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Drinking Water
Engineering and Science
Open Access

MUWS (Microbiology in Urban Water Systems) Science an interdisciplinary approach to study microbial communities in urban water systems
P. Deines1,2,3 , R. Sekar1,2,3 , H. S. Jensen1,2,3 , S. Tait4 , J. B. Boxall2 , A. M. Osborn3 , and C. A. Biggs1
1

Open Access

Earth System

Data

ChELSI Institute, Pennine Water Group, Department of Chemical and Biological Engineering, The University of Sheeld, UK 2 Pennine Water Group, Department of Civil and Structural Engineering, The University of Sheeld, UK 3 Department of Animal and Plant Sciences, The University of Sheeld, Sheeld, UK 4 Pennine Water Group, School of Engineering Design and Technology, The University of Bradford, Bradford, UK
Received: 11 December 2009 Published in Drink. Water Eng. Sci. Discuss.: 22 January 2010 Revised: 23 June 2010 Accepted: 1 July 2010 Published: 12 July 2010

Abstract. Microbiology in Urban Water Systems (MUWS) is an integrated project, which aims to charac-

terize the microorganisms found in both potable water distribution systems and sewer networks. These large infrastructure systems have a major impact on our quality of life, and despite the importance of these systems as major components of the water cycle, little is known about their microbial ecology. Potable water distribution systems and sewer networks are both large, highly interconnected, dynamic, subject to time and varying inputs and demands, and dicult to control. Their performance also faces increasing loading due to increasing urbanization and longer-term environmental changes. Therefore, understanding the link between microbial ecology and any potential impacts on short or long-term engineering performance within urban water infrastructure systems is important. By combining the strengths and research expertise of civil-, biochemical engineers and molecular microbial ecologists, we ultimately aim to link microbial community abundance, diversity and function to physical and engineering variables so that novel insights into the performance and management of both water distribution systems and sewer networks can be explored. By presenting the details and principals behind the molecular microbiological techniques that we use, this paper demonstrates the potential of an integrated approach to better understand how urban water system function, and so meet future challenges.

1 1.1

Introduction The challenges

and social development. Their overall performance can be evaluated by physical, chemical and biological processes. Urban water systems throughout Europe face signicant new challenges to continue to maintain the provision of safe water supplies, hygienic sanitation and good environmental management against the setting of increased urbanisation, ageing infrastructure and changing climate conditions. These changes are expected to have a negative impact on freshwater resources. The important role of urban water systems has been recognized by the EU with the provision of a series of directives (e.g. Urban Wastewater Treatment, Bathing Waters and Water Framework Directive (WFD)), which govern the use of water in order to provide equitable standards of service and improving environmental protection. Unlike earlier directives, which quantied environmental quality by simple physical and chemical parameters, the WFD aims to ensure that good ecological status is attained in all European

Urban water systems (e.g. drinking water distribution and sewers networks, wetlands and urban rivers) are important for millions of people living in urban areas. They are major components of the water cycle and present unique challenges; the systems are large, complex, highly interconnected and dynamic, with variable hydraulics, input sources and behaviour. These large infrastructure systems have a major impact on peoples quality of life by preventing serious disease, protecting/enhancing the environment and reducing ood damage to other infrastructure, thus enabling economic Correspondence to: C. A. Biggs ([email protected])

Published by Copernicus Publications on behalf of the Delft University of Technology.

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water bodies. It is less prescriptive than previous directives and aims to address the management of the water bodies in a more holistic manner.
1.1.1 Water distribution systems

latter aecting the public perception of the sewers (HvitvedJacobsen, 2002).

1.2

The interdisciplinary approach

A technological challenge for the EU and worldwide water industry is the continuous delivery of high quality drinking water to customers taps that meets increasingly stringent standards for aesthetic, bacteriological and chemical water quality. Most distribution systems comprise a complex network of pipes of dierent ages and material types (UKWIR, 2003). Frequently, each system is supplied from a number of dierent treatment works each with dierent source water and treatment systems. Despite the fact that modern water treatment works produce high quality water as it enters the distribution system, the quality of the water is known to deteriorate during transportation within the system (e.g. Furtado et al. (1998) found 7 out of the 10 intestinal disease outbreaks reported in a 3 year study, arose due to contamination occurring within the distribution network). Changes in water quality are due to distribution systems acting as large bio-chemical reactors in which many complex, dynamic, and interrelated hydrodynamic and biochemical processes occur. Water distribution systems harbour microbial consortia, for example, anaerobic bacteria, protozoa, together with meioand macro-fauna such as copepods and nematodes (Evins, 2004; Berry et al., 2006). Additionally the presence of pathogenic bacteria, that are normally undetectable by traditional culture based methods, can represent a potential reservoir for disease outbreaks or long-term illness (Szewzyk et al., 2000).
1.1.2 Sewer systems

