While more than 100 years of research has focused on removing acute and chronic health threats from water, substantially less study has focused on potable water infrastructure and water quality deterioration, monitoring technologies, and relationships between water taste and consumer health. These knowledge–gaps have left infrastructure users, owners, regulators, and public health professionals largely unaware of how premise and buried polymer water pipes deteriorate and sorb/ desorb organic contaminants during normal operations and following water contamination events. These knowledge–gaps also prevent infrastructure managers from producing drinking water that optimizes mineral content for both water taste and health benefits, and employing a monitoring tool capable of immediately detecting water contamination or equipment failures. Research was conducted to address these challenges using analytical chemistry, environmental engineering, food science, polymer chemistry, public health, and material science principles. This work was enhanced by collaborations with sixteen American water utilities and the National Institute for Standards and Technology. These efforts were funded by the National Science Foundation, American Water Works Association, and the Water Research Foundation. Research results are unique and provide important scientific contributions to the public health, potable water, and material science industries. Particular achievements include the: (1) Evaluation of linkages between minerals, water palatability, and health useful for water production and public health decisions; (2) Creation of a novel infrastructure and water quality surveillance tool that has begun water utility implementation in the USA; (3) Development of an accelerated chlorinated water aging method with stable water pH, free chlorine, and alkalinity concentration that enables interpretation of polymer pipe surface and bulk characteristic changes; (4) Discovery that polar compounds are 2–193% more soluble in PEX than HDPE water pipes; (5) Finding that several polymer and contaminant properties can be used to predict contaminant diffusivity and solubility during sorption and desorption in new, lab aged, and water utility PE pipes; and the (6) Discovery that chlorinated water exposure of HDPE and PEX pipes increases polar contaminant diffusivity during sorption by 50–162% and decreases diffusivity during desorption as much as 211%. Outcomes of this work have domestic and global significance, and if engaged, can greatly improve public health protection, potable water infrastructure operations, water quality, sustainability, and regulation. / Ph. D.

The taste of a preferred food, pizza, was adulterated with quinine sulfate and the effects of taste experience on subsequent measures of palatability were measured. The measures of palatability were salivary responses to the thought and presentation of pizza. Additional measures were latency to start eating, amount eaten, meal duration, rate of eating and preference ratings of the pizza's taste, aroma and appearance. Thirty-six subjects received access to regular and/or adulterated pizza over two experimental sessions. The resulting groups of nine subjects each received either adulterated and adulterated, adulterated and regular, regular and regular, or regular and adulterated pizza over the two sessions. In a third session all subjects received regular pizza. In session two, groups which had received regular pizza in session one showed a reliably greater salivary response on the presentation trial than on the thought trial. Groups which had received adulterated pizza showed minimal differences in salivation between these trials. In session three, groups which had received regular pizza in session one yielded reliably greater salivation on the presentation trial than did groups which had received adulterated pizza in session one. Furthermore, nonshift groups, which had received the same pizza condition over sessions one and two, showed a greater difference between thought and presentation trial responses than did shift groups, which had received different pizza conditions over sessions one and two. An approach-avoidance conflict model of behavior was applied to the salivation data. Preference ratings of the pizza's taste, the amount eaten and the rate of eating data yielded reliably greater responses for groups which received regular pizza than for groups which received adulterated pizza in sessions one and two. Positive and negative contrast effects were also evidenced by these data. The meal duration and latency to start eating data yielded highly similar responses among groups over days. / M.S.

