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.     

Resistance of Legionella to disinfection in hot water distribution systems.

The efficiency of various disinfection treatments against Legionella was tested on a hot water distribution system (HWDS) pilot unit. The results demonstrated clearly that most Legionella in the networks were fixed in the biofilm at the surface of the pipe (more than 98% for each loop). Chemical treatments (continuous chlorination, hyperchlorination, hydrogen peroxide and peracetic acid mixing) commonly used for the eradication of Legionella in hot water distribution networks appeared to be inadequate for eradicating the bacteria in the biofilm. Unfortunately, the biofilm contained most of the pathogens in an HWDS whereas legislation is only restricted to the Legionella concentration in the water phase. Thermal treatment appeared to be efficient to disinfect most of the biofilm but seemed to promote the biofilm re-growth as well. It was then concluded that the best solution to prevent Legionella contamination in hot water distribution systems would be to have perfect control of the temperature in the networks (temperature > 55 degrees C at all points). Nevertheless, in many cases it is difficult to have such control, so during the time necessary to modify networks, the best solution to control Legionella proliferation appears to be to apply a treatment shock (thermal or chlorination as a function of pipe characteristics). These treatments must be followed by a continuous chlorination that is totally controlled and equipped with alarm systems. This study demonstrates that biofilm sampling devices must be installed in hot water distribution systems to anticipate Legionella contamination and correctly determine the efficiency of the treatments.     

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.     

Photoreactivation of Escherichia coli following medium-pressure ultraviolet disinfection and its control using chloramination.

Ultraviolet (UV) disinfection is becoming increasingly popular as an alternative disinfection technology to chlorination in recent years. In this study, we investigated the photoreactivation of Escherichia coli following medium-pressure (MP) UV disinfection of synthetic water by a bench-scale collimated beam apparatus. The UV doses ranged from 1.6 -19.7 mWs/cm2 and photoreactivation was investigated for 6 hours under fluorescent light. In addition, chloramination was applied after UV disinfection to investigate its ability to control photoreactivation. It was found that photoreactivation occurred for all UV doses tested and the increase in bacteria numbers ranged from 0.04 to 1.35 log10. However, the degree of photoreactivation decreased with increased UV doses. Chloramination experiments revealed that the addition of 0.5 mg/l of monochloramine resulted in suppression of photoreactivation for 1 hour only. An increased monochloramine dose of 1 mg/l was found to prevent photoreactivation for the entire duration of the experiment. The results of this study have shown that photoreactivation occurs even after MP UV disinfection, although it is of a lesser extent at higher UV doses. This study has also established that secondary chloramination can effectively suppress and eliminate photoreactivation with a chloramine dose of 1 mg/l.     

Exploring the potential synergistic effects of chemical disinfectants and UV on the inactivation of free-living bacteria and treatment of biofilms in a pilot-scale system

The main objective of this study is to explore possible synergistic or additive effects of combinations of chemical disinfectants (sodium hypochlorite, peracetic acid, hydrogen peroxide, chlorine dioxide) and UV in their efficacy in inactivating free-living bacteria and removing biofilms. In contrast to most studies, this study examines disinfection of municipal water in a pilot-scale system using a mixed bacterial suspension, which enables a better simulation of the conditions encountered in actual industrial environments. It was shown that the combination of either hypochlorite, hydrogen peroxide, peracetic acid, or chlorine dioxide with UV yielded additive effects on the inactivation of free-living bacteria. Actual synergy was observed for the combination of UV and 5 ppm hydrogen peroxide. Regarding biofilm treatment, additive effects were observed using the combination of hydrogen peroxide and UV. The promising results obtained in this study indicate that the combination of UV and chemical disinfectants can considerably reduce the amount of chemicals required for the effective disinfection and treatment of biofilms.     

Contamination potential of drinking water distribution network biofilms.

Drinking water distribution system biofilms were investigated for the presence of hygienically relevant microorganisms. Early biofilm formation was evaluated in biofilm reactors on stainless steel, copper, polyvinyl chloride (PVC) and polyethylene coupons exposed to unchlorinated drinking water. After 12 to 18 months, a plateau phase of biofilm development was reached. Surface colonization on the materials ranged between 4 x 10(6) and 3 x 10(7) cells/cm2, with heterotrophic plate count (HPC) bacteria between 9 x 10(3) and 7 x 10(5) colony-forming units (cfu)/cm2. Established biofilms were investigated in 18 pipe sections (2 to 99 years old) cut out from distribution pipelines. Materials included cast iron, galvanized steel, cement and PVC. Colonization ranged from 4 x 10(5) to 2 x 10(8) cells/cm2, HPC levels varied between 1 and 2 x 10(5) cfu/cm2. No correlation was found between extent of colonization and age of the pipes. Using cultural detection methods, coliform bacteria were rarely found, while Escherichia coli, Pseudomonas aeruginosa and Legionella spp. were not detected in the biofilms. In regular operation, distribution system biofilms do not seem to be common habitats for pathogens. However, nutrient-leaching materials like rubber-coated valves were observed with massive biofilms which harboured coliform bacteria contaminating drinking water.     

