As waterborne disease originating in potable water plumbing systems (such as Legionnaires' Disease and Nontuberculous Mycobacterial (NTM) infections) continue to increase, it is important to better understand the cause(s), responsible parties and interventions to prevent disease. This dissertation begins with a literature review characterizing the propensity of building (premise) plumbing to enhance or diminish opportunistic pathogen growth, including Legionella. It then holistically examines the problem at the field, bench and pilot scale by first discovering problems with lead and Legionella in Flint, MI, during an event popularly referred to as the Flint Water Crisis in 2014-2016. Four years were then spent simulating critical factors hypothesized to have triggered the Legionella outbreak in residences and in a large hospital in Flint. In parallel with that work, pilot scale rigs were operated for several years, to examine the important role of water heater system design and operation on energy efficiency, hot water delivery, and Legionella. The first chapter literature review is entitled "Critical Review of the Propensity of Premise Plumbing Pipe Materials to Enhance or Diminish the Growth of Legionella and Other Opportunistic Pathogens." It examines the complex environments found in premise systems, focusing primarily on the role of pipe materials. The effects of metallic (copper, iron) and plastic pipe materials on opportunistic pathogens and Legionella include their effect on nutrient availability, disinfectant levels, and the composition of the broader microbiome. Design, configuration, and operation are also examined in terms of their potential for influencing opportunistic pathogens. This chapter demonstrates that pipe materials have the potential to stimulate or inhibit pathogen growth, dependent on circumstance and water chemistry. This chapter will be submitted to the journal Pathogens. The field study in this work first predicted, discovered and then exposed problems with lead and Legionella in Flint, Michigan. A citizen science project that sampled Flint water in August 2015, demonstrated a city-wide problem with water lead exceeding the EPA limit of 15 µg/L after corrosion control was interrupted. Follow-up sampling events between August 2015 and August 2017 demonstrated that the switch back to the original water source and addition of enhanced corrosion control in October 2015, dramatically reduced lead, copper and iron levels flowing into consumer homes. Entitled "Evaluating Water Lead Levels During the Flint Water Crisis," this work was published in Environmental Science and Technology in 2018. After our Virginia Tech team's work helped expose a Legionnaires' disease outbreak that killed twelve people and sickened nearly one hundred individuals, the started to explore possible links between corrosion control, plumbing materials and disinfection that could help explain the trajectory of disease in Flint and elsewhere. Three separate experiments were performed using bench-scale simulated glass water heaters. Two of the studies attempted to simulate what occurred in Flint homes before, during and after the water crisis in relation to factors that either encouraged or discouraged Legionella growth, while the third examined the more benign Blacksburg tap water and a broader range of influential plumbing conditions. The first study entitled "Copper Pipe, Lack of Corrosion Control, and Uncontrolled pH Influenced the Trajectory of the Flint Legionnaires' Disease Outbreak," determined that the very low pH levels in summer 2015 and interruption of phosphate corrosion control, could cause explosive growth of Legionella in PEX plumbing held at warm temperature, without disinfectant and with constant mixing. Under the same conditions copper pipe had antimicrobial properties that markedly reduced Legionella in our experiments. This work has been submitted for review to Environmental Science and Technology. The second companion study conducted at a higher pH, without mixing and with trace chlorine, found 2.5 log10 lower levels of Legionella compared to the worst-case conditions in the aforementioned study, demonstrating the importance of mixing and traces of chlorine. Higher levels of disinfectant and the presence of copper pipe also enhanced control of Legionella. This manuscript is titled "Interactive Effects of Copper Pipe, Stagnation, Corrosion Control, and Disinfectant Residual Influenced Reduction of Legionella pneumophila during Simulations of the Flint Water Crisis," and it has been published in Pathogens. The third simulated glass water heater study examined the disinfection of opportunistic pathogens in the presence of six different premise plumbing materials or conditions in Blacksburg tap water. Generally speaking, all of the premise plumbing materials reduced disinfection of opportunistic pathogens compared to a control condition with glass surfaces. Chlorine decay was catalyzed by iron pipe, warmer temperature and the presence of organic matter, increasing the persistence of Legionella. Magnesium anodes in particular, encouraged much higher Legionella growth compared to all other materials. This work titled "Chlorine and Chloramine Disinfection of Legionella spp., L. pneumophila, and Acanthamoeba Under Warm Water Premise Plumbing Conditions," has been submitted to Microorganisms. Results of a six-year pilot study titled "Elucidating the Role of Water Heater System Configuration in Energy Efficiency, Consumer Comfort and Legionella Proliferation," examined different types of residential-sized water heater systems with plastic pipes including: a standard tank system with water stagnant between uses, a recirculating tank system with flowing water between uses, and an on-demand system which only heated water and had flow during use. Considering the volume of water in each tank between 38 and 47 ° C as a measure of Legionella growth risk, with a heater setpoint