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Chloramine is used as a disinfectant and due to temperature variations and higher retention times, some remote locations of the distribution network experience reduced bulk water disinfection residuals. The decay of chloramine along distribution systems is well demonstrated in past literature [ 36 ]. The MRD was installed at a location where chloramine residuals measured as total chlorine remained less than 0. As shown in Fig 1 , the MRD was placed in series as a by-pass to the main distribution line.

Biofouling of different pipe materials concrete, HDPE and stainless steel was investigated by inserting coupons 4 cm x 1. The coupons were orientated along the MRD as 14 rings Fig 1. Each ring was composed of 10 coupons labelled a to j of a single pipe material. There were two opposing coupons at any given arrangement of the ring e.

Specifically, a low water flow rate of 0. The Reynold's number, which reflects flow characteristics was calculated using Eq 1. Bulk water and biofilm sampling MRD and KIWA monitor was carried out fortnightly during the first three months and monthly thereafter for another three months spanning from end of Oct to end of Apr. Subsequently, the two opposing coupons of the relevant rings were removed one at a time so as to minimise drying of attached biofilm. Sampling of the attached biofilm was carried out by swabbing one side of the coupon 6.

Swabbing was limited to 10 times and in between swabbing the swab was immersed into a sterile 25 mL Falcon tube containing 5 mL of autoclaved dechlorinated tap water to further moisten the swab and also to dislodge any swabbed biomass. One falcon tube with two glass rings contained 10 mL of autoclaved dechlorinated tap water. The third glass ring contained in another falcon tube was used to analyse iron and manganese deposits. The sampling tap was kept flowing for 5 min prior to collecting the bulk water.

The suspensions were then decanted into new tubes and the biofilm suspension and glass rings were sonicated once more for 3 min in fresh autoclaved dechlorinated tap water 5 mL for swabs and 10 mL for rings. The corresponding suspensions of coupons and glass rings were pooled and an aliquot of each was subjected to an ATP analysis. The remainder of suspensions were filtered 0. An aliquot of each of the suspensions was used for metal analysis and the remainder was used to extract DNA. The concentrated samples were subsequently used to extract DNA.

Quantitative estimation of iron and manganese was given precedence over other metals considering the influence these two metals have on discoloured water events [ 38 ]. Iron, manganese and ATP were analysed in triplicate for each sample and additional details can be found in Ginige et al. ATP provides a quantitative estimation of viable microorganisms present in samples.

Sequencing was carried out utilizing a Roche FLX titanium instrument and reagents. The USEARCH sequence analysis method carried out clustering, chimera checking, denoising and also performed quality checks and filtering of de-multiplexed sequences [ 41 ]. The unprocessed DNA sequences of this study were deposited accession number To explore the impact of pipe material on chemical and biological fouling, a one-way between-groups analysis of variance was conducted using SPSS version 13 SPSS, Chicago.

The samples were divided into 3 groups according to biofilm age 0 to1 month, 2 to 4 months and 5 to 6 months to facilitate this analysis. The percentage abundance of sequences in each OTU calculated based on total sequences of each sample was compared using program PAST [ 44 ] to assess bacterial similarities between samples.

Since the pipes are aboveground they are exposed to the environmental elements e. According to this study the ranking of the pipe material in the reduction of average ATP is as follows:. This suggests that although the overall level of biofilm formation on concrete is lower than that of HDPE, the initial increase of biofilm formation on concrete is higher. Whether the slow increase of biofilm formation on HDPE and on stainless steel is a result of surface characteristics remains unclear.

Biofilms can actively and passively contribute towards deposition of iron and manganese on pipe materials in drinking water systems [ 7 , 45 ]. However, no correlations were observed with concrete. According to Tsai et al.

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However, with stainless steel and HDPE being largely chemically inert, the deposition of iron and manganese may have been triggered upon an initial colonisation of bacteria on these two plumbing materials. The lack of correlation with concrete on the other hand is a result of the inherent ability of concrete to facilitate chemical deposition of iron and manganese [ 46 , 47 ].

In this instance, chemical deposition has far exceeded biologically mediated deposition of iron and manganese on concrete. Overall the ranking of the pipe material in terms of susceptibility to metals deposition iron and manganese, determined based on the mean concentration of metals is as follows:. Overall this study was restricted to a single location of distribution system and since many factors some of which are area dependent determine the fouling biological and chemical potential risk of plumbing materials, further research is required to validate e.

