Deposition of manganese in a drinking water distribution system

The deposition of manganese in a water distribution system with manganese-related "dirty water" problems was studied over a 1-year period. Four monitoring laboratories with Robbins biofilm sampling devices fitted to the water mains were used to correlate the relationship among manganese deposition, the level of manganese in the water, and the chlorination conditions. Manganese deposition occurred by both chemical and microbial processes. Chemical deposition occurred when Mn(II) not removed during water treatment penetrated the filters and entered the distribution system, where it was oxidized by chlorine and chlorine dioxide used for disinfection. Microbial deposition occurred in areas with insufficient chlorination to control the growth of manganese-depositing biofilm. At 0.05 mg of Mn(II) per liter, the chemical deposition rate was much greater than microbial deposition. Significant deposition occurred at 0.03 mg of manganese per liter, and dirty water complaints were not eliminated until manganese levels were continuously less than 0.02 mg/liter and chlorination levels were greater than 0.2 mg/liter. A guideline level of 0.01 mg of manganese per liter is recommended.     

Deposition of manganese in a drinking water distribution system.

The deposition of manganese in a water distribution system with manganese-related "dirty water" problems was studied over a 1-year period. Four monitoring laboratories with Robbins biofilm sampling devices fitted to the water mains were used to correlate the relationship among manganese deposition, the level of manganese in the water, and the chlorination conditions. Manganese deposition occurred by both chemical and microbial processes. Chemical deposition occurred when Mn(II) not removed during water treatment penetrated the filters and entered the distribution system, where it was oxidized by chlorine and chlorine dioxide used for disinfection. Microbial deposition occurred in areas with insufficient chlorination to control the growth of manganese-depositing biofilm. At 0.05 mg of Mn(II) per liter, the chemical deposition rate was much greater than microbial deposition. Significant deposition occurred at 0.03 mg of manganese per liter, and dirty water complaints were not eliminated until manganese levels were continuously less than 0.02 mg/liter and chlorination levels were greater than 0.2 mg/liter. A guideline level of 0.01 mg of manganese per liter is recommended.     

Efficacy of bromochlorodimethylhydantoin againstLegionella pneumophila in industrial cooling water

Summary Free residual chlorine and bromine can be generated in water from bromochlorodimethylhydantoin (BCDMH). Efficacy of chlorine from inorganic sources has been studied extensively, but there is much less information on the efficacy of bromine againstL. pneumophila; only a few efficacy studies of organically-derived. halogen appear in the literature and the results from different studies conflict or are difficult to interpret. This paper describes the efficacy of halogen from BCDMH against planktonic, pure cultureL. pneumophila in an industrial cooling water. There was no difference in efficacy between halogen derived from organic or inorganic sources in controlled laboratory experiments. Effective doses in laboratory studies cannot be translated directly to field applications because of significant differences in the microbiology. However, the data suggest that disinfection (>99.9% reduction in viability within 10 min) of planktonic, pure cultureL. pneumophila can be achieved with about 1 ppm free residual halogen (expressed as chlorine) from BCDMH in a typical industrial cooling water.     

Examination of microbial contaminants of emergency showers and eyewash stations

To evaluate the effects of regular flushing, water from fifty emergency eyewash and shower stations was cultured for the presence of potentially pathogenic protozoa, heterotrophic bacteria, and Legionella species. This study also provided the opportunity to evaluate a commercially available molecular assay for the direct detection of Legionella sp in environmental samples. The Perkin Elmer Legionella EnviroAmp polymerase chain reaction (PCR) kit and culture on buffered charcoal yeast extract agar were used to detect Legionella species in water samples. Chemical and physical parameters of station water measured included: pH, hardness, alkalinity, turbidity, conductivity, total chlorine and assimilable organic carbon. Protozoal isolates were identified by classical identification methods, and isolates from the stations were identified as Hartmannella sp, Vexillifera sp, Vahlkampfia sp, Acanthamoeba sp, and Vanella sp. Heterotrophic plate counts ranged from 10² to 10⁶ CFU ml⁻¹ and acridine orange total counts ranged from 10³ to 10⁶ cells ml⁻¹ after regular flushing. PCR and gene probe analysis showed that 89% of the stations (eyewash and shower) were positive for Legionella species by PCR, while 6% of the samples were culture positive. These results indicate that routine flushing alone is not sufficient to control microbial contamination and disinfection must also be included in a routine maintenance program. In addition, regular maintenance, disinfection, and monitoring of emergency eyewash and shower stations is important in preventing potential secondary microbial infections by either direct inoculation or aerosol transmission. Received 02 September 1997/ Accepted in revised form 29 November 1997     

