Monochloramine Disinfection Research Articles

Selective Reactivity of Monochloramine with Extracellular Matrix Components Affects the Disinfection of Biofilm and Detached Clusters

The efficiency of monochloramine disinfection was dependent on the quantity and composition of extracellular polymeric substances (EPS) in biofilms, as monochloramine has a selective reactivity with proteins over polysaccharides. Biofilms with protein-based (Pseudomonas putida) and polysaccharide based EPS (Pseudomonas aeruginosa), as well as biofilms with varied amount of polysaccharide EPS (wild-type and mutant P. aeruginosa), were compared. The different reactivity of EPS components with monochloramine influenced disinfectant penetration, biofilm inactivation, as well as the viability of detached clusters. Monochloramine transport profiling measured by a chloramine-sensitive microelectrode revealed a broader diffusion boundary layer between bulk and biofilm surface in the P. putida biofilm compared to those of P. aeruginosa biofilms. The reaction with proteins in P. putida EPS multiplied both the time and the monochloramine mass required to achieve a full biofilm penetration. Cell viability in biofilms was also spatially influenced by monochloramine diffusion and reaction within biofilms, showing a lower survival in the surface section and a higher persistence in the middle section of the P. putida biofilm compared to the P. aeruginosa biofilms. While polysaccharide EPS promoted biofilm cell viability by obstructing monochloramine reactive sites on bacterial cells, protein EPS hindered monochloramine penetration by reacting with monochloramine and reduced its concentration within biofilms. Furthermore, the persistence of bacterial cells detached from biofilm (over 70% for P. putida and ∼40% for polysaccharide producing P. aeruginosa) suggested that currently recommended monochloramine residual levels may underestimate the risk of water quality deterioration caused by biofilm detachment.

Monochloramine Use for Prevention of Legionella in Hospital Water Systems.

Eradication of Legionella species from water distribution systems especially in hospital settings has proven to be challenging. Legionella species causes Legionnaire's disease that is a potentially fatal respiratory disease often acquired through the aerosolization of contaminated water. Monochloramine has been used successfully in the municipal water systems to eradicate Legionella and there is currently limited data to support its use in the hospital setting. This technology appears to be affordable, safe and effective at penetrating biofilm in water distribution systems.

Control of Legionella Contamination with Monochloramine Disinfection in a Large Urban Hospital Hot Water System

Control of Legionella Contamination with Monochloramine Disinfection in a Large Urban Hospital Hot Water System

Source water quality effects on monochloramine inactivation of adenovirus, coxsackievirus, echovirus, and murine norovirus

There is a need for more information regarding monochloramine disinfection efficacy for viruses in water. In this study, monochloramine disinfection efficacy was investigated for coxsackievirus B5 (CVB5), echovirus 11 (E11), murine norovirus (MNV), and human adenovirus 2 (HAdV2) in one untreated ground water and two partially treated surface waters. Duplicate disinfection experiments were completed at pH 7 and 8 in source water at concentrations of 1 and 3 mg/L monochloramine at 5 and 15 °C. The Efficiency Factor Hom (EFH) model was used to calculate CT values (mg-min/L) required to achieve 2-, 3-, and 4-log 10 reductions in viral titers. In all water types, monochloramine disinfection was most effective for MNV, with 3-log 10 CT values at 5 °C ranging from 27 to 110. Monochloramine disinfection was least effective for HAdV2 and E11, depending on water type, with 3-log 10 CT values at 5 °C ranging from 1200 to 3300 and 810 to 2300, respectively. Overall, disinfection proceeded faster at 15 °C and pH 7 for all water types. Inactivation of the study viruses was significantly different between water types, but there was no indication that overall disinfection efficacy was enhanced or inhibited in any one water type. CT values for HAdV2 in two types of source water exceeded federal CT value recommendations in the US. The results of this study demonstrate that water quality impacts the inactivation of viruses and should be considered when developing chloramination plans.

