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The general consensus is that the abundance of tap water bacteria is greatly influenced by water purification and distribution. Those bacteria that are released from biofilm in the distribution system are especially considered as the major potential risk for drinking water bio-safety. For the first time, this full-scale study has captured and identified the proportional contribution of the source water, treated water, and distribution system in shaping the tap water bacterial community based on their microbial community fingerprints using the Bayesian “SourceTracker” method. The bacterial community profiles and diversity analyses illustrated that the water purification process shaped the community of planktonic and suspended particle-associated bacteria in treated water. The bacterial communities associated with suspended particles, loose deposits, and biofilm were similar to each other, while the community of tap water planktonic bacteria varied across different locations in distribution system. The microbial source tracking results showed that there was not a detectable contribution of source water to bacterial community in the tap water and distribution system. The planktonic bacteria in the treated water was the major contributor to planktonic bacteria in the tap water (17.7–54.1%). The particle-associated bacterial community in the treated water seeded the bacterial community associated with loose deposits (24.9–32.7%) and biofilm (37.8–43.8%) in the distribution system. In return, the loose deposits and biofilm showed a significant influence on tap water planktonic and particle-associated bacteria, which were location dependent and influenced by hydraulic changes. This was revealed by the increased contribution of loose deposits to tap water planktonic bacteria (from 2.5% to 38.0%) and an increased contribution of biofilm to tap water particle-associated bacteria (from 5.9% to 19.7%) caused by possible hydraulic disturbance from proximal to distal regions. Therefore, our findings indicate that the tap water bacteria could possibly be managed by selecting and operating the purification process properly and cleaning the distribution system effectively.

Easily assimilable organic carbon (AOC) is frequently used for the assessment of the biological stability of drinking water, which is an important consideration in the control of bacterial growth in distribution networks. The first AOC bioassay was developed in 1982 and is based on growth of two bacterial strains (Pseudomonas fluorescens P17 and Spirillum spp. NOX) in drinking water relative to their growth on acetate. Much research was subsequently conducted to modify, simplify and increase the speed of the assay which resulted in a number of alternative AOC assays. Application of these assays raises legitimate questions about the comparison of AOC data from different studies. In the present study, a round-robin test was performed to evaluate the correlation between three established AOC methods. A total of 14 water samples, covering a wide range of AOC concentrations, were analyzed with the original ‘van der Kooij’ method, the ‘Werner & Hambsch’ method and ‘Eawag’ method. Good correlations were found between AOC concentrations measured with the various methods. The data suggest an acceptable compatibility between different AOC methods, although deviations between the methods call for careful interpretation and reporting of AOC data.

Soil column experiments were conducted to investigate the effects of effluent organic matter (EfOM) characteristics on the removal of bulk organic matter (OM) and pharmaceutically active compounds (PhACs) during managed aquifer recharge (MAR) treatment processes. The fate of bulk OM and PhACs during an MAR is important to assess post-treatment requirements. Biodegradable OM from EfOM, originating from biological wastewater treatment, was effectively removed during soil passage. Based on a fluorescence excitation-emission matrix (F-EEM) analysis of wastewater effluent-dominated (WWE-dom) surface water (SW), protein-like substances, i.e., biopolymers, were removed more favorably than fluorescent humic-like substances under oxic compared to anoxic conditions. However, there was no preferential removal of biopolymers or humic substances, determined as dissolved organic carbon (DOC) observed via liquid chromatography with online organic carbon detection (LC-OCD) analysis. Most of the selected PhACs exhibited removal efficiencies of greater than 90% in both SW and WWE-dom SW. However, the removal efficiencies of bezafibrate, diclofenac and gemfibrozil were relatively low in WWE-dom SW, which contained more biodegradable OM than did SW (copiotrophic metabolism). Based on this study, low biodegradable fractions such as humic substances in MR may have enhanced the degradation of diclofenac, gemfibrozil and bezafibrate by inducing an oligotrophic microbial community via long term starvation. Both carbamazepine and clofibric acid showed persistent behaviors and were not influenced by EfOM.

An ever-growing need exists for rapid, quantitative and meaningful methods to quantify and characterize the effect of different treatment steps on the microbiological processes and events that occur during drinking water treatment and distribution. Here we compared cultivation-independent flow cytometry (FCM) and adenosine tri-phosphate (ATP) analysis with conventional cultivation-based microbiological methods, on water samples from two full-scale treatment and distribution systems. The two systems consist of nearly identical treatment trains, but their raw water quality and pre-treatment differed significantly. All of the drinking water treatment processes affected the microbiological content of the water considerably, but once treated, the finished water remained remarkably stable throughout the distribution system. Both the FCM and ATP data were able to describe the microbiology of the systems accurately, providing meaningful process data when combined with other parameters such as dissolved organic carbon analysis. Importantly, the results highlighted a complimentary value of the two independent methods: while similar trends were mostly observed, variations in ATP-per-cell values between water samples were adequately explained by differences in the FCM fingerprints of the samples. This work demonstrates the value of alternative microbial methods for process/system control, optimization and routine monitoring of the general microbial quality of water during treatment and distribution.