at 48 °C (118 °F) the recirculating system had 90% of its volume at risk daily compared to only 24% of the standard system volume. The on-demand system used a minimum of 10% less energy than the standard tank, and 50% less energy than the recirculating tank, and had one tenth of the volume at risk of growing Legionella than either tank system. In fact, it was only by contriving a system to keep distal lines artificially warmed to above room temperature, that Legionella growth could occur in the on-demand system, whereas it rose to 107 L. pneumophila MPN per liter in a normally operating recirculating system. On the other hand, the on-demand heaters were repeatedly subject to mechanical malfunction during the study, and had difficulty delivering water at the desired temperature and flow rates versus traditional tank systems. This manuscript will be submitted to Water Research. / Doctor of Philosophy / Recent water crises in Flint, Michigan and Legionnaires' Disease outbreaks in Flint, New York City, and Quincy, Illinois have demonstrated the need to better understand the cause(s), responsible parties, and interventions required to prevent waterborne diseases. As waterborne disease originating in building plumbing systems (premise plumbing), such as Legionnaires' Disease and Nontuberculous Mycobacterial infections, continue to increase each year, the burden on healthcare systems and impact on public health also grows. In this dissertation, a literature review, a field study of water in Flint, small-scale laboratory studies, and residential-sized water heater systems were examined to study interactions between water chemistry, premise plumbing, and disease-causing opportunistic pathogens (OPs) with a focus on Legionella, the OP which causes Legionnaires' Disease. The first chapter literature review is entitled "Critical Review of the Propensity of Premise Plumbing Pipe Materials to Enhance or Diminish the Growth of Legionella and Other Opportunistic Pathogens." It examines the complex environments found in premise systems, and the important role of pipe material selection. The effects of metallic (copper, iron) and plastic pipe materials on opportunistic pathogens and Legionella include their impact on nutrient availability, disinfectant levels, and the composition of the broader microbiome. Design, configuration, and operation of plumbing systems are also examined in terms of their potential for influencing growth of opportunistic pathogens. This chapter demonstrates that pipe materials have the potential to stimulate or inhibit pathogen growth, dependent on circumstance and water chemistry. The field study in this work exposed problems with lead and Legionella in Flint, Michigan, during an event known in the popular press as the Flint Water Crisis 2014-2016. In August 2015, a citizen science sampling campaign demonstrated a citywide lead in water problem. After a federal emergency was declared, follow-up sampling events between August 2015 and August 2017 demonstrated that the switch back to the original water source, enhanced disinfection, and corrosion control under federal direction had reduced lead levels to half of the EPA limit. Additionally, the pipe material installed between each home and the water main (service lines) affected levels of lead and copper in water, with the lowest lead concentrations measured in homes with copper service lines. After our teams' work in Flint helped expose a Legionnaires' disease outbreak that killed twelve people and sickened nearly one hundred other individuals, we started to explore possible links between corrosion control, plumbing materials, and disinfection that could affect the trajectory of disease in Flint and elsewhere. Three follow-up studies, using small glass bottles to simulate water heaters, provided more specific comparisons between water conditions in premise plumbing and OP occurrence. Two studies expanded on phosphate corrosion control, chlorine (disinfectant) decay, and pH-related research questions, which arose during the Flint water crisis field study. The first determined that properly treated drinking water and some mixing could inhibit Legionella growth in copper pipes. The second study found that without mixing, copper could be antimicrobial and reduce Legionella growth even if the water chemistry was slightly altered with higher pH. The third simulated water heater study examined the reduction of OPs using a chlorine or chlorine plus ammonia disinfectant to reduce Legionella in the presence of six different plumbing conditions. The reduction of Legionella with chlorine was inhibited when carbon was increased and in the presence of a magnesium anode rod, a necessary water heater component. A six-year study using a residential-sized water heater system holistically examined three different types of water heater systems with plastic pipes: one using a standard water heater tank with water stagnant between uses, one using a water heater tank connected to a recirculating pump to provide constantly flowing water, and one tankless (on-demand) heater which only heated water and provided flow during use. Using temperature as an indicator of risk for Legionella growth, the recirculating system at a temperature setpoint of 48 °C (118 °F) would be at high risk for Legionella growth (water volume at 38-47 °C or 100-117 °F) in 90% of the tank volume each day, whereas the standard system would only be at high risk in 24% of the tank each day. The on-demand system provided the safest alternative for hot water distribution with virtually undetectable levels of Legionella risk when the pipes were kept at room temperature as per normal operation. The on-demand system also used at least 50% less energy than the recirculating system and 10% less energy than the standard system; however, we were not successful in finding a reliable on-demand system that could also provide hot water at the desired temperature and flow rate.

