Tesi sul tema "Tracing pollution"

This dissertation addressed questions pertaining to mercury (Hg) fate and transport in aquatic ecosystems by applying stable Hg isotopes as a tracer. Mercury poses a public health burden worldwide. In parts of the developing world, Hg-use during artisanal and small-scale gold mining (ASGM) is pointed at as the source of elevated Hg in the environment. However, Hg from other sources including soil erosion associated with land cover and land-use change (LCLUC) may also contribute to local Hg pollution.

Stable Hg isotope profiles of sediment cores, surface sediments, and soils from two aquatic ecosystems in Amapá, Brazil, one downstream artisanal gold mining (AGM) and one isolated from AGM were assessed. Although previous studies attributed elevated environmental Hg levels in this area to AGM, stable Hg isotopic evidence suggests elevated Hg downstream of AGM sites is dominantly from erosion of soils due to LCLUC.

In contrast, the impact of Hg-use during small-scale gold mining (SGM) in the Southern Andean Region of Portovelo-Zaruma, Ecuador on Hg in the trans-boundary Puyango-Tumbes River was also investigated. By comparing preliminary isotopic Hg signatures from river sediment along the Puyango-Tumbes to soil and sediment from upstream locations along the Puyango tributaries, we suggest Hg-use during SGM in this region is likely responsible for elevated Hg downstream and into Peru. Technical and policy challenges in measuring and responding to gold mining-related cumulative impacts were also reviewed in the context of Portovelo-Ecuador.

Together, the findings not only answer questions of critical importance to preventing Hg pollution in two of the world's most vulnerable ecosystems but also provide information that can be used to better target interventions to reduce environmental Hg levels and subsequent human exposures. Furthermore, the validation and application of the stable Hg isotope method to trace Hg pollution from ASGM in different aquatic ecosystems represents a critical step to the application of stable Hg isotopes to trace pollution in other complex natural environments and to address public health-related questions.

The contemporary world landscape is epitomised by a highly globalised world economy, where neo-liberal agendas push for ultimate competitiveness and much of the manufacturing and production processes have been relocated from core to semi-peripheral countries. To be able to compete in the global arena, many developing nations and newly emerging economies have sacrificed an approach to sustainable development by neglecting social and ecological aspects. A showcase of such unsustainable growth in recent years can be observed in Tirupur, South India. Tirupur’s heavy export orientation of ready-knitted garments transformed it into a major textiles cluster for fashion retailers and buyer groups worldwide. However, the substantial pollution of water resources due to the discharge of raw effluents by processing units into the Noyyal river and the overexploitation of groundwater cast a shadow on the city’s economic expansion. In order to tackle these challenges, a number of regulatory directives were issued from the mid 1990’s onwards, leading to the enforcement of strict Zero Liquid Discharge norms in the cluster. This study investigates whether intervention efforts directed at Tirupur’s textiles sector have been able to address inherent challenges impacting the local environment and population. The findings gathered from an extensive literature review and a field study to Tirupur suggest that while some problems of the past have been attended to, major sustainability issues remain. Serious concerns such as a shift in pollution and economical uncertainty in the cluster have been identified as consequences from the policy interventions.

Following the crash of a Norwegian Hercules aircraft on Rabots glaciär in the Kebnekaise mountain range in 2012, a field campaign was initiated in order to assess the fate of the hydrocarbon pollution in the system. It is hypothesized that soluble components of the aircraft fuel will be transported within the glacial meltwater. This thesis focuses on constraining the likely transit time and dispersion of the meltwater as a proxy for potential pollution pathways. Therefore, the hydrologic configuration of Rabots glaciär was studied during the ablation season 2013 by means of dye tracing experiments and discharge monitoring in the proglacial stream. The analyses of the dye return curves and stream monitoring suggest that Rabots glaciär exhibits a widely efficient drainage system towards the end of the ablation season, but with analyses revealing heterogeneity in the drainage system form. The seasonal evolution of efficiency was also assessed, showing an increase over time, although was hampered by early onset of melting before the field season began. There are different hydrological configurations on the north and south side of the glacier, possibly influenced by shading. The system on the north side is routing meltwater along the glacier bed over a long distance as indicated by the turbid outlet stream. Water routing on the southern side likely occurs through englacial channels. This configuration may be influenced by the thermal regime and distribution of cold surface layers. It has further been revealed that both systems are likely to be disconnected from each other. Pollution that is transported with the meltwater down from the crash site on the southern side does not reach the drainage system on the northern side. Besides revealing potential pathways for soluble hydrocarbon pollutants, this case study contributes to the previously very limited knowledge of Rabots glacial hydrology, and our general understanding of polythermal glacier hydrology.