Comparison of culture methods and an immunofluorescence assay for the detection ofLegionella pneumophila in domestic hot water devices

The objective of this study was to compare an indirect immunofluorescence assay with culture methods for the identification ofLegionella pneumophila serogroups 1 to 6 in hot water samples taken from domestic environments. Hot water samples were obtained from the water heater, the shower heads, and the most frequently used faucet of 211 private houses. Concentrated water samples were inoculated on buffered charcoal yeast extract agar (BCYE) and on a semi-selective culture medium (GPV). Colonies with a morphology similar to that ofLegionellaceae were subcultured on BCYE and on blood agar plates; those that grew on the former but not the latter were further characterized and identified by direct immunofluorescence techniques. The concentrated samples were also smeared on multiple-well microscope slides and tested by indirect immunofluorescence with monoclonal antibodies againstL. pneumophila, serogroups 1 to 6. Of the houses studied, 30% were found to contain culturableL. pneumophila in at least one water sample, whereas 63% were positive by indirect immunofluorescence. The sensitivity of this assay compared with culture varied from 16.7–21.1%, and its specificity was between 76.7% and 88.3% depending on the sample source (water heater, shower heads, or faucet). In the 38 houses with at least one positive sample found by both immunofluorescence and culture, total or partial agreement between serogroups identified by both techniques was only 34%. The results obtained in this study strongly suggest that indirect immunofluorescence is not an adequate alternative for the identification ofL. pneumophila in hot water systems.     

Disinfection characteristics of waterborne pathogenic protozoaGiardia lamblia

Giardia lamblia is a parasitic protozoa which is transmitted in the form of a cyst through untreated water and also treated drinking water. Since its presence in water has led to frequent outbreaks of giardiasis and death in many countries, the removal and disinfection of this protozoan cyst from the water supply are of great concern for public health. This study examined the disinfection characteristics ofG. lamblia cysts isolated from a Korean patient with giardiasis. When using sodium hypochlorite includdig 5 or 10 ppm chlorine, the killing rate was initially rapid however, the disinfection slowed down and a Blog reduction could not be achieved even after 2 h. The disinfection effectiveness was also reduced at a lower temperature, thereby implying that the risk of a giardiasis outbreak will be higher in the winter season. A CT (concentration time) curve was constructed based on the results with sodium hypochlorite for use in designing and predicting disinfection performance. The organic chlorination disinfectant SDIC (sodium dichloroisocyanurate) produced a lower pH and a much higher residual effect than sodium hypochlorote. The disinfection of cysts by SDIC continued steadily throughout 2 h of contact, although the initial killing rate was lower than that with sodium hypochlorite.     

Chlorination-mediated EPS excretion shapes early-stage biofilm formation in drinking water systems

Microbial surface adhesion to surfaces and subsequent biofilm establishment are ubiquitous in drinking water systems, which often contribute to deteriorated water quality. Disinfectants are common agents applied to drinking water controlling microbial propagation, yet the underlying mechanisms of how disinfectants function to regulate microbial activity and thereby biofilm development remains elusive. We experimentally studied the effects of chlorination on extracellular polymeric substance (EPS) production, and its impacts on early-stage biofilm formation in a model drinking water system. Results showed that low-level chlorine (≤ 1.0 mg/L) stimulated microbial EPS (especially of proteins) excretion that favored early-stage biofilm formation. Microbes experiencing higher chlorination (>1.0 mg/L) exhibited clearly suppressed growth associated with reduced EPS release, consequently yielding less biofilm formation. Removal of cell-attached proteins and polysaccharides diminished biofilm formation, which highlighted the critical role of EPS (especially protein components) in biofilm development. A negative correlation between chlorination-mediated microbial protein production and cell surface charge suggested that chlorine disinfection may modify cell surface properties through regulation of microbial EPS excretion and thereby mediate biofilm formation. With these quantitative estimations, this study provides novel insights into how chlorination-mediated EPS excretion shapes early-stage biofilm formation, which is essential for practical functioning of drinking water systems.     

