Disinfection By-products Mixtures Research Articles

Risks of obstructive genitourinary birth defects in relation to trihalomethane and haloacetic acid exposures: expanding disinfection byproduct mixtures analyses using relative potency factors.

Some disinfection byproducts (DBPs) are teratogens based on toxicological evidence. Conventional use of predominant DBPs as proxies for complex mixtures may result in decreased ability to detect associations in epidemiological studies. We assessed risks of obstructive genitourinary birth defects (OGDs) in relation to 12 DBP mixtures and 13 individual component DBPs. We designed a nested registry-based case-control study (210 OGD cases; 2100 controls) in Massachusetts towns with complete quarterly 1999-2004 data on four trihalomethanes (THMs) and five haloacetic acids (HAAs). We estimated temporally-weighted average DBP exposures for the first trimester of pregnancy. We estimated adjusted odds ratios (aORs) and 95% confidence intervals (CIs) for OGD in relation to individual DBPs, unweighted mixtures, and weighted mixtures based on THM/HAA relative potency factors (RPF) from animal toxicology data for full-litter resorption, eye defects, and neural tube defects. We detected elevated aORs for OGDs for the highest of bromodichloromethane (aOR = 1.75; 95% CI: 1.15-2.65), dibromochloromethane (aOR = 1.71; 95% CI: 1.15-2.54), bromodichloroacetic acid (aOR = 1.56; 95%CI: 0.97-2.51), chlorodibromoacetic acid (aOR = 1.97, 95% CI: 1.23-3.15), and tribromoacetic acid (aOR = 1.90; 95%CI: 1.20-3.03). Across unweighted mixture sums, the highest aORs were for the sum of three brominated THMs (aOR = 1.74; 95% CI: 1.15-2.64), the sum of six brominated HAAs (aOR = 1.43; 95% CI: 0.89-2.31), and the sum of nine brominated DBPs (aOR = 1.80; 95% CI: 1.05-3.10). Comparing eight RPF-weighted to unweighted mixtures, the largest aOR differences were for two HAA metrics, which both were higher with RPF weighting; other metrics had reduced or minimally changed ORs in RPF-weighted models.

Multi-omics signatures of exposure to disinfection by-products in swimming

Background and aims: Untargeted HRMS metabolomics provides an agnostic global profiling of small molecules in a biological system capturing an overview of exposure to exogenous compounds, and their effects on metabolic pathways, rendering an essential tool for characterising the exposome. Disinfection by-products (DBPs) are a complex mixture of halogenated organic compounds present in drinking and swimming pool water. Although several epidemiological studies have pointed their potential to induce adverse health outcomes such as bladder cancer, their mixture effect and the downstream molecules perturbed by their exposure have been poorly explored. In this work, we aim to identify molecular signatures of exposures to mixtures of DBPs in individuals with measured exposures. Methods: In PISCINA-II study, metabolomics, transcriptomics, proteomics in serum, and levels of DBPs in exhaled breaths, were measured in 60 subjects before and after swimming. Pre-processing of HRMS data from PISCINA-II study was customised to include more halogenated compounds. Multivariate normal (MVN) and partial least square (PLS) models were used to identify metabolic features associated with DBP exposures. Conditional correlation networks were constructed to understand the holistic relationship amongst the metabolic features and their relationship with proteomics and transcriptomics. Results: 98 metabolic features were identified to be associated with exposure to DBP mixture, of which 39 were not identified in the previous analysis without specific reprocessing and inclusion of halogenated compounds. In conditional network analysis, the 98 features showed cluster structures, suggesting they could contribute to different downstream biological pathways. Multi-omic networks showed that transcriptomic and proteomic signals were differentially connected to the different clusters of the metabolic signatures. Conclusions: New data processing approach allowed identification of novel metabolic signatures of DBP mixture exposure. Conditional correlation network implied the functional proximity of these signatures with other measured omic signals. Keywords: Disinfection by-products, PLS, network analysis, omics, exposome, statistical analysis

Characteristics and chlorine reactivity of biochar-derived dissolved organic matter: Effects of feedstock type and pyrolysis temperature

