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The existence of chlorine-resistant bacteria (CRB) in drinking water supply systems (DWSSs) results in significant challenges to the biological security of drinking water. However, little is known about the intrinsic chlorine-resistant molecular metabolic mechanism of bacteria in DWSSs. This research explored the microbial interactions and the key metabolic pathways that modulate the chlorine resistance of bacteria in full-scale chloraminated DWSSs. The dominant CRB, including Bdellovibrio, Bradyrhizobium, Peredibacter, Sphingomonas, and Hydrogenophaga, strongly interacted with each other to maintain basic metabolism. A total of 4.21% of the bacterial metabolic pathways were key and specific to chlorine-resistant bacteria. Glutaminyl-tRNA biosynthesis was the dominant metabolic pathway of CRB in the target DWSSs. After chloramine disinfection, the relative abundance of glutamate-tRNA ligase (GlnRS) and the related orthologous genes increased by 10.11% and 14.58%, respectively. The inactivation rate of the GlnRS overexpression strain (81.40%) was lower than that of the wild-type strain (90.11%) after exposure to chloramine. Meanwhile, the growth rate of the GlnRS overexpression strain was higher than that of the wild-type strain. Glutaminyl-tRNA biosynthesis can enhance chlorine resistance in DWSSs., 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 © 2022 Elsevier Ltd. All rights reserved.)

Water is an important medium for virus transmission and viral pathogens are increasingly appreciated as a significant water safety issue. However, the effect of pipe biofilms on viral pathogens remains unclear. This research aimed to investigate the dissemination of viruses in a full-scale drinking water supply system (DWSS) and the effect of pipe biofilms on viral pathogens in bulking water. Viral pathogens, pathogenic viral hosts, and viral virulence factors (VFs) were found to disseminate from source water to tap water. The proportion of virus and viral VFs in the biofilm was far less than that in water. The contribution of biofilms in pipe wall to viruses and viral VFs in bulking water was less than 4%, and viruses in the biofilm had no obvious effect on pathogenic viruses in water. Dominant viruses carrying VFs changed from Cyanobacteria virus to Mycobacterium virus after advanced water treatment. Mycobacterium and organics were identified as the key factors influencing composition and abundance of viral VFs, which could explain 41.1% of the variation in viral virulence in the water supply system. Host bacteria and organics may be used as the key targets to control the risk of viruses in DWSSs., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Deterioration of water quality is commonly found in secondary water supply systems (SWSSs), especially the growth of microbes. To explore the metabolic mechanism for rapid microbial regrowth in SWSSs, a regrowth potential assessment, flow cytometry, and quantitative PCR were conducted. Metagenomic and 16S rRNA gene sequencing were performed to better understand the microbial communities and metabolism. It was found that the increased biomass in the SWSS was significantly higher than that in the drinking water distribution system (DWDS). Statistical analysis revealed that ammonia oxidation was the dominant driver of increased biomass in the SWSS. The abundances of ammonia oxidation bacteria, concentration of nitrogen species, and related enzymes demonstrated that ammonia oxidation in the SWSS was more vigorous than that in the DWDS. In the SWSS, the metabolism of the ammonia oxidation cluster was more vigorous, and ammonia-oxidizing bacteria (AOB) were the dominant nitrifying bacteria. Incomplete nitrification products were involved in the metabolism of heterotrophic bacteria and promoted the growth of heterotrophic bacteria in the SWSS. More attention should be given to controlling incomplete nitrification to improve tap water quality., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The mechanism driving the dissemination of antibiotic resistance genes (ARGs) in drinking water supply systems (DWSSs) with multiple barriers remains poorly understood despite several recent efforts. Phosphorothioate (PT) modifications, governed by dndABCDE genes, occur naturally in various bacteria and involve the incorporation of sulfur into the DNA backbone. PT is regarded as a mild antioxidant in vivo and is known to provide protection against bacterial genomes. We combined quantitative polymerase chain reaction, metagenomic, and network analyses for the water treatment process and laboratory-scale experiments for chlorine treatment using model strains to determine if DNA PT modification occurred in DWSS and facilitated the dissemination of mobilized colistin resistance-1 (mcr-1) and New Delhi metallo-β-lactamase-1 (bla NDM-1 ) in DWSS. Our results indicated that the relative abundance of dndB increased in the effluent, compared with the influent, in the water treatment plants. Presence of dndB copies had a positive correlation with the concentration of chloramine disinfectant. Network analysis revealed Bdellovibrio as a potential host for MCR genes, NDM genes, and dndB in the DWSS. E. coli DH10B (Wild-type with the dndABCDE gene cluster and ΔdndB) model strains were used to investigate resistance to chlorine treatment at the concentration range of 0.5-3 mg/L. The resistance of the wild-type strain increased with increasing concentration of chlorine. DNA PT modification protected MCR- and NDM-carrying bacteria from chloramine disinfection during the water treatment process. The higher relative abundance of ARGs in the effluent of the water treatment plants may be due to the resistance of DNA PT modification to chloramine disinfection, thereby causing the enrichment of genera carrying MCR, NDM, and dndB. This study provides a new understanding on the mechanism of ARG dissemination in DWSS, which will help to improve the performance of drinking water treatment to control the risk associated with antibiotic-resistant 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 © 2020 Elsevier Ltd. All rights reserved.)

