Richard J. Miltner

Integrated Disinfection By-Products Mixtures Research: Comprehensive Characterization of Water Concentrates Prepared from Chlorinated and Ozonated/Postchlorinated Drinking Water

This article describes the disinfection by-product (DBP) characterization portion of a series of experiments designed for comprehensive chemical and toxicological evaluation of two drinking-water concentrates containing highly complex mixtures of DBPs. This project, called the Four Lab Study, involved the participation of scientists from four laboratories and centers of the U.S. Environmental Protection Agency (EPA) Office of Research and Development, along with collaborators from the water industry and academia, and addressed toxicologic effects of complex DBP mixtures, with an emphasis on reproductive and developmental effects that are associated with DBP exposures in epidemiologic studies. Complex mixtures of DBPs from two different disinfection schemes (chlorination and ozonation/postchlorination) were concentrated successfully, while maintaining a water matrix suitable for animal studies. An array of chlorinated/brominated/iodinated DBPs was created. The DBPs were relatively stable over the course of the animal experiments, and a significant portion of the halogenated DBPs formed in the drinking water was accounted for through a comprehensive qualitative and quantitative identification approach. DBPs quantified included priority DBPs that are not regulated but have been predicted to produce adverse health effects, as well as those currently regulated in the United States and those targeted during implementation of the Information Collection Rule. New by-products were also reported for the first time. These included previously undetected and unreported bromo- and chloroacids, iodinated compounds, bromo- and iodophenols, and bromoalkyltins.

Research Issues Underlying the Four-Lab Study: Integrated Disinfection By-Products Mixtures Research

Chemical disinfection of drinking water is a major public health triumph of the 20th century, resulting in significant decreases in morbidity and mortality from waterborne diseases. Disinfection by-products (DBP) are chemicals formed by the reaction of oxidizing disinfectants with inorganic and organic materials in the source water. To address potential health concerns that cannot be answered directly by toxicological research on individual DBPs or defined DBP mixtures, scientists residing within the various organizations of the U.S. Environmental Protection Agencys Office of Research and Development (the National Health and Environmental Effects Research Laboratory, the National Risk Management Research Laboratory, the National Exposure Research Laboratory, and the National Center for Environmental Assessment) engaged in joint investigation of environmentally realistic complex mixtures of DBP. Research on complex mixtures of DBP is motivated by three factors: (a) DBP exposure is ubiquitous to all segments of the population; (b) some positive epidemiologic studies are suggestive of potential developmental, reproductive, or carcinogenic health effects in humans exposed to DBP; and (c) significant amounts of the material that makes up the total organic halide portion of the DBP have not been identified. The goal of the Integrated Disinfection Byproducts Mixtures Research Project (the 4Lab Study) is provision of sound, defensible, experimental data on environmentally relevant mixtures of DBP and an improved estimation of the potential health risks associated with exposure to the mixtures of DBP formed during disinfection of drinking water. A phased research plan was developed and implemented. The present series of articles provides the results from the first series of experiments.

Component-Based and Whole-Mixture Techniques for Addressing the Toxicity Of Drinking-Water Disinfection By-Product Mixtures

Chemical disinfection of water is of direct public health benefit as it results in decreased water-borne illness. The chemicals used to disinfect water react with naturally occurring organic matter, bromide, and iodide in the source water, resulting in the formation of disinfection by-products (DBPs). Despite the identification of several hundred DBPs, more than 50% of the mass of total organic halide formed during chlorination remains unidentified. The toxic contribution of the DBPs that are formed and present but not yet chemically identified, the unidentified fraction, has been largely unexplored. A better understanding of the potential for adverse human health consequences associated with exposure to the DBPs present in drinking water will be gained by integration of knowledge on the toxicity of individual DBPs; simple, defined DBP mixtures; complex, environmentally realistic DBP mixtures with partial chemical characterization; and the unidentified fraction.

Pilot-Scale Ozone Inactivation of Cryptosporidium and Other Microorganisms in Natural Water

Abstract A pilot-scale study was conducted to evaluate the inactivation by ozone against Cryptosporidium oocysts, Giardia cysts, poliovirus, and B. subtilis endospores spiked into Ohio River water. The indigenous Ohio River populations of total coliform bacteria, heterotrophic plate count bacteria and endospores of aerobic spore forming bacteria were also evaluated. Endospores were the only organisms found to be more resistant to ozone than Cryptosporidium oocysts. Endospores may serve as an indicator of microbial treatment efficiency. Cryptosporidium oocysts were more resistant than Giardia cysts or poliovirus. Although HPC bacteria were less resistant than Cryptosporidium oocysts, variability limits their usefulness as an indicator of treatment efficiency. Ozone inactivation data generated in a pilot-scale study employing natural surface waters were comparable to inactivation data derived from previously published bench-scale studies using laboratory waters. The ozone requirements for inactivation of Cryptosporidium oocysts may produce elevated levels of bromate and ozone byproducts.