Increasing biochar application worldwide may release more biochar-derived dissolved organic matter (BDOM) to the source water for drinking water supply. However, it is unclear how feedstock types and pyrolysis temperatures for biochar production would affect the characteristics and chlorine reactivity of BDOM. Here, we studied the spectroscopic characteristics of BDOM pyrolyzed from pine needle, wheat straw, walnut shells, alfalfa, pig manure, and sludge derived biochars at 300, 500, and 700 °C, as well as the formation potential of disinfection byproducts (DBPs) and their bulk toxicity after BDOM chlorination. The N/C ratio of biochar was higher for N-rich than C-rich feedstocks. Optical analyses indicated that BDOM aromaticity was highest at 700 °C, while the impact of pyrolysis temperature on the molecular weight of BDOM varied greatly among feedstocks. Increasing pyrolysis temperature caused consistently decreased BDOM reactivity toward haloketone formation but did not show consistent impacts on the other DBPs. Among feedstocks, the N-rich sludge showed the highest specific haloacetonitrile formation potential of BDOM at any given pyrolysis temperature. The DBP formation potential from biochar was consistently highest at 300 °C and was higher for N-rich than C-rich feedstocks. The microtoxicity of DBP mixture was highest for the BDOM derived from sludge produced at 300 °C. This study highlights the high variations in characteristics and chlorination reactivity of BDOM with varying feedstocks and pyrolysis temperatures, which implies that more attention should be paid to the environmental impacts of the intensive application of low-temperature biochar from N-rich feedstock such as sludge.

Toxicological Interactions of Disinfection Byproducts Mixtures of Halogenated Acetamides and Haloacetic Acids on Freshwater Bacteria

Toxicological Interactions of Disinfection Byproducts Mixtures of Halogenated Acetamides and Haloacetic Acids on Freshwater Bacteria

Concentration Addition, Independent Action, and Quantitative Structure-Activity Relationships for Chemical Mixture Toxicities of the Disinfection By products of Haloacetic Acids on the Green Alga Raphidocelis subcapitata.

The potential toxicity of haloacetic acids (HAAs), common disinfection by products (DBPs), has been widely studied; but their combined effects on freshwater green algae remain poorly understood. The present study was conducted to investigate the toxicological interactions of HAA mixtures in the green alga Raphidocelis subcapitata and predict the DBP mixture toxicities based on concentration addition, independent action, and quantitative structure-activity relationship (QSAR) models. The acute toxicities of 6 HAAs (iodoacetic acid [IAA], bromoacetic acid [BAA], chloroacetic acid [CAA], dichloroacetic acid [DCAA], trichloroacetic acid [TCAA], and tribromoacetic acid [TBAA]) and their 68 binary mixtures to the green algae were analyzed in 96-well microplates. Results reveal that the rank order of the toxicity of individual HAAs is CAA > IAA ≈ BAA > TCAA > DCAA > TBAA. With concentration addition as the reference additive model, the mixture effects are synergetic in 47.1% and antagonistic in 25%, whereas the additive effects are only observed in 27.9% of the experiments. The main components that induce synergism are DCAA, IAA, and BAA; and CAA is the main component that causes antagonism. Prediction by concentration addition and independent action indicates that the 2 models fail to accurately predict 72% mixture toxicity at an effective concentration level of 50%. Modeling the mixtures by QSAR was established by statistically analyzing descriptors for the determination of the relationship between their chemical structures and the negative logarithm of the 50% effective concentration. The additive mixture toxicities are accurately predicted by the QSAR model based on 2 parameters, the octanol-water partition coefficient and the acid dissociation constant (pKa ). The toxicities of synergetic mixtures can be interpreted with the total energy (ET ) and pKa of the mixtures. Dipole moment and ET are the quantum descriptors that influence the antagonistic mixture toxicity. Therefore, in silico modeling may be a useful tool in predicting disinfection by-product mixture toxicities. Environ Toxicol Chem 2021;40:1431-1442. © 2021 SETAC.

