Chemical monitoring strategy for the assessment of advanced water treatment plant performance.

A pilot-scale plant was employed to validate the performance of a proposed full-scale advanced water treatment plant (AWTP) in Sydney, Australia. The primary aim of this study was to develop a chemical monitoring program that can demonstrate proper plant operation resulting in the removal of priority chemical constituents in the product water. The feed water quality to the pilot plant was tertiary-treated effluent from a wastewater treatment plant. The unit processes of the AWTP were comprised of an integrated membrane system (ultrafiltration, reverse osmosis) followed by final chlorination generating a water quality that does not present a source of human or environmental health concern. The chemical monitoring program was undertaken over 6 weeks during pilot plant operation and involved the quantitative analysis of pharmaceuticals and personal care products, steroidal hormones, industrial chemicals, pesticides, N-nitrosamines and halomethanes. The first phase consisted of baseline monitoring of target compounds to quantify influent concentrations in feed waters to the plant. This was followed by a period of validation monitoring utilising indicator chemicals and surrogate measures suitable to assess proper process performance at various stages of the AWTP. This effort was supported by challenge testing experiments to further validate removal of a series of indicator chemicals by reverse osmosis. This pilot-scale study demonstrated a simplified analytical approach that can be employed to assure proper operation of advanced water treatment processes and the absence of trace organic chemicals.

Fluorescence monitoring at a recycled water treatment plant and associated dual distribution system--implications for cross-connection detection.

Dual distribution systems are becoming increasingly common in greenfield housing developments in Australia for the redistribution of recycled water to households for non-potable use. Within such schemes there exists the potential for cross-connections between recycled and drinking water systems. Due to the high level of recycled water treatment, these events are unlikely to lead to outbreaks of illness in the community. Nonetheless, they do represent a breach of the recycled water risk management strategy and therefore an elevated level of risk to consumers. Furthermore, cross-connection events have the potential to undermine public confidence in these types of water recycling. A rapid, highly sensitive method of cross-connection detection may therefore provide an additional level of confidence in these schemes. The aim of this research was to determine the potential for using fluorescence spectroscopy as a monitoring tool in water treatment plants and dual distribution systems. Samples from both the water recycling plant and dual distribution system were collected on a weekly basis over 12 weeks. Fluorescence excitation-emission matrix (EEM) spectra and water quality parameters including dissolved organic carbon, UV(254), pH, conductivity, free chlorine and turbidity were obtained for each sample. The fluorescence EEM spectra of recycled and drinking water were distinctly different and exhibited low variability throughout the course of the sampling program, indicating a degree of stability of the fluorescent components within the organic matter. A ten-fold difference in mean fluorescence intensity was observed for recycled water compared to drinking water, which was greater than the difference observed for the other measured water quality parameters. Probabilistic analysis was used to determine the reliable detection limit of recycled water contamination of drinking water. Accounting for the inherent variability of both recycled water and drinking water, a 45% contamination of recycled water in drinking water could be detected with a signal-to-noise ratio greater than 3 for more than 95% of individual random sample pairs. Greater sensitivity can be assured by averaging numerous samples. In comparison, a 70% contamination of recycled water in drinking water was required for the same detection using conductivity.

Biofilms in an urban water distribution system: measurement of biofilm biomass, pathogens and pathogen persistence within the Greater Stockholm Area, Sweden.