Mcr-1 and bla NDM-1 antibiotic resistance genes (ARGs) confer resistance to colistins and carbapenems, which are often antibiotics used as a last resort in tertiary care hospitals. Dissemination of these two ARGs in drinking water supply systems and their effect on healthy gut bacteria are poorly studied. In this study, the dissemination of mcr-1 and bla NDM-1 in a drinking water supply system, and their effect on the antibiotic resistance of mouse gut bacteria are explored. Metagenome analysis revealed that source water (Taipu river and Jinze reservoir) was polluted with ARGs. Mcr-1 and bla NDM-1 can be disseminated through the water distribution system. Even advanced water treatments (ozone and biological activated carbon (BAC)) could not effectively remove mcr-1 and bla NDM-1 . Low concentrations of chloramine disinfectants in the water distribution system were not effective at limiting ARG abundance. Mobile genetic elements were also found to play a major role in the dissemination of ARGs via horizontal gene transfer (HGT) throughout the water supply system. Statistical analysis revealed that there was no effect of temperature on the abundance of mcr-1 and bla NDM-1 throughout the water supply system. A last resort ARG, mcr-1 can disseminate from drinking water to the healthy mouse gut. The presence of mcr-1 in a strain belonging to Enterococcus hirae, which is different from the strain belonging to the Bacillus cereus group isolated from drinking water, strongly supports the phenomena of HGT inside the gut. This research provides novel insights into the role of drinking water in disseminating ARGs to the gut and strongly suggests that drinking water may also play a major role apart from other factors known to be involved in the prevalence of last resort ARGs in the gut., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Two waterworks, with source water derived from the Huangpu or Yangtze River in Shanghai, were investigated, and the effluents were plate-screened for antibiotic-resistant bacteria (ARB) using five antibiotics: ampicillin (AMP), kanamycin (KAN), rifampicin (RFP), chloramphenicol (CM) and streptomycin (STR). The influence of water treatment procedures on the bacterial antibiotic resistance rate and the changes that bacteria underwent when exposed to the five antibiotics at concentration levels ranging from 1 to 100 μg/mL were studied. Multi-drug resistance was also analyzed using drug sensitivity tests. The results indicated that bacteria derived from water treatment plant effluent that used the Huangpu River rather than the Yangtze River as source water exhibited higher antibiotic resistance rates against AMP, STR, RFP and CM but lower antibiotic resistance rates against KAN. When the antibiotic concentration levels ranged from 1 to 10 μg/mL, the antibiotic resistance rates of the bacteria in the water increased as water treatment progressed. Biological activated carbon (BAC) filtration played a key role in increasing the antibiotic resistance rate of bacteria. Chloramine disinfection can enhance antibiotic resistance. Among the isolated ARB, 75% were resistant to multiple antibiotics. Ozone oxidation, BAC filtration and chloramine disinfection can greatly affect the relative abundance of bacteria in the community., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study investigates the effect of chloramine residual on bacteria growth and regrowth and the relationship between heterotrophic plate counts (HPCs) and the concentration of chloramine residual in the Shanghai drinking water distribution system (DWDS). In this study, models to control HPCs in the water distribution system and consumer taps are also developed. Real-time ArcGIS was applied to show the distribution and changed results of the chloramine residual concentration in the pipe system by using these models. Residual regression analysis was used to get a reasonable range of the threshold values that allows the chloramine residual to efficiently inhibit bacteria growth in the Shanghai DWDS; the threshold values should be between 0.45 and 0.5 mg/L in pipe water and 0.2 and 0.25 mg/L in tap water. The low residual chloramine value (0.05 mg/L) of the Chinese drinking water quality standard may pose a potential health risk for microorganisms that should be improved. Disinfection by-products (DBPs) were detected, but no health risk was identified.