氯胺消毒对卤乙酸类消毒副产物的控制研究

对影响氯胺消毒副产物的因子进行了研究,结果表明:降低氯与氨氮的比值能够降低卤乙酸消毒副产物生成总量,当氯与氨氮比降至5:1时,氯胺消毒产生卤乙酸的总量比氯消毒能够减少69.1%;消毒副产物的生成量与氯胺的投加量呈很好的线性关系;延长接触时间对氯胺消毒副产物的生成量影响小;pH升高能够减少卤乙酸类消毒副产物的生成量;升高温度对卤乙酸生成量的影响很小。采用氯胺取代氯消毒能够很好控制卤乙酸类消毒副产物。     

Chemical disinfection of Legionella in hot water systems biofilm: a pilot-scale 1 study

Legionella bacteria encounter optimum growing conditions in hot water systems and cooling towers. A pilot-scale 1 unit was built in order to study the biofilm disinfection. It consisted of two identical loops, one used as a control and the other as a 'Test Loop'. A combination of a bio-detergent and a biocide (hydrogen peroxide + peracetic acid) was applied in the Test Loop three times under the same conditions at 100 and 1,000 mg/L with a contact time of 24 and 3-6 hours, respectively. Each treatment test was preceded by a three week period of biofilm re-colonization. Initial concentrations of culturable Legionella into biofilm were close to 10(3) CFU/cm2. Results showed that culturable Legionella spp. in biofilm were no longer detectable three days following each treatment. evertheless, initial Legionella spp. concentrations were recovered 7 days after the treatments (in two cases). Before the tests, Legionella spp. and L. pneumophila PCR counts were both about 10(4) GU/cm2 in biofilm and they both decreased by 1 to 2 log units 72 hours after each treatment. The three tests had a good but transient efficiency on Legionella disinfection in biofilm.     

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.     

Wastewater reclamation systems in small communities.

The demands established in the rules and regulations by the administration in Catalonia seem to exclude small communities from wastewater reclamation and reuse, due to the comparatively high costs associated with the practice at small scale. In the framework of the DRAC project (Demonstration on Wastewater Reclamation and Reuse in Catalonia) two different pre-treatment systems, one extensive (infiltration-percolation) and another intensive (ring filter), each one followed by chlorine dioxide disinfection, were tested in order to be applied for small communities wastewater reclamation and reuse. The results of this study show that infiltration-percolation systems remove very efficiently physico-chemical contaminants and microorganisms. The ring filter system does not show a significant removal rate of contaminants, The use of infiltration-percolation as a pre-treatment for advanced chemical disinfection allows reducing the dose of disinfectant and the contact time needed to achieve a specific water quality, and diminishes disinfection byproducts (DBPs) generation. Therefore, this reclamation line is suitable for small communities due to its efficiency and low cost. However, further studies are needed in relation to the removal mechanisms of microorganisms, organic compounds in IP systems and the possible DBPs formation using chlorine dioxide.     

Correlations between surrogate nitrogenous organic precursors and C-, N-DBP formation

Nitrosamines have been emerging as disinfection byproducts in drinking water using source water impacted with domestic wastewaters. Nitrogenous organic compounds are suggested as precursors of nitrosamines, but many of them have not been identified. This study investigated the correlations between nine selected nitrogenous organic compounds with different characteristics and corresponding disinfection by-product formation potentials (nitrosamines, trihalomethanes (THMs), and haloacetic acids (HAAs)) from their reactions with free chlorine, chlorine dioxide and monochloramine. Besides dimethylamine, the well-known precursor of nitrosamines, 3-(N,N-dimethyloctylammonio)propanesulfonate (3-N,N-DAPSIS) inner salt and benzyldimethyltetradecylamine (benzalkonium chloride, BKC) were suggested as important nitrosamine precursors. 3-N,N-DAPSIS could form about 1,000 ng/L of N-nitrosodimethylamine (NDMA) and more than 1,000 μg/L of THMs. More than 150,000 ng/L of NDMA were observed when BKC was treated with monochloramine, and high levels of THMs (up to 2,700 μg/L) were also yielded. As expected, free chlorine produced higher levels of THMs and HAAs, and chlorine dioxide generated minor levels of traditional DBPs. Nitrosamines were mainly formed when the precursors were treated with monochloramine.     