Answers to key questions, such as which microorganisms are present?, what are they doing?, and how can we use their outputs and manage their activity to achieve better system outcomes? are important for understanding the physical, chemical and biological interactions in urban water systems. The long term objective of the project Microbiology in Urban Water Systems (MUWS) is to assess the impact of microorganisms, due to their presence, diversity and response to various environmental conditions on aspects of system performance within drinking water distribution systems and sewer networks. Using the research expertise of civil- biochemical engineers and molecular microbial ecologists, the MUWS project aims to address these key questions across dierent length scales of the urban water systems. Combining engineering and biolgoical disciplines to address environmental engineering challenges is not necessarily a new concept, and has been discussed previously (e.g. Daims et al., 2006; Rittmann et al., 2006; McMahon et al., 2007), however these studies primarily focus on biotechnology/treatment type processes, whereas an interdisciplinary approach to study urban water infrastructure systems has received far less attention. The aim of this paper is to describe the work carried out in the MUWS project so far to permit the application of advanced microbial methods specically in drinking water distribution systems and sewer networks, thus highlighting the potential of our integrated approach.

A recent study in the UK indicated that if no remedial measures were adopted, the discharge of excess volumes from sewer systems during rain fall events to the environment could increase by up to 250% based on expected climate and urbanization changes (Evans et al., 2004), therefore increasing their future environment impact. Recent studies have also provided strong evidence that sewer ow quality can be strongly inuenced by microbiological activity within sewer deposits (Tait et al., 2003). In cases of system failure, the discharge of untreated wastewater and sediments can degrade the water quality of the receiving water body, and may also be a risk to public health. In this respect the microorganisms released with the wastewater may be particularly important, due to the potential release of pathogens. In addition to these acute risks, the activity of the microorganisms in sewers changes the composition of the wastewater. This can, for example, lead to septic wastewater, which is associated with problems such as the formation of toxic gases and malodorous gases; the rst posing an acute risk for sewer workers, the
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Methods

In the MUWS project, variations in the microbial community of drinking water distribution systems and sewer networks are characterized by using molecular microbiological techniques, rather than culture based techniques. The culture based techniques are currently used by the water industry for evaluation of the performance of the urban water systems. The molecular microbiological techniques, which have previously been successfully applied to freshwater and marine plankton samples, sediments and soil samples (Gelsomino et al., 1999; Moeseneder et al., 1999; Wagner et al., 2003; Smalla et al., 2007) are further developed specically for the analysis of drinking water, wastewater, biolms and sewer sediment microbial communities (Fig. 1). In this section we introduce two key methods. The data obtained from these methods has allowed us to gain insight into how a microbial community changes under dierent conditions at a variety of scales within urban water systems.
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SEWER SYSTEMS planktonic/biofilm/sediment

WATER DISTRIBUTION SYSTEMS planktonic/biofilm

Fixation

Nucleic acid extraction PCR amplification

Membrane filtration and/or spread plate/ pour plate method

Molecular probes

DGGE profiling

Fluorescence in situ hybridization (FISH)

Sequencing & comparative analysis

Heterotrophic plate count (HPC)

Total cell counts & species abundance

Detailed community composition

Estimates number of heterotrophic bacteria

current industry practice

Figure 1. Scheme for the Figure 1 in the molecular analysis of major steps

bacterial communities from water distribution and sewer systems and the respective results obtained. In comparison, the right hand side shows the current procedure for monitoring water quality.

