No abstract

Phosphate dosing is used by water utilities to prevent plumbosolvency in water supply networks. However, there is a lack of knowledge regarding biofilm formation on lead and plastic materials when phosphate concentrations are modified in drinking water systems. In this study, biofilms were grown over lead coupons and PVC tubes in bioreactors supplied with local drinking water treated to provide different phosphate doses (below 1, 1 and 2 mg/L) over a period of 28 days. A range of commercial iron pellets (GEH104 and WARP) were tested aiming to maintain phosphate levels below the average 1 mg/L found in drinking water. Changes in biofilm community structure in response to three different phosphate treatments were characterised by Illumina sequencing of the 16S rRNA gene for bacteria and the ITS2 gene for fungi. Scanning electron microscopy was used to visualise physical differences in biofilm development in two types of materials, lead and PVC. The experimental results from the kinetics of phosphate absorption showed that the GEH104 pellets were the best option to, in the long term, reduce phosphate levels while preventing undesirable turbidity increases in drinking water. Phosphate-enrichment promoted a reduction of bacterial diversity but increased that of fungi in biofilms. Overall, higher phosphate levels selected for microorganisms with enhanced capabilities related to phosphorus metabolism and heavy metal resistance. This research brings new insights regarding the influence of different phosphate concentrations on mixed-species biofilms formation and drinking water quality, which are relevant to inform best management practices in drinking water treatment.

Water utilities treat drinking water by adding phosphate to prevent metal dissolution from water pipe work systems and particularly lead poisoning. Phosphate can be a limiting nutrient for microbial biofilms in DWDS, yet its effects on these microbial consortia are not well understood. This research presents results from phosphate dosing experiments using a real scale chlorinated DWDS, comparing standard phosphate concentrations of United Kingdom drinking water (1 mgP/L) with a double dose (2 mgP/L) commonly used in plumbosolvency treatment. Biofilm development during phosphate treatment experiments was monitored using a holistic approach by combining metagenomics analysis, flow cytometry and SEM characterisation. The increase of phosphate levels in drinking water, reduced biofilm cell numbers and promoted the presence of poorly distributed biofilms on inner pipe surfaces. Metagenomics analysis using genetic markers (16S rRNA and ITS2) showed that phosphate influenced biofilm community structure, particularly fungal composition. Whole metagenome sequencing showed that phosphate enrichment favoured the presence of sequencing reads associated to ATPases, ion transporters and DNA-interacting proteins, whilst reads associated to nitrogen metabolism were predominant in control samples. This research brings new knowledge regarding the influence of phosphate treatment on the composition and structure of biofilms within DWDS, and the implications that this might have for the management of these systems.

This is a repository copy of The microbial ecology of a Mediterranean chlorinated drinking water distribution systems in the city of Valencia (Spain).

Drinking water distribution systems (DWDS) can host pathogenic amoebae, but the role of biofilms in supporting the occurrence of these organisms needs to be fully explored in the UK systems. The presence of amoebae and associated bacteria in biofilms attached to inner pipe surfaces was studied in an experimental full-scale chlorinated distribution system in the UK. Quantitative polymerase change reaction (qPCR) was used to identify and quantify amoebae, whilst the bacterial communities in the biofilms were characterised by sequencing the 16S rRNA gene. Despite the maintenance of a chlorine residual in the network (free chlorine ≥ 0.24 mg/L), several species of amoebae belonging to the genera Acanthamoeba, Vermamoeba, and Naegleria were identified in 30-day-old biofilm samples; however, no amoebae were detected in the water samples analysed. The dominant bacterial communities present in the biofilm samples were Variovorax, Pseudomonas, and Aquabacterium. These results indicate that the biofilm samples contained potential pathogenic amoebae and bacteria, such as Acanthamoeba and Pseudomonas, respectively, which implies a potential public health risk if the biofilms are mobilised into the bulk water. Several of the amoebae identified in this study are able to support the presence of resistant bacteria that can remain viable within these prokaryotic organisms until they reach people’s taps. The identification of the microorganisms associated with the pathogenic amoeba species in biofilms could be used to improve the surveillance of DWDS in order to protect public health.

This paper uses a two-fold multi-criteria decision-making (MCDM) approach applied for the first time to the field of microbial management of drinking water distribution systems (DWDS). Specifically, the decision-making trial and evaluation laboratory (DEMATEL) was applied removing the need for reliance on expert judgement, and analysed interdependencies among water quality parameters and microbiological characteristics of DWDS composed of different pipe materials. In addition, the fuzzy technique for order preference by similarity to ideal solution (FTOPSIS) ranked the most common bacteria identified during trials in a DWDS according to their relative abundance while managing vagueness affecting the measurements. The novel integrated approach presented and proven here for an initial real world data set provides new insights in the interdependence of environmental conditions and microbial populations. Specifically, the application shows as the bacteria having associated the most significant microbial impact may not be the most abundant. This offers the potential for integrated management strategies to promote favourable microbial conditions to help safeguard drinking water quality.

Amoeba-related diseases have been related with the presence of certain amoebas in domestic water, including drinking water. Biofilms in drinking water distribution systems are able to support amoeba growth by providing a food source and protecting them against disinfectants. Additionally, amoeba growth can be favoured by warm temperatures and climate change appears to contribute to its geographic spread. The presence of amoeba and its association with potential pathogenic bacteria was studied in a real-scale chlorinated DWDS. The test facility comprised three independent pipe loops fed with water from the local water supply and for this study a varied flow hydraulic profile was applied based on daily patterns observed in real UK distribution networks. The daily regime was repeated for a biofilm growth phase of 30 days. Amoeba viability was tested by a culture-based method, non-nutrient agar (NNA)-E. coli plates, and then confirm by qPCR using specific primers to detect species of amoeba including Naegleria and Acanthamoeba. Amplicon sequencing of the 16s rRNA gene was used to characterise the biofilm and planktonic bacterial communities. Several amoeba species belonging to the genera Acanthamoeba, Vermamoeba and Naegleria were identified in 30-day old biofilm samples. While the bacterial communities in biofilms were dominated by Variovorax, Pseudomonas and Aquabacterium. This study yielded new insights on the dynamics of amoeba and bacterial communities in DWDS. However, more research is required to accurately establish the impact of these inter-kingdom associations on human health.

Phosphate is added to drinking water by UK water companies as a treatment to prevent the corrosion and metal leaching, like lead, in pipes. However, phosphate is a nutrient for microorganisms, and it can favour biofilm formation in Drinking Water Distribution Systems (DWDS), which can alter the water quality and safety. This study analyses the effect of phosphate addition on biofilm formation over different materials and its consequences for drinking water quality by i) using controlled experimental pipeline facility representative of a real-scale DWDS with high-density polyethylene coupons and ii) using a small-scale DWDS biofilm reactors with lead coupons. Biofilms developed over one month were exposed to the effect of different phosphate dosing and compare with UK normal water phosphate concentrations. During the experiment, physico-chemical analysis of water and microbial analysis of biofilms was carried out. Sequencing analysis of the 16s rRNA gene, from extracted DNA obtained from biofilms, provided information on any bacterial changes, and Scanning Electron Microscopy gave information about the biofilm organization. The results indicate that microorganisms find more difficult to establish and develop biofilms under high phosphate dosing, resulting in biofilms with less cells. Also, some physico-chemical parameter seems to be affected by phosphate dosing, like chlorine and lead. It is expected that differences in the biofilm community will be found depending on phosphate dosing. This study will provide information on the effect of phosphate on biofilm development in different pipes materials, which will facilitate to adjust an optimal phosphate dose to prevent plumbosolvency in DWDS.