Disinfection by-products and microbial contamination in the treatment of pool water with granular activated carbon.

For swimming pools, it is generally agreed that free chlorine levels have to be maintained to guarantee adequate disinfection. Recommended free chlorine levels can vary between 0.3 and 0.6 mg/L in Germany and up to 3 mg/L in other countries. Bathers introduce considerable amounts of organic matter, mainly in the form of such as urine and sweat, into the pool water. As a consequence, disinfection byproducts (DBPs) are formed. Regulations in Germany recommend levels of combined chlorine of less than 0.2 mg/L and levels of trihalomethanes (THMs) of less than 20 microg/L. Haloacetic acids (HAAs), haloacetonitriles (HANs), chloropicrin and chloral hydrate are also detected in considerable amounts. However, these compounds are not regulated yet. Swimming pool staff and swimmers, especially athletes, are primarily exposed to these byproducts by inhalation and/or dermal uptake. In Germany, new regulations for swimming pool water treatment generally require the use of activated carbon. In this project, three different types of granular activated carbon (GAC) (one standard GAC, two catalytic GACs) are compared for their long time behaviour in pool water treatment. In a pilot plant operated with real swimming pool water, production and removal of disinfection byproducts (THMs, HAAs, AOXs), of biodegradable substances (AOC), of bacteria (Pseudomonas aeruginosa, Legionella, coliforms, HPC) as well as the removal of chlorine and chloramines are monitored as function of GAC bed depth. Combined chlorine penetrates deeper in the filter bed than free chlorine does. However, both, free and combined chlorine removal efficiencies decrease over the time of filter operation. The decreases of removal efficiencies are also observed for parameters such as dissolved organic carbon, spectral absorption coefficient, adsorbable organic carbon and most of the disinfection byproducts. However, THMs, especially chloroform are produced in the filter bed. The GAC beds were contaminated microbially, especially with P. aeruginosa. The contamination was not removable by backwashing with chlorine concentrations up to 2 mg/l free chlorine.     

农村地区饮用水消毒技术的应用

饮用水的消毒是给水工程的一项主要水处理技术,人们为此做了大量的工作,对新的消毒技术和方法进行了不断的探索.目前除了常用的氯消毒外,还有很多新的消毒方法,这些方法也克服或减少了氯消毒时产生的副作用.但这些新方法也存在生产制造成本较高、应用条件尚不够成熟等问题,还需要进一步研究改进.     

Advanced treatment for taste and odour control in drinking water: case study of a pilot scale plant in Seoul, Korea.

Taste and odour problems of tap water in Seoul are attributed to 2-methylisoborneol (2-MIB) and trans-1,10-dimethyl-trans-9-decalol (geosmin), which are the result of metabolism of algae and chlorine for disinfection. This study was carried out to measure 2-MIB and geosmin in the raw water from the Han River, to investigate removal efficiency of GAC and BAC integrated with post-ozonation, and to minimise and quantify the required chlorine concentration as a final disinfectant through the candidate process.     

次氯酸钠与液氯消毒在农村供水中的应用效果对比研究

为明确次氯酸钠消毒在农村供水工程中的实际应用效果,考察了南方某省以同一地表水为水源、水处理工艺相同、分别采用液氯和次氯酸钠消毒的具有一定代表性的两处农村供水工程(T水厂和H水厂)的余氯衰减、消毒效果和副产物生成情况。结果显示:在有效氯投加量分别为1.0 mg/L和2.0 mg/L时,2.4 h后H水厂出厂水的余氯值衰减率比T水厂分别低6.53%和3.30%,两者的消毒效果均较好,H水厂的管网末梢水菌落总数值比T水厂低40.62%,微生物安全性更高;当有效氯投加量由1.0 mg/L增至2.0 mg/L时,T水厂和H水厂出厂水的三氯甲烷生成量分别增长97.36%和59.35%,三卤甲烷分别增长122.64%和91.43%,而二氯乙酸和三氯乙酸生成量增长率相差不大。研究结果表明,次氯酸钠的稳定性及消毒效果略好于液氯,且消毒副产物三氯甲烷和三卤甲烷的生成量较小。由于两水厂在水源类型、水源水质特征、工程规模及水处理工艺方面具有一定代表性,经与其他研究进行对比分析认为,次氯酸钠消毒在农村供水中具有较好的应用前景。     

Sensory aspects of drinking water in contact with epoxy lined copper pipe.

