Survival, biofilm formation, and growth potential of environmental and enteric escherichia coli strains in drinking water microcosms

E. coli is the most commonly used indicator for faecal contamination in a drinking water distribution system (WDS). The assumption is that E. coli are of enteric origin and cannot persist for long outside their host, therefore acting as indicators of recent contamination events. This study investigates the fate of E. coli in drinking water; specifically addressing survival, biofilm formation under shear stress, and regrowth in a series of laboratory-controlled experiments. We show the extended persistence of three E. coli strains (two enteric and one soil isolate) in sterile and non-sterile drinking water microcosms, at 8 and 17°C, with T90 values ranging from 17.4 ± 1.8 to 149 ± 67.7 days, using standard plate counts and a series of (RT)-Q-PCR assays targeting 16S rRNA, tuf, uidA, and rodA genes and transcripts. Furthermore, each strain was capable of attaching to a surface and replicating to form biofilm in the presence of nutrients under a range of shear stress values (0.6, 2.0, and 4.4 dyn cm-2; BioFlux, Fluxion); however, cell numbers did not increase when drinking water was flowed over (t-test; p > 0.05). Finally, E. coli regrowth within drinking water microcosms containing PE-100 pipe-wall material was not observed in the biofilm or water phase using a combination of culturing and Q-PCR methods for E. coli. The results of this work highlight that when E. coli enters drinking water it has the potential to survive and attach to surfaces but that regrowth within drinking water or biofilm is unlikely

Influence of filter age on Fe, Mn and NH4+ removal in dual media rapid sand filters used for drinking water production

Rapid sand filtration is a common method for removal of iron (Fe), manganese (Mn) and ammonium (NH4+) from anoxic groundwaters used for drinking water production. In this study, we combine geochemical and microbiological data to assess how filter age influences Fe, Mn and NH4+ removal in dual media filters, consisting of anthracite overlying quartz sand, that have been in operation for between ∼2 months and ∼11 years. We show that the depth where dissolved Fe and Mn removal occurs is reflected in the filter medium coatings, with ferrihydrite forming in the anthracite in the top of the filters ( 1 m). Removal of NH4+ occurs through nitrification in both the anthracite and sand and is the key driver of oxygen loss. Removal of Fe is independent of filter age and is always efficient (> 97% removal). In contrast, for Mn, the removal efficiency varies with filter age, ranging from 9 to 28% at ∼2–3 months after filter replacement to 100% after 8 months. After 11 years, removal reduces to 60–80%. The lack of Mn removal in the youngest filters (at 2–3 months) is likely the result of a relatively low abundance of mineral coatings that adsorb Mn2+ and provide surfaces for the establishment of a microbial community. 16S rRNA gene amplicon sequencing shows that Gallionella, which are known Fe2+ oxidizers, are present after 2 months, yet Fe2+ removal is mostly chemical. Efficient NH4+ removal (> 90%) establishes within 3 months of operation but leakage occurs upon high NH4+loading (> 160 µM). Two-step nitrification by Nitrosomonas and Candidatus Nitrotoga is likely the most important NH4+ removal mechanism in younger filters during ripening (2 months), after which complete ammonia oxidation by Nitrospira and canonical two-step nitrification occur simultaneously in older filters. Our results highlight the strong effect of filter age on especially Mn2+but also NH4+ removal. We show that ageing of filter medium leads to the development of thick coatings, which we hypothesize leads to preferential flow, and breakthrough of Mn2+. Use of age-specific flow rates may increase the contact time with the filter medium in older filters and improve Mn2+ and NH4+ removal

Public participation in science : The future and value of citizen science in the drinking water research

This paper explores the value of involving citizens in the generation of knowledge in drinking water research. To this end, the significance of the 'Freshness of Water' citizen science project on the microbiological stability of drinking water was analyzed, supplemented with a series of expert interviews. In this project, citizens of Amsterdam participated in taking samples from their own kitchen tap and testing the water using test strips. The subsequent monitoring of bacteria revealed that the total number of bacterial species in all of the Amsterdam drinking water samples was high. For the participants, the presence of ten thousands of bacterial species in their drinking water, as well as the interpretation that this is perfectly normal and not a health concern, was obviously new. However, instead of causing concern or worry, this transparency clearly functioned as a strong confidence-inducing signal. A majority of the citizen scientists state that, as a result of their participation, their confidence in the quality of drinking water and the water company has increased. This study suggests that citizen science can raise the participant's water awareness and that, with the appropriate support, non-professionals can make a valuable contribution to scientific drinking water research