The influence of chlorination timing and concentration on microbial communities in labyrinth channels: implications for biofilm removal

Chlorination is an effective method to control biofilm formation in enclosed pipelines. To date, very little is known about how to control biofilms at the mesoscale in complex pipelines through chlorination. In this study, the dynamic of microbial communities was examined under different residual chlorine concentrations on the biofilms attached to labyrinth channels for drip irrigation using reclaimed water. The results indicated that the microbial phospholipid fatty acids, extracellular polymeric substances, microbial dynamics, and the ace and Shannon microbial diversity indices showed a gradual decrease after chlorination. However, chlorination increased microbial activity by 0.5–19.2%. The increase in the relative abundances of chloride-resistant bacteria (Acinetobacter and Thermomonas) could lead to a potential risk of chlorine resistance. Thus, keeping a low chlorine concentration (0.83?mg l^?1 for 3?h) is effective for controlling biofilm formation in the labyrinth channels.     

Microbial challenges for domestic heating oil storage tanks

Microbial growth on hydrocarbons is common in nature and used in bioremediation of contaminated sites, whereas in fuel storage tanks this phenomenon can affect the stability of the fuel and the tank. The impact of microbial growth and produced metabolites on materials, which are used in the construction of storage tanks, were analyzed. In contrast to metal tank components, polymeric materials did not affect or were influenced by microorganisms. Zinc was highly corroded by microbial growth, most likely due to the formation of organic acids that were produced during microbial growth on hydrocarbons. A contaminated water phase in a storage tank of a heating system was simulated with a self‐constructed pump test bench. Microbial growth began in the water phase of the storage tank and microbes were distributed throughout the tank system, through water‐in‐oil microemulsions. No microbial growth was observed in oil that was previously contaminated, indicating that essential nutrients had been depleted. The identification and removal of these essential nutrients from fuels could minimize or prevent microbial contamination. The results are discussed with regard to developing recommendations for the design and operation of domestic heating oil storage tanks to lower the risk of technical failure due to microbial contamination.     

Acanthamoeba castellanii: Cellular changes induced by chlorination

Chlorination is a well-known disinfection method, used in water treatment to inactivate various microorganisms, it induces numerous cellular changes. Even though Acanthamoebae are frequently found in water, the cellular changes induced in Acanthamoebae have not been described in the literature. Acanthamoebae are pathogenic amoebae and may provide a reservoir for pathogenic bacteria such as Legionellapneumophila; it is consequently important to understand the response of this amoeba to chlorination, and our study was indeed aimed at examining cellular changes in Acanthamoebae following chlorination. Acanthamoeba trophozoites were treated at various chlorine concentrations (1-5 mg/L). A 3-log reduction in Acanthamoebae population was achieved with 5 mg/L of free chlorine. Confocal microscopy and flow cytometry experiments indicated that chlorination induced cell permeabilization, size reduction and likely intracellular thiol concentration. Our data show that among the non-cultivable cells some remained impermeabilized (negative staining with propidium iodide), thereby suggesting that these cells might remained viable. A similar state is described in other microorganisms as a VBNC (viable but not cultivable) state. Electron microscopy observations illustrate drastic morphological changes: the pseudopods disappeared and subcellular components, such as mitochondrion, were pronouncedly affected. In conclusion, depending on the concentration used, chlorination leads to many cellular effects on Acanthamoeba that could well arise in cell inactivation.     

