The first pandemic wave of coronavirus disease 2019 (COVID-19) induced a considerable increase in several antivirals and antibiotics in surface water. The common symptoms of COVID-19 are viral and bacterial infections, while comorbidities (e.g., hypertension and diabetes) and mental shock (e.g., insomnia and anxiety) are nonnegligible. Nevertheless, little is known about the long-term impacts of comorbidities and mental shock on organic micropollutants (OMPs) in surface waters. Herein, we monitored 114 OMPs in surface water and wastewater treatment plants (WWTPs) in Wuhan, China, between 2019 and 2021. The pandemic-induced OMP pollution in surface water was confirmed by significant increases in 26 OMP concentrations. Significant increases in four antihypertensives and one diabetic drug suggest that the treatment of comorbidities may induce OMP pollution. Notably, cotinine (a metabolite of nicotine) increased 155 times to 187 ng·L⁻¹, which might be associated with increased smoking. Additionally, the increases in zolpidem and sulpiride might be the result of worsened insomnia and depression. Hence, it is reasonable to note that mental-health protecting drugs/behavior also contributed to OMP pollution. Among the observed OMPs, telmisartan, lopinavir, and ritonavir were associated with significantly higher ecological risks because of their limited WWTP-removal rate and high ecotoxicity. This study provides new insights into the effects of comorbidities and mental shock on OMPs in surface water during a pandemic and highlights the need to monitor the fate of related pharmaceuticals in the aquatic environment and to improve their removal efficiencies in WWTPs.

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Pipe materials appear to play an important role in the development of biofilms in drinking water distribution systems. However, there is controversy as to whether pipe materials shape the composition and diversity of bacterial communities in biofilms. To investigate the long-term effects of pipe materials on biofilms, triplicate samples of mature biofilms on unplasticized polyvinyl chloride (PVC-U), grey cast iron and asbestos cement (pipe age > 40 years) were obtained from three areas of an unchlorinated drinking water distribution system in the Netherlands. Illumina sequencing was performed and 773 OTUs (730 OTUs-814 OTUs) were detected within the biofilms on the three pipe materials, all of which were dominated by Proteobacteria (36.2%-46.1%). Both the alpha and beta diversity results showed that the bacterial communities of the biofilms formed on different pipe materials were highly similar. The neutral community model revealed that the assembly of the biofilm communities was governed by environmental selection rather than neutral processes. Among the 142 shared OTUs between the water and biofilm samples, there were 25 enriched OTUs (e.g., OTU7, assigned as Nitrospira spp.), which accounted for 62.6% of the total sequences, while 16 OTUs were disadvantaged (e.g., OTU14 and OTU40, assigned as Hyphomicrobiaceae), accounting for 2.2% of the sequences. Based on the findings, we propose and discuss a harmonisation process by which biofilms with significant differences due to the pipe material harmonize over time resulting in biofilms with similar bacterial communities. Our findings provide valuable insights into long-term biofilm development, bridging an essential gap in our current understanding of the influence of pipe materials on biofilm communities. These findings also highlight the importance of long-term studies and point to a potentially masked harmonizing process during biofilm development over years/decades.

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Biofilm detachment contributes to water quality deterioration. However, the contributions of biofilm detachment from different pipes have not been quantified or compared. Following the introduction of partial reverse osmosis (RO) in drinking water production, this study analyzed particles at customers’ ends and tracked their origins to water distribution mains and service lines. For doing so, filter bags were installed in front of water meters to capture upstream detached particles, while biofilm from water main and service line were sampled by cutting pipe specimens. The results showed that elemental concentrations of the biofilm in mains were higher than those of service lines (54.3–268.5 vs. 27.1–44.4 μg/cm2), both dominated by Ca. Differently, filter bags were dominated by Fe/Mn (77.5–98.1%). After introducing RO, Ca significantly decreased in biofilms of mains but not service lines, but the released Fe/Mn rather than Ca arrived at customers’ ends. The ATP concentrations of service lines were higher than mains, which decreased on mains but increased in service lines after introducing RO. For the core ASVs, 13/24 were shared by service lines (17), mains (21), and filter bags (17), which were assigned mainly to Nitrospira spp., Methylomagnum spp., Methylocytis spp., and IheB2–23 spp. According to source tracking results, service lines contributed more than mains to the particulate material collected by filter bags (57.6 ± 13.2% vs. 13.0 ± 11.6%). To the best of our knowledge, the present study provides the first evidence of service lines’ direct and quantitative contributions to potential water quality deterioration at customers’ ends. This highlights the need for the appropriate management of long-neglected service line pipes, e.g., regarding material selection, length optimization, and proper regulation.@en

The aperiodic changes in the quantity and community of planktonic and particle-associated bacteria have hampered the understanding and management of microbiological water quality in drinking water distribution systems. In this study, online sampling was combined with the microbial fingerprint-based SourceTracker2 to capture and trace the spatiotemporal variations in planktonic and particle-associated bacteria in an unchlorinated distribution system. The results showed that spatially, the particle load significantly increased, while in contrast, the quantity of particle-associated bacteria decreased sharply from the treatment plant to the distribution network. Similar to the trend of particle-associated bacterial diversity, the number of observed OTUs first slightly decreased from the treatment plant to the transportation network and then sharply increased from the transportation network to the distribution network. The SourceTracker2 results revealed that the contribution of particle-associated bacteria from the treatment plant decreased along the distribution distance. The spatial results indicate the dominant role of sedimentation of particles from the treatment plant, while the observed increases in particles and the associated bacteria mainly originated from the distribution network, which were confirmed directly by the increased contributions of loose deposits and biofilm. Temporally, the daily peaks of particle-associated bacterial quantity, observed OTU number, and contributions of loose deposits and biofilms were captured during water demand peaks (e.g., 18–21 h). The temporal results reveal clear linkages between the distribution system harboring bacteria (e.g., within loose deposits and biofilms) and the planktonic and particle-associated bacteria flowing through the distribution system, which are dynamically connected and interact. This study highlights that the spatiotemporal variations in planktonic and particle-associated bacteria are valuable and unneglectable for the widely on-going sampling campaigns required by water quality regulations and/or drinking water microbiological studies.

