Biofilms In Distribution Systems Research Articles

Establishment and inactivation of mono-species biofilm in a semipilot-scale water distribution system using nanocomposite of silver nanoparticles/montmorillonite loaded cationic chitosan

This study presents a novel approach in the synthesis and characterization of nanocomposites comprising cationic chitosan (CCS) blended with varying concentrations of silver nanoparticles/montmorillonite (AgNPs/MMT). AgNPs/MMT was synthesized using soluble starch as a reducing and stabilizing agent. Subsequently, nanocomposites, namely CCS/AgMMT-0, CCS/AgMMT-0.5, CCS/AgMMT-1.5, and CCS/AgMMT-2.5, were developed by blending 2.5 g of CCS with 0, 0.5, 1.5, and 2.5 g of AgNPs/MMT, respectively, and the corresponding nanocomposites were prepared using ball milling technique. Transmission electron microscopy (TEM) analysis revealed the formation of nanocomposites that exhibiting nearly spherical morphologies. Dynamic light scattering (DLS) measurements displayed average particle sizes of 1183 nm, 131 nm, 140 nm, and 188 nm for CCS/AgMMT-0, CCS/AgMMT-0.5, CCS/AgMMT-1.5, and CCS/AgMMT-2.5, respectively. The narrow polydispersity index (~0.5) indicated uniform particle size distributions across the nanocomposites, affirming monodispersity. Moreover, the zeta potential values exceeding 30 mV across all nanocomposites that confirmed their stability against agglomeration. Notably, CCS/AgMMT-2.5 nanocomposite exhibited potent antibacterial and antibiofilm properties against diverse pipeline materials. Findings showed that after 15 days of incubation, the highest populations of biofilm cells, Pseudomonas aeruginosa biofilm, developed over UPVC, MDPE, DCI, and SS, with corresponding HPCs of 4.79, 6.38, 8.81, and 7.24 CFU/cm2. The highest cell densities of Enterococcus faecalis biofilm in the identical situation were 4.19, 5.89, 8.12, and 6.9 CFU/cm2. The nanocomposite CCS/AgMMT-2.5 exhibited the largest measured zone of inhibition (ZOI) against both P. aeruginosa and E. faecalis, with measured ZOI values of 19 ± 0.65 and 17 ± 0.21 mm, respectively. Remarkably, the research indicates that the youngest biofilm exhibited the most notable rate of inactivation when exposed to a dose of 150 mg/L, in comparison to the mature biofilm. These such informative findings could offer valuable insights into the development of effective antibiofilm agents and materials applicable in diverse sectors such as water treatment facilities, medical devices, and industrial pipelines.

Modelling drinking water biofilms: Bacterial adhesion and Legionella pneumophila necrotrophic growth

We propose a mathematical model to simulate the establishment and growth of a drinking water distribution system biofilm, focusing on the influence of ionic strength on bacterial adhesion and persistence of Legionella pneumophila. Specifically, we consider how ionic strength affects interaction energies during the initial phase of biofilm formation and we include the dependence of the attachment flux, modelled as a linear rate with respect to free floating cells, on it. The model also incorporates a novel necrotrophic kinetics to simulate the Legionella pneumophila metabolism. The biofilm is modelled as a 1D free boundary domain, and its evolution is governed by hyperbolic–parabolic PDEs. The initial attachment phase is modelled by considering a vanishing initial value for the free boundary. The model is investigated numerically highlighting the impact of the necrotrophic kinetic parameters, the influence of ionic strength on the initial stage of biofilm formation through bacterial attachment, and how variations in nutrient levels affect system dynamics. The numerical results demonstrate that: the ionic strength mainly governs bacterial adhesion for young biofilms and affects the biofilm ecology also in presence of a biocide; the necrotrophic metabolism of Legionella pneumophila favours its persistence in oligotrophic biofilms.