Accumulation and Fate of Microorganisms and Microspheres in Biofilms Formed in a Pilot-Scale Water Distribution System

The accumulation and fate of model microbial “pathogens” within a drinking-water distribution system was investigated in naturally grown biofilms formed in a novel pilot-scale water distribution system provided with chlorinated and UV-treated water. Biofilms were exposed to 1-μm hydrophilic and hydrophobic microspheres, Salmonella bacteriophages 28B, and Legionella pneumophila bacteria, and their fate was monitored over a 38-day period. The accumulation of model pathogens was generally independent of the biofilm cell density and was shown to be dependent on particle surface properties, where hydrophilic spheres accumulated to a larger extent than hydrophobic ones. A higher accumulation of culturable legionellae was measured in the chlorinated system compared to the UV-treated system with increasing residence time. The fate of spheres and fluorescence in situ hybridization-positive legionellae was similar and independent of the primary disinfectant applied and water residence time. The more rapid loss of culturable legionellae compared to the fluorescence in situ hybridization-positive legionellae was attributed to a loss in culturability rather than physical desorption. Loss of bacteriophage 28B plaque-forming ability together with erosion may have affected their fate within biofilms in the pilot-scale distribution system. The current study has demonstrated that desorption was one of the primary mechanisms affecting the loss of microspheres, legionellae, and bacteriophage from biofilms within a pilot-scale distribution system as well as disinfection and biological grazing. In general, two primary disinfection regimens (chlorination and UV treatment) were not shown to have a measurable impact on the accumulation and fate of model microbial pathogens within a water distribution system.     

Effects of Assimilable Organic Carbon and Free Chlorine on Bacterial Growth in Drinking Water

Assimilable organic carbon (AOC) is one of the most important factors affecting the re-growth of microorganisms in drinking water. High AOC concentrations result in biological instability, but disinfection kills microbes to ensure the safety of drinking water. Free chlorine is an important oxidizing agent used during the disinfection process. Therefore, we explored the combined effects of AOC and free chlorine on bacterial growth in drinking water using flow cytometry (FCM). The initial AOC concentration was 168 μg.L^-1 in all water samples. Without free chlorine, the concentrations of intact bacteria increased but the level of AOC decreased. The addition of sodium hypochlorite caused an increase and fluctuation in AOC due to the oxidation of organic carbon. The concentrations of intact bacteria decreased from 1.1×10⁵ cells.mL^-1 to 2.6×10⁴ cells.mL^-1 at an initial free chlorine dose of 0.6 mg.L^-1 to 4.8×10⁴ cells.mL^-1 at an initial free chlorine dose of 0.3 mg.L^-1 due to free chlorine originating from sodium hypochlorite. Additionally, free chlorine might be more obviously affected AOC concentrations than microbial growth did. These results suggested that AOC and free chlorine might have combined effects on microbial growth. In this study, our results showed concentrations determined by FCM were higher than those by HPC, which indicated that some E. coli detected by FCM might not be detected using HPC in drinking water. The level of free chlorine might restrain the consumption of AOC by inhibiting the growth of E. coli; on the other hand, chlorination might increase the level of AOC, thereby increase the potential for microbial growth in the drinking water network.     

Membrane biofouling by chlorine resistant Bacillus spp.: effect of feedwater chlorination on bacteria and membrane biofouling

In this study, bacteria isolated from a lake were characterised for their chlorine resistivity and the effects of chlorination on growth, mortality, protein expression and attachment propensity towards membranes. Biofouling and membrane performance were analysed. All isolated chlorine resistant strains, characterised by 16s rRNA gene sequencing, belonged to the genus Bacillus. Chlorination caused limited effects on bacterial growth and mortality. B. safensis and B. lechinoformis suffered the maximum effects due to chlorination. Live-to-dead ratios immediately after chlorination were above 1.3, with some exceptions. The membrane pure water flux recovery was highly strain dependent. Irreversible membrane fouling was observed with B. aquimaris. Membrane flux decreased substantially during ultrafiltration of water containing chlorine resistant bacteria.     