Exposure Misclassification: Temporal and Spatial Variability in Estimates of Individual Species and Mixtures of Disinfection Byproducts

Disinfection of drinking water is a public health advance, but the potential risk of unintended consequences of disinfection byproduct (DBP) formation remains unclear. Over 600 DBPs have been identified to date, although most have not been analytically quantified in disinfected drinking water. Some DBP exposures occur through multiple routes which can complicate exposure assessment in epidemiological studies especially based on aggregate data derived from routinely-collected monitoring data. Therefore it is important to quantify the spatiotemporal variability in occurrence to minimize measurement error and exposure misclassification. In the absence of biomonitoring data, concentration-based exposure scores based on routinely-collected monitoring data can be spatial water system averages or location-specific estimates. However, intra-water system variation in DBP concentrations results from formation and degradation processes occurring over space and residence time. Temporal variability can also occur from diurnal variation to longer-term trends such as seasonality noted for trihalomethanes (THMs) and haloacetic acids (HAAs). Temporally averaged data may estimate lifetime exposure for cancer outcomes or trimester-specific windows for developmental effects, but they ill fully account for the potential impact of peak exposures. Although the DBP surrogacy issue is beyond the scope here, data availability limitations may preclude examination and delineation of potential risk for individual DBPs or DBP mixtures especially when the measurement error structures can vary due to use of aggregate exposure estimates. With few DBPs regulated or routinely measured, this review will characterize strengths and limitations of existing approaches based on HAAs and THMs and will offer evidence from simulations and sensitivity analyses and offer recommendations to help address the aforementioned challenges. Disclaimer: The views expressed in this abstract are those of the author and do not necessarily reflect the views or policies of the U.S. EPA. Keywords: Water, water quality, disinfection byproducts, environmental epidemiology

Long-term biochar addition alters the characteristics but not the chlorine reactivity of soil-derived dissolved organic matter

Biochar is widely and increasingly applied to farmlands. However, it remains unclear how long-term biochar addition alters the characteristics and chlorine reactivity of soil-derived dissolved organic matter (DOM), an important terrestrial disinfection byproduct (DBP) precursor in watersheds. Here, we analyzed the spectroscopic and molecular-level characteristics of soil-derived DOM and the formation and toxicity of DBP mixtures from DOM chlorination for two long-term (5 and 11 years) biochar addition experimental farmlands. As indicated by spectroscopic indices and Fourier transform ion cyclotron resonance mass spectrometry analyses, 11 years of biochar addition could increase the humic-like and aromatic and condensed aromatic DOM and decrease the microbial-derived DOM, while 5 years of biochar addition at the other site did not. The response of condensed aromatic dissolved black carbon did not increase with increasing cumulative biochar dose but appeared to be affected by biochar aging time. Despite the possible increase in aromatic DOM, biochar addition neither increased the reactivity of DOM in forming trihalomethanes, haloacetonitriles, chloral hydrates, or haloketones nor significantly increased the microtoxicity or genotoxicity of the DBP mixture. This study indicates that biochar addition in watersheds may not deteriorate the drinking water quality via the export of terrestrial DBP precursors like wildfire events.

Effects of ascorbate and carbonate on the conversion and developmental toxicity of halogenated disinfection byproducts during boiling of tap water

Chlorine disinfection inactivates pathogens in drinking water, but meanwhile it causes the formation of halogenated disinfection byproducts (DBPs), which may induce adverse health effects. Humans are unavoidably exposed to halogenated DBPs via tap water ingestion. Boiling of tap water has been found to significantly reduce the concentrations of halogenated DBPs. In this study, we found that compared with boiling only, adding ascorbate (vitamin C) or carbonate (baking soda) to tap water and then boiling the water further reduced the level of total organic halogen (a collective parameter for all halogenated DBPs) by up to 36% or 28%, respectively. Adding ascorbate removed the chlorine residual in tap water and thus prevented the formation of more halogenated DBPs in the boiling process. Adding carbonate elevated pH of tap water and consequently enhanced the hydrolysis (dehalogenation) of halogenated DBPs or led to the formation of more trihalomethanes that might volatilize to air during the boiling process. The comparative developmental toxicity of the DBP mixtures in the water samples was also evaluated. The results showed that adding a tiny amount of sodium ascorbate or carbonate (2.5–5.0 mg/L) to tap water followed by boiling for 5 min reduced the developmental toxicity of tap water to a substantially lower level than boiling only. The addition of sodium ascorbate or carbonate to tap water in household could be realized by preparing them in tiny pills. This study suggests simple and effective methods to reduce the adverse effects of halogenated DBPs on humans through tap water ingestion.