Determination of the Effects of Medium Composition on the Monochloramine Disinfection Kinetics of Nitrosomonas europaea by the Propidium Monoazide Quantitative PCR and Live/Dead BacLight Methods

Various medium compositions (phosphate, 1 to 50 mM; ionic strength, 2.8 to 150 meq/liter) significantly affected Nitrosomonas europaea monochloramine disinfection kinetics, as determined by the Live/Dead BacLight (LD) and propidium monoazide quantitative PCR (PMA-qPCR) methods (lag coefficient, 37 to 490 [LD] and 91 to 490 [PMA-qPCR] mg·min/liter; Chick-Watson rate constant, 4.0 × 10(-3) to 9.3 × 10(-3) [LD] and 1.6 × 10(-3) to 9.6 × 10(-3) [PMA-qPCR] liter/mg·min). Two competing effects may account for the variation in disinfection kinetic parameters: (i) increasing kinetics (disinfection rate constant [k] increased, lag coefficient [b] decreased) with increasing phosphate concentration and (ii) decreasing kinetics (k decreased, b increased) with increasing ionic strength. The results support development of a standard medium for evaluating disinfection kinetics in drinking water.

Inactivation of Adenoviruses, Enteroviruses, and Murine Norovirus in Water by Free Chlorine and Monochloramine

Inactivation of infectious viruses during drinking water treatment is usually achieved with free chlorine. Many drinking water utilities in the United States now use monochloramine as a secondary disinfectant to minimize disinfectant by-product formation and biofilm growth. The inactivation of human adenoviruses 2, 40, and 41 (HAdV2, HAdV40, and HAdV41), coxsackieviruses B3 and B5 (CVB3 and CVB5), echoviruses 1 and 11 (E1 and E11), and murine norovirus (MNV) are compared in this study. Experiments were performed with 0.2 mg of free chlorine or 1 mg of monochloramine/liter at pH 7 and 8 in buffered reagent-grade water at 5 degrees C. CT values (disinfectant concentration x time) for 2- to 4-log(10) (99 to 99.99%) reductions in virus titers were calculated by using the efficiency factor Hom model. The enteroviruses required the longest times for chlorine inactivation and MNV the least time. CVB5 required the longest exposure time, with CT values of 7.4 and 10 mg x min/liter (pH 7 and 8) for 4-log(10) inactivation. Monochloramine disinfection was most effective for E1 (CT values ranged from 8 to 18 mg x min/liter for 2- and 3-log(10) reductions, respectively). E11 and HAdV2 were the least susceptible to monochloramine disinfection (CT values of 1,300 and 1,600 mg-min/liter for 3-log(10) reductions, respectively). Monochloramine inactivation was most successful for the adenoviruses, CVB5, and E1 at pH 7. A greater variation in inactivation rates between viruses was observed during monochloramine disinfection than during chlorine disinfection. These data will be useful in drinking water risk assessment studies and disinfection system planning.

Monochloramine Disinfection Kinetics of Nitrosomonas europaea by Propidium Monoazide Quantitative PCR and Live/Dead BacLight Methods

Monochloramine disinfection kinetics were determined for the pure-culture ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718) by two culture-independent methods, namely, Live/Dead BacLight (LD) and propidium monoazide quantitative PCR (PMA-qPCR). Both methods were first verified with mixtures of heat-killed (nonviable) and non-heat-killed (viable) cells before a series of batch disinfection experiments with stationary-phase cultures (batch grown for 7 days) at pH 8.0, 25 degrees C, and 5, 10, and 20 mg Cl(2)/liter monochloramine. Two data sets were generated based on the viability method used, either (i) LD or (ii) PMA-qPCR. These two data sets were used to estimate kinetic parameters for the delayed Chick-Watson disinfection model through a Bayesian analysis implemented in WinBUGS. This analysis provided parameter estimates of 490 mg Cl(2)-min/liter for the lag coefficient (b) and 1.6 x 10(-3) to 4.0 x 10(-3) liter/mg Cl(2)-min for the Chick-Watson disinfection rate constant (k). While estimates of b were similar for both data sets, the LD data set resulted in a greater k estimate than that obtained with the PMA-qPCR data set, implying that the PMA-qPCR viability measure was more conservative than LD. For N. europaea, the lag phase was not previously reported for culture-independent methods and may have implications for nitrification in drinking water distribution systems. This is the first published application of a PMA-qPCR method for disinfection kinetic model parameter estimation as well as its application to N. europaea or monochloramine. Ultimately, this PMA-qPCR method will allow evaluation of monochloramine disinfection kinetics for mixed-culture bacteria in drinking water distribution systems.