Classiquement le transfert des contaminants dans le milieu souterrain est modélisé par un couplage des processus de transport physiques (écoulements contrôlés par les structures géologiques poreuses) et des processus de dégradation ou d'immobilisation chimiques et biologiques. Tant sur les structures géologiques que sur la chimie et la physique, les modèles sont de plus en plus détaillés mais de plus en plus difficiles à calibrer sur des données toujours très parcellaires. Dans cette thèse, nous développons une approche alternative basée sur des modèles parcimonieux sous la forme d’un simple graphe de compartiments interconnectés généralisant les modèles d’interaction de continuums (MINC) ou de transfert à taux multiples (MRMT). Nous montrons que ces modèles sont particulièrement adaptés aux milieux dans lesquels la diffusion de solutés occupe un rôle prépondérant par rapport à l’advection, tels les sols ou les aquifères très hétérogènes comme les aquifères fracturés. L'homogénéisation induite par la diffusion réduit les gradients de concentration, accélère les mélanges entre espèces et fait de la distribution des temps de résidence un excellent proxy de la réactivité. En effet, ces structures simplifiées reconstituées à partir d’informations de temps de résidence se révèlent également pertinentes pour des réactions chimiques non linéaires (e.g. sorption, précipitation/dissolution). Nous montrons finalement comment ces modèles peuvent être adaptés automatiquement à des observations d’essais de traceurs ou de réactions de biodégradation. Ces approches parcimonieuses présentent de nombreux avantages dont la simplicité de développement et de mise en œuvre. Elles permettent d’identifier les déterminants majeurs des échanges entre zones advectives et diffusives ou entre zones inertes et réactives, et d’extrapoler des processus de réactivité à des échelles plus larges. L’utilisation de données de fractionnement isotopique est proposée pour améliorer la dissociation entre l’effet des structures et de la réactivité
Modelling of contaminant transfer in the subsurface classically relies on a detailed representation of transport processes (groundwater flow controlled by geological structures) coupled to chemical and biological reactivity (immobilization, degradation). Calibration of such detailed models is however often limited by the small amount of available data on the subsurface structures and characteristics. In this thesis, we develop an alternative approach of parsimonious models based on simple graphs of interconnected compartments, taken as generalized multiple interacting continua (MINC) and multiple rate mass transfer (MRMT). We show that this approach is well suited to systems where diffusion-like processes are dominant over advection, like for instance in soils or highly heterogeneous aquifers like fractured aquifers. Homogenization induced by diffusion reduces concentration gradients, speeds up mixing between chemical species and makes residence time distributions excellent proxies for reactivity. Indeed, simplified structures calibrated solely from transit time information prove to provide consistent estimations of non-linear reactivity (e.g. sorption and precipitation/dissolution). Finally, we show how these models can be applied to tracer observations and to biodegradation reactions. Two important advantages of these parsimonious approaches are their facility of development and application. They help identifying the major controls of exchanges between advective and diffusive zones or between inert and reactive zones. They are also amenable to extrapolate reactive processes at larger scale. The use of isotopic fractionation data is proposed to help discriminating between structure-induced effects and reactivity