2.1

Denaturing gradient gel electrophoresis (DGGE)

DGGE analysis (Muyzer et al., 1993) can be used to investigate mixed microbial communities from various environments. The method is based on the molecular separation of DNA fragments when migrating through a DGGE gel, which results in a specic banding pattern. Each individual discrete band refers to a unique sequence type or phylotype (van Hannen et al., 1999) which can further be analyzed by sequencing for taxonomic identication. The similarities between banding patterns from dierent samples can then be analyzed using multivariate analysis such as cluster analysis (Fromin et al., 2002). To demonstrate the DGGE proling technique, the method was applied to both planktonic samples from a drinking water distribution system and sewer biolms. The key steps are outlined in Fig. 1 with specic detail relating to the individual samples listed below.
2.1.1 Drinking water

of water was ltered through a 0.22 m polycarbonate membrane lter (diameter 47 mm; Millipore Ltd., UK) and the lters were kept at 80 C until further analysis. The membrane lters were cut in to halves under sterile conditions and placed directly into the bead solution tubes of the MoBio Ultra Clean Soil DNA Isolation Kit (Cambio Ltd., Cambridge, UK). The DNA was extracted as per the manufacturers protocol and then used for polymerase chain reaction (PCR) amplication of 16S rRNA genes (Saiki et al., 1988). A direct-PCR, nested-PCR approach was used for comparative analysis of the culture independent drinking water samples. After performing all the DNA extractions, 16S rRNA gene fragments were amplied by direct-PCR, using the universal bacterial primers 338F with a GC-clamp and 530R (Whiteley and Bailey, 2000). For the same water samples, a rst round PCR was performed with the bacterial primers 27F and 1492R (Lane, 1991) followed by a second PCR amplication using the primers mentioned above. This approach is called nested-PCR and it can improve the sensitivity of the PCR. For the cultivation-dependent approach one half of individual membrane lters were sonicated in R2A medium (Reasoner and Geldreich, 1985) and 100 l of those samples were used to inoculate 50 ml of R2A medium and cultured at 20 C for 48 h. Two ml of the culture were used for DNA extraction as described above. After performing all the extractions, 16S rRNA gene fragments were amplied by directPCR, using the universal bacterial primers 338F with a GCclamp and 530R as mentioned above. The PCR products were then loaded on an 8% polyacrylamide gel with a denaturant gradient ranging from 40 to 70%. DGGE analysis was performed using the Bio-Rad DCode System (Bio-Rad, Hertfordshire, UK). The gel was run at 100 V at 60 C for 16 h in 1x TAE (Tris-AcetateEDTA) running buer and the gels were stained with SYBR Gold prior to image acquisition.
2.1.2 Sewer biolms

Planktonic samples were collected from a water distribution system (domestic cold water tap) in Sheeld in February 2008. Water samples, rather than biolms, were collected for the drinking water distribution system to provide direct comparison with the culture based methods currently used by water companies. In brief, after a 1 min ush, water samples (3 2 L) were collected in sterile bottles and transported to the laboratory on ice and processed immediately. Two liters
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Sewer biolms were collected from two dierent sewer systems, one in Nantes (France) and the other in Frejlev (Denmark). Two sampling sites were used in Nantes and one site in Frejlev. Two sewer systems were chosen to demonstrate the applicability of the molecular techniques across spatial variability in sewer networks. The sampling site at Frejlev is run by the Environmental Engineering Group at Aalborg University. The samples were recovered from a 300 mm diameter combined sewer that served an 87 ha catchment with mainly residential inputs. The samples are collected downstream of the town of Frejlev. The site was established as a research station in 1996. The sampling sites in Nantes, France are run by the LCPC, Division Eau et Environnement, Bouguenais, France. The samples were collected from two sites, in combined sewers, located in the central part of Nantes. Both sites were positioned in large
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Drinking Water Culture Independent nested-PCR direct-PCR Drinking Water Culture Dependent direct-PCR a b a b

egg-shaped collectors and are part of ongoing studies carried out by LCPC. Sewer biolm samples were collected in Nantes in November 2008 and in Frejlev in February 2009. All samples were collected in triplicates. The biolm samples were scraped o the sewer pipe surfaces directly above the waterline using sterile 15 ml Falcon tubes. The samples were stored at 20 C until further processing. DNA was extracted from 0.2 g (wet weight) of biolms using the same DNA isolation kit as mentioned above for drinking water. To minimize interference with humic substances DNA extracts were diluted (1:10) before performing PCR amplications. For the amplication of 16S rRNA gene fragments, the same primer pair with the GCclamp as mentioned above was used (direct-PCR approach). PCR products were visualized by gel electrophoresis followed by ethidium bromide staining to ensure that the correct size fragment was amplied. DGGE analysis of the PCR fragments was performed as described above for the drinking water samples.
2.2 Fluorescence in situ hybridization (FISH)