Effect of exposure to UV-C irradiation and monochloramine on adenovirus serotype 2 early protein expression and DNA replication.

The mechanisms of adenovirus serotype 2 inactivation with either UV light (with a narrow emission spectrum centered at 254 nm) or monochloramine were investigated by assessing the potential inhibition of two key steps of the adenovirus life cycle, namely, E1A protein synthesis and viral genomic replication. E1A early protein synthesis was assayed by using immunoblotting, while the replication of viral DNA was analyzed by using slot blotting. Disinfection experiments were performed in phosphate buffer solutions at pH 8 and room temperature (UV) or 20 degrees C (monochloramine). Experimental results revealed that normalized E1A levels at 12 h postinfection (p.i.) were statistically the same as the corresponding decrease in survival ratio for both UV and monochloramine disinfection. Normalized DNA levels at 24 h p.i. were also found to be statistically the same as the corresponding decrease in survival ratio for monochloramine disinfection. In contrast, for UV disinfection, genomic DNA levels were much lower than E1A or survival ratios, possibly as a result of a delay in DNA replication for UV-treated virions compared to that for controls. Future efforts will determine the pre-E1A synthesis step in the adenovirus life cycle affected by exposure to UV and monochloramine, with the goal of identifying the viral molecular target of these two disinfectants.

Legionella reduction after conversion to monochloramine for residual disinfection

Previous studies have shown that monochloramine disinfection of municipal water supplies is associated with decreased risk of healthcare‐associated Legionnaires— disease. A twoyear, prospective environmental study was conducted to determine whether converting from chlorine to monochloramine for water disinfection would decrease Legionella colonization of hot water systems. Water and biofilm samples were collected from 53 buildings. Prevalence ratios comparing Legionella colonization before and after monochloramine disinfection were adjusted for water system characteristics. Legionella colonized 60% of the hot water systems before monochloramine conversion versus 4% after conversion. The median number of colonized sites per building declined significantly with monochloramine disinfection. Increased prevalence of Legionella colonization was associated with water heater temperatures below 50oC, buildings taller than 10 stories, and interruptions in water service. Although no outbreaks of Legionnaires— disease were found, the study provides support that use of monochloramine in water supplies may reduce Legionella transmission and incidence of Legionnaires— disease.

Induction of Viable but Nonculturable (VBNC) state and the effect of multiple subculturing on the survival ofLegionella pneumophila strains in the presence of monochloramine

The aim of the study was to evaluate the response of two differentLegionella pneumophila strains with their 20 times subcultured passages, regarding VBNC induction and to test the effect of multiple subculturing on cell vulnerability in the presence of monochloramine. A freshly opened ofL. pneumophila ATCC 33152, a first subculture of environmental isolate ofL. pneumophila and the 20th subcultures of both strains were tested for survival in the presence of different doses of monochloramine (24 hours). Besides culture method, live-dead cells were visualised. It was found that multiple subcultured strains were significantly more susceptible to monochloramine and not capable of entering to VBNC state in comparison with freshly opened/isolated strains. Environmental isolate was survived at 2 ppm dose of monochloramine after 24 h, whereas 20th subculture of this strain failed to survive. Multiple subculturedL. pneumophila strains were lost their culturability and viability significantly. This phenomenon should be considered while working with laboratory strains. After monochloramine disinfection,L. pneumophila bacteria can completely lose their cultivability but do not lose viability, which remains responsible for serious outbreaks worldwide. Even after completely loosing cultivability, it is possible to find live cells in network water in the VBNC state.