• Bacteria can enter an intermediate injured but viable state by chlorine disinfection. • Viable injured bacteria can be acquired by FCM cell sorting and cultivation. • DNA injury rather than membrane injury was first repaired before resuscitation. • NHEJ pathway played an important role in repairing DNA double-strand breaks. Chlorine disinfection for the treatment of drinking water can cause injury to the membrane and DNA of bacterial cells and may induce the surviving injured bacteria into a viable but non-culturable (VBNC) state. It is difficult to monitor viable injured bacteria by heterotrophic plate counting (HPC), and their presence is also easily miscalculated in flow cytometry intact cell counting (FCM-ICC). Viable injured bacteria have a potential risk of resuscitation in drinking water distribution systems (DWDSs) and pose a threat to public health when drinking from faucets. In this study, bacteria with injured membranes were isolated from chlorinated drinking water by FCM cell sorting. The culture rate of injured bacteria varied from 0.08% to 2.6% on agar plates and 0.39% to 6.5% in 96-well plates. As the dominant genus among the five identified genera, as well as an opportunistic pathogen with multiple antibiotic resistance, Achromobacter was selected and further studied. After treatment with chlorine at a concentration of 1.2 mg/L, Achromobacter entered into the intermediate injured state on the FCM plot, and the injury on the bacterial surface was observed by electron microscopy. However, the CTC respiratory activity assay showed that 75.0% of the bacteria were still physiologically active, and they entered into a VBNC state. The injured VBNC Achromobacter in sterile drinking water were resuscitated after approximately 25 h. The cellular repair behavior of injured bacteria was studied by Fourier transform infrared attenuated total reflectance (FTIR-ATR) and comet assays. It was found that DNA injury rather than membrane injury was repaired first. The expression of Ku and ligD increased significantly during the DNA repair period, indicating that non-homologous end-joining (NHEJ) played an important role in repairing DNA double-strand breaks. This study deepened the understanding of the effect of chlorine disinfection on bacterial viability in drinking water and will provide support for the improvement of the chlorine disinfection process for the treatment of drinking water. [Display omitted] [ABSTRACT FROM AUTHOR]

Beta-lactamases (bla) are the largest class of antibiotic resistance genes (ARGs) and can survive drinking water treatment processes. However, the key influencing factors of bla genes and the control measures in drinking water supply systems (DWSSs) remain unclear. Quantitative PCR, metagenomic, and 16S rRNA gene sequencing assays were performed to investigate the bla genes in the DWSS. The main carriers of bla genes are genera Caenimonas and Sphingomonas , which were the dominant bacterial genera in the DWSS. The abundance of bla genes was associated with bacterial community assembly events. When the heterotrophic plate count (HPC) in water was below 500 CFU/mL, stochastic assembly was the major contributor to bacterial community assembly and the bacterial community was less affected by the environment. When the HPC in water was above 500 CFU/mL, deterministic assembly played an important role and decreasing the organic matter improved the efficiency of bla genes control. Organic matter can affect bla genes by shifting the diversity of the microbial community, and less organic matter appeared to be beneficial to reducing the bacterial niche width in the DWSS. Nanofiltration (NF) can decrease the effluent organic matter in waterworks and slow the dissemination of bacteria carrying bla genes in the DWSS. [Display omitted] • Less organic matter was beneficial to reducing the bacterial niche width in the DWSS. • It is more effective to control bla in high HPC water with deterministic bacterial assembly. • Organic matter influences bla genes by shifting microbial community diversity. • Nanofiltration is effective for the control of organics and bla genes in the DWDS. [ABSTRACT FROM AUTHOR]