Thirteen volunteer panelists were trained according to <i>Standard Method</i> 2170, flavor profile analysis (FPA). Following training these panelists underwent triangle test screening to determine whether or not they could detect the odorants used in this study. Following triangle testing, panelists underwent directional difference testing to determine if temperature affected odor perception when presented with two water samples. Following directional difference testing, panelists used FPA and evaluated water samples that contained odorants at either 25°C or 45°C. Samples containing geosmin cooled to 5°C were also evaluated. Sensory analyses experiments indicate that odor intensity is a function of both aqueous concentration and water temperature for geosmin, MIB, nonadienal, n-hexanal, free chlorine, and 1-butanol. The higher water temperature resulted in an increase in odor intensity for some, but not all, concentrations of geosmin, 2-methylisoborneol, <i>trans-2,</i> <i>cis-6-</i>nonadienal, n-hexanal, free chlorine, and 1-butanol. Additionally, above 400 ng/L of geosmin, 400 ng/L of MIB, and 100 ng/L the odor intensity was equal to or less than the odor intensity at 600, 600, and 200 ng/L, respectively. Henry's Law should predict that an increase in concentration would increase the amount of odorant the panelist comes into contact with; however, results demonstrated that at specific aqueous odorant concentrations odor perception did not follow Henry's Law. Odor response to drinking water containing isobutanal was affected by concentration but not water temperature. / Master of Science

River ecosystems across the US and globally face numerous stressors that impact both ecological function and water quality. In 2015-16, municipalities along the Dan River in southern Virginia experienced repeated taste and odor (TandO) issues in their drinking water that originated from the river source water. Given that the source of TandO issues during these events were unknown, this research aimed to identify patterns in the distributions of river microorganisms that could help identify potential biological causes. Monthly water, sediment, and periphyton samples were collected for a full year from the Smith and Dan Rivers to quantify actinomycete, fungi, and chlorophyll a concentrations, which have historically been linked to TandO problems, and to characterize changes in microbial community structure. Although no significant TandO event occurred during the study period, the work produced unique and valuable data that describe patterns of microbial populations and communities in a river ecosystem. Results from the study show the abundances of actinomycetes, fungi, and chlorophyll a expressing seasonal and regional variation by habitat. From a broader ecological perspective, microbial communities sampled from water, sediment, and periphyton were each unique from each other regardless of river reach and season sampled. Overall, this research adds to our understanding of river ecology by detailing the microbial abundance and diversity in three river habitats, including periphyton, that can be used to predict sources of river TandO in future events, and offers new questions regarding how microbial diversity changes over space and time. / Master of Science / In 2015-16, cities along the Dan River in Virginia experienced multiple taste and odor (T&O) events that led to earthy and musty odors in drinking water. As part of a larger project looking at a range of possible chemical and biological sources of T&O, this research aimed to identify changes in abundance of river microorganisms that might indicate potential biological causes to T&O events. Monthly samples of water, sediment, and algal growth were collected for a year from 12 sites on the Smith and Dan Rivers. Samples were analyzed for abundances of three known T&O causing groups of organisms—actinomycetes, fungi, and photoautotrophs—and to characterize changes in total microbial communities as an indicator of ecological change occurring along the rivers. Although a significant T&O event did not occur during the study period, the research produced valuable descriptions of how important microorganisms change in a freshwater ecosystem. Actinomycetes elevated in the lower Dan River during fall, fungi elevated during the spring, and chlorophyll a was highest in the upper Smith River during winter, suggesting that photoautotrophic growth was more likely to be linked to previous T&O events. The diversity and makeup of the microbial communities in the rivers was primarily dependent on where they were growing (water, sediment, or periphyton) and secondarily on the season or the river reach. Combined, these results will help to identify causes of future T&O events in the Dan River and also provide new insights into ecological patterns of microorganisms in river ecosystems.