Integrated Disinfection By-Products Mixtures Research: Disinfection of Drinking Waters by Chlorination and Ozonation/Postchlorination Treatment Scenarios

This article describes disinfection of the same source water by two commonly used disinfection treatment scenarios for purposes of subsequent concentration, chemical analysis, and toxicological evaluation. Accompanying articles in this issue of the Journal of Toxicology and Environmental Health describe concentration of these finished waters by reverse osmosis techniques, chemical characterization of the resulting disinfection by-product (DBP) concentrates, in vivo and in vitro toxicological results, and risk assessment methods developed to analyze data from this project. This project, called the “Four Lab Study,” involved participation of scientists from four laboratories/centers of the U.S. Environmental Protection Agency Office of Research and Development as well as extramural collaborators from the water industry and academia. One of the two finished waters was prepared by conventional treatment and disinfected by chlorination. The other finished water was also prepared by conventional treatment and disinfected by ozonation followed by chlorination (ozonation/postchlorination). Chlorination conditions of dose, time and temperature were similar for both treatment scenarios, allowing for a comparison. Both finished waters had acceptably low levels of particulates and bacteria, representative pH and chlorine levels, and contained numerous DBP. Known effects of ozonation were observed in that, relative to the water that was chlorinated only, the ozonated/postchlorinated water had lower concentrations of total organic halogen, trihalomethanes (THM), haloacetic acids (HAA), and higher concentrations of bromate, and aldehydes.

Integrated disinfection by-products mixtures research: concentration by reverse osmosis membrane techniques of disinfection by-products from water disinfected by chlorination and ozonation/postchlorination.

To conduct the health-effect studies described in subsequent articles in this series, concentrated aqueous mixtures of disinfection by-products were required for the two water treatment trains described in the preceding article (Miltner et al., 2008). To accomplish this, the finished drinking waters from each treatment train were sent through cation-exchange resin columns to remove hardness and free chlorine. Reverse osmosis membranes were then used to concentrate approximately 2400 L of each finished water down to approximately 18 L. The resulting volumetric concentration factors for the chlorinated and ozonated/postchlorinated waters were 136- and 124-fold, respectively. The concentrates were spiked with select disinfection by-products (DBPs) that were lost during the concentration effort. The results, along with the rationale for choosing the method of concentration, are presented. After reintroduction of a select list of lost DBPs, the concentration methodology used herein was able to produce concentrates that retained large percentages of the DBPs that were in the initial finished drinking waters. Further, the distributions of the DBPs in the concentrates matched those found in the finished drinking waters.

Effectiveness of Pre- and Intermediate Ozonation on the Enhanced Coagulation of Disinfection By-product Precursors in Drinking Water

This study evaluated the impact of pre- and intermediate ozonation coupled with enhanced coagulation in controlling halogenated disinfection by-product formation in drinking water. Raw waters from utilities representing each of the nine elements of the enhanced coagulation matrix presented in Table I were examined. All testing was completed using bench-scale, batch experimental procedures. The various waters were analyzed for turbidity, total organic carbon, dissolved organic carbon, ultraviolet absorbance, trihalomethane formation potential, and haloacetic acid formation potential before and after ozonation. The results indicated that formation of trihalomethanes and haloacetic acids following enhanced coagulation decreased with both pre- and intermediate ozonation applications relative to the decreases observed by enhanced coagulation alone. The amount of trihalomethanes and haloacetic acids formed were lower for the waters that were pre-ozonated and then coagulated compared to those that were coagulated first and then ozonated. This comparison must be tempered by the fact that the settled waters treated by intermediate ozonation were not subjected to subsequent biofiltration which is commonly used in water treatment practice to remove additional DBP precursors. Strong correlations between disinfection by-product formation potentials and ultraviolet absorbance at 254 nm were observed for enhanced coagulation with and without pre- and intermediate ozonation.

