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      Long-Term Bacterial Dynamics in a Full-Scale Drinking Water Distribution System

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          Abstract

          Large seasonal variations in microbial drinking water quality can occur in distribution networks, but are often not taken into account when evaluating results from short-term water sampling campaigns. Temporal dynamics in bacterial community characteristics were investigated during a two-year drinking water monitoring campaign in a full-scale distribution system operating without detectable disinfectant residual. A total of 368 water samples were collected on a biweekly basis at the water treatment plant (WTP) effluent and at one fixed location in the drinking water distribution network (NET). The samples were analysed for heterotrophic plate counts (HPC), Aeromonas plate counts, adenosine-tri-phosphate (ATP) concentrations, and flow cytometric (FCM) total and intact cell counts (TCC, ICC), water temperature, pH, conductivity, total organic carbon (TOC) and assimilable organic carbon (AOC). Multivariate analysis of the large dataset was performed to explore correlative trends between microbial and environmental parameters. The WTP effluent displayed considerable seasonal variations in TCC (from 90 × 10 3 cells mL -1 in winter time up to 455 × 10 3 cells mL -1 in summer time) and in bacterial ATP concentrations (<1–3.6 ng L -1), which were congruent with water temperature variations. These fluctuations were not detected with HPC and Aeromonas counts. The water in the network was predominantly influenced by the characteristics of the WTP effluent. The increase in ICC between the WTP effluent and the network sampling location was small (34 × 10 3 cells mL -1 on average) compared to seasonal fluctuations in ICC in the WTP effluent. Interestingly, the extent of bacterial growth in the NET was inversely correlated to AOC concentrations in the WTP effluent (Pearson’s correlation factor r = -0.35), and positively correlated with water temperature (r = 0.49). Collecting a large dataset at high frequency over a two year period enabled the characterization of previously undocumented seasonal dynamics in the distribution network. Moreover, high-resolution FCM data enabled prediction of bacterial cell concentrations at specific water temperatures and time of year. The study highlights the need to systematically assess temporal fluctuations in parallel to spatial dynamics for individual drinking water distribution systems.

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          Flow-cytometric total bacterial cell counts as a descriptive microbiological parameter for drinking water treatment processes.

          There are significantly more microbial cells in drinking water than what can be cultured on synthetic growth media. Nonetheless, cultivation-based heterotrophic plate counts (HPCs) are used worldwide as a general microbial quality parameter in drinking water treatment and distribution. Total bacterial cell concentrations are normally not considered during drinking water treatment as a design, operative or legislative parameters. This is mainly because easy and rapid methods for quantification of total bacterial cell concentrations have, up to now, not been available. As a consequence, the existing lack of data does not allow demonstrating the practical value of this parameter. In this study, we have used fluorescence staining of microbial cells with the nucleic acid stain SYBR((R)) Green I together with quantitative flow cytometry (FCM) to analyse total cell concentrations in water samples from a drinking water pilot plant. The plant treats surface water (Lake Zürich) through sequential ozonation, granular active carbon (GAC) filtration and membrane ultrafiltration (UF). The data were compared with adenosine tri-phosphate (ATP) measurements and conventional HPCs performed on the same water samples. We demonstrated that the impact of all three major treatment steps on the microbiology in the system could accurately be described with total cell counting: (1) ozonation caused chemical destruction of the bacterial cells; (2) GAC filtration facilitated significant regrowth of the microbial community; and (3) membrane UF physically removed the bacterial cells from the water. FCM typically detected 1-2 log units more than HPC, while ATP measurements were prone to interference from extracellular ATP released during the ozonation step in the treatment train. We have shown that total cell concentration measured with FCM is a rapid, easy, sensitive and importantly, a descriptive parameter of several widely applied drinking water treatment processes.
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            Monitoring microbiological changes in drinking water systems using a fast and reproducible flow cytometric method.

