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Biofilms are the dominant form of microbial loading (and organic material) within drinking water distribution systems (DWDS), yet our understanding of DWDS microbiomes is focused on the more easily accessible bulk-water. Disinfectant residuals are commonly provided to manage planktonic microbial activity in DWDS to safeguard water quality and public health, yet the impacts on the biofilm microbiome are largely unknown. We report results from a full-scale DWDS facility used to develop biofilms naturally, under one of three chlorine concentrations: Low, Medium, or High. Increasing the chlorine concentration reduced the bacterial concentration within the biofilms but quantities of fungi were unaffected. The chlorine regime was influential in shaping the community structure and composition of both taxa. There were microbial members common to all biofilms but the abundance of these varied such that at the end of the Growth phase the communities from each regime were distinct.  Alpha-, Beta- , and  Gamma-proteobacteria  were the most abundant bacterial classes;  Sordariomycetes, Leotiomycetes , and  Microbotryomycetes  were the most abundant classes of fungi. Mechanical cleaning was shown to immediately reduce the bacterial and fungal concentrations, followed by a lag effect on the microbiome with continued decreases in quantity and ecological indices after cleaning. However, an established community remained, which recovered such that the microbial compositions at the end of the Re-growth and initial Growth phases were similar. Interestingly, the High-chlorine biofilms showed a significant elevation in bacterial concentrations at the end of the Re-growth (after cleaning) compared the initial Growth, unlike the other regimes. This suggests adaptation to a form a resilient biofilm with potentially equal or greater risks to water quality as the other regimes. Overall, this study provides critical insights into the interaction between chlorine and the microbiome of DWDS biofilms representative of real networks, implications are made for the operation and maintenance of DWDS disinfectant and cleaning strategies.

Biofilm Microbiome (Re)Growth Dynamics in Drinking Water Distribution Systems Are Impacted by Chlorine Concentration