Inorganic chloramines: a critical review of the toxicological and epidemiological evidence as a basis for occupational exposure limit setting.

Inorganic chloramines are not commercially available, but monochloramine is produced in situ for disinfection or for use in chemical synthesis. Inorganic chloramines are also formed when free chlorine reacts with nitrogen containing substances, e.g. ammonia and urea, present in chlorinated water sources. Occupational exposure may, therefore, occur in e.g. swimming pool facilities and the food processing industry. Monochloramine is soluble and stable in water and the dominating inorganic chloramine in chlorinated water sources. No clinical effects were seen in healthy volunteers given monochloramine in drinking water during 4 or 12 weeks in doses of 0.043 or 0.034 mg/kg bw/day, respectively. Limited data indicate that monochloramine is weakly mutagenic in vitro but not genotoxic in vivo. One drinking water study indicated equivocal evidence of carcinogenicity in female rats but not in male rats and mice. No reproductive or developmental effects were shown in rodents in the few studies located. Dichloramine is soluble but unstable in water. In the only study located, mild histological effects in kidneys, thyroid and gastric cardia were observed in rats administered dichloramine in drinking water for 13 weeks. Trichloramine is immiscible with water and evaporates easily from water into air. Therefore, the primary exposure route of concern in the occupational setting is inhalation. Occupational exposure to trichloramine has been demonstrated in indoor swimming pool facilities and in the food processing industry where chlorinated water is used for disinfection. Exposure-response relationships between airborne levels and self-reported ocular and upper airway irritation have been shown in several studies. Exposure to trichloramine may aggravate asthma symptoms in individuals with existing asthma. The risk of developing asthma following long-term exposure to trichloramine cannot be evaluated at present. No data on genotoxic, carcinogenic, reproductive or developmental effects were located. The toxicological data for mono- and dichloramine are insufficient to recommend health-based occupational exposure limits (OELs).As regard trichloramine, the critical effect is judged to be irritation observed in several studies on pool workers, starting at approximately 0.4 mg/m3 (stationary sampling). Based on these data, a health-based OEL of 0.1 mg/m3 (8-h time-weighted average) is recommended. This corresponds to 0.2 mg/m3 for stationary measurements in swimming pool facilities. No short-term exposure limit (STEL) is recommended.

Evaluation of biocidal efficacy of Chloramine T trihydrate on planktonic and sessile bacteria in a model cooling tower water system.

A model cooling tower system was experimentally seeded with Legionella pneumophila and real industrial cooling tower (CT) water has been run at the closest to full-scale system operating conditions. The water/biofilm samples were taken from the model system monthly, and the effectiveness of the different concentrations of Chloramine T trihydrate biocide was evaluated in terms of its ability to control both planktonic/sessile microbial populations. Although Chloramine T is a recommended commercial formulation for disinfecting CTs, there is a lack of published data on the efficacy of this compound against both planktonic and sessile populations in the cooling tower. Biocide response in both sessile/planktonic bacteria counts varied according to months. Tested biocide concentrations provided the clean tower conditions by reducing the concentration of heterotrophic plate count (HPC) below <104 cfu mL-1, L. pneumophila <10 cfu mL-1 and of adenosine triphosphate (ATP) values <300 relative light units (RLU), after 1, 3 and 24 h of exposure, during a 6-month period. There were no statistically significant differences in efficacy between concentrations in terms of reduction in the number of bacteria, decrease in ATP value and viability. The results revealed that Chloramine T can effectively control biofouling in cooling systems according to the limit values of the successful control program.

Haemolysis in haemodialysis.