The use of polymer pipes is now very common in home plumbing and other parts of the drinking water distribution system. Many taste and odor complaints in drinking water are known to originate from contact of water with materials. The ability of polymer pipes used in home plumbing to affect drinking water quality and aesthetics was investigated using the Utility Quick Test (UQT). Analysis of water quality and aesthetics were conducted in the absence of disinfectant and the presence of either chlorine or chloramines. A Flavor Profile Analysis (FPA) panel was trained according to Standard Methods 2170B to evaluate the organoleptic characteristics of the samples. Laboratory analyses were performed to determine levels of total organic carbon (TOC), disinfectant residual, pH and to attempt to identify specific volatile organics. The first part of this study investigated HDPE, cPVC and epoxy lined copper pipes using the UQT method. Both HDPE and epoxy-lined copper had significant effects on water quality and aesthetics during the approximately 10 day exposure of the UQT. HDPE and epoxy-lined copper leached significant amounts of TOC and consumed more disinfectant than controls, but in different amounts. cPVC was the most inert of the materials tested and had the least impact on water quality and did not contribute any significant odor. None of the pipes tested appeared to contribute trihalomethanes to drinking water, but further analysis revealed that the compounds may actually be generated and subsequently sorbed into pipe walls. These data show the effects of newly installed polymeric pipe materials and their potential to impact water quality in differing capacities. The second part of this study compared the results of the UQT on HDPE, epoxy lined copper, cPVC, PEX-a and PEX-b with the goal of comparing and contrasting how five different polymeric plumbing materials can impact drinking water quality. Results demonstrated the short-term ability of all pipe materials except cPVC to impact water quality and aesthetics. This data could potentially be useful in generating selection criteria for homeowners and plumbing professionals regarding the impact of newly installed plumbing materials. / Master of Science

Rats received access to either distilled water, sucrose, quinine, hydrochloric acid, saline, or the three flavors of sucrose, quinine, and hydrochloric followed one or 28-days later by testing for neophobia and training and testing for conditioned taste aversion to saline. At the one-day interval, preexposures to saline, hydrochloric, or multiple flavors attenuated neophobia to saline while only preexposures to saline attenuated the conditioned aversion to saline. At the 28-day interval, none of the preexposure conditions attenuated neophobia to saline while preexposures to saline attenuated the conditioned aversion to saline. / M.S.

Yes / In recent years, diverse scholars have addressed the issue of the chemosensory perceptions associated with traditional medicines, nevertheless there is still a distinct lack of studies grounded in the social sciences and conducted from a cross-cultural, comparative perspective. In this urban ethnobotanical field study, 254 informants belonging to the Gujarati, Kashmiri and English ethnic groups and living in Western Yorkshire in Northern England were interviewed about the relationship between taste and medicinal perceptions of five herbal drugs, which were selected during a preliminary study. The herbal drugs included cinnamon (the dried bark of Cinnamomum verum, Lauraceae), mint (the leaves of Mentha spp., Lamiaceae), garlic (the bulbs of Allium sativum, Alliaceae), ginger (the rhizome of Zingiber officinale, Zingiberaceae), and cloves (the dried flower buds of Syzygium aromaticum, Myrtaceae). The main cross-cultural differences in taste perceptions regarded the perception the perception of the spicy taste of ginger, garlic, and cinnamon, of the bitter taste of ginger, the sweet taste of mint, and of the sour taste of garlic. The part of the study of how the five selected herbal drugs are perceived medicinally showed that TK (Traditional Knowledge) is widespread among Kashmiris, but not so prevalent among the Gujarati and especially the English samples. Among Kashmiris, ginger was frequently considered to be helpful for healing infections and muscular-skeletal and digestive disorders, mint was chosen for healing digestive and respiratory troubles, garlic for blood system disorders, and cinnamon was perceived to be efficacious for infectious diseases. Among the Gujarati and Kashmiri groups there was evidence of a strong link between the bitter and spicy tastes of ginger, garlic, cloves, and cinnamon and their perceived medicinal properties, whereas there was a far less obvious link between the sweet taste of mint and cinnamon and their perceived medicinal properties, although the link did exist among some members of the Gujarati group. Data presented in this study show how that links between taste perceptions and medicinal uses of herbal drugs may be understood as bio-cultural phenomena rooted in human physiology, but also constructed through individual experiences and culture, and that these links can therefore be quite different across diverse cultures.