Pipe relining via in situ epoxy lining is used to remediate corroded plumbing or distribution systems. This investigation examined the effects on odour, TOC, THM formation and disinfectant demand in water exposed to epoxy-lined copper pipes used for home plumbing. The study was conducted in accordance with the Utility Quick Test, a migration/leaching method for utilities to conduct sensory analysis of materials in contact with drinking water. The test was performed using water with no disinfectant and levels of chlorine and monochloramines representative of those found in the distribution system. Panelists repeatedly and consistently described a "plastic/adhesive/putty" odour in the water from the pipes. The odour intensity remained relatively constant for each of two subsequent flushes. Water samples stored in the epoxy-lined pipes showed a significant increase in the leaching of organic compounds (as TOC), and this TOC was demonstrated to react with free chlorine to form trichloromethane. Water stored in the pipes also showed a marked increase in disinfectant demand relative to the water stored in glass control flasks. A study conducted at a full scale installation at an apartment demonstrated that after installation and regular use, the epoxy lining did not yield detectable differences in water quality.     

Application of HACCP principles as a management tool for monitoring and controlling microbiological hazards in water treatment facilities.

HACCP (hazard analysis and critical control points) principles were applied to evaluate the effectiveness of two water treatment facilities to continually produce potable water free of microbiological health hazards. This paper reports a hazard analyses protocol (microbiological hazards based on faecal coliforms (FC) and turbidity (TBY) as indicators) for critical control points (CCPs) within each facility. The CCPs were raw resource water, sedimentation, filtration and chlorine-disinfection. The aim was to determine the effectiveness of each CCP to remove the indicators from the water under treatment. Arbitrary critical performance limit targets (CPLTs) were set up for each CCP to determine to what extent each contributed to effective removal and to predict what the effect would be if any of the CCPs should fail. Health-related water quality guideline limits for expected health effects were applied and compliance measured at the 90th percentile. The raw resource river water used at both treatment facilities complied with raw resource water extraction CPLTs. The treated potable water complied with health-related drinking water guidelines. Sedimentation removed the largest proportion of the indicators from the raw water, but showed failure potential that could overload the consequent system. Filtration effectiveness at both treatment facilities showed potential to break down the overall effectiveness of the entire treatment facility, since the filter systems failed to meet their respective CPLTs. This left the disinfection phase to remove the remaining portion of indicators. Faecal coliforms appeared to be completely removed from post-chlorination samples. This indicated that both chlorine disinfection phases were 100% effective in meeting their disinfection CPLTs, despite having to "clean up" the indicator organisms that spilt over from the upstream CCPs. This, nevertheless, implied a risk of unsafe water release into distribution. CCPs at these treatment facilities had some difficulties in reducing the health-related risks to meet their respective CPLTs. Applying a HACCP programme would minimise the risk of contaminated water distribution in cases of system component failure.     

Two-dimensional cellular automaton model for mixed-culture biofilm.

Structural and microbial heterogeneity occurs in almost any type of biofilm system. General approaches for the design of biofilm systems consider biofilms as homogeneous and of constant thickness. In order to improve the design of biofilms systems, models need to incorporate structural heterogeneity and the effect of inert microbial mass. We have improved a 2D biofilm model based on cellular automata (CA) and used it to simulate multidimensional biofilms with active and inert biomass including a self-organizing development. Results indicate that the presence of inert biomass within biofilm structures does not change considerably the substrate flux into the biofilm because the active biomass is located at the surface of the biofilm. Long-term simulations revealed that although the biofilm system is highly heterogeneous and the microstructure is continuously changing, the biofilm reaches a dynamic steady-state with prediction of biofilm thickness and substrate flux stabilizing on a delimited range.     

Microsensor measurement of oxygen concentration in biofilms: from one dimension to three dimensions.

In this study, we measured oxygen concentration in biofilms in one dimension in field conditions and in three dimensions in laboratory conditions by using a robust oxygen microsensor in combination with an automation and data acquisition system. The biofilms were on the discs of rotating biological contactors treating domestic wastewater. The results of this study provide experimental evidence on oxygen distribution in wastewater biofilms and on biofilm structure. (1) The three dimensional measurements of oxygen concentration in biofilms revealed "pockets" of oxygen in deep sections of biofilms. In these isolated "pockets," located 600-760 microm from the biofilm surface, dissolved oxygen concentration was as high as 1 mg/L. This depth of oxygen diffusion is deeper than what was determined based on one dimensional measurements. (2) The heterogeneity of oxygen distribution was related to the surface structure of biofilms. The structure of the biofilm surface affected the diffusion boundary layer over the surface and, in turn, the oxygen diffusion and distribution inside biofilms. (3) Oxygen concentration in biofilms changed generally from a high degree of heterogeneity near the biofilm surface to a low degree of heterogeneity in deep sections of biofilms, indicating a cell-clusters-like structure near the surface and a more compact base layer close to the substratum.