Distribution pipe biofilms can provide sites for the concentration of a wide range of microbial pathogens, thereby acting as a potential source of continual microbial exposure and furthermore can affect the aesthetic quality of water. In a joint project between Stockholm Water, the MISTRA "Sustainable Urban Water" program, the Swedish Institute for Infectious Disease Control and the Royal Technical University, Stockholm, the aim of the current study was to investigate biofilms formed in an urban water distribution system, and quantify the impact of such biofilms on potential pathogen accumulation and persistence within the Greater Stockholm Area, Sweden. When used for primary disinfection, ultra-violet (UV) treatment had no measurable influence on biofilm formation within the distribution system when compared to conventional chlorination. Biofilms produced within a model pilot-plant were found to be representative to those that had formed within the larger municipal water distribution system, demonstrating the applicability of the novel pilot-plant for future studies. Polystyrene microspheres (1.0 microm) and Salmonella bacteriophages demonstrated their ability to accumulate and persist within the model pilot-plant system, where the means of primary disinfection (UV-treatment, chlorination) had no influence on such phenomena. With the exception of aeromonads, potential pathogens and faecal indicators could not be detected within biofilms from the Stockholm water distribution system. Results from this investigation may provide information for water treatment and distribution management strategies, and fill key data gaps that presently hinder the refinement of microbial risk models.

Detection, integration and persistence of aeromonads in water distribution pipe biofilms.

The occurrence of Aeromonas spp. within biofilms formed on stainless steel (SS), unplasticized polyvinyl chloride (uPVC) and glass (GL) substrata was investigated in modified Robbins Devices (MRD) in potable (MRD-p) and recycled (MRD-r) water systems, a Biofilm Reactor (BR) and a laboratory-scale pipe loop (PL) receiving simulated recycled wastewater. No aeromonads were isolated from the MRD-p whereas 3-10% of SS and uPVC coupons (mean 3.85 CFU cm(-2) and 12.8 CFU cm(-2), respectively) were aeromonad-positive in the MRD-r. Aeromonads were isolated from six SS coupons (67%) (mean 63.4 CFU cm(-2)) and nine uPVC coupons (100%) (mean 6.50x 10(2) CFU cm(-2)) in the BR fed with recycled water and from all coupons (100%) in the simulated recycled water system (PL). Mean numbers of aeromonads on GL and SS coupons were 5.83 x 10(2) CFU cm(-2) and 8.73 x 10(2) CFU cm(-2), respectively. No isolate was of known human health significance (i.e. Aeromonas caviae, A. hydrophila or A. veronii), though they were confirmed as Aeromonas spp. by PCR and fluorescence in situ hybridization (FISH). Challenging the PL biofilms with a slug dose of A. hydrophila (ATCC 14715) showed that biofilm in the PL accumulated in the order of 10(3)-10(4) A. hydrophila cm(-2), the number of which decreased over time, though could not be explained in terms of conventional 1st order decay kinetics. A sub-population of FISH-positive A. hydrophila became established within the biofilm, thereby demonstrating their ability to incorporate and persist in biofilms formed within distribution pipe systems. A similar observation was not made for culturable aeromonads, though the exact human health significance of this remains unknown. These findings, however, further question the adequacy of culture-based techniques and their often anomalous discrepancy with direct techniques for the enumeration of bacterial pathogens in environmental samples.

The fate of legionellae within distribution pipe biofilms: measurement of their persistence, inactivation and detachment.

Distribution pipe biofilms present a currently unquantified public health risk to consumers receiving water for domestic potable and non-potable use. The aim of this study was to quantify the numbers of legionellae, used here as model bacterial pathogens, that may accumulate, persist within and detach from distribution pipe biofilms. L. pneumophila recovered by standard culture from an 8 week-old biofilm formed within a novel pilot-scale water distribution system represented 1% of those present in the adjacent bulk water. A combined chlorine concentration exceeding 0.2 mg x L(-1) eliminated culturable sessile legionellae altogether, though the reduction in FISH-positive cells represented just 75+/-25% of the original amount, compared to a 5-log reduction in culturable cells during the same period. Where there was < 0.1 mg x L(-1) combined chlorine, an exponential decay/loss of sessile L. pneumophila was observed (k = 0.37 - 0.41) over the course of a 38-day experimental period. The inoculation of the system with 1 microm fluorescent microspheres and legionellae demonstrated that removal of the latter was dominated by chemical disinfection, with erosion and biological grazing playing lesser roles. Under turbulent (Re approximately 5000) conditions, larger clusters of biofilm become detached from substrata, with more than 90% of sessile legionellae mobilised into the bulk water phase. Interaction with both biofilms and a thermophilic Acanthamoeba isolate reduced the susceptibility of legionellae to thermal inactivation by between one and two orders of magnitude, though it increased their sensitivity to chemical (free and combined chlorine) disinfection.