Studies involving social learning have shown that social interactions are influential in directing an individual's behavior toward relatively neutral stimuli. The present study investigated the possibility that social interactions direct an individual's behavior toward aversive stimuli. Following aversion conditioning to a visual (red water) or taste (3% vinegar) CS, 80 chicks individually observed an audience of two conspecifics ingest the aversive CS or observed a nondrinking audience in the presence of the CS. Observation of a drinking audience reduced the magnitude of the aversion to the visual CS but not to the taste CS. This effect was demonstrated in latency to respond and in log intake. The differences in observational training effects found for the visual CS but not for the taste CS may have been due to differences in visual appearance between red water and vinegar. Percent intake data revealed no differences in strength of conditioning between red water and vinegar. Subjects were retested five days following the last day of initial testing. No evidence was found for observational training effects in retention. Findings were interpreted by the classical conditioning model. / Ph. D.

Die Wahrnehmung von Geschmacksempfindungen beruht auf dem Zusammenspiel verschiedener Sinneseindrücke wie Schmecken, Riechen und Tasten. Diese Komplexität der gustatorischen Wahrnehmung erschwert die Beantwortung der Frage wie Geschmacksinformationen vom Mund ins Gehirn weitergeleitet, prozessiert und kodiert werden. Die Analysen zur neuronalen Prozessierung von Geschmacksinformationen erfolgten zumeist mit Bitterstimuli am Mausmodell. Zwar ist bekannt, dass das Genom der Maus für 35 funktionelle Bitterrezeptoren kodiert, jedoch war nur für zwei unter ihnen ein Ligand ermittelt worden. Um eine bessere Grundlage für tierexperimentelle Arbeiten zu schaffen, wurden 16 der 35 Bitterrezeptoren der Maus heterolog in HEK293T-Zellen exprimiert und in Calcium-Imaging-Experimenten funktionell charakterisiert. Die Daten belegen, dass das Funktionsspektrum der Bitterrezeptoren der Maus im Vergleich zum Menschen enger ist und widerlegen damit die Aussage, dass humane und murine orthologe Rezeptoren durch das gleiche Ligandenspektrum angesprochen werden. Die Interpretation von tierexperimentellen Daten und die Übertragbarkeit auf den Menschen werden folglich nicht nur durch die Komplexität des Geschmacks, sondern auch durch Speziesunterschiede verkompliziert. Die Komplexität des Geschmacks beruht u. a. auf der Tatsache, dass Geschmacksstoffe selten isoliert auftreten und daher eine Vielzahl an Informationen kodiert werden muss. Um solche geschmacksstoffassoziierten Stimuli in der Analyse der gustatorischen Kommunikationsbahnen auszuschließen, sollten Opsine, die durch Licht spezifischer Wellenlänge angeregt werden können, für die selektive Ersetzung von Geschmacksrezeptoren genutzt werden. Um die Funktionalität dieser angestrebten Knockout-Knockin-Modelle zu evaluieren, die eine Kopplung von Opsinen mit dem geschmacksspezifischen G-Protein Gustducin voraussetzte, wurden Oozyten vom Krallenfrosch Xenopus laevis mit dem Zwei-Elektroden-Spannungsklemm-Verfahren hinsichtlich dieser Interaktion analysiert. Der positiven Bewertung dieser Kopplung folgte die Erzeugung von drei Mauslinien, die in der kodierenden Region eines spezifischen Geschmacksrezeptors (Tas1r1, Tas1r2, Tas2r114) Photorezeptoren exprimierten. Durch RT-PCR-, In-situ-Hybridisierungs- und immunhistochemische Experimente konnte der erfolgreiche Knockout der Rezeptorgene und der Knockin der Opsine belegt werden. Der Nachweis der Funktionalität der Opsine im gustatorischen System wird Gegenstand zukünftiger Analysen sein. Bei erfolgreichem Beleg der Lichtempfindlichkeit von Geschmacksrezeptorzellen dieser Mausmodelle wäre ein System geschaffen, dass es ermöglichen würde, gustatorische neuronale Netzwerke und Hirnareale zu identifizieren, die auf einen reinen geschmacks- und qualitätsspezifischen Stimulus zurückzuführen wären. / Taste impression is based on the interaction of taste, smell and touch. To evaluate the nutritious content of food mammals possess five distinct taste qualities: sweet, bitter, umami (taste of amino acids), sour and salty. For bitter, sweet, and umami compounds taste signaling is initiated by binding of tastants to G protein-coupled receptors. The interactions of taste stimuli, usually watersoluble chemicals, with their cognate receptors lead to the activation of the G protein gustducin, which, in turn, initiates a signal resulting in the activation of gustatory afferents. However, details of gustatory signal transmission and processing as well as neural coding are only incompletely understood. This is partly due to the property of some tastants to elicit several sensations simultaneously, unspecific effects caused by the temperature, viscosity, osmolarity, and pH of the solvents, as well as by mechanical stimulation of the tongue during stimulus application. The analysis of gustatory processing of taste information are mainly based on mouse models after stimulation with bitter taste stimuli. Even though it is known that the mouse genome codes for 35 bitter taste receptor genes only few of them had been analysed so far. For better understanding and interpretation of animal experiments 16 mouse bitter receptors had been analysed by Calcium Imaging experiments with HEK293T cells. The data reveal that mouse bitter taste receptors are more narrow tuned than human bitter taste receptors, proving that the ligand spectra of murine and human orthologous receptors are not complient. In order to avoid the disturbing effects of solvents and stimulus application on the analysis of gustatory information transfer and processing, I employ an optogenetical approach to address this problem. For this purpose I generated three strains of gene-targeted mice in which the coding regions of the genes for the umami receptor subunit Tas1r1, the sweet receptor subunit Tas1r2 or the bitter taste receptor Tas2r114 have been replaced by the coding sequences of different opsins (photoreceptors of visual transduction) that are sensitive to light of various wavelengths. In these animals I should be able to activate sweet, bitter, or umami signalling by light avoiding any solvent effects. In initial experiments of this project I demonstrated that the various visual opsins indeed functionally couple to taste signal transduction pathway in oocyte expression system, generating basic knowledge and foundation for the generation of the gene-targeted animals. The knockout-knockin strategies have been successfully realized in the case of all three mouse models, revealed by RT-PCR, in situ hybridization and immunohistochemical analysis of taste papillae. All data confirm that the particular taste receptors have been replaced by the different opsins in taste cells. Further analysis concerning the functional consequences of opsin knockin and taste receptor knockout are part of prospective work.