Initiating guidance values for novel biological stability parameters in drinking water to control regrowth in the distribution system

Nine novel biological stability parameters for drinking water have been developed recently. Here, we report data for these nine parameters in treated water from 34 treatment plants in the Netherlands to deduce guidance values for these parameters. Most parameters did not show a strong correlation with another biological stability parameter in the same sample, demonstrating that most parameters hold different information on the biological stability of drinking water. Furthermore, the novel biological stability parameters in treated water varied considerably between plants and five parameters in treated water were significantly lower for drinking water produced from groundwater than surface water. The maximum biomass concentration (MBC7), cumulative biomass potential (CBP14) from the biomass production potential test (BPP-W) and the total organic carbon concentration in treated water from groundwater were predictive parameters for HPC22 and Aeromonas regrowth in the distribution system. Guidance values of 8.6 ng ATP L−1, 110 d·ng ATP L−1 and 4.1 mg C L−1 were deduced for these parameters, under which the HPC22 and Aeromonas numbers remain at regulatory level. The maximum biomass growth (MBG7) from the BPP-W test, the particulate and/or high molecular organic carbon and the iron accumulation rate in treated water from surface water were predictive parameters for HPC22 and Aeromonas regrowth in the distribution system. Deduced guidance values for these biological stability parameters were 4.5 ng ATP L−1, 47 μg C L−1 and 0.34 mg Fe m−2 day−1, respectively. We conclude from our study that a multiple parameter assessment is required to reliable describe the biological stability of drinking water, that the biological stability of drinking water produced from groundwater is described with other parameters than the biological stability of drinking water produced from surface water, and that guidance values for predictive biological stability parameters were inferred under which HPC22 and Aeromonas regrowth is under control

The presence and growth of Legionella species in thermostatic shower mixer taps: an exploratory field study

Legislation in the Netherlands requires routine analysis of drinking water samples for cultivable Legionella species from high-priority installations. A field study was conducted to investigate the presence of Legionella species in thermostatic shower mixer taps. Water samples and the interior of ten thermostatic shower mixer taps were investigated for cultivable Legionella species. In seven cases, Legionella species was found in at least one of the samples. In four cases, Legionella species was detected in the biofilm on the thermostatic shower mixer taps interior, with the highest values on rubber parts, and in five cases in the cold supply water. These results show that thermostatic shower mixer taps can play a role in exceeding the threshold limit for cultivable Legionella species, but the cold supply water can also be responsible. Practical implications: This study showed that contamination of thermostatic shower mixer taps (TSMTs) with Legionella spp. was frequently observed in combination with contamination of the water system. Consequently, a combined focus is necessary to prevent the proliferation of cultivable Legionella spp. in TSMTs. In addition, the results also demonstrated that biofilms on rubbers inside the TSMT had high numbers of Legionella spp., probably because rubber contains relatively high concentrations of biodegradable substrates. Therefore, improvement of the rubber materials is necessary to reduce the proliferation of cultivable Legionella spp. in TSMTs

Legionella in thermostatische douchemengkranen

Prioritaire instellingen dienen in het bezit te zijn van een beheersplan legionellapreventie. Desondanks worden in de praktijk geregeld te hoge aantallen legionellabacteriën aangetroffen in monsters genomen uit thermostatische douchemengkranen. Het is niet bekend of de thermostatische douchemengkraan zelf de oorzaak is van deze overschrijdingen. Daarom is onderzoek uitgevoerd aan douchemengkranen die afkomstig zijn uit de praktijk. De resultaten laten zien dat de douchemengkraan een rol kan spelen bij een overschrijding, maar dat ook andere onderdelen van de leidingwaterinstallatie de oorzaak kunnen zijn

Effects of cold recovery technology on the microbial drinking water quality in unchlorinated distribution systems