FISH with rRNA-targeted oligonucleotide probes is used for detection and quantication of microorganisms without prior cultivation (Amann et al., 1995). The method has been applied widely to dierent environmental samples including drinking water systems (Manz et al., 1993). To overcome problems of low detection limit and uorescence intensity faced in oligotrophic environmental samples (such as drinking water), we have optimized and applied the CARD-FISH (catalysed reported deposition-uorescence in situ hybridization) method. This method was originally developed for studying bacterioplankton in marine samples (Pernthaler et al., 2002). Again to demonstrate the technique the planktonic samples from a drinking water distribution system and sewer biolms were analysed using CARD-FISH (Fig. 1).
2.2.1 Drinking water

Figure 2. DGGE proles of PCR-amplied 16S rRNA gene frag-

ments derived from direct- and nested-PCR approach for culture inFigure 2 dependent drinking water samples, and a direct-PCR approach for culture dependent drinking water samples. Arrows indicate the two dominant phylotypes present using the culture dependent approach. a and b refer to biological replicates.

pus Ltd., UK). Quantication of cell numbers of the drinking water samples was performed by imaging and counting the DAPI stained cells.
2.2.2 Sewer biolms

The water samples (50 ml) were xed in 2% (v/v) nal concentration of formalin for less than 24 h. The samples were then ltered on to 0.22 m pore size white polycarbonate membrane lters (diameter 47 mm, Millipore Ltd., UK) and stored at 20 C until further processing. The samples were permeabilised with lysozyme and achromopeptidase as described previously (Pernthaler et al., 2002; Sekar et al., 2003). The hybridization was done with the HRP labeled eubacterial oligonucleotide probes (EUB338). The hybridization, washing and tyramide signal amplication with FITClabeled tyramides were done as per the protocol described in Pernthaler et al. (2002). The preparations were counterstained with the DNA specic uorescent stain, DAPI, and observed under an Olympus BX51 epiuorescence microscope (Olympus Ltd., UK). The images (30 per triplicate sample) were captured using CellB imaging software (OlymDrink. Water Eng. Sci., 3, 9199, 2010

Triplicate sewer samples were xed either with 2% (v/v) formalin or 1:1 PBS/Ethanol on the day of sampling. The sewer biolm samples collected from France and Denmark were gently vortexed, mixed with low gelling agarose (0.2% w/vol) and 10 l of the samples were pipetted into the wells of a standard type multi-well Epoxy slide (Carl Roth GmbH + Co, Karlsruhe, Germany). The permeabilization, hybridization with the eubacterial probes (EUB338) and the tyramide signal amplication was done as described above for the water samples. The preparations were observed under an epiuorescence microscope as described above.

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P. Deines et al.: MUWS (Microbiology in Urban Water Systems) 3 3.1 Results and discussion Bacterial community proling of drinking water samples

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Sewer biofilms
Dk Fr 1 Fr 2

Figure 2 shows the DGGE patterns for drinking water samples based on a culture-independent and culture dependent approach. The dierences in DGGE patterns of the directPCR approach for both samples show that with the culturedependent approach we target a specic bacterial community that responds to the imposed enrichment. One would conclude from Fig. 2, that by using the culture-dependent approach, two phylotypes dominate the bacterial community in the drinking water collected. (NB: Often the brightest bands in the prole represent the dominant members of the community, but be aware of potential biases (Forney et al., 2004)). This is signicant as one method that is widely used to assess the general microbial water quality of drinking water by water distributors is the cultivation-dependent method of heterotrophic plate counts (HPCs) (see Sartory, 2004). The method is highly variable since the cultivation medium, incubation temperature, incubation time, origin, season of the year, and age of the water sample have a signicant eect on the fraction of the total bacterial cells that grow and hence will be detected (Allen et al., 2004). Despite the discrepancy between total bacterial concentrations and cultivable cell concentrations (HPCs) in aquatic samples as shown by Staley and Konopka (1985), HPCs are still used for routine monitoring applications in a quantitative way (Standard Methods for the Examination of Water and Wastewater, 20th Edn.). Pepper et al. (2004), for example, quantied the concentration of HPC bacteria within water from the source to the consumers tap. Their study showed that the number of HPC bacteria increased dramatically from the distribution system to the consumers tap, but they did not quantify whether the bacterial community changed as well. There are only a few studies providing qualitative data about the HPC community composition and/or population dynamics (e.g. Kalmbach et al., 1997; Norton and LeChevallier, 2000). For the enumeration of bacteria in drinking water, lownutrient media are commonly used such as R2A (Reasoner and Geldreich, 1985). It was designed specically as a lownutrient, low-ionic strength formulation to isolate bacteria that have a water-based lifestyle (Reasoner, 1990). Our results support previous observations that media used for HPC are selective for those bacteria that can grow under the specic conditions used. Comparing these results to the DGGE proles of either direct- or nested-PCR approach (no cultivation step involved) reveals considerable dierences in the banding pattern observed. Several unique bands that were not visible in the culture dependent approach were seen, supporting that the culture dependent method underestimated the number of phylotypes present in the sample and that when the culture dependent method is used for assessment of microbial contamination in drinking water distribution systems
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Figure 3. DGGE proles of PCR-amplied 16s rRNA gene frag-