            Flow cytometry (FCM) is a rapid, cultivation-independent tool to assess and evaluate bacteriological quality and biological stability of water. Here we demonstrate that a stringent, reproducible staining protocol combined with fixed FCM operational and gating settings is essential for reliable quantification of bacteria and detection of changes in aquatic bacterial communities. Triplicate measurements of diverse water samples with this protocol typically showed relative standard deviation values and 95% confidence interval values below 2.5% on all the main FCM parameters. We propose a straightforward and instrument-independent method for the characterization of water samples based on the combination of bacterial cell concentration and fluorescence distribution. Analysis of the fluorescence distribution (or so-called fluorescence fingerprint) was accomplished firstly through a direct comparison of the raw FCM data and subsequently simplified by quantifying the percentage of large and brightly fluorescent high nucleic acid (HNA) content bacteria in each sample. Our approach enables fast differentiation of dissimilar bacterial communities (less than 15 min from sampling to final result), and allows accurate detection of even small changes in aquatic environments (detection above 3% change). Demonstrative studies on (a) indigenous bacterial growth in water, (b) contamination of drinking water with wastewater, (c) household drinking water stagnation and (d) mixing of two drinking water types, univocally showed that this FCM approach enables detection and quantification of relevant bacterial water quality changes with high sensitivity. This approach has the potential to be used as a new tool for application in the drinking water field, e.g. for rapid screening of the microbial water quality and stability during water treatment and distribution in networks and premise plumbing. Copyright © 2013 Elsevier Ltd. All rights reserved.
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              Biological Stability of Drinking Water: Controlling Factors, Methods, and Challenges

              Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order to maintain good drinking water microbial quality up to consumer’s tap. A new definition and methodological approach for biological stability is proposed.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                28 October 2016
                2016
                : 11
                : 10
                : e0164445
                Affiliations
                [1 ]Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands
                [2 ]Center for Microbial Communities, Department of Chemistry and Bioscience, Aalborg University, Fredrik Bajers Vej 7, 9220 Aalborg, Denmark
                [3 ]Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, CH-8600 Dübendorf, Switzerland
                [4 ]King Abdullah University of Science and Technology (KAUST), Water Desalination and Reuse Center (WDRC), Division of Biological and Environmental Science and Engineering (BESE), Thuwal 23955–6900, Saudi Arabia
                [5 ]Wetsus, European Centre of Excellence for Sustainable Water Technology, Oostergoweg 9, 8911 MA Leeuwarden, The Netherlands
                Universidade Federal do Estado do Rio de Janeiro, BRAZIL
                Author notes

                Competing Interests: The presented work has been funded by a commercial source (Evides Waterbedrijf). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                • Conceptualization: EIP FH MCMvL JSV.

                • Data curation: EIP DGW.

                • Formal analysis: EIP DGW.

                • Funding acquisition: JSV.

                • Investigation: EIP.

                • Methodology: EIP FH.

                • Project administration: JSV.

                • Resources: JSV.

                • Supervision: FH MCMvL JSV.

                • Validation: EIP DGW.

                • Visualization: DGW FH MCMvL JSV.

                • Writing – original draft: EIP.

                • Writing – review & editing: EIP DGW FH MCMvL JSV.

                Article
                PONE-D-15-31048
                10.1371/journal.pone.0164445
                5085035
                27792739
                810e65da-8242-458c-957c-c0cbfd918c7e
                © 2016 Prest et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 15 July 2015
                : 26 September 2016
                Page count
                Figures: 7, Tables: 0, Pages: 20
                Funding
                Funded by: Evides Waterbedrijf
                The presented work was funded by Evides Waterbedrijf, who also contributed to sample collection and part of routine analysis.
                Categories
                Research Article
                Engineering and Technology
                Environmental Engineering
                Water Management
                Ecology and Environmental Sciences
                Limnology
                Effluent
                Earth Sciences
                Marine and Aquatic Sciences
                Limnology
                Effluent
                Ecology and Environmental Sciences
                Natural Resources
                Water Resources
                Biology and Life Sciences
                Organisms
                Bacteria
                Biology and Life Sciences
                Organisms
                Bacteria
                Aeromonas
                Biology and Life Sciences
                Developmental Biology
                Microbial Growth and Development
                Bacterial Growth
                Biology and Life Sciences
                Microbiology
                Microbial Physiology
                Microbial Growth and Development
                Bacterial Growth
                Earth Sciences
                Seasons
                Seasonal Variations
                Earth Sciences
                Marine and Aquatic Sciences
                Water Quality
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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                Uncategorized

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