Residual disinfection is often used to suppress biological growth in drinking water distribution systems (DWDSs), but not without undesirable side effects. In this study, water-main biofilms, drinking water, and bacteria under corrosion tubercles were analyzed from a chloraminated DWDS (USA) and a no-residual DWDS (Norway). Using quantitative real-time PCR, we quantified bacterial 16S rRNA genes and ammonia monooxygenase genes (amoA) of Nitrosomonas oligotropha and ammonia-oxidizing archaea—organisms that may contribute to chloramine loss. PCR-amplified 16S rRNA genes were sequenced to assess community taxa and diversity. The chloraminated DWDS had lower biofilm biomass (P=1×10−6) but higher N. oligotropha-like amoA genes (P=2×10−7) than the no-residual DWDS (medians =4.7×104 and 1.1×103amoA copies cm−2, chloraminated and no residual, respectively); archaeal amoA genes were only detected in the no-residual DWDS (median =2.8×104 copies cm−2). Unlike the no-residual DWDS, biofilms in the chloraminated DWDS had lower within-sample diversity than the corresponding drinking water (P<1×10−4). Chloramine was also associated with biofilms dominated by the genera, Mycobacterium and Nitrosomonas (≤91.7% and ≤39.6% of sequences, respectively). Under-tubercle communities from both systems contained corrosion-associated taxa, especially Desulfovibrio spp. (≤98.4% of sequences). Although residual chloramine appeared to decrease biofilm biomass and alpha diversity as intended, it selected for environmental mycobacteria and Nitrosomonas oligotropha—taxa that may pose water quality challenges. Drinking water contained common freshwater plankton and did not resemble corresponding biofilm communities in either DWDS; monitoring of tap water alone may therefore miss significant constituents of the DWDS microbiome. Corrosion-associated Desulfovibrio spp. were observed under tubercles in both systems but were particularly dominant in the chloraminated DWDS, possibly due to the addition of sulfate from the coagulant alum.

The disease burden from Legionella spp. infections has been increasing in many industrialized countries and, despite decades of scientific advances, ranks amongst the highest for waterborne diseases. We review here several key research areas from a multidisciplinary perspective and list critical research needs to address some of the challenges of Legionella spp. management in engineered environments. These include: (i) a consideration of Legionella species diversity and cooccurrence, beyond Legionella pneumophila only; (ii) an assessment of their environmental prevalence and clinical relevance, and how that may affect legislation, management, and intervention prioritization; (iii) a consideration of Legionella spp. sources, their definition and prioritization; (iv) the factors affecting Legionnaires’ disease seasonality, how they link to sources, Legionella spp. proliferation and ecology, and how these may be affected by climate change; (v) the challenge of saving energy in buildings while controlling Legionella spp. with high water temperatures and chemical disinfection; and (vi) the ecological interactions of Legionella spp. with other microbes, and their potential as a biological control strategy. Ultimately, we call for increased interdisciplinary collaboration between multiple research domains, as well as transdisciplinary engagement and collaboration across government, industry, and science as the way toward controlling and reducing Legionella-derived infections.

Flushing is a common corrective action recommended by Legionella management guidelines to remove stagnant water and replenish disinfectant. Due to water age and different local regulations, buildings may receive water with low or no residual disinfectant. In such situations, the evidence for flushing efficacy is often anecdotal, and the benefits are poorly quantified. Using a pilot-scale premise plumbing system, flushed shower outlets were evaluated against non-flushed outlets during simulated periods of both active and low water demand. Water and biofilm concentrations of total bacteria, Legionella spp., and Vermamoeba vermiformis were quantified using real-time quantitative PCR. Even after all outlets returned to active water demand, flushed shower outlets continued to have lower quantities of Legionella compared to non-flushed outlets for several days, but the decrease was of little practical benefit (<0.5 log₁₀[copies/L]). During prolonged periods of low water demand, however, there was no apparent benefit to flushing. Total bacteria grew to stationary phase within 3 ± 1 days, while Legionella spp. took 12 ± 6 days, regardless of whether the pipes had recently been flushed or not. Flushing with hot or cold water had little to no effect on the concentrations of total bacteria and Legionella in pipe wall biofilms. Flushing with cold water (9.6 and 13.2°C) decreased total bacteria concentrations in the water by 0.96 and 1.00 log₁₀[copies/L], respectively and by 1.27 and 1.74 log₁₀[copies/L] with hot water (49 and 60°C, respectively) but the difference in Legionella spp. concentrations between flushing with cold (1.55 log₁₀[copies/L] for both) and hot water (1.32 to 1.88 log₁₀[copies/L], respectively) was negligible. This suggests that hot water flushing, even at 60°C, provides little or no added benefit for managing Legionella in non-chlorinated building plumbing systems versus cold water flushing. Finally, the immediate benefits of flushing in terms of reductions in total bacteria and Legionella in the water were comparable as well as the rates at which those populations recovered during post-flush or post-shower stagnation.