Haemolysis in haemodialysis, although rare in current times, is associated with significant mortality and morbidity. As such prompt recognition, treatment, analysis of root cause and correction of underlying causative factors is crucial. Dialysate, extracorporeal circuit and patient related factors all contribute to haemolysis risk. Haemolysis can manifest with non-specific signs and symptoms including but not restricted to hypertension, nausea, pain (abdominal, chest, back) and dyspnoea. It may present acutely during the dialysis session or may take a protracted course. Potential life threating consequences include; hyperkalaemia induced cardiac arrhythmias, profound anaemia and associated acute coronary events and respiratory distress, and severe necrotizing pancreatitis. Chronic haemolysis results in impaired endothelial function thus contributing to the long-term cardiovascular risk profile in haemodialysis patients. Stringent national and international standards, technological advancements in membrane and dialysis equipment design, dialyser purification methods and water treatment systems have greatly reduced the incidence of haemolysis. Despite these improvements recognition of haemolysis risk and ongoing clinical vigilance is important.

Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells.

We evaluated the antifungal and anti-biofilm activity, mechanism of action and cytotoxicity of chloramine T trihydrate (CAT) against Candida spp. The Minimum Inhibitory and Fungicidal Concentrations (MIC/MFC) of CAT were determined. Changes in CAT-treated C. albicans growth kinetics and micromorphology were evaluated, as well as the mechanism of action, and its effects on biofilm. Cytotoxicity was assessed by the hemolysis method. The data were analyzed by inferential statistics (p ≤ 0.05). CAT showed antifungal activity against all strains, with MIC values ranging between 1.38 and 5.54 mmol/L (MIC75%: 2.77 mmol/L). CAT demonstrated an immediate and sustained action on C. albicans growth kinetics, particularly at 2 × MIC. This compound likely acts on the cell wall and membrane permeability simultaneously and was found to cause changes in C. albicans micromorphology. Tha antibiofilm activity of CAT was similar to that of sodium hypochlorite (p > 0.05) against mature biofilms. CAT was more effective than NaOCl in reducing mature biofilm upon 1-min exposure at 2 × MIC (24 h) and 4 × MIC (48 h) (p < 0.05). Toxicological analysis revealed that CAT had hemolytic activity between 61 and 67.7% as compared to 100% by NaOCl. CAT has antifungal and anti-biofilm properties, probably acting on both cell wall and membrane permeability, and showed low toxicity in vitro.

Comparative mammalian cell cytotoxicity of wastewater with elevated bromide and iodide after chlorination, chloramination, or ozonation.

Recycling wastewater is becoming more common as communities around the world try to better control their water resources against an increased frequency of either prolonged droughts or intense flooding. For communities in coastal areas, wastewaters may contain elevated levels of bromide (Br-) and iodide (I-) from seawater intrusion or high mineral content of source waters. Disinfection of such wastewater is mandatory to prevent the spread of pathogens, however little is known about the toxicity of wastewater after disinfection in the presence of Br- and I-. In this study we compared the induction of chronic cytotoxicity in mammalian cells in samples of municipal secondary wastewater effluent amended with elevated levels of Br-/I- after disinfection by chlorine, chloramines or ozone to identify which disinfection process generated wastewater with the lowest level of adverse biological response. Chlorination increased mammalian cell cytotoxicity by 5 times as compared to non-disinfected controls. Chloramination produced disinfected wastewater that expressed 6.3 times more cytotoxicity than the non-disinfected controls and was 1.3 times more cytotoxic than the chlorinated samples. Ozonation produced wastewater with cytotoxicity comparable to the non-disinfected controls and was at least 4 times less cytotoxic than the chlorine disinfected wastewaters. These results indicate that compared to chlorination and chloramination, ozonation of wastewater with high Br-/I- levels yielded the lowest mammalian cell cytotoxicity, suggesting its potential as a more favorable method to disinfect wastewater with minimizing the biological toxicity in mind.

Evaluating Evidence for Association of Human Bladder Cancer with Drinking-Water Chlorination Disinfection By-Products.