Integrated Disinfection By-Products Research: Assessing Reproductive and Developmental Risks Posed by Complex Disinfection By-Product Mixtures

This article presents a toxicologically-based risk assessment strategy for identifying the individual components or fractions of a complex mixture that are associated with its toxicity. The strategy relies on conventional component-based mixtures risk approaches such as dose addition, response addition, and analyses of interactions. Developmental toxicity data from two drinking-water concentrates containing disinfection by-products (DBP) mixtures were used to illustrate the strategy. The results of this study showed that future studies of DBP concentrates using the Chernoff–Kavlock bioassay need to consider evaluating DBP that are concentrated more than 130-fold and using a rat strain that is more sensitive to chemically-induced pregnancy loss than Sprague-Dawley rats. The results support the planned experimental design of a multigeneration reproductive and developmental study of DBP concentrates. Finally, this article discusses the need for a systematic evaluation of DBP concentrates obtained from multiple source waters and treatment types. The development of such a database could be useful in evaluating whether a specific DBP concentrate is sufficiently similar to tested combinations of source waters and treatment alternatives so that health risks for the former may be estimated using data on the latter.

Developmental Toxicity Evaluations of Whole Mixtures of Disinfection By-products using Concentrated Drinking Water in Rats: Gestational and Lactational Effects of Sulfate and Sodium

A developmental toxicity bioassay was used in three experiments to evaluate water concentrates for suitability in multigenerational studies. First, chlorinated water was concentrated 135-fold by reverse osmosis; select lost disinfection by-products were spiked back. Concentrate was provided as drinking water to Sprague-Dawley and F344 rats from gestation day 6 to postnatal day 6. Maternal serum levels of luteinizing hormone on gestation day 10 were unaffected by treatment for both strains. Treated dams had increased water consumption, and increased incidences of polyuria, diarrhea, and (in Sprague-Dawley rats) red perinasal staining. Pup weights were reduced. An increased incidence of eye defects was seen in F344 litters. Chemical analysis of the concentrate revealed high sodium (6.6 g/l) and sulfate (10.4 g/l) levels. To confirm that these chemicals caused polyuria and osmotic diarrhea, respectively, Na₂SO₄ (5-20 g/l) or NaCl (16.5 g/l) was provided to rats in drinking water. Water consumption was increased at 5- and 10-g Na₂SO₄/l and with NaCl. Pup weights were reduced at 20-g Na₂SO₄/l. Dose-related incidences and severity of polyuria and diarrhea occurred in Na₂SO₄-treated rats; perinasal staining was seen at 20 g/l. NaCl caused polyuria and perinasal staining, but not diarrhea. Subsequently, water was concentrated ∼120-fold and sulfate levels were reduced by barium hydroxide before chlorination, yielding lower sodium (≤1.5 g/l) and sulfate (≤2.1 g/l) levels. Treatment resulted in increased water consumption, but pup weight and survival were unaffected. There were no treatment-related clinical findings, indicating that mixtures produced by the second method are suitable for multigenerational testing.

Disinfection byproduct formation in reverse-osmosis concentrated and lyophilized natural organic matter from a drinking water source.

Drinking water treatment and disinfection byproduct (DBP) research can be complicated by natural organic matter (NOM) temporal variability. NOM preservation by lyophilization (freeze-drying) has been long practiced to address this issue; however, its applicability for drinking water research has been limited because the selected NOM sources are atypical of most drinking water sources. The purpose of this research was to demonstrate that reconstituted NOM from a lyophilized reverse-osmosis (RO) concentrate of a typical drinking water source closely represents DBP formation in the original NOM. A preliminary experiment assessed DBP formation kinetics and yields in concentrated NOM, which demonstrated that chlorine decays faster in concentrate, in some cases leading to altered DBP speciation. Potential changes in NOM reactivity caused by lyophilization were evaluated by chlorination of lyophilized and reconstituted NOM, its parent RO concentrate, and the source water. Bromide lost during RO concentration was replaced by adding potassium bromide prior to chlorination. Although total measured DBP formation tended to decrease slightly and unidentified halogenated organic formation tended to increase slightly as a result of RO concentration, the changes associated with lyophilization were minor. In lyophilized NOM reconstituted back to source water TOC levels and then chlorinated, the concentrations of 19 of 21 measured DBPs, constituting 96% of the total identified DBP mass, were statistically indistinguishable from those in the chlorinated source water. Furthermore, the concentrations of 16 of 21 DBPs in lyophilized NOM reconstituted back to the RO concentrate TOC levels, constituting 86% DBP mass, were statistically indistinguishable from those in the RO concentrate. This study suggests that lyophilization can be used to preserve concentrated NOM without substantially altering the precursors to DBP formation.