Biofilms, thermophilic amoebae and Legionella pneumophila--a quantitative risk assessment for distributed water.

A simplistic quantitative microbial risk assessment (QMRA) based on the maximum risk curve (r = 1) was developed for Legionella within a water distribution system. Both biofilms and a thermophilic isolate of acanthamoebae were shown to increase the resistance of Legionella to conventional thermal disinfection by between one and two logs respectively. The level of risk presented to consumers was shown to exceed the USEPA 10(-4) benchmark in many cases tested. This was caused, in part, by the sensitivity of the risk model but also through a lack of reliable dose-response data for Legionella. Not withstanding this, the current study provided comparative information on the efficacy of conventional disinfection against Legionella. Combined chlorine was shown to reduce the risk of infection by as much as 1-log when compared to free chlorine, although thermal disinfection provided the most effective means of risk reduction. Biofilm detachment and the interaction of Legionella with acanthamoebae were two important ecological factors that significantly increased the risk of legionellosis, and thus should be further considered in the refinement of QMRA models.

Enteric virions and microbial biofilms--a secondary source of public health concern?

Through their many sorption sites, microbial biofilms can accumulate both organic and inorganic particulate and colloidal material from bulk water environments. An application of such first principles to the ecology of "biocolloidal" enteric virions would suggest that they too may be concentrated by biofilms in a similar way. Though previous studies have isolated human gastrointestinal (enteric) virions from microbial biofilms, the exact human health significance of this has been neither fully investigated nor completely understood. Through an assessment of the location, accumulation and persistence of model enteric virion (phiX174, MS2 and B40-8 bacteriophages as well as 20 nm fluorescent latex microspheres) within biofilms, the aim of the current study was to investigate whether the interaction of enteric virions with distribution pipe biofilms could provide a secondary source of public health concern to consumers. Model enteric virions were found to be incorporated into biofilms at concentrations representing 1% of those present in the adjacent bulk water environment. A sub-population (0.01%) of these persisted throughout an experimental period of 30 days, inferring their potential to accumulate over time. Furthermore, model enteric virions were partitioned into bacterial microcolonies, environments where biofilm bacteria can persist and re-grow, even in the presence of "acceptable" levels of disinfection. A risk model for enteric virion accumulation and release from distribution pipe biofilms suggested that associated risks may exceed USEPA benchmark values. These findings could have wide-reaching implications in water treatment and distribution strategies, and necessitate a re-appraisal of current water guideline values.

Persistence of two model enteric viruses (B40-8 and MS-2 bacteriophages) in water distribution pipe biofilms.

The persistence of two model enteric virions (Bacteroides fragilis phage B40-8 and coliphage MS-2) within pipe biofilms was investigated in situ in an urban distribution system. Biofilms were allowed to develop on uPVC and stainless steel (SS) coupons in a modified Robbins' device for 70 d within a 150 mm uPVC reticulation main. Coupons were then placed in annular reactors and slug dosed with B40-8 and MS-2 phages (10(8) pfu/mL). Pipe water velocity, pH and free chlorine were recorded during the experimental period. Biofilms on uPVC were generally more abundant (based on total bacterial counts, HPCs, total protein and total carbohydrate). Both B40-8 and MS-2 were incorporated into biofilms formed on uPVC and SS coupons (> 10(4) and > 10(3) pfu/microgram protein respectively) and persisted for > 30 d and 6 d respectively, reflecting biofilm biomass on the two pipe surfaces. Virion loss/inactivation from biofilm followed an initial rapid phase, followed by a very slow phase representing approximately 0.01% of the original virion population. Virions, therefore, have the potential to accumulate within distribution biofilm and problems could arise when clusters of biofilm-associated enteric virions become detached from the substrata by hydrodynamic forces or sudden changes in disinfection regime.