ments derived from direct-PCR approach for sewer biolm samples from one site in Denmark (Dk) and two sampling sites in France (Fr1, Fr2).

Figure 3

there is a signicant risk that microbial contaminations can exist but cannot be detected with current practice. Figure 2 shows that dierences can be seen between the nested-PCR approach, which includes an additional PCR step, and the direct-PCR approach. Bands that appear at the same position have changed their intensity and new bands become visible. Therefore, in certain cases the nested-PCR approach may need to be applied to increase sensitivity and to complete the overall presentation of the taxonomic diversity present in the sample.
3.2 Bacterial community proling of sewer biolm samples

Figure 3 shows the DGGE proling for a sampling site from Denmark and two sites from France. A total of 26 discrete bands were detected on the gel. A total of 18 bands were present in the samples from Denmark where as 19 and 9 bands were found in France sampling site 1 and 2, respectively. This analysis suggests similar levels of diversity (although not necessary the same composition) between sewer biolms from Denmark and France sampling site 1; and that sampling site 2 in France showed lower diversity. The overall results indicate that 4 bands were common to all three sites and 12 were found in two sites and 10 were unique to one site. Identication of common bands (and further taxonomic
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(a) (b)

P. Deines et al.: MUWS (Microbiology in Urban Water Systems)

Photomicrographs of drinking water bacterioplankton (a) and sewer biolm bacteria (b) hybridized with eubacterial probes.
Figure 4.

identication) could be used to identify indicator microorganisms that are present across diverse spatial distributions within the sewer networks. Unique bands can also be used for ngerprinting microorganisms related to distinctive netFigure 4 works, or spatial distribution within the network if comparison between euent, biolm and sediment samples is made. Hence the DGGE technique can be used to monitor changes in bacterial communities within urban water systems, taking spatial and temporal variation into account (see Fig. 1). This variability can be investigated in terms of dierences in the presence or absence of specic bands and/or in changes in their relative abundance (band intensity). Clustering techniques can be applied to identify samples, which generate similar patterns (Boon et al., 2002). Multivariate analysis also allows the interpretation of DGGE patterns in relation to environmental variables (e.g. McCaig et al., 2001). Hence, when conducted in combination with relevant physical and chemical measurements, DGGE provides the opportunity to study the changes in microbial diversity relevant to conditions within the urban water system.

al., 2008) and is already being routinely used by the Zurich Waterworks (Egli et al., 2008). Figure 4b demonstrates the application of CARD-FISH for sewer biolms collected from France. Whilst it is not possible to easily count the number of cells that are present in the biolm, as found with Fig. 4a, the CARD-FISH method with eubacterial probes enables the morphology of the biolm to be visualized. Comparison of CARD-FISH images across spatial and or temporal samples could therefore provide insight into the morphological development of biolms within urban water systems. An added functionality of CARD-FISH is the ability to target specic microorganisms (see Fig. 1) which will reveal the abundance of dierent phylogenetic or functional bacterial and other microbial groups within biolms or water samples. CARD-FISH in combination with epiuorescence microscropy or confocal laser scanning microscopy and digital image analysis can be combined into quantitative polyphasic approach to study the microbes in urban water systems. Microorganisms in the source water, drinking water, wastewater and in biolms can be studied as well as their seasonal and successional changes. Particularly, the CARDFISH technique is very suitable for studying bacteria (and other microbes) in highly oligotrophic environments such as water distribution systems because of their increased sensitivity and uorescence intensity. In combination with microautoradigraphy (MAR) or stable isotope probing (SIP), this will provide both structural and functional characteristics of targeted microorganisms (Teira et al., 2004; Sintes and Herndl, 2006; Wagner et al., 2006).
4 Future interdisciplinary approach