There has been a recent increase in the adoption of point-of-use (POU) household water filters as an alternative to untreated tap water or bottled water. POU filters certified for lead removal have recently been distributed by the hundreds of thousands in communities amid water lead crises, as a temporary solution to protect consumers from elevated water lead levels. This thesis rigorously examines the efficacy of POU lead certified filters in removing lead under a wide range of conditions, and evaluates premature clogging due to iron and associated impacts on the cost analysis of using filters instead of bottled water. In testing ten brands of POU devices against up to four different waters for lead removal, most devices consistently removed lead to below the 5 µg/L FDA bottled water standard. However, several failures were documented, including manufacturing flaws, premature clogging, and inconsistency between duplicate filters. When waters containing more difficult to treat lead particulates were synthesized, treated water often had lead concentrations greater than the 5 µg/L bottled water standard and sometimes were even over the 15 µg/L EPA action level. In some cases, less than 50% of the particulate lead was removed by the filter, thereby replicating some problems with these devices identified in the field. While POUs usually reduced water lead concentrations by at least 80%, a combination of manufacturing issues and difficult to treat waters can cause treated water to exceed expectations. Consumers often purchase POU devices to remove particles and lead in waters that also contain high iron, prompting studies to examine the role of iron on filter performance. When we exposed two brands of pour-through POUs to waters with both high lead and iron, lead removal performance was generally not compromised, as treated water typically had lead concentrations less than 5 µg/L. One case was observed in which lead passed through a set of filters at high levels in association with iron, confirming expectations that in some waters iron could cause formation of lead particulates that are difficult to remove. High levels of iron sometimes rapidly clogged the POU filters, preventing them from reaching their rated capacity and increasing operational costs and time to filter water. Specifically, 50% (3/6) of the filters tested clogged prematurely at an iron concentration of 0.37 mg/L, 66% (4/6) at 1 mg/L and 100% (6/6) at 20 mg/L. A cost analysis for POUs vs. bottled water demonstrated that in waters with higher iron, store-brand bottled water was often the more cost-effective option, especially when iron levels were significantly higher than the EPA Secondary Maximum Contaminant Level (0.3 mg/L). The lower costs of bottled water in these situations was even more apparent if consumer time was factored into the analysis. / Master of Science / There has been a recent increase in the use of household water filters as an alternative to tap water or bottled water. Filters that are certified for lead removal have recently been distributed by the hundreds of thousands in communities amid water lead crises, as a temporary solution to protect consumers from elevated water lead levels. This thesis rigorously examines the effectiveness of these filters under a wide range of conditions. When tested against up to four different waters for lead removal, most filters consistently reduced lead to below the concentrations allowed in bottled water. In cases where the filters did not perform as expected, several filter failure modes were identified, including manufacturing flaws, filter clogging, and inconsistency between duplicate filters. In addition to these failures, when a water that contained particulate lead that was difficult to filter, as little as 50% of the lead was removed. While household filters often significantly reduce water lead concentrations, a combination of manufacturing issues and difficult to treat waters can cause poor performance. In many cases, consumers purchase filters to remove particles or lead in waters that also contain iron, which caused us to investigate the effect of iron on filter performance. When two brands of pour-through filters were tested against waters with both lead and iron, lead removal performance was generally not compromised. One exceptional case was observed where both high levels of lead and iron passed through the filters, leading us to believe that iron in some waters could create conditions where lead is more difficult to remove. In many cases, the presence of iron caused filters to dramatically slow down or clog. Premature clogging due to iron prevented filters from reaching their rated capacity and, in doing so, significantly increased cost and filter times. A cost analysis for filters vs. bottled water demonstrated that in waters with higher iron, store-brand bottled water was often the more cost-effective option, especially in waters with higher levels of iron. The lower costs of bottled water in these situations was even more apparent if consumer time was factored into the analysis.