Inactivation of biofilm-bound bacterial cells using irradiation across UVC wavelengths

Opportunistic pathogens (OPs), such as Pseudomonas spp., Legionella spp., and mycobacteria, have been detected in biofilms in drinking water distribution systems and water storage tanks and pose potential risks to finished drinking water quality and safety. Emerging UV technologies, such as UV light emitting diodes (LEDs) and krypton chloride (KrCl*) excimers, could provide an alternative to chemical-based secondary disinfection (i.e., chlorine or chloramines) for controlling biofilm-bound OPs. UV systems offer long lifetimes, ability to select wavelength, small size with high power density, and limited-to-no disinfection by-product formation. In this study, inactivation of biofilm-bound Pseudomonas aeruginosa cells across different maturities was investigated using five UVC devices with different peak emission wavelengths, including a KrCl* excimer (222 nm), a low pressure mercury vapor lamp (254 nm), and three UV LEDs (260 nm, 270 nm, and 282 nm). The UV transmittance and absorbance through the biofilm structure was also documented for the first time using a unique approach. Our results show all UVC devices can inactivate biofilm-bound P. aeruginosa cells up to a point, among which the UV LED with peak emission at 270 nm provided the best disinfection performance. UV sensitivities of biofilm-bound cells decreased with biofilm maturity and while initial rates of inactivation were high, no more than 1.5-2.5 log reduction was possible. Re-suspended biofilm bacteria in aqueous solution were highly sensitive to UV, reaching greater than 6 log reduction. UV shielding by biofilm constituents was observed and was likely one of the reasons for UV resistance but did not fully explain the difference in UV sensitivity between biofilm-bound cells versus planktonic cells. This study improves upon fundamental knowledge and provides guidance for innovative designs using emerging UV technologies for biofilm and pathogen control in water distribution systems.

Water and biofilm in drinking water distribution systems in the Netherlands

To keep the high quality of drinking water in the future for non-chlorinated drinking water systems, knowledge about the variables that most strongly affect this quality is necessary in order to know where to focus on and possibly even change aspects of drinking water production and distribution. Therefore, the aim of this study was to investigate which variables (source of drinking water, growth potential and pipe material type) have the biggest influence on bacterial community composition and biomass concentration of drinking water and biofilm in distribution systems. Ten different distribution systems were sampled for water and biofilm, obtained from four different pipe materials, throughout the Netherlands. The distribution systems are supplied either with drinking water produced from groundwater or surface water, and differ in drinking water quality parameters such as the growth potential. We found a significant relationship for growth potential and ATP concentration in water, but for the ATP in the biofilm none of the parameters showed a significant effect. Furthermore, the source of the drinking water and the pipe material did not significantly affect the ATP concentration in water and biofilm. The bacterial composition of in both water and biofilm was significantly different between distribution systems delivering water with low and high growth potential and between drinking water produced from groundwater or surface water. In contrast, the different pipe materials did not significantly affect composition of biofilm-associated communities. We conclude from these results that the growth potential of the treated water best explains the variation in biomass and bacterial composition in water and biofilm of non-chlorinated drinking water distribution systems followed by the drinking water source, whereas pipe materials seem to be of lesser importance.

Inhibiting the increase of antibiotic resistance genes during drinking water distribution by superior microbial interface using Fe modified granular activated carbon

The effects of biological activated carbon treatment using Fe2O3-modified granular activated carbon in inhibiting antibiotic resistance genes in simulated drinking water distribution systems was compared with unmodified granular activated carbon as a reference. Fe2O3-modified biofiltration resulted in a sustained inhibition of resistance genes during drinking water chlorination and distribution (relative abundance in simulated tap water after the modified and unmodified filtration was 1.31% and 9.40%, respectively). A new electron transfer pathway occurring in attached biofilms on Fe2O3-modified granular activated carbon surface, which was identified using X-ray photoelectron spectroscopy and the phenanthroline spectrophotometric method, enhanced the extracellular electron transfer rate and weakened the pressure of organic micropollutants on microorganisms. Hence, the relative abundance of resistance genes (36.32%) and integron (8.79%) on modified carbon was considerably lower than that on unmodified carbon (115.59% and 13.85%, respectively). Meanwhile, the secreted extracellular polymeric substances on modified carbon presented higher flocculating efficiency and better mechanical stability, resulting in the suspended extracellular polymeric substances in downstream water exhibiting stronger electrostatic repulsion. The particle-attached biofilms in downstream distribution systems consistently failed to form larger aggregates, inhibiting horizontal gene transfer, and overall microbial metabolism. Based on network analysis, 11 OTUs in the water samples from raw water to simulated tap water formed an extremely interrelated module with no links to target resistance genes and integron. Therefore, a range of microbial variations triggered by the microbial interface on modified carbon successfully controlled the transfer of antibiotic resistance genes-associated risk from biological activated carbon effluent to tap water. Our findings revealed that enhancing the microbial interface using Fe2O3-modified granular activated carbon is a promising option for inhibiting the antibiotic resistance genes increase in tap water.