Eboigbodin et al. Considering the device was exposed to reduce flow velocities 0. Further, a significant drop of ATP Fig 2B was noted over time between 0 to 1 month and 2 to 4 months.

With ATP measurements only reflecting the active biomass fraction, it is unclear whether the dead biomass fraction on roughened glass increased with time and biofouling still resembled coupons of MRD. While other operational strategies e. Specifically iron deposition on glass was 21 times higher than on concrete, 31 times higher than on HDPE and 70 times higher than on stainless steel.

Similarly manganese deposition was 52 times higher than on concrete, times higher than on HDPE and times higher than on stainless steel. Adsorption of cations, onto glass has been extensively studied [ 49 ] and stronger binding was observed with bivalent and trivalent cations e. Accordingly, the presence of excessive quantities of iron and manganese on roughened glass of the KIWA monitor is not surprising.

Similar to glass, concrete and HDPE adsorb cations, and if the KIWA monitor is to reflect metal deposition on pipe material, the glass monitoring rings are best replaced with the same pipe material of the distribution system. While this change may contribute towards a positive outcome, other operational strategies may also need to be explored to successfully retrofit the use of the KIWA monitor to quantitatively estimate iron and manganese build up in distribution systems.

In this study, the biofilms on coupons of MRD continued to mature during the experimental period.


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The main distribution pipeline upstream of MRD, however, contained a mature biofilm and bulk water characteristics monitored reflect changes taking place in the larger distribution system. Hence, making an attempt to correlate bulk water changes to changes occurring on an immature biofilm on coupons of MRD is unrealistic. High water flow velocities during summer [ 50 ] may have resulted in this observation.

However, the increase of bulk water iron and manganese during summer did not coincide with a release of deposited metal from coupons of MRD. The bacterial diversity of the collected samples spread across a total 21 bacterial phyla. With Proteobacteria outnumbering other bacterial phyla in all samples examined, this study together with other studies [ 51 — 55 ] confirm the dominance of Proteobacteria in drinking water distribution systems. Other bacterial phyla such as Chloroflexi, Gemmatimonadetes, Actinobacteria, Firmicutes, Verrucomicrobia, Acidobacteria were observed, but their abundance was only 3.

These bacterial phyla also have been noted in drinking water environments, and factors such as the source water quality pH, organic matter, nitrogen etc.

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Irrespective of the type of media that facilitated biofilm growth, bacterial species richness Chao1 , phylogenetic diversity PD and bacterial diversity Shannon index were highest in bulk water Table 2. As a complete contrast to our study, Douterelo et al. Srinivasan et al. This highlights the influence of the disinfection residual towards abundance and diversity of bacteria in bulk water and in biofilm. While past research could provide some insight to the observed differences of Douterelo et al. As the biofilm on pipe material aged, the bacterial diversity indices Cho1, Shannon and PD decreased Table 2 and a notable difference was evident between ages 0 to 1 month and 5 to 6 month.

However, an opposite trend was observed in biofilm of KIWA monitor. Liu et al. In addition, an elemental analysis has revealed significant concentrations of aluminium, calcium, iron, magnesium, manganese, and arsenic like metals in loose deposits [ 59 ]. Our study revealed higher deposition of metals on roughened glass of the KIWA monitor and characteristics of glass was assumed to have contributed towards this observation.

Accordingly, it is unclear whether higher metal concentrations contributed towards the higher bacterial diversity on roughened glass of the KIWA monitor. The pipe material did not significantly contribute towards bacterial diversity differences in pipe wall biofilms Table 2 , Fig 3. However, according to the cluster analysis Fig 4B , some differences were noted amongst biofilms on different pipe materials of similar age.

The above observation is further consolidated when bacterial phyla were compared on pipe materials at different ages of biofilms. The abundance of some bacterial phyla increased while some decreased with increasing age of biofilm. The differences in abundance were found to be more pronounced at a genus level S2 Table. Biofilm management in a distribution system is reliant on a disinfectant residual that is effective towards the bacterial diversity on pipe wall biofilms [ 60 ]. Fig 3 demonstrates a relatively uniform bacterial community across all pipe materials specifically during early age of biofilm.