Drinking water distribution systems (DWDSs) are used to supply hygienically safe and biologically stable water for human consumption. The potential of thermal energy recovery from drinking water has been explored recently to provide cooling for buildings. Yet, the effects of increased water temperature induced by this “cold recovery” on the water quality in DWDSs are not known. The objective of this study was to investigate the impact of cold recovery from DWDSs on the microbiological quality of drinking water. For this purpose, three pilot distribution systems were operated in parallel for 38 weeks. System 1 has an operational heat exchanger, mimicking the cold recovery system by maintaining the water temperature at 25 °C; system 2 operated with a non-operational heat exchanger and system 3 run without heat exchanger. The results showed no significant effects on drinking water quality: cell numbers and ATP concentrations remained around 3.5 × 105 cells/ml and 4 ng ATP/l, comparable observed operational taxonomic units (OTUs) (~470–490) and similar Shannon indices (7.7–8.9). In the system with cold recovery, a higher relative abundance of Pseudomonas spp. and Chryseobacterium spp. was observed in the drinking water microbial community, but only when the cold recovery induced temperature difference (ΔT) was higher than 9 °C. In the 38 weeks’ old biofilm, higher ATP concentration (475 vs. 89 pg/cm2), lower diversity (observed OTUs: 88 vs. ≥200) and a different bacterial community composition (e.g. higher relative abundance of Novosphingobium spp.) were detected, which did not influence water quality. No impacts were observed for the selected opportunisitic pathogens after introducing cold recovery. It is concluded that cold recovery does not affect bacterial water quality. Further investigation for a longer period is commended to understand the dynamic responses of biofilm to the increased temperature caused by cold recovery.</p

Recovery of microbial biomass and purification performance after scraping of full-scale slow sand filters

Slow sand filters (SSFs) are widely used in drinking water production to improve microbial safety and biological stability of water. Full-scale SSFs are maintained by scraping the biomass-rich top layers of sand. The period of downtime required for filter recovery after scraping is a major challenge due to limited knowledge of the re-stabilisation of purification processes. This study examined the recovery of microbial biomass, and removal of dissolved organic carbon (DOC) and ammonium (NH4+) in water phase and/or on sand along the depth of a scraped full-scale SSF. Scraping reduced microbial biomass on sand in the top layers, while the main prokaryotic taxa remained unaltered. Cellular ATP (cATP) and intact cell counts (ICC) in water sampled from the top layers increased, indicating a temporary disruption in functionality for 37 days. However, stable concentrations of cATP and ICC and similar microbial community composition in the effluent after scraping revealed that deeper layer biofilms offset any scraping effect. Consistent DOC and NH4+ removal after scraping showed that deeper layers effectively performed the role of the top layer. These findings highlight the resilience and robustness of microbial communities in mature full-scale SSFs and their contribution to water treatment efficiency after disturbances caused by scraping.Sanitary Engineerin

Identifying Eukaryotes and Factors Influencing Their Biogeography in Drinking Water Metagenomes

The biogeography of eukaryotes in drinking water systems is poorly understood relative to that of prokaryotes or viruses, limiting the understanding of their role and management. A challenge with studying complex eukaryotic communities is that metagenomic analysis workflows are currently not as mature as those that focus on prokaryotes or viruses. In this study, we benchmarked different strategies to recover eukaryotic sequences and genomes from metagenomic data and applied the best-performing workflow to explore the factors affecting the relative abundance and diversity of eukaryotic communities in drinking water distribution systems (DWDSs). We developed an ensemble approach exploiting k-mer- and reference-based strategies to improve eukaryotic sequence identification and identified MetaBAT2 as the best-performing binning approach for their clustering. Applying this workflow to the DWDS metagenomes showed that eukaryotic sequences typically constituted small proportions (i.e., <1%) of the overall metagenomic data with higher relative abundances in surface water-fed or chlorinated systems with high residuals. The α and β diversities of eukaryotes were correlated with those of prokaryotic and viral communities, highlighting the common role of environmental/management factors. Finally, a co-occurrence analysis highlighted clusters of eukaryotes whose members’ presence and abundance in DWDSs were affected by disinfection strategies, climate conditions, and source water types