Intermittent Water Supply Impacts on Distribution System Biofilms and Water Quality

Intermittent water supplies (IWS) are routinely experienced by drinking water distribution systems around the world, either due to ongoing operational practices or due to one off interruptions. During IWS events changing conditions may impact the endemic biofilms leading to hydraulic mobilisation of organic and inorganic materials attached to pipes walls with a resulting degradation in water quality. To study the impact of IWS on the microbiological and physico-chemical characteristics of drinking water, an experimental full-scale chlorinated pipe facility was operated over 60 days under realistic hydraulic conditions to allow for biofilm growth and to investigate flow resumption behaviour post-IWS events of 6, 48 and 144 hours.Turbidity and metal concentrations showed significant responses to flow restarting, indicating biofilm changes, with events greater than 6 hours generating more turbidity responses and hence discolouration risk. The increase in pressure when the system was restarted showed a substantial increase in total cell counts, while the subsequent increases in flow led to elevated turbidity and metals concentrations. SUVA254 monitoring indicated that shorter times of non-water supply increased the risk of aromatic organic compounds and hence risk of disinfection-by-products formation. DNA sequencing indicated that increasing IWS times resulted in increased relative abundance of potential pathogenic microorganisms, such as Mycobacterium, Sphingomonas, and the fungi Penicillium and Cladosporium.Overall findings indicate that shorter IWS result in a higher proportion of aromatic organic compounds, which can potentially react with chlorine and increase risk of disinfection-by-products formation. However, by minimising IWS times, biofilm-associated impacts can be reduced, yet these are complex ecosystems and much remains to be understood about how microbial interactions can be managed to best ensure continued water safe supply.

From farm-scale to lab-scale: The characterization of engineered irrigation water distribution system biofilm models using an artificial freshwater source

Contaminants in freshwater environments, as well as the associated negative impacts on agricultural produce, have emerged as a critical theme of the water–energy–food nexus affecting food safety and irrigation management. Agricultural produce exposed to irrigation with questionable freshwater can internalize and concentrate pollutants. However, the potential risks posed by the ubiquitous presence of biofilms within irrigation water distribution systems (IWDS) remains overlooked, even though such biofilms may harbor and spread pathogenic, chemical, and other environmental pollutants. Our limited knowledge about the role and functional attributes of IWDS biofilms can be blamed mostly to experimental challenges encountered during attempted studies of these biofilms in their natural environments. Hence, a laboratory-based experimental system designed to simulate a freshwater environment was combined with a biofilm reactor capable of recreating the piping environments in water distribution systems. This experimental system was then tested to assess the robustness and repeatability of experimental early-stage biofilms with respect to physical structure and microbial community, using state-of-the-art confocal microscopy and next-generation sequencing, respectively. The results demonstrated the suitability of this laboratory-based experimental system for studying the impacts of selected pollutants on irrigation water distribution systems.

Bacterial release from pipe biofilm in a full-scale drinking water distribution system

Safe drinking water is delivered to the consumer through kilometres of pipes. These pipes are lined with biofilm, which is thought to affect water quality by releasing bacteria into the drinking water. This study describes the number of cells released from this biofilm, their cellular characteristics, and their identity as they shaped a drinking water microbiome. Installation of ultrafiltration (UF) at full scale in Varberg, Sweden reduced the total cell count to 1.5 × 103 ± 0.5 × 103 cells mL−1 in water leaving the treatment plant. This removed a limitation of both flow cytometry and 16S rRNA amplicon sequencing, which have difficulties in resolving small changes against a high background cell count. Following installation, 58% of the bacteria in the distributed water originated from the pipe biofilm, in contrast to before, when 99.5% of the cells originated from the treatment plant, showing that UF shifts the origin of the drinking water microbiome. The number of bacteria released from the biofilm into the distributed water was 2.1 × 103 ± 1.3 × 103 cells mL−1 and the percentage of HNA (high nucleic acid) content bacteria and intact cells increased as it moved through the distribution system. DESeq2 analysis of 16S rRNA amplicon reads showed increases in 29 operational taxonomic units (OTUs), including genera identified as Sphingomonas, Nitrospira, Mycobacterium, and Hyphomicrobium. This study demonstrated that, due to the installation of UF, the bacteria entering a drinking water microbiome from a pipe biofilm could be both quantitated and described.