Evaluation of the sources and cycles of water quality contaminants in watersheds is critical for effective surface water resource management. In particular, iron (Fe) and manganese (Mn), commonly found in rocks and sediments, have adverse impacts on water quality. However, controlling Fe and Mn in surface water systems is often complex and requires careful consideration of the hydrologic and biogeochemical factors that influence the speciation and mobility of these metals. This dissertation investigates the sources and cycles of Fe and Mn in surface waters designated for human use. Here, I present the findings from three field- and laboratory-based studies conducted at sites in western Virginia, United States. The first study examines the impacts of reservoir-derived and watershed-derived metals on water quality along the 180 km reach of the Roanoke River downgradient from Leesville Dam. The results from this study showed strong temporal influences on river water quality immediately downgradient of the dam, resulting from seasonal reservoir dynamics. Further downgradient in the Roanoke River, water quality was strongly tied to hydrologic conditions resulting from influences generated in the watershed. The second study investigated the effects of increasing dissolved oxygen (DO) concentrations in the hypolimnion of stratified drinking water reservoir on Fe and Mn oxidation and removal. Results from a whole-ecosystem experiment showed that increasing DO concentrations through hypolimnetic oxygenation was effective for preventing the accumulation of soluble Fe in the water column. Although Mn oxidation increased under well-oxygenated conditions, soluble Mn still accumulated in the hypolimnion. Results from a laboratory experiment demonstrated that the oxidation of Mn was strongly tied to the activity of Mn oxidizing microbes. The third study examined the relative contribution of external and internal metal loadings to the exchange of metals between sediments and the water column and the source/sink behavior of a seasonally stratified reservoir under varying hydrologic conditions in the inflows and outflows and redox conditions in the reservoir hypolimnion. Results from this study showed that redox conditions strongly influenced the exchange of metals between the sediment and aqueous phase, but had little effect on the source/sink behavior of the reservoir, while external tributary loadings had little effect on internal redox cycles, but was a strong indicator for whether the reservoir behaved as a net metal source or sink. Overall, the findings from these studies exemplify the value of characterizing the hydrologic and biogeochemical drivers of Fe and Mn cycles for managing the water quality effects of these metals in surface water supplies. / Ph. D.

Scientific explorations can lead to life changing discoveries or light the path for new discoveries as scientists continue to carry or pass on the torch of knowledge to current and future generations. This torch of knowledge radiates in many directions, as the path of discovery often demands a multidimensional perspective. This research explored the many aspects of metallic flavor in drinking water through applications of sensory science, medicine, health, and the environment. Humans interact with their environment through the five senses and are often exposed to contaminants through multiple routes; oral intake of trace metal contaminants through drinking water is a likely source. The biochemical mechanism by which humans are able to detect the flavor of strongly metallic agents such as iron has been previously elucidated, but little is known about population variability in the ability to sense metallic flavors. This research evaluated sensory thresholds and biochemical indicators of metallic flavor perception in healthy adults for ferrous iron in drinking water; 61 subjects aged 19 – 84 years, participated. The findings demonstrated an age-dependent sensitivity to iron indicating as people age they are less sensitive to metallic perception; impairment of olfactory functions is a contributing factor. Unlike in healthy adults, where human senses are often protective of overexposure to contaminants, and supportive of sensations of everyday life's pleasures, cancer patients often suffer from chemosensory dysfunctions. Metallic phantom taste is a commonly experienced sensation, yet very little studied aspect of this debilitating disorder. Impact of cancer therapy on chemosensory functions of patients with malignant brain tumors undergoing combined modality treatment (CMT) was explored. The results indicated that chemosensory dysfunctions of the patients can range from minimal to moderate impairment with maximum impairment developing during the 6-week CMT. Study of salivary constituents may provide clues on to the causes of chemosensory dysfunctions. On health aspects, implication of individual sensitivity to metallic flavor on beverage choices and overall water consumption was assessed in 33 healthy adults through self-reported beverage questionnaire. The results indicated that among the elderly reduced intake of drinking water coincided with reduced sensitivity to metallic flavor. The findings have important health implications in terms of hydration status and beverage choices. Finally, with environmental exposure relevance, preliminary findings on sensory properties of zerovalent iron nanoparticles (nZVI) indicated that oral exposure to nZVI may induce sensory properties different from that of ferrous iron, likely predictive of a diminished detection of metallic flavor by humans. Further research is warranted in this area. / Ph. D.