Exposure to chlorination disinfection by-products (CxDBPs) is prevalent in populations using chlorination-based methods to disinfect public water supplies. Multifaceted research has been directed for decades to identify, characterize, and understand the toxicology of these compounds, control and minimize their formation, and conduct epidemiologic studies related to exposure. Urinary bladder cancer has been the health risk most consistently associated with CxDBPs in epidemiologic studies. An international workshop was held to (1) discuss the qualitative strengths and limitations that inform the association between bladder cancer and CxDBPs in the context of possible causation, (2) identify knowledge gaps for this topic in relation to chlorine/chloramine-based disinfection practice(s) in the United States, and (3) assess the evidence for informing risk management. Epidemiological evidence linking exposures to CxDBPs in drinking water to human bladder cancer risk provides insight into causality. However, because of imprecise, inaccurate, or incomplete estimation of CxDBPs levels in epidemiologic studies, translation from hazard identification directly to risk management and regulatory policy for CxDBPs can be challenging. Quantitative risk estimates derived from toxicological risk assessment for CxDBPs currently cannot be reconciled with those from epidemiologic studies, notwithstanding the complexities involved, making regulatory interpretation difficult. Evidence presented here has both strengths and limitations that require additional studies to resolve and improve the understanding of exposure response relationships. Replication of epidemiologic findings in independent populations with further elaboration of exposure assessment is needed to strengthen the knowledge base needed to better inform effective regulatory approaches.

Swimming facilities and work-related asthma.

BACKGROUND: Exposure to chlorinated water in swimming facilities may aggravate preexisting asthma or cause new onset asthma. This may be a particular problem for individuals who work and therefore spend prolonged time at swimming facilities. Chloramines formed by the interaction of chlorine-based disinfection products with the nitrogen in water from human sweat, urine and skin cells are the suspected causal agents. METHODS: Cases were reviewed from the state surveillance systems in California (CA), Michigan (MI) and New Jersey (NJ) to identify individuals with confirmed work-related asthma (WRA) attributed to exposures in swimming pools, water parks or hydrotherapy spas. A standardized method was used to confirm cases. RESULTS: A total of 44 confirmed cases of WRA were identified; 17 from 1994 to 2011 in CA, 15 from 1991 to 2012 in MI and 12 from 1990 to 2011 in NJ. A majority (52.2%) of the cases were new onset; 31.8% secondary to an acute exposure incident and 20.4% to repeated exposure. These represented 0.3-1.6% of all confirmed cases of WRA received during these time periods. Maintenance workers (34.9%) and lifeguards (31.8%) were the most common occupations. CONCLUSIONS: Swimming pool workers were identified from three states where the pool environment was either a trigger of preexisting asthma or associated with new onset of WRA. Regulations to require air monitoring and improvements in ventilation are recommended to reduce exposure levels of chloramines, the presumed etiologic agents. Clinical assessment of patients with asthma should include consideration of the effect on respiratory symptoms from exposures in a swimming pool environment.

Toxic impact of bromide and iodide on drinking water disinfected with chlorine or chloramines.

Disinfectants inactivate pathogens in source water; however, they also react with organic matter and bromide/iodide to form disinfection byproducts (DBPs). Although only a few DBP classes have been systematically analyzed for toxicity, iodinated and brominated DBPs tend to be the most toxic. The objectives of this research were (1) to determine if monochloramine (NH2Cl) disinfection generated drinking water with less toxicity than water disinfected with free chlorine (HOCl) and (2) to determine the impact of added bromide and iodide in conjunction with HOCl or NH2Cl disinfection on mammalian cell cytotoxicity and genomic DNA damage induction. Water disinfected with chlorine was less cytotoxic but more genotoxic than water disinfected with chloramine. For both disinfectants, the addition of Br(-) and I(-) increased cytotoxicity and genotoxicity with a greater response observed with NH2Cl disinfection. Both cytotoxicity and genotoxicity were highly correlated with TOBr and TOI. However, toxicity was weakly and inversely correlated with TOCl. Thus, the forcing agents for cytotoxicity and genotoxicity were the generation of brominated and iodinated DBPs rather than the formation of chlorinated DBPs. Disinfection practices need careful consideration especially when using source waters containing elevated bromide and iodide.

Differential resistance of drinking water bacterial populations to monochloramine disinfection.