Mycobacterial metabolic characteristics in a water meter biofilm revealed by metagenomics and metatranscriptomics

Mycobacteria represent one of the most persistent bacterial populations in drinking water distribution system (DWDS) biofilm communities; however, mycobacterial in situ metabolic profiles are largely unknown. In this study, the metabolic characteristics of mycobacteria in a household water meter biofilm were unveiled using a coupled metagenomic/metatranscriptomic approach. The water meter biofilm appeared to express nitrogenase genes (nifDKH) and a full complement of genes coding for several carbon-fixation pathways, especially the Calvin cycle, suggesting the CO2 sequestration and dinitrogen fixation potential of the biofilm. These findings indicate that it may be difficult to prevent the formation of DWDS biofilms simply by controlling the availability of organic carbon or nitrogen. The composite genome of mycobacteria (CG-M) was reconstructed based on the obtained omics data. CG-M shared similar genome phylogeny and virulence-factor profiles with Mycobacterium avium complex, suggesting that population CG-M might represent a member of mycobacteria with pathogenicity. According to the gene expression patterns, population CG-M showed the metabolic potential to assimilate CO2 via the Calvin cycle and/or anaplerotic reactions, and even to grow autotrophically with CO as the sole carbon and energy source. This suggests that organic carbon may not be a limiting factor for mycobacterial growth in DWDSs. Moreover, our results suggest that mycobacterial aromatic degradation is primarily achieved through the catechol meta-cleavage pathway, and biofilm mycobacteria could prefer phosphate as the phosphorus source.

Biofilm and planktonic bacterial communities in a drinking water distribution system supplied with untreated groundwater.

It is known that both disinfection and water quality can influence the bacterial communities in a drinking water distribution system (DWDS). Here, we hypothesized that bacterial communities in a DWDS with untreated groundwater with no prior purification and disinfection might differ from those in a DWDS with disinfected surface water. The present study applied Illumina MiSeq sequencing to investigate biofilm and planktonic bacterial communities in a DWDS fed with untreated groundwater (receiving no prior purification and disinfection). Considerable differences in bacterial richness (Chao1 richness estimator: 389-745 for water and 392-485 for biofilm), diversity (Shannon diversity index: 2.70-3.77 for water and 2.53-3.66 for biofilm) and community structure existed among both DWDS waters and biofilms. Biofilm and planktonic bacterial communities had distinct structures. The service time of DWDS could affect biofilm bacterial richness, diversity and community structure. Moreover, planktonic bacterial diversity and community structure might be influenced by NO2- concentration, while planktonic bacterial richness was related to NO3- concentration. Proteobacteria dominated in both biofilm and planktonic bacterial communities. Higher concentrations of NO2- favored the deltaproteobacterial proportion, but lowered the gammaproteobacterial proportion in drinking water. Overall, our study indicates that bacterial communities in a DWDS could be influenced by a variety of factors, such as habitats (water or biofilm), DWDS service time, and water chemistry.

Changes in bacterial composition of biofilm in a metropolitan drinking water distribution system

This study examined the development of bacterial biofilms within a metropolitan distribution system. The distribution system is fed with different source water (i.e. groundwater, GW and surface water, SW) and undergoes different treatment processes in separate facilities. The biofilm community was characterized using 16S rRNA gene clone libraries and functional potential analysis, generated from total DNA extracted from coupons in biofilm annular reactors fed with onsite drinking water for up to 18months. Differences in the bacterial community structure were observed between GW and SW. Representatives that explained the dissimilarity were associated with the classes Betaproteobacteria, Alphaproteobacteria, Actinobacteria, Gammaproteobacteria and Firmicutes. After 9months the biofilm bacterial community from both GW and SW were dominated by Mycobacterium species. The distribution of the dominant operational taxonomic unit (OTU) (Mycobacterium) positively correlated with the drinking water distribution system (DWDS) temperature. In this study, the biofilm community structure observed between GW and SW were dissimilar, while communities from different locations receiving SW did not show significant differences. The results suggest that source water and/or the water quality shaped by their respective treatment processes may play an important role in shaping the bacterial communities in the distribution system. In addition, several bacterial groups were present in all samples, suggesting that they are an integral part of the core microbiota of this DWDS. These results provide an ecological insight into biofilm bacterial structure in chlorine-treated drinking water influenced by different water sources and their respective treatment processes.