Corrosion of drinking water distribution systems can cost water utilities and homeowners tens of billions of dollars each year in infrastructure damage, adversely impacting public health and causing water loss through leaks. Often, seemingly innocuous choices made by utilities, plumbers, and consumers can have a dramatic impacts on corrosion and pipeline longevity. This work demonstrated that brass pipe connectors used in partial lead service line replacements (PLSLR) can significantly influence galvanic corrosion between lead and copper pipes. Galvanic crevice corrosion was implicated in a fourfold increase in lead compared to a traditional direct connection, which was previously assumed to be a worst-case connection method. In field sampling conducted in two cities, a new sampling method designed to detect particulate lead risks demonstrated that the choice of flow rate has a substantial impact on lead-in-water hazards. On average, lead concentrations detected in water at high flow without stagnation were at least 3X-4X higher than in traditional regulatory samples with stagnation, demonstrating a new 'worst case' lead release scenario due to detachment of lead particulates. Although galvanized steel was previously considered a minor lead source, it can contain up to 2% lead on the surface, and elevated lead-in-water samples from several cities were traced to galvanized pipe, including the home of a child with elevated blood lead. Furthermore, if both galvanized and copper pipe are present, as occurs in large buildings, deposition corrosion is possible, leading to both increased lead exposure and pipe failures in as little as two years. Systematic laboratory studies of deposition corrosion identified key factors that increase or decrease its likelihood; soluble copper concentration and flow pattern were identified as controlling factors. Because of the high copper concentrations and continuous flow associated with mixed-metal hot water recirculating systems, these systems were identified as a worst-case scenario for galvanic corrosion. Deposition corrosion was also confirmed as a contributing mechanism to increased lead release, if copper pipe is placed before a lead pipe as occurs in partial service line replacements. Dump-and-fill tests confirmed copper solubility as a key factor in deposition corrosion impacts, and a detailed analysis of lead pipes from both laboratory studies and field tests was consistent with pure metallic copper deposits on the pipe surface, especially near the galvanic junction with copper. Finally, preliminary experiments were conducted to determine whether nanoparticles from novel water treatment techniques could have a negative impact on downstream drinking water pipeline infrastructure. Although increases in the corrosion of iron, copper, and stainless steel pipes in the presence of silver and carbon nanomaterials were generally small or non-existent, in one case the presence of silver nanoparticles increased iron release from stainless steel by more than 30X via a localized corrosion mechanism, with pitting rates as high as 1.2 mm/y, implying serious corrosion consequences are possible for stainless steel pipes if nanoparticles are present. / Ph. D.

Lead is a common additive in plumbing components despite its known adverse health effects. Recent research has attributed cases of elevated blood lead levels in children and even fetal death with the consumption of drinking water containing high levels of lead. Although the federal Environmental Protection Agency (USEPA) strives to minimize lead exposure from water utilities through the Lead and Copper Rule (LCR), an estimated 47 million U.S. residents reliant on private unregulated water systems (generally individual and rural) are not protected. Detection, evaluation, and mitigation of lead in private systems is challenging due to lack of monitoring data, appropriate sampling protocols, and entities to fund research. Through a statewide sampling survey, over 2,000 homeowners submitted water samples for analysis. This survey documented that 19% of households had lead concentrations in the first draw sample (i.e., 250 mL sample collected after 6+ hours of stagnation) above the EPA action level of 15, with concentrations as high as 24,740. Due to the high incidence observed, this research focused on identifying system and household characteristics that increased a homeowner's susceptibility of lead in water. However, 1% of households had elevated lead concentrations after flushing for five minutes, which highlighted potential sources of lead release beyond the faucet. Therefore, a follow-up study was conducted to investigate sources and locations of lead release throughout the entire plumbing network. Using profiling techniques (i.e., sequential and time series sampling), three patterns of waterborne lead release were identified: no elevated lead or lead elevated in the first draw of water only (Type I), erratic spikes of particulate lead mobilized from plumbing during periods of water use (Type II), and sustained detectable lead concentrations (>1 ) even with extensive flushing (Type III). Lastly, emphasis was given to understand potential lead leaching from NSF Standard 61 Section 9 certified lead-free plumbing components as the synthetic test water is not representative of water quality observed in private water systems. Overall, this dissertation research provides insight into a population that is outside the jurisdiction of many federal agencies. / Ph. D.