3.3

FISH of microorganisms in drinking water and sewer biolms

In this study, we have applied CARD-FISH methods to detect and quantify the microorganisms in drinking water (Fig. 4a), and sewer biolms (Fig. 4b). The CARD-FISH images, with the eubacterial probes, provide an opportunity to quantify the number of bacteria within the water samples (Fig. 4a). Unlike HPC however, this technique is not based on a prior basis of culturable bacteria. Counter staining the samples with a DNA stain like DAPI (results not shown), conrms that CARD-FISH is able to detect, and therefore condently account for, more than 90 to 93% of the bacteria present in drinking water sample. Hence the CARD-FISH method provides more condence in enumerating the number of bacteria in water samples over the routinely used HPC method. Another new rapid and reliable method for cell enumeration of drinking water has also been developed based on ow cytometry (Berney et al., 2008; Hammes et al., 2008; Siebel et
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The aim of this paper was to present a conceptual outline to water practitioners on the types of culture independent molecular microbiological techniques available, and the level of information achievable from these techniques, when applied to urban water systems. Through this, we aimed to show the benets of a partnership with microbial ecologists, specically looking at urban water infrastructure, as better quantication of bacterial communities and their temporal or spatial changes in urban water systems, through an integrated approach will lead to further understanding of their associated biological processes. The benet of an interdisciplinary approach comes in a variety of dierent aspects. For example, existing engineering knowledge and/or computer models can provide insight into choosing the most appropriate sampling locations within drinking water distribution systems or sewer networks that enable specic research questions to be addressed e.g. inuence of spatial or temporal changes on systems performance due to changes in microbial diversity. Sampling protocols across the dierent disciplines also need to be integrated to ensure the safe collection of representative samples within
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Laboratory scale
isolation of bacterial strains from drinking water and sewers

Field studies

3 5 1 2

studying interspecies interactions in biofilm formation metabolic fingerprinting cell surface properties

planktonic and biofilm microbial community composition in urban systems water age and microbial diversity flow rate, temperature and diversity seasonal variation

Potable water distribution

nutrient stress triggers aggregation multicellular structures

low-nutrient environment

planktonic biofilm
cells

Urban Water Systems

Challenges

Sewer networks
Pilot scale rig
community profiling of planktonic and biofilm microbial communities in response to changing conditions studying biofilm characteristics

Determine and predict the impact of: hydraulics temperature water quality nutrient status flowrate (e.g. storm events) pathogen detection and abundance

on microbial assemblages and urban water system function

Figure 5. Overview of the MUWS (Microbiology in Urban Water Systems) project showing the challenges, the approaches and the dierent

departments involved.

Figure 5 and behaviour of microbial assemblages and their the urban water system in terms of both biological repropresence ducibility as well as engineering relevance. potential release into the environment, with asset characteristics, operation (hydraulics and cleaning/disinfection regimes) Also, in an integrated approach, molecular microbiologiand water quality (i.e. linking biological function with engical analysis of water and biolm/sediment samples, as preneering performance). sented here, should be conducted at the same time as the measurement of the physical and chemical properties of urAcknowledgements. Funding was provided by FP6 EU Marie ban water systems. Multivariate analysis of the dierent paCurie Transfer of Knowledge programme (No. 42444). CAB also rameters, will then allow future interpretation of changes in thanks the EPSRC for the provision of an Advanced Research Felbiological diversity to specic environmental variables, hylowship (EP/E053556/01) and ChELSI funding (GR/S84347/01). draulic conditions etc. This will provide the fundamental The authors are grateful to Professor Vollertsen at Aalborg knowledge to ultimately develop biological management University, Denmark and Dr Larrarte at LCPC, Division Eau et tools that will aid system operators to achieve improved levEnvironnement, Bouguenais, France, for biolm samples. They also thank Professor Amann, Max Planck Institute for Marine els of environmental and public health protection without Microbiology, Germany, for providing FISH probes. resorting to the need for additional infrastructure or energy intensive treatment processes. To achieve this, the MUWS Edited by: J. Vreeburg project specically operates across the length scales from laboratory to pilot and eld studies (Fig. 5) and draws on the expertise of civil-, biochemical engineers and molecular microbial ecologists to address key challenges. This interdisciplinary and multi-scale approach provides a unique opportunity to develop and understand relationships between the
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