Inactivation kinetics of adenovirus serotype 2 with monochloramine

The effect of pH (6–10), temperature (10–30 °C), disinfectant concentration (1–11 mg/l as Cl 2), and ammonia nitrogen-to-chlorine molar ratio (1.3–52) on the inactivation kinetics of adenovirus serotype 2 with monochloramine was investigated by performing batch-reactor experiments with synthetic 0.01 M buffer (phosphate or borate) solutions. The inactivation kinetics was independent of monochloramine concentration and ammonia nitrogen-to-chlorine molar ratio but had strong pH dependence, with the rate of inactivation decreasing with increasing pH. The kinetics at pH 6 and 8 were consistent with pseudo-first-order kinetics, while curves at pH 10 were characterized by a lag phase followed by a pseudo-first-order phase. The rate of inactivation increased with increasing temperature-activation energies of 56.5 kJ/mole (pH 8) and 72.6 kJ/mole (pH 10). The results obtained in this study revealed that monochloramine disinfection might not generally provide adequate control of adenoviruses in drinking water at high pH and low temperature.

Inactivation of environmental and reference strains of heterotrophic bacteria and Escherichia coli O157:H7 by free chlorine and monochloramine

Reference cultures from culture collections are often used in disinfection experiments instead of indigenous environmental bacteria, but they may not accurately represent the organisms found in drinking water distribution systems due to physiological differences. This may explain why disinfectant concentrations and contact times, determined under typical laboratory conditions, are not always sufficient to control microbial growth or survival in distribution systems. The objective of this study was to investigate the effect of chlorine or monochloramine disinfection on Escherichia coli O157:H7 and other heterotrophic bacteria obtained from a culture collection and strains isolated from various environments, including disinfected water. It was hypothesized that previous exposure of the environmental strains to sublethal concentrations of disinfectant may allow them to develop greater resistance. Accordingly, it was observed that environmental strains of E. coli O157:H7 were either equally or less susceptible to chlorine and monochloramine than the reference strain. In contrast however, environmental strains of Brevundimonas vesicularis, Pseudomonas fluorescens, and Sphingomonas paucimobilis were either equally or more susceptible to free chlorine or monochloramine than their corresponding reference strains. This was counterintuitive because the environmental strains were able to survive disinfection in the pipes from which they were isolated. It is hypothesized that upon culturing in the laboratory the environmental strains may have become susceptible to low levels of disinfectant. Other researchers have suggested that changes in culture conditions may impact disinfectant sensitivity by affecting cell permeability, cell composition, or growth rate. This emphasizes the importance of designing laboratory studies to mimic environmental conditions as closely as possible to accurately represent environmental inactivation kinetics, including proper handling of organisms (subculturing, media transfers, etc.) before their use in inactivation studies.

Blending membrane treated WTP waste residuals with finished water: impacts to water quality and biofilm formation

The purpose of this study was to evaluate the impact of blending filtered water with treated waste residuals streams from a water treatment plant on finished water quality and biofilm formation in a modelled distribution system. A mixed waste residual stream composed of an equalized blend of filter backwash water (FBWW) and clarifier sludge blowdown was treated with a pilot-scale ultrafiltration (UF) membrane. The UF treated residuals stream was blended with plant finished water to model a recycle return location to the clearwell of a drinking water treatment train. Field evaluations involved the use of bench-scale annular reactors (ARs) to investigate two parallel model distribution systems: (i) a control system representing 100% plant filtered water (AR-filtered) and (ii) a blended system representing 10% by volume of the UF permeate blended with plant filtered water (AR-blended). To evaluate the impact of chemical disinfection on bacterial regrowth and the fate of inorganic constituents, free chlorine, monochloramine and chlorine dioxide were fed to the ARs during three distinct disinfection trials. An initial disinfection trial consisted of a control run during which no disinfectant was added. The results of the field experiments showed that blending 10% by volume of the UF permeate with plant finished water did not significantly impact regrowth of heterotrophic bacteria in the bulk or biofilm phases. Throughout each disinfection trial, the largest difference in HPCs noted between the two systems was 1 log in the bulk and biofilm samples. Free chlorine and monochloramine disinfection resulted in the greatest mitigation of microbial biofilm in both AR-filter and AR-blended systems. Aluminum, manganese, and iron in the bulk phase were found to be below Canadian Drinking Water Guideline levels. However, the detection of manganese-oxidizing bacteria (MOB) and elevated manganese in biofilm samples from AR-blended could hold potential implications for blending UF treated residual streams on biofilm development and inorganic post-precipitation occurrences in systems treating source waters containing elevated concentrations of manganese.