Mestrado em Biologia Aplicada
As águas marinhas costeiras são suscetíveis a contaminação fecal, tanto por fontes pontuais, como por fontes difusas, que podem ter contribuições de fontes individuais pertencentes a animais selvagens, animais domésticos e seres humanos. Os inputs de fontes difusas no ambiente são dispersos e esporádicos, o que torna a sua deteção difícil. A distinção entre a contaminação fecal de origem humana e não-humana tem vindo a tornar-se, nos últimos anos, um objetivo global crucial, uma vez que tem impacto na saúde humana e na economia local. Uma vez que a qualidade das águas superficiais é relevante para a saúde pública devido à sua ampla utilização, especialmente em atividades de lazer e consumo de marisco, a avaliação das fontes de poluição fecal primárias torna-se, assim, uma medida prioritária. Apesar da contaminação fecal por animais selvagens ser considerada de baixo risco para a saúde humana quando comparada com a poluição fecal de origem humana, as fezes de animais selvagens podem também transportar microrganismos patogénicos para humanos. Nos últimos anos, um problema de contaminação fecal foi detetado na água da praia da Ilha da Berlenga. No sentido de esclarecer qual a origem desta contaminação surgiu este estudo, tendo como principal objetivo a determinação e identificação da(s) fonte(s) de poluição fecal responsáveis pela contaminação da água detetada na Ilha da Berlenga. Este objetivo foi alcançado utilizando a metodologia de “Microbial Source Tracking”, através de tipagem molecular (BOXPCR) de isolados de Escherichia coli provenientes da água da praia, de fezes de gaivotas e de um efluente de origem humana e da análise dos dendrogramas resultantes. Para além disso, outros aspetos foram analisados, nomeadamente, a abundância relativa, a saturação de amostragem e índices de diversidade. Tendo em conta os dados resultantes do presente estudo, é possível concluir que: (i) as gaivotas podem ser consideradas o principal responsável pela poluição fecal da água praia; (ii) o método de amostragem e a estratégia da análise dos resultados obtidos podem ser considerados eficientes, para este tipo de ambiente e isolados; e (iii) o esforço de amostragem não foi suficiente para atingir toda a diversidade das populações de E. coli durante amostragem permitindo, no entanto, concluir quanto à principal fonte de contaminação fecal neste ambiente.
Coastal marine waters are often susceptible to fecal contamination from a range of point and nonpoint sources, with potential contributions from many individual sources belonging to wildlife, domesticated animals, and humans. These nonpoint source inputs into the environment are dispersed and sporadic, which makes their detection difficult. The distinction between human and non-human fecal contamination is becoming an important worldwide purpose, in light of the impact of fecal pollution on human health and economic affairs. Since quality of surface waters is relevant to public health due its wide use, particularly for recreational activities and seafood consumption, accurate assessment of primary sources of fecal pollution is clearly a priority measure. While fecal contamination from wildlife sources is often believed to present low human health risks compared to sewage, wildlife species are believed to carry human pathogens that may pose a health risk to humans as well. In the last few years a problem of fecal contamination has been detected in the beach of the Berlenga Island. Thus, this study has emerged having as major aim the determination and identification of which sources of fecal pollution are the responsible for the water contamination detected in the Berlenga Island. This aim was achieved using a Microbial Source Tracking methodology through molecular typing (BOX-PCR) of Escherichia coli isolates from contaminated water, seagull feces and a human-derived effluent and analysis of the resulting clustering. In addition, relative abundance, sampling saturation and diversity indices were analyzed. Taking into account the data resulting from the present study, it is possible to conclude that: (i) the seagulls can be considered the main responsible for the fecal pollution of the beach water; (ii) the sampling method and the analysis methodology can be considered efficient to this type of environment and isolates; (iii) the sampling efforts were not enough to achieve all the diversity of the E. coli populations sampled allowing, however, the determination of the dominant source of fecal pollution in this environment.