Distribution of microbial activity and specific microorganisms across sediment size fractions and pipe wall biofilm in a drinking water distribution system

The aim of our study was to identify the dominant environmental niche (pipe wall and different sediment size fractions) in the distribution system for biological activity, Aeromonas and the occurrence of several microbial groups or specific opportunistic pathogens. The pipe wall and sediment fractions of three locations in a distribution system were sampled and different microbiological parameters were determined. The results demonstrated that most of the active biomass in the distribution system is located in the pipe wall biofilm and the smallest sediment size fraction. Fungi were more dominantly present in the pipe wall biofilm than in the sediment, whereas Aeromonas spp. has its dominant niche in the sediment. The highest numbers of the opportunistic pathogens Pseudomonas aeruginosa and Aspergillus fumigatus were observed in the biofilm. In contrast, Stenotrophomonas maltophilia was only observed in the sediment. The results from our study demonstrate that: (i) microbial activity in the distribution system is high on the pipe wall and sediment and (ii) different microbial groups or microbial species have different dominant environmental niches in the distribution system.

Research highlights: functions of the drinking water microbiome – from treatment to tap

We highlight determinants and functions of the drinking water microbiome in water treatment processes, distribution system biofilms, and premise plumbing components.

Characteristics of corrosion sales and biofilm in aged pipe distribution systems with switching water source

Red water has occasionally been observed in the water distribution system of Beijing following a change in the city's water source. Aged cast iron pipes from both affected and unaffected areas were used to establish experimental drinking water distribution systems under different disinfection conditions. The scale deposits formed under different conditions were characterized by X-ray diffraction and scanning electron microscopy, and the bacterial characteristics of the corrosion scales were determined using several molecular methods. The water quality changed less in pipe samples from the unaffected areas than pipes from the affected areas. The pipe samples from the unaffected area included predominantly α-FeOOH, Fe3O4, and γ-FeOOH, and 24.2% nitrate-reducing bacteria (NRB) were predominant with fewer iron-oxidizing bacteria (IOB) being observed. The composition of corrosion products did not exhibit large variation over time although their crystallinity exhibited a slight decrease throughout the duration of the experiment. In contrast, in the pipe samples from the affected, α-FeOOH was the main compound with less γ-FeOOH and Fe3O4, and 3.3% IOB and 12% NRB with few IRB were observed. However, after transporting surface water containing disinfectants, the Fe3O4 and dense corrosion layer formed, inhibiting the release of iron. Moreover, IRB and NRB became dominant. 22.6% and 34.3% NRB appeared in the pipes after chlorination and chloramination, respectively. The synergistic interaction of IRB and NRB with the corrosion layer played a large role in the release of iron from a cast iron pipe following changes in water quality.

Effect of Disinfectants on Bacterial Quorum Sensing in Drinking-Water Distribution System

Several kinds of disinfectants were investigated in this paper for their ability to inhibit quorum sensing activities in water in distribution system. Among these disinfectants, free chlorine is the strongest inhibitor for QS activities and all disinfectants have inhibiting effect on QS activities in water. The results offered a novel approach for preventing formation of biofilm in distribution system.

Inactivation of bacterial biothreat agents in water, a review.

Water supplies and water distribution systems have been identified as potential targets for contamination by bacterial biothreat agents. Since the 2001 Bacillus anthracis bioterrorist attacks, additional efforts have been aimed at research to characterize biothreat organisms in regards to their susceptibility to disinfectants and technologies currently in use for potable water. Here, we present a review of research relevant to disinfection of bacteria with the potential to pose a severe threat to public health and safety, and their potential surrogates. The efficacy of chlorine, monochloramine, chlorine dioxide, and ultraviolet light to inactivate each organism in suspension is described. The complexities of disinfection under varying water conditions and when the organisms are associated with biofilms in distribution systems are discussed.

Biofilm and Helicobacter pylori: from environment to human host.