Assessing Additivity of Cytotoxicity Associated with Disinfection Byproducts in Potable Reuse and Conventional Drinking Waters.

Recent studies used the sum of the measured concentrations of individual disinfection byproducts (DBPs) weighted by their Chinese hamster ovary (CHO) cell cytotoxicity LC50 values to estimate the DBP-associated cytotoxicity of disinfected waters. This approach assumed that cytotoxicity was additive rather than synergistic or antagonistic. In this study, we evaluated whether this assumption was valid for mixtures containing DBPs at the concentration ratios measured in authentic disinfected waters. We examined the CHO cell cytotoxicity of defined DBP mixtures based on the concentrations of 43 regulated and unregulated DBPs measured in eight drinking and potable reuse waters. The hypothesis for additivity was supported using three experimental approaches. First, we demonstrated that the calculated additive toxicity (CAT) and bioassay-based calculated additive toxicity (BCAT) of the DBP mixtures agree within 12% on a median basis. We also found an additive toxicity response (CAT ≈ BCAT) between the regulated and unregulated DBP classes. Finally, the empirical biological cytotoxicity of the DBP subset mixtures, independent of the calculated toxicity, was additive. These results support the validity of using the sum of cytotoxic potency-weighted DBP concentrations as an estimate of the CHO cell cytotoxicity associated with known DBPs in real disinfected waters.

Volatile DBPs contributed marginally to the developmental toxicity of drinking water DBP mixtures against Platynereis dumerilii

Halogenated disinfection byproducts (DBPs) are formed during chlorine disinfection of drinking water. The complicated natural organic matter in source water causes the formation of an even more complicated mixture of DBPs. To evaluate the toxicity of a DBP mixture in a disinfected water sample, the sample needs to be pretreated in order to attain an observable acute adverse effect in the toxicity test. During sample pretreatment, volatile DBPs including trihalomethanes, haloacetonitriles and haloketones may be lost, which could affect the toxicity evaluation of the DBP mixture. In this study, we intentionally prepared “concentrated” simulated drinking water samples, which contained sufficiently high levels of volatile and nonvolatile DBPs and thus enabled directly evaluating the toxicity of the DBP mixtures without sample pretreatment. Specifically, the natural organic matter and bromide concentrations and the chlorine dose in the concentrated water samples were 250 times higher than those in a typical drinking water sample. Each concentrated water sample was divided into two aliquots, and one of them was nitrogen sparged to eliminate volatile DBPs; then, both aliquots were used directly in a well-established developmental toxicity test. No significant difference (p > 0.10) was found between the developmental toxicity indexes of each concentrated water sample without and with nitrogen sparging, indicating that the contribution of volatile DBPs to the developmental toxicity of the DBP mixture might be marginal. A reasonable interpretation is that nonvolatile halogenated DBPs (especially the aromatic ones) in the DBP mixture could be the major developmental toxicity contributor that warrants more attention.

Effects of dechlorination conditions on the developmental toxicity of a chlorinated saline primary sewage effluent: Excessive dechlorination is better than not enough.

Chlorine-disinfected sewage effluents are typically dechlorinated by using NaHSO3, Na2SO3, or Na2S2O3, as chlorine residual could be harmful to aquatic organisms upon discharge of sewage effluents into receiving marine water. In this study, we systematically investigated the effects of dechlorination-related factors on the developmental toxicity of a chlorinated saline primary sewage effluent, via direct exposure of the embryos of a marine polychaete to the effluent. The results showed that dechlorination ratio (i.e., the ratio of the dosed amount to the requisite stoichiometric amount of a dechlorination agent) and mixing condition were critical factors affecting the toxicity of the effluent. The toxicity of the effluent under insufficient dechlorination conditions was mainly caused by residual chlorine, especially monochloramine. Although the three dechlorination agents generally performed similarly, dechlorination with Na2S2O3 required a more vigorous mixing condition than that with NaHSO3 or Na2SO3, as the relatively high density of Na2S2O3 might affect the mixing efficiency. Under insufficient mixing conditions, a prolonged dechlorination time was beneficial to achieving complete dechlorination and thus lowered the toxicity of the effluent. Moreover, because disinfection byproducts (DBPs) may have chronic effects on aquatic organisms, the developmental toxicity of the DBP mixtures in the chlorinated effluent in different dechlorination scenarios was also evaluated. The results indicated that increasing the dechlorination ratio reduced the developmental toxicity of the DBP mixture in the chlorinated saline sewage effluent, which might be ascribed to the decrease of the levels of overall brominated and iodinated DBPs; the dechlorination agent (NaHSO3 or Na2S2O3) might act as a nucleophile in the nucleophilic substitution and cause the substitution of bromine or iodine atoms in brominated and iodinated DBPs. The results from this study might aid in the design and operation of dechlorination facilities in sewage treatment plants.

