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.

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.

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.

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.

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.

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).

Premise plumbing plays an essential role in determining the final quality of drinking water consumed by customers. However, little is known about the influences of plumbing configuration on water quality changes. This study selected parallel premise plumbing in the same building with different configurations, i.e., laboratory and toilet plumbing. Water quality deteriorations induced by premise plumbing under regular and interrupted water supply were investigated. The results showed that most of the water quality parameters did not vary under regular supply, except Zn, which was significantly increased by laboratory plumbing (78.2 to 260.7 µg/l). For the bacterial community, the Chao1 index was significantly increased by both plumbing types to a similar level (52 to 104). Laboratory plumbing significantly changed the bacterial community, but toilet plumbing did not. Remarkably, water supply interruption/restoration led to serious water quality deterioration in both plumbing types but resulted in different changes. Physiochemically, discoloration was observed only in laboratory plumbing, along with sharp increases in Mn and Zn. Microbiologically, the increase in ATP was sharper in toilet plumbing than in laboratory plumbing. Some opportunistic pathogen-containing genera, e.g., Legionella spp. and Pseudomonas spp., were present in both plumbing types but only in disturbed samples. This study highlighted the esthetic, chemical, and microbiological risks associated with premise plumbing, for which system configuration plays an important role. Attention should be given to optimizing premise plumbing design for managing building water quality.

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.