Future research could explore whether higher phylogenetic similarity of young biofilms on pipe materials Fig 4A , could be exploited to better manage disinfection residuals of distribution systems having concrete, HDPE, stainless steel or a mixture of the above pipe materials. The phylogenetic similarity between bulk water bacteria and biofilms of pipe materials decreased as the biofilms aged Figs 3 and 4. Accordingly, if a pipe wall biofilm is young e.

Consistent with the cluster analysis, the shared OTUs gradually decreased with increasing biofilm age of pipe materials Fig 5A. When similar age biofilms e. This further suggests that pipe materials have an influence on the bacterial diversity of a biofilm. Specifically pipe composition, age, surface roughness was found to impact biomass deposition on pipe material [ 19 , 61 ].

Based on this understanding, van der Kooij et al. The inert glass and Teflon surfaces were specifically chosen to remove the effects of pipe materials on biomass accumulation, e. However, as the use of glass in KIWA monitors failed to phylogenetically reflect bacterial diversities in both the bulk water and pipe material Figs 3B and 5B , whether the use of a KIWA monitor fitted with inert glass could be extended to reflect microbial quality of bulk water remains questionable.

To examine whether pipe wall biofilms could be monitored through alternative means, offline biofilm a KIWA monitor and bulk water monitoring were employed. Biofouling was examined at community level using pyrosequencing. The findings of the study can be summarized as follows:. The authors would like to acknowledge Dr. Geoffrey Puzon, Dr. Matthew Morgan, Dr. Naomi McSweeney, and Dr. Suzy Rea for their valuable comments.

Supervision: MPG. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field. Abstract A Modified Robbins Device MRD was installed in a full-scale water distribution system to investigate biofouling and metal depositions on concrete, high-density polyethylene HDPE and stainless steel surfaces. Introduction Drinking water comes into contact with different surfaces made from a variety of materials during its travel from source to consumer.

Materials and Methods Biofilm monitoring setup on the full-scale distribution system A MRD length: cm and diameter: Download: PPT. Fig 1. Sampling of biofilm and bulk water Bulk water and biofilm sampling MRD and KIWA monitor was carried out fortnightly during the first three months and monthly thereafter for another three months spanning from end of Oct to end of Apr.

Sample processing and analysis Processing of coupons, glass rings and bulk water. Iron, manganese and ATP analysis. Bacterial community analysis using sequencing. Statistical analysis To explore the impact of pipe material on chemical and biological fouling, a one-way between-groups analysis of variance was conducted using SPSS version 13 SPSS, Chicago. Fig 2. Details of monitored parameters during the study period.

Plumbing material has an influence on biofilm formation. Plumbing material and biofilm influence metal deposition. Table 1. Relationship between ATP and deposited metals on pipe material. Whether bulk water could be used to make predictions remains inconclusive. Bacterial community compositions in bulk water, pipe material and KIWA monitor pyrosequencing was performed to examine the bacterial community composition and diversity in bulk water, different pipe materials and on KIWA monitor.

Physicochemical parameters determine the extent of bacterial diversity. Table 2. Bacterial diversity indices of biofilms on pipe materials, glass and in bulk water. Diversity variations are based on biofilm age and not on colonization material. Fig 3. Comparison of the relative abundances of major bacterial classes found in biofilms on pipe materials and bulk water during three different sampling periods.

Fig 4. Fig 5. The findings of the study can be summarized as follows: HDPE pipe surfaces were more susceptible towards biofouling while concrete surfaces were towards metals deposition. KIWA monitors under operational conditions used in this study do not reflect biological and chemical fouling of pipe surfaces.


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Roughened glass of KIWA influenced abiotic deposition of metals. Bacterial diversity and phylogenetic similarity decreased with age of biofilm on concrete, HDPE and stainless steel pipe surfaces. However, bacterial diversity increased on roughened glass of KIWA monitor. Supporting Information.

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S1 Table. S2 Table. Bacterial community composition at genus level observed in biofilms on pipe materials, glass and bulk water at different ages of biofilm and bulk water temperature. S1 Fig. S2 Fig. Comparison of the relative abundances of major bacterial phyla at different age of biofilm and bulk water temperatures on pipe materials, bulk water and KIWA monitor.

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