In an era of pervasive water stress caused by population growth, urbanization, drought, and climate change, limiting the dissemination of microbial contaminants of emerging concern (MCECs) is of the utmost importance for the protection of public health. In this dissertation, two important subsets of MCECs, opportunistic pathogens (OP) and antibiotic resistant genes (ARG), are studied across several compartments of the urban water cycle, including surface water, stormwater, wastewater, recycled water, and potable water. Collectively, this dissertation advances knowledge about the occurrence of OPs and ARGs across these water systems and highlights trends that may be of value in developing management strategies for limiting their regrowth and transmission. Field studies of two surface water catchments impacted by stormwater runoff demonstrated the prevalence of ARGs in urban stormwater compared to pristine, unimpacted sites, or to days when no precipitation was recorded. The role of wastewater reuse in transmitting OPs and ARGs was also investigated. Traditional tertiary wastewater treatment plants producing water for non-potable use were found to be largely ineffective at removing ARGs, but plants using advanced oxidation processes or ozonation paired with biofiltration to produce direct potable reuse water were highly effective at removing ARGs. Non-potable reclaimed water consistently had greater quantities of sul1, a sulfonamide ARG, and Legionella and Mycobacterium, two OPs of significant public health concern, present than corresponding potable systems. Limited regrowth of OPs and ARGs did occur in simulated premise (i.e., building) plumbing systems operated with direct potable reuse waters, but regrowth was comparable to that observed in systems fed with potable water derived from surface or groundwater. Advancements were also made in understanding the role of several hypothesized driving forces shaping the antibiotic resistome in natural and engineered water systems: selection by antimicrobials and other compounds, horizontal gene transfer, and microbial community composition. Finally, whole-genome and metagenomic characterization were applied together towards profiling L. pneumophila in clinical and water samples collected from Flint, Michigan, where an economically-motivated switch to an alternative water source created conditions favorable for growth of this organism and likely triggered one of the largest Legionnaires' Disease outbreaks in U.S. history. / PHD

Opportunistic pathogens (OPs) are of special concern for immunocompromised populations and are known to grow in both drinking water and reclaimed water (i.e., non-potable recycled water) distribution systems, with aerosol inhalation and other non-ingestion exposures that are not addressed by existing regulatory frameworks. Factors enabling the growth of OPs in water distribution and premise (i.e., building) plumbing systems distributing reclaimed and other water sources systems are poorly understood especially for the emerging OP, Naegleria fowleri (i.e. brain-eating amoeba). Three phases of investigation were carried out to identify factors that facilitate the growth of OPs in main distribution and premise plumbing systems, with particular attention on reclaimed water systems, aging water mains, and private well systems. Phase one examined the role of biological treatment to remove organic carbon and disinfectant type on the occurrence of OPs during distribution of reclaimed water. Laboratory-scale simulated reclaimed water distribution systems were employed to systematically examine the effects of prior granular activated carbon (GAC) biofiltration of the water; chlorine, chloramines, or no disinfectant, and water ages ranging up to 5 days. The second and third phases of research explored the role of nitrification, iron corrosion, and disinfectant on the growth of N. fowleri both in municipal drinking water from a city grappling with aging water infrastructure and untreated private well water. Results from the simulated reclaimed water distribution systems suggested that biologically-active GAC filtration may unintentionally select for specific OPs, contrary to expectations and experiences with oligotrophic conditions in potable water systems. While GAC biofiltration was associated with lower total bacteria and Legionella spp. gene markers, there were no apparent benefits in terms of other OPs analyzed. Similarly, disinfectant treatments successful for controlling OPs in potable water were either ineffective or associated with increased levels of OPs, such as Mycobacterium spp. and Acanthamoeba spp., in the reclaimed water examined. In the potable water study, it was possible to recreate conditions associated