ReducingLegionellaColonization of Water Systems with Monochloramine

Monochloramine disinfection of municipal water supplies is associated with decreased risk for Legionnaires' disease. We conducted a 2-year, prospective, environmental study to evaluate whether converting from chlorine to monochloramine for water disinfection would decrease Legionella colonization of hot water systems. Water and biofilm samples from 53 buildings were collected for Legionella culture during 6 intervals. Prevalence ratios (PRs) comparing Legionella colonization before and after monochloramine disinfection were adjusted for water system characteristics. Legionella colonized 60% of the hot water systems before monochloramine versus 4% after conversion (PR 0.07, 95% confidence interval 0.03-0.16). The median number of colonized sites per building decreased with monochloramine disinfection. Increased prevalence of Legionella colonization was associated with water heater temperatures <50 degrees C, buildings taller than 10 stories, and interruptions in water service. Increasing use of monochloramine in water supplies throughout the United States may reduce Legionella transmission and incidence of Legionnaires' disease.

Disinfection of Burkholderia pseudomallei in potable water

The effect of chlorine, monochloramine and UV disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed. Persistence of B. pseudomallei was verified by MPN involving a one-step recovery procedure. Chlorine proved the most effective disinfectant with a 99.99% reduction of a 10(6) CFU/mL pure bacterial culture followed by 99.9% reduction by monochloramine and 99% reduction by UV. Co-culture of B. pseudomallei with Acanthamoeba astronyxis was found to greatly enhance survival of B. pseudomallei in the presence of all disinfecting agents tested. For example, when amoebae were present 100 times more monochloramine was required to maintain the disinfectant efficacy. Given the results obtained from these co-culture experiments, more research is needed to investigate the role of amoeba and biofilms in survival of B. pseudomallei in potable water.

Precursors of N-Nitrosodimethylamine in Natural Waters

Monochloramine disinfection of drinking water can result in the formation of the potent carcinogen N-nitrosodimethylamine (NDMA). To assess NDMA precursor concentrations in natural waters, samples from lakes, reservoirs, groundwaters, and isolated natural organic matter were exposed to relatively high concentrations of monochloramine. After 1 week, the concentration of NDMA ranged from approximately 0.1 to 0.8 nM. Application of a sensitive new GC/MS/MS technique and amendment of a sample with the known NDMA precursor dimethylamine (DMA) prior to chloramination indicated that only a small fraction of the NDMA produced during chloramination could be attributable to DMA. Chloramination of isolated natural organic matter and application of solid-phase extraction prior to chloramination suggested that natural organic matter accounts for a significant fraction of the precursors. However, the concentrations of NDMA precursors were higher in the epilimnion than the hypolimnion of two stratified reservoirs despite similar concentrations of dissolved organic carbon throughout the water column. Experiments performed to simulate the formation of NDMA during conditions comparable to those encountered in water treatment and distribution systems that use chloramine indicated that the concentration of NDMA produced from naturally occurring precursors usually will be less than 0.1 nM. The results indicate that chloramination of natural organic matter alone is unlikely to produce NDMA levels exceeding the current regulations.