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The presence of newly emerging pollutants in the aquatic environment poses great challenges for drinking water treatment plants. Due to their low concentrations and unknown characteristics, emerging pollutants cannot be efficiently removed by conventional water treatment processes, making technically, economically, and environmentally friendly water purification technologies increasingly important. This article introduces a one-step reverse osmosis (OSRO) concept consisting of riverbank filtration (RBF) and reverse osmosis (RO) for drinking water treatment. The OSRO concept combines the relatively low-cost natural pretreatment of river water with an advanced engineered purification system. RBF provides a continuous natural source of water with stable water quality and a robust barrier for contaminants. With the pre-removal of particles, organic matter, organic micro-pollutants (OMPs), and microbes, RBF becomes an ideal source for a purification system based on RO membranes, in comparison with the direct intake of surface water. OSRO treatment removes almost 99.9% of the particles, pathogens, viruses, and OMPs, as well as the vast majority of nutrients, and thus meets the requirements for the chlorine-free delivery of drinking water with high biostability. The OSRO treatment is cost effective compared with the standard conventional series of purification steps involving sprinkling filters, softening, and activated carbon. Artificial bank filtration (ABF), which functions as an artificial recharge in combination with a sand filtration system, is proposed as an alternative for RBF in the OSRO concept to supply drinking water from locally available resources. It is also suggested that the OSRO concept be implemented with wind power as an alternative energy source in order to be more sustainable and renewable. An OSRO-based decentralized water system is proposed for water reclaiming and reuse. It is suggested that future water treatment focus on the combination of natural and engineered systems to provide drinking water through technically efficient, financially feasible, resource reusable, and environmentally relevant means.

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Drinking water quality deteriorates from treatment plant to customer taps, especially in the plumbing system. There is no direct evidence about what the differences are contributed by plumbing system. This study compared the water quality in the water main and at customer tap by preparing a sampling tap on the water main. The biomass was quantified by adenosine triphosphate (ATP) and the microbial community was profiled by 454 pyrosequencing. The results showed that in distribution pipes, biofilm contributed >94% of the total biomass, while loose deposits showed little contribution (< 2%) because of the low amount of loose deposits. The distribution of biological stable water had minor effects on the microbiocidal water quality regarding both quantity (ATP 1 ng/L vs. 1.7 ng/L) and community of the bacteria. Whereas the plumbing system has significant contribution to the increase of active biomass (1.7 ng/L vs. 2.9 ng/L) and the changes of bacterial community. The relative abundance of Sphingomonas spp. at tap (22%) was higher than that at water main (2%), while the relative abundance of Pseudomonas spp. in tap water (15%) was lower than that in the water from street water main (29%). Though only one location was prepared and studied, the present study showed that the protocol of making sampling tap on water main offered directly evidences about the impacts of plumbing system on tap water quality, which makes it possible to distinguish and study the processes in distribution system and plumbing system separately.

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Biodegradation plays an important role in the removal of organic micropollutants (OMPs) during riverbank filtration (RBF) for drinking water production. The ability of ammonia-oxidizing microorganisms (AOM) to remove OMPs has attracted increasing attention. However, the distribution of AOM in RBF and its role in the degradation of OMPs remains unknown. In this study, the behavior of 128 selected OMPs and the distribution of AOM and their roles in the degradation of OMPs in RBF were explored by column and batch experiments simulating the first meter of the riverbank. The results showed that the selected OMPs were effectively removed (82/128 OMPs, >70% removal) primarily by biodegradation and partly by adsorption. Inefficiently removed OMPs tended to have low molecular weights, low log P, and contain secondary amides, secondary sulfonamides, secondary ketimines, and benzyls. In terms of the microbial communities, the relative abundance of AOM increased from 0.1%–0.2% (inlet_-sand) to 5.3%–5.9% (outlet_-sand), which was dominated by ammonia-oxidizing archaea whose relative abundance increased from 23%–72% (inlet_-sand) to 97% (outlet_-sand). Comammox accounted for 23%–64% in the inlet_-sand and 1% in the outlet_-sand. The abundances of AOM amoA genes kept stable in the inlet_-sand of control columns, while decreased by 78% in the treatment columns, suggesting the inhibition effect of allylthiourea (ATU) on AOM. It is observed that AOM played an important role in the degradation of OMPs, where its inhibition led to the corresponding inhibition of 32 OMPs (5/32 were completely suppressed). In particular, OMPs with low molecular weights and containing primary amides, secondary amides, benzyls, and secondary sulfonamides were more likely to be removed by AOM. This study reveals the vital role of AOM in the removal of OMPs, deepens our understanding of the degradation of OMPs in RBF, and offers valuable insights into the physiochemical properties of OMPs and their AOM co-metabolic potential.

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