Helicobacter pylori (H. pylori) is a Gram negative pathogen that selectively colonizes the human gastric epithelium. Over 50% of the world population is infected with H. pylori reaching up to 90% of infected individuals in developing countries. Nonetheless the increased impact upon public health care, its reservoir and the transmission pathway of the species has not been clearly established yet. Molecular studies allowed the detection of H. pylori in various aquatic environments, even forming biofilm in tap water distribution systems in several countries, suggesting a role of water as a possible reservoir of the pathogen. The persistence of human infection with H. pylori and the resistance of clinical isolates to commonly used antibiotics in eradication therapy have been related to the genetic variability of the species and its ability to develop biofilm, demonstrated both in vivo and in vitro experiments. Thus, during the last years, experimental work with this pathogen has been focused in the search for biofilm inhibitors and biofilm destabilizing agents. However, only two anti- H. pylori biofilm disrupting agents have been successfully used: Curcumin - a natural dye - and N-acetyl cysteine - a mucolytic agent used in respiratory diseases. The main goal of this review was to discuss the evidences available in the literature supporting the ability of H. pylori to form biofilm upon various surfaces in aquatic environments, both in vivo and in vitro. The results published and our own observations suggest that the ability of H. pylori to form biofilm may be important for surviving under stress conditions or in the spread of the infection among humans, mainly through natural water sources and water distribution systems.

Monochloramine Use for Prevention of Legionella in Hospital Water Systems.

Eradication of Legionella species from water distribution systems especially in hospital settings has proven to be challenging. Legionella species causes Legionnaire's disease that is a potentially fatal respiratory disease often acquired through the aerosolization of contaminated water. Monochloramine has been used successfully in the municipal water systems to eradicate Legionella and there is currently limited data to support its use in the hospital setting. This technology appears to be affordable, safe and effective at penetrating biofilm in water distribution systems.

Microbial community structures in a closed raw water distribution system biofilm as revealed by 454-pyrosequencing analysis and the effect of microbial biofilm communities on raw water quality

This is the first report on the characterization of the microbial biofilm community structure and water quality change along a closed and stable raw water distribution system. 454-pyrosequencing was employed to investigate the microbial communities in four biofilm samples. 25,426 optimized sequences were obtained. Results showed Proteobacteria was the dominant phylum in each biofilm sample. The abundance of Nitrospiraes in M6 biofilm, Firmicutes in M8 biofilm, Actinobacteria in M9 biofilm were higher by comparing with other three biofilm samples. The M6 microbial biofilm community structure was similar to that of M7, dissimilar to that of M9. Dissolved oxygen and nitrogen was probably major factors to influence the microbial biofilm communities. Nitrospiraes in M6 biofilm and Firmicutes in M8 biofilm were crucial to remove ammonia nitrogen and nitrate in raw water. How to enrich functional microbes in biofilm to pretreat raw water is an important area of future research.

Comparison of microbial quality of irrigation water delivered in aluminum and PVC pipes

Microbial quality of irrigation water attracts substantial attention due to the increased incidence of gastrointestinal illness caused by contaminated produce. Little is known about the changes in microbial quality of water during its delivery to crops. Studies were conducted to compare the biofilm formation and changes in microbial water quality in aluminum and PVC irrigation pipes. Irrigation events were conducted weekly and sections of irrigation pipe (coupons) analyzed for total protein, for total and fecal coliform bacteria, and for Escherichia coli. Coliform and E. coli concentrations along with nitrate, orthophosphate, and total organic carbon were monitored in the intake surface water, output irrigation water, and measured in residual standing water in pipes just prior to each irrigation event. Proteins accumulated to a greater extent in aluminum-associated biofilms than in plastic-associated biofilms. Numbers of total coliforms associated with aluminum coupons increased with time while numbers of total coliforms associated with plastic coupons fluctuated. Nitrates disappeared in standing water after one week in aluminum pipes and remained present in plastic. No detectable E. coli remained in standing water in both types of pipe. There was a high probability that coliform concentrations in output irrigation water were different from the intake concentrations in plastic pipes but not in aluminum pipes. Further research is required to evaluate how pipe material may affect the potential of biofilms in irrigation distribution systems to serve as reservoirs of pathogens that can be disseminated to crops during irrigation.