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      Impact of stagnation and sampling volume on water microbial quality monitoring in large buildings

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          Abstract

          Microbial drinking water quality can be altered in large buildings, especially after stagnation. In this study, bacterial profiles were generated according to the stagnation time and the volume of water collected at the tap. Successive volumes of cold and hot water were sampled after controlled stagnation periods. Bacterial profiles revealed an important decline (> 2 log) in culturable cells in the first 500 mL sampled from the hot and cold water systems, with a steep decline in the first 15 mL. The strong exponential correlation (R 2 ≥ 0.97) between the culturable cell counts in water and the pipe surface-to-volume ratio suggests the biofilm as the main contributor to the rapid increase in suspended culturable cells measured after a short stagnation of one-hour. Results evidence the contribution of the high surface-to-volume ratio at the point of use and the impact of short stagnation times on the increased bacterial load observed. Simple faucets with minimal internal surface area should be preferred to minimize surface area. Sampling protocol, including sampling volume and prior stagnation, was also shown to impact the resulting culturable cell concentration by more than 1000-fold. Sampling a smaller volume on first draw after stagnation will help maximize recovery of bacteria.

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          Most cited references31

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          The importance of the viable but non-culturable state in human bacterial pathogens

          Many bacterial species have been found to exist in a viable but non-culturable (VBNC) state since its discovery in 1982. VBNC cells are characterized by a loss of culturability on routine agar, which impairs their detection by conventional plate count techniques. This leads to an underestimation of total viable cells in environmental or clinical samples, and thus poses a risk to public health. In this review, we present recent findings on the VBNC state of human bacterial pathogens. The characteristics of VBNC cells, including the similarities and differences to viable, culturable cells and dead cells, and different detection methods are discussed. Exposure to various stresses can induce the VBNC state, and VBNC cells may be resuscitated back to culturable cells under suitable stimuli. The conditions that trigger the induction of the VBNC state and resuscitation from it are summarized and the mechanisms underlying these two processes are discussed. Last but not least, the significance of VBNC cells and their potential influence on human health are also reviewed.
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            LIVE/DEAD BacLight : application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water.

            A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit (BacLight) was applied to estimate both viable and total counts of bacteria in drinking water. BacLight is composed of two nucleic acid-binding stains: SYTO 9 and propidium iodide. SYTO 9 penetrates all bacterial membranes and stains the cells green, while propidium iodide only penetrates cells with damaged membranes, and the combination of the two stains produces red fluorescing cells. Optimal incubation conditions were found to be 15 to 20 min, at room temperature in the dark. Total (red + green) and viable (green) cells can hence be counted simultaneously. Factors affecting the staining procedure were tested (addition of glutaraldehyde, staining time, chlorine impact). In the absence of stress, BacLight viable counts were comparable and to 5-cyano-2,3-ditolyl tetrazolium (CTC) counts. BacLight total counts were comparable to acridine orange counts (differing by <0.1 log/ml). However, the increase in environmental stresses (chlorine, growth rate or temperature) induced a decrease in viability that was more pronounced for CTC and plate counts than for BacLight viable counts.
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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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                Author and article information

                Contributors
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 June 2018
                2018
                : 13
                : 6
                : e0199429
                Affiliations
                [1 ] Department of Civil Engineering, Polytechnique Montréal, Montréal, QC, Canada
                [2 ] INRS-Institut Armand-Frappier, Laval, QC, Canada
                [3 ] Department of Microbiology and Immunology (Infection control), CHU Ste-Justine, Université de Montréal, Montréal, QC, Canada
                Nanjing University, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-1447-929X
                Article
                PONE-D-18-03891
                10.1371/journal.pone.0199429
                6013212
                29928013
                16072c3f-77f0-48c6-b338-451d106aa213
                © 2018 Bédard 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
                : 5 February 2018
                : 7 June 2018
                Page count
                Figures: 6, Tables: 0, Pages: 14
                Funding
                This study was supported by NSERC through the Industrial Chair on Drinking Water and the E. Bedard Vanier Canada Graduate Scholarship. Eric Déziel holds a Canada Research Chair in socio-microbiology.The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Microbiology
                Bacteriology
                Bacterial Biofilms
                Biology and Life Sciences
                Microbiology
                Biofilms
                Bacterial Biofilms
                Research and Analysis Methods
                Cell Enumeration Techniques
                Viable Cell Counting
                Research and Analysis Methods
                Biological Cultures
                Cell Culturing Techniques
                Biofilm Culture
                Engineering and Technology
                Environmental Engineering
                Water Management
                Earth Sciences
                Marine and Aquatic Sciences
                Water Quality
                Biology and Life Sciences
                Organisms
                Bacteria
                Physical Sciences
                Chemistry
                Chemical Elements
                Chlorine
                Research and Analysis Methods
                Cell Enumeration Techniques
                Total Cell Counting
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

                Uncategorized
                Uncategorized

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