Effect of monochloramine disinfection of municipal drinking water on risk of nosocomial Legionnaires'disease

Many Legionella infections are acquired through inhalation or aspiration of drinking water. Although about 25% of municipalities in the USA use monochloramine for disinfection of drinking water, the effect of monochloramine on the occurrence of Legionnaires' disease has never been studied. We used a case-control study to compare disinfection methods for drinking water supplied to 32 hospitals that had had outbreaks of Legionnaires' disease with the disinfection method for water supplied to 48 control-hospitals, with control for selected hospital characteristics and water treatment factors. Hospitals supplied with drinking water containing free chlorine as a residual disinfectant were more likely to have a reported outbreak of Legionnaires' disease than those that used water with monochloramine as a residual disinfectant (odds ratio 10.2 [95% CI 1.4-460]). This result suggests that 90% of outbreaks associated with drinking water might not have occurred if monochloramine had been used instead of free chlorine for residual disinfection (attributable proportion 0.90 [0.29-1.00]). The protective effect of monochloramine against legionella should be confirmed by other studies. Chloramination of drinking water may be a cost-effective method for control of Legionnaires' disease at the municipal level or in individual hospitals, and widespread implementation could prevent thousands of cases.

Reduction of Norwalk virus, poliovirus 1 and coliphage MS2 by monochloramine disinfection of water

The reduction of Norwalk virus (NV) by a 2 mg/L dose of pre-formed monochloramine was determined at pH 8 and 5°C in bench-scale, batch disinfection experiments using quantitative RT-PCR for NV assays. Two other enteric viruses, poliovirus 1 (PV1) and coliphage MS2, were included for comparison and assayed by infectivity as well as RT-PCR. After 3h, reductions of PV1 and MS2 by infectivity assays were about 1 log10 but there were no reductions of these viruses by RT-PCR assays. Hence, RT-PCR underestimated virus inactivation by monochloramine. However, NV reduction by monochloramine was about 1 log10 by RT-PCR assay, suggesting that it is more susceptible to monochloramine than the other two viruses tested. Based on RT-PCR titre reduction, the CT99 value for NV was about 775 mg-min/L. If the reduction of NV infectivity by monochloramine is ever greater than the reduction of RT-PCR signals, the CT99 value would be smaller. However, the results of this study indicate that NV and the other enteric viruses tested are not rapidly and extensively reduced by disinfection with pre-formed monochloramine.

Reduction of Norwalk virus, poliovirus 1 and coliphage MS2 by monochloramine disinfection of water

The reduction of Norwalk virus (NV) by a 2 mg/L dose of pre-formed monochloramine was determined at pH 8 and 5°C in bench-scale, batch disinfection experiments using quantitative RT-PCR for NV assays. Two other enteric viruses, poliovirus 1 (PV1) and coliphage MS2, were included for comparison and assayed by infectivity as well as RT-PCR. After 3h, reductions of PV1 and MS2 by infectivity assays were about 1 log10 but there were no reductions of these viruses by RT-PCR assays. Hence, RT-PCR underestimated virus inactivation by monochloramine. However, NV reduction by monochloramine was about 1 logl0 by RT-PCR assay, suggesting that it is more susceptible to monochloramine than the other two viruses tested. Based on RT-PCR titre reduction, the CT99 value for NV was about 775 mg-min/L. If the reduction of NV infectivity by monochloramine is ever greater than the reduction of RT-PCR signals, the CT99 value would be smaller. However, the results of this study indicate that NV and the other enteric viruses tested are not rapidly and extensively reduced by disinfection with pre-formed monochloramine.

Comparison of respiratory activity and culturability during monochloramine disinfection of binary population biofilms

Biofilm bacteria challenged with monochloramine retained significant respiratory activity, even though they could not be cultured on agar plates. Microbial colony counts on agar media declined by approximately 99.9% after 1 h of disinfection, whereas the number of bacteria stained by a fluorescent redox dye experienced a 93% reduction. Integrated measures of biofilm respiratory activity, including net oxygen and glucose utilization rates, showed only a 10 to 15% reduction. In this biofilm system, measures of microbial respiratory activity and culturability yielded widely differing estimates of biocide efficacy.