Cytotoxic comparison of macrolide antibiotics and their chlorinated disinfection byproduct mixtures

Erythromycin (ERY), azithromycin (AZI) and telithromycin (TEL) are widely-used macrolide antibiotics that are frequently detected in various water environments, including resource water and drinking water. In the performed chlorination disinfection process, at least 10, 20 and 200 new disinfection byproducts of ERY, AZI and TEL, respectively, were observed (the mixtures of the disinfection byproducts of ERY, AZI and TEL were named ERY-M, AZI-M and TEL-M, respectively). There is limited information available regarding their comparative toxicities, and their potential health risks are still unknown. In this study, the Jurkat cell line was used to compare the toxicities of the disinfection byproduct mixtures and their precursor compounds. The cell viability results indicated that the toxicity of ERY-M may not be enhanced after disinfection by chlorination. In contrast, at the same concentrations, AZI-M and TEL-M induced more significant inhibitory effects on cell viability than their parent compounds. Additionally, the total antioxidant capacity (T-AOC) and cell cytokine release (including interleukin-2, interleukin-8 and tumor necrosis factor-α) analyses of AZI-M and TEL-M further verified these results. Our findings demonstrate that the cytotoxicity of AZI and TEL was enhanced during the chlorination disinfection process. This investigation will provide substantial new details related to the toxicity of the mixed disinfection byproducts (DBPs) of ERY, AZI and TEL generated in the chlorination disinfection process.

Disinfection Byproducts Bind Human Estrogen Receptor-α.

Disinfection byproducts are formed during most drinking water treatment and presently number >800, some of which are implicated in human health outcomes including bladder cancer and infertility, with unknown mechanisms of action. In particular, it is not yet understood whether these compounds can disrupt the estrogen-signaling pathway through binding to the human estrogen receptor (ER). In the present study, 21 disinfection byproducts, selected for their predicted involvement in endocrine-related diseases and their structural diversity, were individually evaluated for their binding affinity to the human ER and in silico, and then a subset of these chemicals was studied in binary mixtures with the known weak estrogen, 4-n-nonylphenol. Individually, 9 of the 21 disinfection byproducts were able to weakly bind to the ER, with affinities ranging from log median inhibitory concentration values of -3.83 to -2.19 M. In binary mixtures, the chemicals followed concentration addition, with their weak binding affinities having little contribution to the overall mixture affinity. These results demonstrate the variety of small-molecule disinfection byproduct structures that are capable of binding to the ER, and that their weak binding can still be of importance when overall human exposure to mixtures of disinfection byproducts in disinfected drinking water is considered. Environ Toxicol Chem 2019;9999:1-9. © 2019 SETAC.

Conversion of haloacid disinfection byproducts to amino acids via ammonolysis

Haloacetic acids (HAAs) are the major disinfection byproducts (DBPs) that are formed during chlorination of drinking water. In this paper, the conversion of HAAs to amino acids (e.g., glycine) via ammonolysis was studied. First, a new and sensitive method for detecting glycine was developed by setting selected ion recording m/z 76 in positive electrospray ionization mass spectrometry coupled with ultra performance liquid chromatography. Second, among the mono-HAAs under the same test conditions, iodoacetic acid (49.3%) showed a considerably higher conversion to glycine during ammonolysis than chloroacetic acid (4.2%) and bromoacetic acid (27.7%). The conversion of iodoacetic acid to glycine increased with increasing temperature, increasing reaction time, or decreasing the ratio of (NH4)2CO3 to NH3·H2O in the aminating agent. Hydrolysis of iodoacetic acid to glycolic acid was also observed during ammonolysis, and it accounted for at most 50% of the iodoacetic acid conversion. The conversion to amino acids and the hydrolysis were the two major pathways during ammonolysis of HAAs. Third, compared with the iodoacetic acid sample and the simulated tap water sample without ammonolysis, the developmental toxicity of the corresponding samples with ammonolysis decreased by up to 10.4% and 32.1%, respectively. The ammonolysis was thus demonstrated to be a detoxification process for both individual HAAs and DBP mixture in chlorinated tap water. In practice, the ammonolysis of haloacid DBPs in tap water may be realized by simply adding an appropriate amount of an aminating agent during cooking.