The mechanism driving the dissemination of antibiotic resistance genes (ARGs) in drinking water supply systems (DWSSs) with multiple barriers remains poorly understood despite several recent efforts. Phosphorothioate (PT) modifications, governed by dndABCDE genes, occur naturally in various bacteria and involve the incorporation of sulfur into the DNA backbone. PT is regarded as a mild antioxidant in vivo and is known to provide protection against bacterial genomes. We combined quantitative polymerase chain reaction, metagenomic, and network analyses for the water treatment process and laboratory-scale experiments for chlorine treatment using model strains to determine if DNA PT modification occurred in DWSS and facilitated the dissemination of mobilized colistin resistance-1 (mcr-1) and New Delhi metallo-β-lactamase-1 (bla NDM-1) in DWSS. Our results indicated that the relative abundance of dndB increased in the effluent, compared with the influent, in the water treatment plants. Presence of dndB copies had a positive correlation with the concentration of chloramine disinfectant. Network analysis revealed Bdellovibrio as a potential host for MCR genes, NDM genes, and dndB in the DWSS. E. coli DH10B (Wild-type with the dndABCDE gene cluster and ΔdndB) model strains were used to investigate resistance to chlorine treatment at the concentration range of 0.5–3 mg/L. The resistance of the wild-type strain increased with increasing concentration of chlorine. DNA PT modification protected MCR- and NDM -carrying bacteria from chloramine disinfection during the water treatment process. The higher relative abundance of ARGs in the effluent of the water treatment plants may be due to the resistance of DNA PT modification to chloramine disinfection, thereby causing the enrichment of genera carrying MCR , NDM , and dndB. This study provides a new understanding on the mechanism of ARG dissemination in DWSS, which will help to improve the performance of drinking water treatment to control the risk associated with antibiotic-resistant bacteria. Image 1 • dndB positively correlates with the concentration of chloramine disinfectants. • dndB positively correlates with MCR and NDM in water supply system. • Bdellovibrio is the core genera carrying MCR , NDM and dndB. • DNA PT modification can protect ARB escape chloramine disinfection. Main finding: DNA phosphorothioate modification mediates the dissemination of antibiotic resistance genes, mcr-1 and bla NDM-1 , in drinking water supply system. [ABSTRACT FROM AUTHOR]

Mcr-1 and bla NDM-1 antibiotic resistance genes (ARGs) confer resistance to colistins and carbapenems, which are often antibiotics used as a last resort in tertiary care hospitals. Dissemination of these two ARGs in drinking water supply systems and their effect on healthy gut bacteria are poorly studied. In this study, the dissemination of mcr-1 and bla NDM-1 in a drinking water supply system, and their effect on the antibiotic resistance of mouse gut bacteria are explored. Metagenome analysis revealed that source water (Taipu river and Jinze reservoir) was polluted with ARGs. Mcr-1 and bla NDM-1 can be disseminated through the water distribution system. Even advanced water treatments (ozone and biological activated carbon (BAC)) could not effectively remove mcr-1 and bla NDM-1. Low concentrations of chloramine disinfectants in the water distribution system were not effective at limiting ARG abundance. Mobile genetic elements were also found to play a major role in the dissemination of ARGs via horizontal gene transfer (HGT) throughout the water supply system. Statistical analysis revealed that there was no effect of temperature on the abundance of mcr-1 and bla NDM-1 throughout the water supply system. A last resort ARG, mcr-1 can disseminate from drinking water to the healthy mouse gut. The presence of mcr-1 in a strain belonging to Enterococcus hirae , which is different from the strain belonging to the Bacillus cereus group isolated from drinking water, strongly supports the phenomena of HGT inside the gut. This research provides novel insights into the role of drinking water in disseminating ARGs to the gut and strongly suggests that drinking water may also play a major role apart from other factors known to be involved in the prevalence of last resort ARGs in the gut. Image 1 • mcr-1 and bla NDM-1 were found to disseminate from source to tap in water supply system. • Advanced water treatment process could not effectively remove mcr-1 and bla NDM-1. • Chloramine disinfection had no effect on elimination of mcr-1 and bla NDM-1. • Last resort ARG mcr-1 could disseminate from drinking water to the gut via HGT. • Drinking water is one of the reasons of last resort ARGs transfer in a healthy gut. mcr-1 and bla NDM-1 can disseminate through drinking water supply systems and mcr-1 was found to disseminate from drinking water to the mouse gut. [ABSTRACT FROM AUTHOR]

It is important to comprehensively assess and monitor drinking water quality to ensure a safe and clean drinking water supply. A real-time comprehensive assessment based on modeling and visualized monitoring of urban drinking water quality is discussed in this paper. A weighted aggregative index, a method for evaluating problems with multiple indexes, is used to evaluate the drinking water quality in Shanghai city. Considering the disadvantages of the subjective and objective weighted methods, a combined weighted index method called the geometric mean weighted method is proposed to assign weights to the indexes of drinking water in order to make the weight distribution more scientific, reasonable and robust. Then, the drinking water quality is displayed visually by a distribution map of drinking water quality based on a geographic information system (GIS). This real application shows that the proposed methods are effective and promising for the assessment and monitoring of water quality. [ABSTRACT FROM AUTHOR]