Chemoreception plays an important role in predator-prey interactions and feeding dynamics. While the chemoreception of attractant or pleasant tasting compounds has been well studied, aversive chemoreceptive signaling has been difficult to investigate behaviorally in an ecological context because these interactions are species- and context- specific and deterrent compounds vary among prey. Using the coral reef system, this thesis explores on a molecular level the deterrent mechanism underlying detection by fish predators of an aversive compound, in order to gain a greater understanding of predator-prey interactions in this community. Like other organisms that are sessile or slow-moving, marine sponges have special mechanisms for defense from predation, commonly containing aversive-tasting compounds that defend these organisms from predation. To this end, we sought to identify and characterize a fish chemoreceptor that detects one or more of these compounds. We isolated a single cDNA clone encoding RAMP-like triterpene glycoside receptor (RL-TGR), a novel co-receptor involved in the signaling of triterpene glycosides. This co-receptor appears to be structurally and functionally related to receptor activity-modifying proteins (RAMPs), a family of co-receptors that physically associate with and modify the activity of G protein-coupled receptors (GPCRs). Expression in Xenopus oocytes showed that it responds to triterpene glycosides in a receptor-mediated manner and requires co-expression of a GPCR to enable signaling in oocytes; both of these receptors may be components of a larger signaling complex. A 40 bp portion of the gene is conserved across multiple fish species, but is not found in any other organism with a sequenced genome, suggesting that the expression of this receptor is limited to fish species. RL-TGR is the first identified gene encoding a co-receptor that responds to a chemical defense. This finding may lead the way for the identification of many other receptors that mediate chemical defense signaling in both marine and terrestrial environments, as this protein has the potential to represent the first of an entire family of co-receptors that respond to aversive compounds.