In this study, we collected water from different locations in 32 drinking water distribution networks in the Netherlands and analysed the spatial and temporal variation in microbial community composition by high-throughput sequencing of 16S rRNA gene amplicons. We observed that microbial community compositions of raw source and processed water were very different for each distribution network sampled. In each network, major differences in community compositions were observed between raw and processed water, although community structures of processed water did not differ substantially from end-point tap water. End-point water samples within the same distribution network revealed very similar community structures. Network-specific communities were shown to be surprisingly stable in time. Biofilm communities sampled from domestic water metres varied distinctly between households and showed no resemblance to planktonic communities within the same distribution networks. Our findings demonstrate that high-throughput sequencing provides a powerful and sensitive tool to probe microbial community composition in drinking water distribution systems. Furthermore, this approach can be used to quantitatively compare the microbial communities to match end-point water samples to specific distribution networks. Insight in the ecology of drinking water distribution systems will facilitate the development of effective control strategies that will ensure safe and high-quality drinking water.

The objective of our study was to explore microbiological parameters that are suitable as indicators for regrowth in distribution systems that receive unchlorinated drinking water in the Netherlands. Treated water and distributed water at two locations in the distribution system of 28 treatment plants were analyzed for a range of biological parameters. The results demonstrated that Aeromonas, HPCs, and Mycobacterium were significantly (more often) higher and ATP was significantly (more often) lower in the distributed water than in the treated water, whereas cell numbers remained stable during distribution. Average fungal 18S rRNA gene copies did not differ significantly between the treated and the distributed water, but fungal numbers were significantly more often higher in the distributed water than in the corresponding treated water. These results demonstrate that the distribution system has a clear effect on HPCs, Aeromonas, Mycobacterium, ATP and, to a lesser extent, on fungi in drinking water, but not on cell numbers. Furthermore, we noticed that ATP in the treated water is a better predictor for the regrowth level in the distribution system than total, membrane-intact or HNA cell numbers. In contrast to previous studies, we conclude that ATP and cell numbers seem to be poor indicators for regrowth in the drinking water environment, whereas HPCs, Aeromonas, Mycobacterium and, to a lesser extent, fungi appear to be more reliable indicators to monitor regrowth.