The impact of monochloramine disinfection on the complex bacterial community structure in drinking water systems was investigated using culture-dependent and culture-independent methods. Changes in viable bacterial diversity were monitored using culture-independent methods that distinguish between live and dead cells based on membrane integrity, providing a highly conservative measure of viability. Samples were collected from lab-scale and full-scale drinking water filters exposed to monochloramine for a range of contact times. Culture-independent detection of live cells was based on propidium monoazide (PMA) treatment to selectively remove DNA from membrane-compromised cells. Quantitative PCR (qPCR) and pyrosequencing of 16S rRNA genes was used to quantify the DNA of live bacteria and characterize the bacterial communities, respectively. The inactivation rate determined by the culture-independent PMA-qPCR method (1.5-log removal at 664 mg·min/L) was lower than the inactivation rate measured by the culture-based methods (4-log removal at 66 mg·min/L). Moreover, drastic changes in the live bacterial community structure were detected during monochloramine disinfection using PMA-pyrosequencing, while the community structure appeared to remain stable when pyrosequencing was performed on samples that were not subject to PMA treatment. Genera that increased in relative abundance during monochloramine treatment include Legionella, Escherichia, and Geobacter in the lab-scale system and Mycobacterium, Sphingomonas, and Coxiella in the full-scale system. These results demonstrate that bacterial populations in drinking water exhibit differential resistance to monochloramine, and that the disinfection process selects for resistant bacterial populations.

Integrated chemical and toxicological investigation of UV-chlorine/chloramine drinking water treatment.

As the use of alternative drinking water treatment increases, it is important to understand potential public health implications associated with these processes. The objective of this study was to evaluate the formation of disinfection byproducts (DBPs) and cytotoxicity of natural organic matter (NOM) concentrates treated with chlorine, chloramine, and medium pressure ultraviolet (UV) irradiation followed by chlorine or chloramine, with and without nitrate or iodide spiking. The use of concentrated NOM conserved volatile DBPs and allowed for direct analysis of the treated water. Treatment with UV prior to chlorine in ambient (unspiked) samples did not affect cytotoxicity as measured using an in vitro normal human colon cell (NCM460) assay, compared to chlorination alone when toxicity is expressed on the basis of dissolved organic carbon (DOC). Nitrate-spiked UV+chlorine treatment produced greater cytotoxicity than nitrate-spiked chlorine alone or ambient UV+chlorine samples, on both a DOC and total organic halogen basis. Samples treated with UV+chloramine were more cytotoxic than those treated with only chloramine using either dose metric. This study demonstrated the combination of cytotoxicity and DBP measurements for process evaluation in drinking water treatment. The results highlight the importance of dose metric when considering the relative toxicity of complex DBP mixtures formed under different disinfection scenarios.

Impacts of chloramine-T treatment on antioxidant enzyme activities and genotoxicity in rainbow trout, Oncorhynchus mykiss (Walbaum).

Juvenile rainbow trout Oncorhynchus mykiss (Walbaum) were exposed to therapeutic, and higher concentrations of chloramine-T (Cl-T) to assess the effects of this chemical on the antioxidant enzyme system and genetic structure. Red blood cells acetylcholinesterase, ∆-aminolevulinic acid dehydratase, paraoxonase and liver glutathione S-transferase activity were increased at 10 and 20 mg L(-1) Cl-T-exposed fish, while they were decreased at 30 mg L(-1) Cl-T-exposed fish. On the other hand, liver catalase activity and liver protein levels increased at 10 mg L(-1) and decreased at 20 and 30 mg L(-1) concentrations of Cl-T. Liver super-oxide dismutase activity decreased at 10 mg L(-1) and 20 mg L(-1) Cl-T and increased at 30 mg L(-1) of Cl-T. Compared to control, comet assay indicated that Cl-T did not cause significant DNA damage to red blood cells of the fish. Results indicate that 10 or 20 mg L(-1) Cl-T can be safely used to prevent or treat external parasitic and bacterial infection of rainbow trout.

Cytogenotoxic potential and toxicity in adult Danio rerio (zebrafish) exposed to chloramine T.

