This study investigates the influence of pipe characteristics on the bacteriological composition of material mobilised from a drinking water distribution system (DWDS) and the impact of biofilm removal on water quality. Hydrants in a single UK Distribution Management Area (DMA) with both polyethylene and cast iron pipe sections were subjected to incremental increases in flow to mobilise material from the pipe walls. Turbidity was monitored during these operations and water samples were collected for physico-chemical and bacteriological analysis. DNA was extracted from the material mobilised into the bulk water before and during flushing. Bacterial tag-encoded 454 pyrosequencing was then used to characterize the bacterial communities present in this material. Turbidity values were high in the samples from cast iron pipes. Iron, aluminium, manganese and phosphate concentrations were found to correlate to observed turbidity. The bacterial community composition of the material mobilised from the pipes was significantly different between plastic and cast iron pipe sections (p < 0.5). High relative abundances of Alphaproteobacteria (23.3%), Clostridia (10.3%) and Actinobacteria (10.3%) were detected in the material removed from plastic pipes. Sequences related to Alphaproteobacteria (22.8%), Bacilli (16.6%), and Gammaproteobacteria (1.4%) were predominant in the samples obtained from cast iron pipes. The highest species richness and diversity were found in the samples from material mobilised from plastic pipes. Spirochaeta spp., Methylobacterium spp. Clostridium spp. and Desulfobacterium spp., were the most represented genera in the material obtained prior to and during the flushing of the plastic pipes. In cast iron pipes a high relative abundance of bacteria able to utilise different iron and manganese compounds were found such as Lysinibacillus spp., Geobacillus spp. and Magnetobacterium spp.

The majority of biomass within water distribution systems is in the form of attached biofilm. This is known to be central to drinking water quality degradation following treatment, yet little understanding of the dynamics of these highly heterogeneous communities exists. This paper presents original information on such dynamics, with findings demonstrating patterns of material accumulation, seasonality, and influential factors. Rigorous flushing operations repeated over a 1-year period on an operational chlorinated system in the United Kingdom are presented here. Intensive monitoring and sampling were undertaken, including time-series turbidity and detailed microbial analysis using 16S rRNA Illumina MiSeq sequencing. The results show that bacterial dynamics were influenced by differences in the supplied water and by the material remaining attached to the pipe wall following flushing. Turbidity, metals, and phosphate were the main factors correlated with the distribution of bacteria in the samples. Coupled with the lack of inhibition of biofilm development due to residual chlorine, this suggests that limiting inorganic nutrients, rather than organic carbon, might be a viable component in treatment strategies to manage biofilms. The research also showed that repeat flushing exerted beneficial selective pressure, giving another reason for flushing being a viable advantageous biofilm management option. This work advances our understanding of microbiological processes in drinking water distribution systems and helps inform strategies to optimize asset performance.IMPORTANCE This research provides novel information regarding the dynamics of biofilm formation in real drinking water distribution systems made of different materials. This new knowledge on microbiological process in water supply systems can be used to optimize the performance of the distribution network and to guarantee safe and good-quality drinking water to consumers.

Disinfection residuals in drinking water protect water quality and public heath by limiting planktonic microbial regrowth during distribution. However, we do not consider the consequences and selective pressures of such residuals on the ubiquitous biofilms that persist on the vast internal surface area of drinking water distribution systems. Using a full scale experimental facility, integrated analyses were applied to determine the physical, chemical and biological impacts of different free chlorine regimes on biofilm characteristics (composition, structure and microbiome) and water quality. Unexpectedly, higher free chlorine concentrations resulted in greater water quality degredation, observable as elevated inorganic loading and greater discolouration (a major cause of water quality complaints and a mask for other failures). High-chlorine concentrations also reduced biofilm cell concentrations but selected for a distinct biofilm bacterial community and inorganic composition, presenting unique risks. The results challenge the assumption that a measurable free chlorine residual necessarily assures drinking water safety.

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High quality drinking water exits modern treatment works, yet water quality degradation such as discolouration continues to occur within drinking water distribution systems (DWDS). Discolouration is observed globally, suggesting a common process despite variations in source, treatment, disinfection and network configurations. The primary cause of discolouration has been identified as mobilisation of particulate material from pipe walls and the verified Prediction of Discolouration in Distribution Systems (PODDS) model uses measurable network hydraulics to simulate this response.In this paper the cohesive properties of discolouration material are explored and it is hypothesised that in simulating the turbidity response, the PODDS model is actually describing the development and cohesive strength behaviour of biofilms. Applying this concept can therefore facilitate a rapid and simple assessment of DWDS biofilm activity. A review of the findings from PODDS studies conducted internationally is presented, focussing on the macro or observable aspects of discolouration. These are compared and contrasted with associated biofilm studies which consider discolouration material at the micro-scale. Combining the results from these (past) studies to improve the understanding of interactions between microbial ecology and discolouration are discussed with a view to DWDS operational strategies that safeguard and optimise drinking water supply.