Evaluating the Comparative Toxicity of DBP Mixtures from Different Disinfection Scenarios: A New Approach by Combining Freeze-Drying or Rotoevaporation with a Marine Polychaete Bioassay.

The unintended formation of disinfection byproducts (DBPs) may compromise the safety of drinking water. Since no specified DBPs have been found to be responsible for the overall adverse effects and over half of total organic halogen (TOX) remains unidentified, DBP mixture toxicity is gaining increasing interest as a potential indicator of how risky drinking water might be. In this study, a new approach to evaluating the toxicity of drinking water DBP mixtures was developed by combining freeze-drying or rotoevaporation pretreatment with an in vivo high-salinity-tolerance bioassay with the embryos of a marine polychaete Platynereis dumerilii. The DBP recoveries by freeze-drying or rotoevaporation were compared with those by commonly applied liquid-liquid-extraction (LLE). For drinking water subjected to typical disinfection processes (i.e., chlorination, chloramination, chlorine dioxide treatment, and ozonation with or without postchlorination), LLE led to the lowest TOX recovery (11-18%) and the loss of all inorganic DBPs, while freeze-drying and rotoevaporation recovered 28-58% and 35-61% of TOX, respectively, and effectively recovered 81-99% and 85-104% of inorganic DBPs, respectively. Thus, LLE caused an underestimation of the toxicity of DBP mixtures compared with freeze-drying and rotoevaporation. Besides, the comparative toxicity varied significantly for water samples pretreated with different methods due to the effect of inorganic DBPs and a synergistic effect of organic and inorganic DBPs. The new approach revealed that the bromide-rich source water disinfected with ozone caused the highest developmental toxicity, followed by those disinfected with chlorine, chlorine dioxide, and chloramine in that order.

Chemical characterization and relative toxicity assessment of disinfection byproduct mixtures in a large drinking water supply network

In order to reduce the formation of disinfection by-products (DBPs) during potabilization of water it is necessary to explore the potential of the source water and the applied treatment to generate these chemicals. This is actually more challenging in large drinking water networks that use different source waters to satisfy drinking water demand. In this regard, this work investigated the formation of DBPs in water matrices that are commonly supplied to the city of Barcelona and its metropolitan area. The regulated trihalomethanes and haloacetic acids were the most abundant DBP classes in these waters, followed by haloacetamides and haloacetonitriles or trihalogenated acetaldehydes (THALs). On the contrary, the formation potential of iodo-DBPs was minor. Mixing of drinking water treatment plant finished waters with desalinated water decreased the overall DBP formation potential of the water but resulted in the increased formation of brominated DBPs after long chlorine contact time. The formation of most DBPs was enhanced at high water temperatures (except for Br-THALs) and increasing residence times. Potential cytotoxicity and genotoxicity of the DBP mixtures were mainly attributed to the presence of nitrogen-containing DBPs and HAAs.

Predicted Impact of Aeration on Toxicity From Trihalomethanes and Other Disinfection Byproducts

The removal by aeration of a mixture of disinfection byproducts (DBPs) was modeled, and the resulting impact on overall cytotoxicity or genotoxicity was calculated. Both the aeration model and the toxicity model were simplistic, but all assumptions were made to err on the side of overestimating DBP volatility to try to demonstrate a possible health benefit. The results predicted that aeration designed to remove 90% of chloroform would leave the concentrations of many other DBPs largely unaffected; since these other DBPs are predicted to contribute to the majority of the cytotoxicity and genotoxicity, the models used in the current research predicted only a 4% reduction in both cytotoxicity and genotoxicity. Although this study was a preliminary screening limited by simplistic models, it underscores a need to conduct more research in this area.