Background: Chloramine-T (CL-T) is a synthetic sodium salt used as a disinfectant in fish farms to combat bacterial infections in fish gills and skin. While its efficacy in pathogen control is well-established, its reactivity with various functional groups has raised concerns. However, limited research exists on the toxicity of disinfection by-products to aquatic organisms. Therefore, this study aims to assess the sublethal effects of CL-T on adult zebrafish by examining biomarkers of nucleus cytotoxicity and genotoxicity, acetylcholinesterase (AChE) inhibition, and histopathological changes. Methods: Male and female adult zebrafish (wildtype AB lineage) specimens were exposed to 70, 140, and 200 mg/L of CL-T and evaluated after 96 h. Cytotoxic and genotoxic effects were evaluated by estimating the frequencies of nuclear abnormalities (NA), micronuclei (MN), and integrated optical density (IOD) of nuclear erythrocytes. Histopathological changes in the gills and liver were assessed using the degree of tissue changes (DTC). AChE activity was measured in brain samples. Results and conclusions: At a concentration of 200 mg/L, NA increased, indicating the cytogenotoxic potential of CL-T in adult zebrafish. Morphological alterations in the nuclei were observed at both 70 and 200 mg/L concentrations. Distinct IOD profiles were identified across the three concentrations. There were no changes in AChE activity in adult zebrafish. The DTC scores were high in all concentrations, and histological alterations suggested low to moderate toxicity of CL-T for adult zebrafish.

Identification of proteins susceptible to thiol oxidation in endothelial cells exposed to hypochlorous acid and N-chloramines.

Hypochlorous acid (HOCl) is a potent oxidant produced by the enzyme myeloperoxidase, which is released by neutrophils under inflammatory conditions. Although important in the immune system, HOCl can also damage host tissue, which contributes to the development of disease. HOCl reacts readily with free amino groups to form N-chloramines, which also cause damage in vivo, owing to the extracellular release of myeloperoxidase and production of HOCl. HOCl and N-chloramines react readily with cellular thiols, which causes dysfunction via enzyme inactivation and modulation of redox signaling processes. In this study, the ability of HOCl and model N-chloramines produced on histamine and ammonia at inflammatory sites, to oxidize specific thiol-containing proteins in human coronary artery endothelial cells was investigated. Using a proteomics approach with the thiol-specific probe, 5-iodoacetamidofluorescein, we show that several proteins including peptidylprolyl isomerase A (cyclophilin A), protein disulfide isomerase, glyceraldehyde-3-phosphate dehydrogenase and galectin-1 are particularly sensitive to oxidation by HOCl and N-chloramines formed at inflammatory sites. This will contribute to cellular dysfunction and may play a role in inflammatory disease pathogenesis.

Occupational asthma in professional cleaning work: a clinical study.

BACKGROUND: Several epidemiological studies have reported an increased risk of asthma among professional cleaners. To date, however, no analysis of large patient series from clinic of occupational medicine has been published. AIMS: To describe the cases of occupational asthma (OA) diagnosed at the Finnish Institute of Occupational Health (FIOH) during the period 1994-2004 in workers employed in professional cleaning work. METHODS: OA was diagnosed according to patient history, lung function examinations and specific challenge tests with measurements of the forced expiratory volume in 1 second and peak expiratory flow values. RESULTS: Our series comprised 20 patients, all female, with a mean age of 48.8 years (range 27-60 years). The mean duration of cleaning work before the onset of the respiratory symptoms was 14.3 years (range 1-36 years), and the mean duration of cleaning work before the FIOH examinations was 18.6 years (range 3-38 years). OA was triggered by chemicals in 9 cases (45%) and by moulds in 11 cases (55%). The chemicals were cleaning chemicals (wax-removing substances containing ethanolamines in five cases and a cleaning agent containing chloramine-T in one case) and chemicals used in the industrial processes at workplaces (three cases). Of the moulds, the most frequently associated with OA was Aspergillus fumigatus (nine cases). CONCLUSIONS: OA was attributed not only to cleaning chemicals but also to other chemicals used in work environments. Moulds are presented as a new cause of OA in cleaners.

Monochloramine-induced toxicity and dysregulation of intracellular Zn2+ in parietal cells of rabbit gastric glands.