Natural water treatment systems such as bank filtration have been recognized as providing effective barriers in the multi-barrier approach for attenuation of organic micropollutants for safe drinking water supply. In this study, the role of biodegradation in the removal of selected pharmaceutically active compounds (PhACs) during soil passage was investigated. Batch studies were conducted to investigate the removal of 13 selected PhACs from different water sources with respect to different sources of biodegradable organic matter. Neutral PhACs (phenacetine, paracetamol, and caffeine) and acidic PhACs (ibuprofen, fenoprofen, bezafibrate, and naproxen) were removed with efficiencies greater than 88% from different organic matter water matrices during batch studies (hydraulic retention time (HRT): 60 days). Column experiments were then performed to differentiate between biodegradation and sorption with regard to the removal of selected PhACs. In column studies, removal efficiencies of acidic PhACs (e.g., analgesics) decreased under conditions of limited biodegradable carbon. The removal efficiencies of acidic PhACs were found to be less than 21% under abiotic conditions. These observations were attributed to sorption under abiotic conditions established by a biocide (20 mM sodium azide), which suppresses microbial activity/biodegradation. However, under biotic conditions, the removal efficiencies of these acidic PhACs were found to be greater than 59%. This is mainly attributed to biodegradation. Moreover, the average removal efficiencies of hydrophilic (polar) neutral PhACs (paracetamol, pentoxifylline, and caffeine) with low octanol/water partition coefficients (log Kow less than 1) were low (11%) under abiotic conditions. However, under biotic conditions, removal efficiencies of the neutral PhACs were greater than 98%. In contrast, carbamazepine persisted and was not easily removed under either biotic or abiotic conditions. This study indicates that biodegradation represents an important mechanism for the removal of PhACs during soil passage.

Reduction of bulk organic matter, nitrogen, and pharmaceutically active compounds from primary effluent during managed aquifer recharge was investigated using laboratory-scale batch reactors. Biologically stable batch reactors were spiked with different concentrations of sodium azide to inhibit biological activity and probe the effect of microbial activity on attenuation of various pollutants of concern. The experimental results obtained revealed that removal of dissolved organic carbon correlated with active microbial biomass. Furthermore, addition of 2 mM of sodium azide affected nitrite-oxidizing bacteria leading to accumulation of nitrite-nitrogen in the reactors while an ammonium-nitrogen reduction of 95.5 % was achieved. Removal efficiencies of the hydrophilic neutral compounds phenacetin, paracetamol, and caffeine were independent of the extent of the active microbial biomass and were >90 % in all reactors, whereas removal of pentoxifylline was dependent on the biological stability of the reactor. However, hydrophobic ionic compounds exhibited removal efficiency >80 % in batch reactors with the highest biological activity as evidenced by high concentration of adenosine triphosphate.

Water quality regulations commonly place quantitative limits on the number of organisms (e.g., heterotrophic plate count and coliforms) without considering the presence of multiple cells per particle, which is only counted as one regardless how many cells attached. Therefore, it is important to quantify particle-associated bacteria (PAB), especially cells per particle. In addition, PAB may house (opportunistic) pathogens and have higher resistance to disinfection than planktonic bacteria. It is essential to know bacterial distribution on particles. However, limited information is available on quantification and identification of PAB in drinking water. In the present study, PAB were sampled from the unchlorinated drinking water at three treatment plants in the Netherlands, each with different particle compositions. Adenosine triphosphate (ATP) and total cell counts (TCC) with flow cytometry were used to quantify the PAB, and high-throughput pyrosequencing was used to identify them. The number and activity of PAB ranged from 1.0 to 3.5 × 10(3) cells ml(-1) and 0.04-0.154 ng l(-1) ATP. There were between 25 and 50 cells found to be attached on a single particle. ATP per cell in PAB was higher than in planktonic bacteria. Among the identified sequences, Proteobacteria were found to be the most dominant phylum at all locations, followed by OP3 candidate division and Nitrospirae. Sequences related to anoxic bacteria from the OP3 candidate division and other anaerobic bacteria were detected. Genera of bacteria were found appear to be consistent with the major element composition of the associated particles. The presence of multiple cells per particle challenges the use of quantitative methods such as HPC and Coliforms that are used in the current drinking water quality regulations. The detection of anoxic and anaerobic bacteria suggests the ecological importance of PAB in drinking water distribution systems.