In vitro studies on the tumorigenic potential of the halonitromethanes trichloronitromethane and bromonitromethane

Epidemiological data indicate that chronic exposure to water disinfection by-products (DBPs) may result in increased risk of cancer. However, the real carcinogenic potential of individual DBPs is not well known. In this study, we assessed the in vitro carcinogenic potential of trichloronitromethane (TCNM) and bromonitromethane (BNM), two halonitromethanes (HNMs) commonly found in DBPs' mixtures at comparably high concentrations. Human lung BEAS-2B cells were exposed for 8weeks to TCNM and BNM, and the acquisition of different in vitro cancer-like features was evaluated. The results indicate that long-term exposure to non-cytotoxic doses of TCNM and BNM did not cause carcinogenic transformation as indicated by the absence of morphological changes, no effects on cell growth, no changes in the level of matrix metalloproteinases (MMPs) secretion, and no increased anchorage-independent cell growth capacity. Furthermore, TCNM- and BNM-exposed BEAS-2B cells were unable to enhance tumour growth directly or by indirect influence of the surrounding stroma. Our results indicate that the carcinogenic effects of DBP mixtures cannot be attributed to the evaluated HNMs. This is the first study evaluating the cell transformation effects of TCNM and BNM under a long-term exposure scenario using suitable hallmarks of the cancer process.

Method to assess component contribution to toxicity of complex mixtures: Assessment of puberty acquisition in rats exposed to disinfection byproducts

A method based on regression modeling was developed to discern the contribution of component chemicals to the toxicity of highly complex, environmentally realistic mixtures of disinfection byproducts (DBPs). Chemical disinfection of drinking water forms DBP mixtures. Because of concerns about possible reproductive and developmental toxicity, a whole mixture (WM) of DBPs produced by chlorination of a water concentrate was administered as drinking water to Sprague–Dawley (S–D) rats in a multigenerational study. Age of puberty acquisition, i.e., preputial separation (PPS) and vaginal opening (VO), was examined in male and female offspring, respectively. When compared to controls, a slight, but statistically significant delay in puberty acquisition was observed in females but not in males. WM-induced differences in the age at puberty acquisition were compared to those reported in S–D rats administered either a defined mixture (DM) of nine regulated DBPs or individual DBPs. Regression models were developed using individual animal data on age at PPS or VO from the DM study. Puberty acquisition data reported in the WM and individual DBP studies were then compared with the DM models. The delay in puberty acquisition observed in the WM-treated female rats could not be distinguished from delays predicted by the DM regression model, suggesting that the nine regulated DBPs in the DM might account for much of the delay observed in the WM. This method is applicable to mixtures of other types of chemicals and other endpoints.

Formation of iodo-trihalomethanes, iodo-haloacetic acids, and haloacetaldehydes during chlorination and chloramination of iodine containing waters in laboratory controlled reactions

Iodine containing disinfection by-products (I-DBPs) and haloacetaldehydes (HALs) are emerging disinfection by-product (DBP) classes of concern. The former due to its increased potential toxicity and the latter because it was found to be the third most relevant DBP class in mass in a U.S. nationwide drinking water study. These DBP classes have been scarcely investigated, and this work was performed to further explore their formation in drinking water under chlorination and chloramination scenarios. In order to do this, iodo-trihalomethanes (I-THMs), iodo-haloacetic acids (I-HAAs) and selected HALs (mono-HALs and di-HALs species, including iodoacetaldehyde) were investigated in DBP mixtures generated after chlorination and chloramination of different water matrices containing different levels of bromide and iodide in laboratory controlled reactions. Results confirmed the enhancement of I-DBP formation in the presence of monochloramine. While I-THMs and I-HAAs contributed almost equally to total I-DBP concentrations in chlorinated water, I-THMs contributed the most to total I-DBP levels in the case of chloraminated water. The most abundant and common I-THM species generated were bromochloroiodomethane, dichloroiodomethane, and chlorodiiodomethane. Iodoacetic acid and chloroiodoacetic acid contributed the most to the total I-HAA concentrations measured in the investigated disinfected water. As for the studied HALs, dihalogenated species were the compounds that predominantly formed under both investigated treatments.