Monochloramine (NH(2)Cl) is a potent, thiol-directed oxidant capable of oxidizing thiol (S-H) residues in a wide variety of proteins. Generated in the stomach by the interaction of bacterial and host products, monochloramine has been shown to dysregulate Ca(2+) homeostasis and disrupt mucosal integrity. In this report, we show that monochloramine also leads to disturbances in intracellular free zinc concentration ([Zn(2+)](i)) in the gastric gland of the rabbit and that the increased Zn(2+) within the cell causes an independent decrease in cell viability. Changes in [Zn(2+)](i) were measured by using the fluorescent reporter FluoZin-3, whereas cell viability was assessed by measuring the conversion of calcein-AM to fluorescent calcein, an assay that is not affected by intracellular oxidation state. Cell death was confirmed using propidium iodide and YO-PRO-1 dye uptake measurements. Our experiments demonstrate that [Zn(2+)](i) is increased in gastric glands exposed to NH(2)Cl and that elevated [Zn(2+)](i) decreases cell viability. Chelation of Zn(2+) with tetrakis-(2-pyridylmethyl) ethylenediamine decreases the toxicity of NH(2)Cl, but only when administered concurrently. These findings suggest that the toxic effect of thiol oxidants present during chronic gastritis is partially due to dysregulation of [Zn(2+)](i) early in the process and that zinc chelation can protect, but not rescue, gastric glands exposed to toxic doses of NH(2)Cl.

Ability of hypochlorous acid and N-chloramines to chlorinate DNA and its constituents.

Myeloperoxidase is a heme enzyme released by activated phagocytes that is responsible for the generation of the strong oxidant hypochlorous acid (HOCl). Although HOCl has potent bactericidal properties and plays an important role in the human immune system, this oxidant also causes damage to tissues, particularly under inflammatory conditions. There is a strong link between chronic inflammation and the incidence of many cancers, which may be associated with the ability of HOCl and related oxidants such as N-chloramines to damage DNA. However, in contrast to HOCl, little is known about the reactivity of N-chloramines with DNA and its constituents. In this study, we examine the ability of HOCl and various N-chloramines to form chlorinated base products on nucleosides, nucleotides, DNA, and in cellular systems. Experiments were performed with N-chloramines formed on Nalpha-acetyl-histidine (His-C), Nalpha-acetyl-lysine (Lys-C), glycine (Gly-C), taurine (Tau-C), and ammonia (Mono-C). Treatment of DNA and related materials with HOCl and His-C resulted in the formation of 5-chloro-2'-deoxycytidine (5CldC), 8-chloro-2'-deoxyadenosine (8CldA) and 8-chloro-2'-deoxyguanosine (8CldG). With the nucleosides, 8CldG was the favored product in each case, and HOCl was the most efficient chlorinating agent. 5Cl(d)C was the most abundant product on exposure of the nucleotides and DNA to HOCl and His-C, with only low levels of chlorinated products observed with Lys-C, Gly-C, Tau-C, and Mono-C. 5CldC was also formed on exposure of smooth muscle cells to either HOCl or His-C. Cellular RNA was also a target for HOCl and His-C, with evidence for the formation of 5-chloro-cytidine (5ClC). This study shows that HOCl and the model N-chloramine, His-C, are able to chlorinate cellular genetic material, which may play a role in the development of various inflammatory cancers.

Monochloramine produces reactive oxygen species in liver by converting xanthine dehydrogenase into xanthine oxidase.