Population diversity in model potable water biofilms receiving chlorine or chloramine residual

Abstract

Most water utilities use chlorine or chloramine to produce potable water. These disinfecting agents react with water to produce residual oxidants within a water distribution system (WDS) to control bacterial growth. While monochloramine is considered more stable than chlorine, little is known about the effect it has on WDS biofilms. Community structure of 10-week old WDS biofilms exposed to disinfectants was assessed after developing model biofilms from unamended distribution water. Four biofilm types were developed on polycarbonate slides within annular reactors while receiving chlorine, chloramine, or inactivated disinfectant residual. Eubacteria were identified through 16S rDNA sequence analysis. The model WDS biofilm exposed to chloramine mainly contained Mycobacterium and Dechloromonas sequences, while a variety of alpha- and additional beta-proteobacteria dominated the 16S rDNA clone libraries in the other three biofilms. Additionally, bacterial clones distantly related to Legionella were found in one of the biofilms receiving water with inactivated chlorine residual. The biofilm reactor receiving chloraminated water required increasing amounts of disinfectant after 2 weeks to maintain chlorine residual. In contrast, free chlorine residual remained steady in the reactor that received chlorinated water. The differences in bacterial populations of potable water biofilms suggest that disinfecting agents can influence biofilm development. These results also suggest that biofilm communities in distribution systems are capable of changing in response to disinfection practices.     

Incidence of nontuberculous mycobacteria in four hot water systems using various types of disinfection

The objective of this study was to determine the incidence of nontuberculous mycobacteria (NTM) in hot water systems of 4 selected hospital settings. The hospitals provided the following types of disinfection for their hot water systems: hydrogen peroxide and silver, thermal disinfection, chlorine dioxide, and no treatment (control). In each building, 6 samples were collected from 5 sites during a 3 month period. NTM were detected in 56 (46.7%) of 120 samples; the CFU counts ranged from 10 to 1625 CFU/L. The detected NTM species were the pathogens Mycobacterium kansasii, Mycobacterium xenopi, and Mycobacterium fortuitum and the saprophyte Mycobacterium gordonae. The most common to be isolated was M. xenopi, which was present in 51 samples. The hot water systems differed significantly in the incidence of NTM. NTM were not detected in the system treated by thermal disinfection, and a relatively low incidence (20% positive samples) was found in the system disinfected with chlorine dioxide. However, a high incidence was found in the control system with no additional disinfection (70% positives) and in the system using hydrogen peroxide and silver (97% positives). Water temperatures above 50 degrees C significantly limited the occurrence of NTM.     

Amoebae in domestic water systems: resistance to disinfection treatments and implication in Legionella persistence

AIMS: Monitoring of microbial changes during and after application of various disinfection treatments in a model domestic water system. METHODS AND RESULTS: A pilot-scale domestic water system consisting of seven galvanized steel re-circulation loops and copper dead legs was constructed. Culture techniques, confocal laser scanning microscopy after fluorescent in situ hybridization and viability staining with the BacLight LIVE/DEAD kit were used for planktonic and biofilm flora monitoring. Before starting the treatments, the system was highly contaminated with Legionella pneumophila and biofilm populations mainly consisted of beta-proteobacteria. In the water and the biofilm of the loops, continuous application of chlorine dioxide (0.5 mg l(-1)), or chlorine (2.5 mg l(-1)) were very effective in reducing the microbial flora, including L. pneumophila. Heterotrophic bacteria, although strongly reduced, were still detectable after ozone application (0.5 mg l(-1)), whereas with monochloramine (0.5 mg l(-1)) and copper-silver ionization (0.8/0.02 mg l(-1)), the contamination remained significantly higher. Monochloramine and copper-silver did not remove the biofilm. During copper-silver application, Legionella re-growth was observed. Only chlorine dioxide led to detectable effects in the dead leg. Amoebae could not be eliminated, and after interrupting the treatments, L. pneumophila quickly recovered their initial levels, in all cases. CONCLUSIONS: Chlorine dioxide, applied as a continuous treatment, was identified in this study as the most efficient for controlling L. pneumophila in a domestic water system. Chlorine dioxide showed a longer residual activity, leading to improved performance in the dead leg. Amoebae resisted to all the treatments applied and probably acted as reservoirs for L. pneumophila, allowing a quick re-colonization of the system once the treatments were interrupted. SIGNIFICANCE AND IMPACT OF THE STUDY: Control of microbial contamination requires maintenance of a constant disinfectant residual throughout the water system. Treatment strategies targeting free-living amoebae should lead to improved control of L. pneumophila. Such treatment strategies still have to be investigated.     