Pellet softening reactors are used in centralized and decentralized drinking water treatment plants for the removal of calcium (hardness) through chemically induced precipitation of calcite. This is accomplished in fluidized pellet reactors, where a strong base is added to the influent to increase the pH and facilitate the process of precipitation on an added seeding material. Here we describe for the first time the opportunistic bacterial colonization of the calcite pellets in a full-scale pellet softening reactor and the functional contribution of these colonizing bacteria to the overall drinking water treatment process. ATP analysis, advanced microscopy, and community fingerprinting with denaturing gradient gel electrophoretic (DGGE) analysis were used to characterize the biomass on the pellets, while assimilable organic carbon (AOC), dissolved organic carbon, and flow cytometric analysis were used to characterize the impact of the biological processes on drinking water quality. The data revealed pellet colonization at concentrations in excess of 500 ng of ATP/g of pellet and reactor biomass concentrations as high as 220 mg of ATP/m 3 of reactor, comprising a wide variety of different microorganisms. These organisms removed as much as 60% of AOC from the water during treatment, thus contributing toward the biological stabilization of the drinking water. Notably, only a small fraction (about 60,000 cells/ml) of the bacteria in the reactors was released into the effluent under normal conditions, while the majority of the bacteria colonizing the pellets were captured in the calcite structures of the pellets and were removed as a reusable product.

AIM: Identification of the predominating cultivable bacteria in granular activated carbon (GAC) filters used in a variety of water treatment plants for selecting representative strains to study the role of bacteria in the removal of dissolved organic matter. METHODS AND RESULTS: Bacterial isolates were collected from 21 GAC filters in nine water treatment plants treating either ground water or surface water with or without oxidative pretreatment. Enrichment of samples in dilute liquid medium improved culturability of the bacteria by approximately log unit, to 9% up to 70% of the total cell counts. Genomic fingerprinting and 16S rDNA sequence analysis revealed that most (68%) of the isolates belonged to the Betaproteobacteria and 25% were identified as Alphaproteobacteria. The number of different genera within the Betaproteobacteria was higher in the GAC filters treating ozonated water than in the filters treating nonozonated water. Polaromonas was observed in nearly all of the GAC filters (86%), and the genera Hydrogenophaga, Sphingomonas and Afipia were observed in 43%, 33% and 29% of the filter beds, respectively. AFLP analysis revealed that the predominating genus Polaromonas included a total of 23 different genotypes. CONCLUSIONS: This study is the first to demonstrate that Polaromonas, which has mainly been observed in ultraoligotrophic freshwater environments, is a common component of the microbial community in GAC filters used in water treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The predominance of ultraoligotrophic bacteria in the GAC filters indicates that very low concentrations of substrates are available for microbial growth. Polaromonas species are suited for further studies on the nutritional versatility and growth kinetics enabling the modelling of biodegradation processes in GAC filters

Understanding the fate of effluent organic matter (EfOM) and natural organic matter (NOM) through riverbank filtration is essential to assess the impact of wastewater effluent on the post treatment requirements of riverbank filtrates. Furthermore, their fate during drinking water treatment can significantly determine the process design. The objective of this study was to characterise bulk organic matter which consists of EfOM and NOM during riverbank filtration using a suite of innovative analytical tools. Wastewater effluent-derived surface water and surface water were used as source waters in experiments with soil columns. Results showed the preferential removal of non-humic substances (i.e. biopolymers) from wastewater effluent-derived surface water. The bulk organic matter characteristics of wastewater effluent-derived surface water and surface water were similar after 5 m soil passage in laboratory column experiment. Humic-like organic matter in surface water and wastewater effluent-derived surface water persisted through the soil passage. More than 50% of total dissolved organic carbon (DOC) removal with significant reduction of dissolved oxygen (DO) was observed in the top 50 cm of the soil columns for both surface water and wastewater effluent-derived surface water. This was due to biodegradation by soil biomass which was determined by adenosine triphosphate (ATP) concentrations and heterotrophic plate counts. High concentrations of ATP in the first few centimeters of infiltration surface reflect the highest microbial activity which correlates with the extent of DOC reduction. Good correlation of DOC removal with DO and biomass development was observed in the soil columns.

A method for determining the concentration of active microbial biomass in granular activated carbon (GAC) filters used in water treatment was developed to facilitate studies on the interactions between adsorption processes and biological activity in such filters. High-energy sonication at a power input of 40 W was applied to GAC samples for the detachment of biomass which was measured as adenosine triphosphate (ATP)A method for determining the concentration of active microbial biomass in granular activated carbon (GAC) filters used in water treatment was developed to facilitate studies on the interactions between adsorption processes and biological activity in such filters. High-energy sonication at a power input of 40 W was applied to GAC samples for the detachment of biomass which was measured as adenosine triphosphate (ATP). Modelling of biomass removal indicated that a series of six to eight sonication treatments of 2 min each yielded more than 90% of the attached active biomass. The ATP concentrations in 30 different GAC filters at nine treatment plants in The Netherlands ranged from 25 to 5000 ng ATP cm(-3) GAC, with the highest concentrations at long filter run times and pretreatment with ozone. A similar concentration range was observed in nine rapid sand (RS) filters. ATP concentrations correlated significantly (p I mum diameter is included in these calculations. The quantitative biomass analysis with ATP enables direct comparisons with biofilm concentrations reported for spiral wound membranes used in water treatment, for distribution system pipes and other aquatic environments. (C) 2004 Elsevier Ltd. All rights reserved.