In the present study, we assessed the influence of monochloramine (NH(2)Cl) on the conversion of xanthine dehydrogenase (XD) into xanthine oxidase (XO) in rat liver in vitro. When incubated with the partially purified cytosolic fraction from rat liver, NH(2)Cl (2.5-20 microM) dose-dependently enhanced XO activity concomitant with a decrease in XD activity, implying that NH(2)Cl can convert XD into the reactive oxygen species (ROS) producing form XO. The NH(2)Cl (5 microM)-induced XD/XO interconversion in the rat liver cytosol was completely inhibited when added in combination with an inhibitor of NH(2)Cl methionine (25 microM). A sulfhydryl reducing agent, dithiothreitol at concentrations of 0.1, 1 and 5 mM also dose-dependently reversed the NH(2)Cl (5 microM)-induced XD/XO interconversion. These imply that NH(2)Cl itself acts on the XD/XO interconversion, and that this conversion occurs at the cysteine residues in XD. Furthermore, using the fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate, it was found that NH(2)Cl could increase ROS generation in the cytoplasm of rat primary hepatocyte cultures, and that this increase might be reversed by an XO inhibitor, allopurinol. These results suggest that NH(2)Cl has the potential to convert XD into XO in the liver, which in turn may induce the ROS generation in this region.

Chemical wastes in the sea: new forms of marine pollution.

Drums containing chemical wastes have been found in and along the North Sea. The wastes were analyzed and were found to include lower chlorinated aliphatic compounds, vinyl esters, chlorinated aromatic amines and nitrocompounds, and the insecticide endosulfan. Because dropping these drums into the sea endangers the environment and results in damage to fishing operations, measures should be taken to stop this practice.

Removal of disinfection byproduct precursors and reduction in additive toxicity of chlorinated and chloraminated waters by ozonation and up-flow biological activated carbon process.

The variations of disinfection byproduct (DBP) precursors and DBPs-associated toxic potencies were evaluated by ozonation, followed by a up-flow biological activated carbon (O3/UBAC) filter treating two reconstituted water samples, featuring either high bromide (105.3 µg/L) or dissolved organic nitrogen (0.73 mg N/L) concentration, respectively. Ozonation contributed to ∼20% decrease in dissolved organic carbon (DOC) concentration at a dosage of 0.7 mg of O3/mg of DOC, but no further reduction in DOC level was observed with an increased dose of 1.0 mg of O3/mg of DOC. When chlorine or preformed monochloramine was used as a disinfectant, UBAC process led to ∼40% reduction in the sum of detected DBP formation potential (FP) due to the removal of precursors at a feasible empty bed contact time of 15 min. The integrated effect of ozonation and UBAC biofiltration decreased the sum of DBP FP by ∼50% including halonitromethanes (THNMs), N-nitrosamines (NAs), and bromate, which increased in the effluent of ozonation. Chloramination produced less DBPs by weight as well as DBPs-associated additive toxic potencies than chlorination. The reduction in additive toxic potencies was generally lower than the removal efficiency of DBP FP after chlor(am)ination of treated waters by O3/UBAC, indicating that the removal of DBPs-associated additive toxic potencies should be focused to better understand on the residual risk to public health in controlling DBP precursors.

Mechanisms of hypochlorite injury of target cells.

HOCl, which is produced by the action of myeloperoxidase during the respiratory burst of stimulated neutrophils, was used as a cytotoxic reagent in P388D1 cells. Low concentrations of HOCl (10-20 microM) caused oxidation of plasma membrane sulfhydryls determined as decreased binding of iodoacetylated phycoerythrin. These same low concentrations of HOCl caused disturbance of various plasma membrane functions: they inactivated glucose and aminoisobutyric acid uptake, caused loss of cellular K+, and an increase in cell volume. It is likely that these changes were the consequence of plasma membrane SH-oxidation, since similar effects were observed with para-chloromercuriphenylsulfonate (pCMBS), a sulfhydryl reagent acting at the cell surface. Given in combination pCMBS and HOCl showed an additive effect. Higher doses of HOCl (greater than 50 microM) led to general oxidation of -SH, methionine and tryptophan residues, and formation of protein carbonyls. HOCl-induced loss of ATP and undegraded NAD was closely followed by cell lysis. In contrast, NAD degradation and ATP depletion caused by H2O2 preceded cell death by several hours. Formation of DNA strand breaks, a major factor of H2O2-induced injury, was not observed with HOCl. Thus targets of HOCl were distinct from those of H2O2 with the exception of glyceraldehyde-3-phosphate dehydrogenase, which was inactivated by both oxidants.