Formation of natural biofilms during chlorine dioxide and u.v. disinfection in a public drinking water distribution system

AIMS: The influence of two disinfection techniques on natural biofilm development during drinking water treatment and subsequent distribution is compared with regard to the supply of a high-quality drinking water. METHODS AND RESULTS: The growth of biofilms was studied using the biofilm device technique in a real public technical drinking water asset. Different pipe materials which are commonly used in drinking water facilities (hardened polyethylene, polyvinyl chloride, steel and copper) were used as substrates for biofilm formation. Apart from young biofilms, several months old biofilms were compared in terms of material dependence, biomass and physiological state. Vital staining of biofilms with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and the DNA-specific 4',6-diamidino-2-phenylindole (DAPI) staining resulted in a significant difference in physiological behaviour of biofilm populations depending on the disinfection technique. Compared with chlorine dioxide disinfection (0.12-0.16 mg l-1), the respiratory activities of the micro-organisms were increased on all materials during u.v. disinfection (u.v.254; 400 J m-2). The biofilm biocoenosis was analysed by in situ hybridization with labelled oligonucleotides specific for some subclasses of Proteobacteria. Using PCR and additional hybridization techniques, the biofilms were also tested for the presence of Legionella spp., atypical mycobacteria and enterococci. The results of the molecular-biological experiments in combination with cultivation tests showed that enterococci were able to pass the u.v. disinfection barrier and persist in biofilms of the distribution system, but not after chlorine dioxide disinfection. CONCLUSIONS: The results indicated that bacteria are able to regenerate and proliferate more effectively after u.v. irradiation at the waterworks, and chlorine dioxide disinfection appears to be more applicative to maintain a biological stable drinking water. SIGNIFICANCE AND IMPACT OF THE STUDY: As far as the application of u.v. disinfection is used for conditioning of critical water sources for drinking water, the efficiency of u.v. irradiation in natural systems should reach a high standard to avoid adverse impacts on human health.     

Attachment as a factor in the protection of Enterobacter cloacae from chlorination

Enterobacter cloacae attached to drinking water distribution particles was subjected to chlorination. Attachment resulted in the protection of these organisms from disinfection. This effect was found to be dependent upon both the level of chlorine in the system and attachment time. The results obtained in this study indicate that attached organisms may play an important role in coliform outbreaks.     

Attachment as a factor in the protection of Enterobacter cloacae from chlorination.

Enterobacter cloacae attached to drinking water distribution particles was subjected to chlorination. Attachment resulted in the protection of these organisms from disinfection. This effect was found to be dependent upon both the level of chlorine in the system and attachment time. The results obtained in this study indicate that attached organisms may play an important role in coliform outbreaks.     

Necessity and Effect of Combating Legionella pneumophila in Municipal Shower Systems

The objective was to obtain research-based, holistic knowledge about necessity and effect of practiced measures against L. pneumophila in municipal shower systems in Stavanger, Norway. The effects of hot water treatment and membrane-filtering were investigated and compared to no intervention at all. The studies were done under real-world conditions. Additionally, a surveillance pilot study of municipal showers in Stavanger was performed. The validity of high total plate count (TPC) as an indication of L. pneumophila was evaluated. A simplified method, named “dripping method”, for detection and quantification of L. pneumophila was developed. The sensitivity of the dripping method is 5 colony-forming units of L. pneumophila/ml. The transference of L. pneumophila from shower water to aerosols was studied. Interviews and observational studies among the stakeholders were done in order to identify patterns of communication and behavior in a Legionella risk perspective. No substantial effects of the measures against L. pneumophila were demonstrated, except for a distally placed membrane filter. No significant positive correlation between TPC and L. pneumophila concentrations were found. L. pneumophila serogroup 2–14 was demonstrated in 21% of the 29 buildings tested in the surveillance pilot. Relatively few cells of L. pneumophila were transferred from shower water to aerosols. Anxiety appeared as the major driving force in the risk governance of Legionella. In conclusion, the risk of acquiring Legionnaires'' disease from municipal shower systems is evaluated as low and uncertain. By eliminating ineffective approaches, targeted Legionella risk governance can be practiced. Risk management by surveillance is evaluated as appropriate.