with growth of N. fowleri in the aged main distribution system from where the water for the experiment was collected; including corroding iron mains, nitrification, and disinfectant decay. While the effects of nitrification could not be confirmed, there was a clear association of iron corrosion with N. fowleri proliferation. The role of iron was explored further in what, to the author's knowledge, was the first study of N. fowleri in private wells. Analysis of 40 wells found correlations between N. fowleri and stagnant iron levels, further supporting the hypothesis that iron corrosion or iron encourages the growth of N. fowleri, and, because wells are not routinely disinfected, not necessarily as a result of promoting disinfectant decay. As this study took place following a major flooding event, it provided insight not only into how surface water contamination may influence private well water microbial communities, but also added to the understanding that current recommendations for disinfecting private wells are inadequate and standards should be implemented to aid homeowners in the event of flooding. This exploratory research illuminated several factors influencing the OP growth in a range of water systems. Identifying key variables that control growth is crucial to improving the safety of these systems. / MS

In recent years, communities that source raw water from the Dan River experienced two severe and unprecedented outbreaks of unpleasant tastes and odors in their drinking water. During both TandO events strong 'earthy', 'musty' odors were reported, but the source was not identified. The first TandO event began in early February, 2015 and coincided with an algal bloom in the Dan River. The algal bloom was thought to be the cause, but after the bloom dissipated, odors persisted until May 2015. The second TandO in October, 2015 did not coincide with observed algal blooms. On February 2, 2014 approximately 39,000 tons of coal ash from a Duke Energy coal ash pond was spilled into the Dan River near Eden, NC. As there were no documented TandO events before the spill, there is concern the coal ash adversely impacted water quality and biological communities in the Dan River leading to the TandO events. In addition to the coal ash spill, years of industrial and agricultural activity in the Dan River area may have contributed to the TandO events. The purpose of this research was to elucidate causes of the two TandO events and provide guidance to prevent future problems. Monthly water samples were collected from August, 2016 to September, 2017 from twelve sites along the Dan and Smith Rivers. Multivariate analyses were applied to look for underlying factors, spatial or temporal trends in the data. There were no reported TandO events during the project but sensory analysis, Flavor Profile Analysis, characterized earthy/musty odors present. No temporal or spatial trends of odors were observed. Seven earthy/musty odorants commonly associated with TandO events were detected. Odor intensity was mainly driven by geosmin, but no relationship between strong odors and odorants was observed. / Master of Science

Past research regarding complexed iron has focused on the resistance to and kinetics of oxidation by O₂(aq) and the extent of stabilization. The 0.45 um filter was typically used to differentiate between dissolved and particulate iron. This research investigated Fe(II) oxidation by free chlorine and ClO₂ in the presence of DOC by varying the pH, DOC to Fe ratios, DOC sources, oxidant dosages, and contact time. Complexed iron removal by alum coagulation with and without oxidant addition was also examined. Particulate, colloidal, and soluble iron were differentiated by the use of 0.2 um filters and 100K ultrafilters. Ultrafiltration and oxidation studies revealed that, at the DOC-to-iron ratios used for this research, not all of the Fe(II) in solution was actually complexed. Thus, oxidation studies represented the oxidation of uncomplexed Fe(II) to Fe(III), which was then complexed by the higher molecular weight DOC. Results indicated that particulate iron formation (as defined as retention by a 0.2 um filter) was a function of the DOC source and oxidant used for testing. The formation of colloidal iron (as defined by retention on 100K ultrafilter) due to oxidation was dependent upon the initial DOC-to-iron ratio and the DOC source. A correlation between DOC adsorption to iron oxide solids and the solution pH, initial DOC-to-iron ratio, and the oxidant used was also evident. Complexed Fe(II) was removed from solution by alum coagulation. Oxidant addition to alum coagulation was necessary to effectively remove uncomplexed Fe(II) (in the presence of DOC) from solution. / Master of Science