Avec une utilisation mondiale croissante pour des applications variées (nouvelles technologies, énergies vertes, etc.), les terres rares (REEs pour ‘rare earth elements’) sont devenues des polluants émergents. Ces pollutions peuvent/pourront être d’origine industrielle (ex. rejets dans l’environnement en sortie d’usine), minière, ou causées par le stockage inapproprié de produits industriels (ex. aimants à base de néodyme (Nd) fragiles à la corrosion). Dans ce contexte, ce projet de thèse a cherché à développer un nouvel outil : l’étude du fractionnement dépendant de la masse des isotopes du Nd (δNd en ‰). Les objectifs de ce projet ont été i) d’apporter un maximum d’éléments pour aider à tracer les pollutions anthropiques de Nd dans l’environnement et ii) d’aider à mieux comprendre comment fonctionne l’accumulation des REEs par les plantes, en particulier les fougères hyperaccumulatrices Dicranopteris linearis, dans le but d’appuyer les études de phytoremédiation des sites pollués. Le δNd de matériels anthropiques (solutions synthétiques de Nd pur et aimants industriels Nd2Fe14B) a été mesuré (gamme de variation de 1,45 ‰ en δ148Nd, littérature incluse) et comparé à la signature des roches terrestres de la littérature (gamme de 0,66 ‰). Les résultats montrent que l’utilisation combinée i) du rapport de l’isotope radiogénique 143Nd/144Nd (ε143Nd, traceur de sources), ii) de la composition des isotopes stables du Nd (δNd, traceur de sources et processus) et, iii) du type de fractionnement (cinétique ou à l’équilibre thermodynamique) des isotopes stables du Nd, peut permettre à l’avenir de distinguer le Nd naturel du Nd anthropique et de tracer ce Nd anthropique dans l’environnement. La mesure du δNd de 5 spécimens de fougères D. linearis, ainsi que de 3 standards biologiques (lichen, feuilles de pommes et lentilles d’eau) a permis de déterminer de façon évidente que la biologie peut fractionner les isotopes stables du Nd. Les valeurs extrêmes en δ148Nd, parmi les échantillons biologiques, sont -0,415 ± 0,060 ‰ et -0,011 ± 0,022 ‰ (2σmean), respectivement pour l’une des pétioles des fougères et pour les lentilles d’eau. Trois résultats principaux en découlent : i) le fractionnement des isotopes stables (δNd) est corrélé au fractionnement des REEs légères par rapport aux REEs lourdes pour tous les échantillons mesurés (fougères, lichen, feuilles de pomme, lentilles d’eau et tous les sols) ; ii) le transport et la distribution du Nd (et des REEs) dans les différentes parties des fougères sont corrélés à ceux du manganèse (Mn), suggérant un mécanisme de transport similaire des REEs et du Mn, pour les fougères mais aussi chez d’autres végétaux comme les pommiers ; iii) la dégradation de l’argile, sur laquelle les REEs sont massivement adsorbées dans les sols étudiés, peut induire le fractionnement des isotopes stables du Nd des sols, que ce soit une dégradation causée par des processus naturels (ex. biologie) ou anthropiques (extraction des REEs avec des acides miniers)
With the worldwide growing demand for various applications (new technologies, green energies, etc.), rare earth elements (REEs) are now considered as emerging pollutants. These pollutions are/will be of industrial origin (e.g. industrial wastes), of mining origin, or caused by the inappropriate storage of industrial products (e.g. neodymium (Nd) magnets fragile against corrosion). In this context, this PhD project aims to develop a new tool: the study of the mass-dependent isotopic fractionation of Nd (δNd in ‰). The goals of this work were to i) bring the maximum of information in order to help to identify and trace anthropogenic Nd in the environment, and ii) help the understanding of how plants accumulate REEs, especially the hyperaccumulator Dicranopteris linearis ferns, in order to support the studies about the phytoremediation of polluted areas. The δNd of anthropogenic materials (pure Nd synthetic solutions, Nd2Fe14B industrial magnets; δ148Nd range of 1.45 ‰, literature included) was compared to the signature of natural terrestrial rocks of the literature (δ148Nd range of 0.66 ‰). Our results show that the use together of i) the 143Nd radiogenic isotope (ε143Nd, tracer of sources), ii) the stable Nd isotopic composition of Nd (δNd, tracer of sources and processes) and, iii) the kind of stable Nd isotopic fractionation (kinetic or thermodynamic equilibrium), could allow the distinction of the natural from the anthropogenic Nd, and to trace the anthropogenic Nd in the environment. The measurement of the δNd of 5 D. linearis fern specimens, and of 3 biological standards (lichen, apple leaves and duck weed) allowed discovering that biology can fractionate the stable Nd isotopes. Extreme δ148Nd values are -0.415 ± 0.060 ‰ and -0.011 ± 0.022 ‰ (2σmean), respectively for one of the fern petiole and the duck weed. Three principal results were obtained: i) the δNd fractionation is correlated to the fractionation of the light REEs than the heavy REEs for all the studied samples (ferns, lichen, apple leaves, duck weed and all soil materials); ii) the distribution and transport of Nd (and REEs) in the different parts of ferns are correlated to those of manganese (Mn), suggesting similar transport mechanisms for REEs and Mn, for ferns but also for other plants as apple trees; iii) the degradation of clay minerals, on which REEs are mostly adsorbed in the studied soils, can induce the fractionation of stable Nd isotopes, from natural processes (e.g. biological) or anthropogenic processes (REEs mining acidic extraction)

Sources of E. coli were determined from impaired waterways in the Appomattox River watershed (in the lower Piedmont and South-Central Virginia) for the development of bacterial Total Maximum Daily Loads (TMDLs). The Appomattox River watershed is primarily undeveloped with 70.8% of the land forested, 17.0% used for agriculture (mainly livestock production), and 7.7% classified as water, wetland or barren land. The remaining 4.5% is developed for residential, commercial, and industrial land uses (mainly within the city of Petersburg). Using Antibiotic Resistance Analysis, a known source library of 1,280 E. coli isolates (320 isolates per source) was constructed. Water samples were collected monthly for between eleven and fourteen months (11/02-12/03) from 40 locations throughout the Appomattox watershed and analyzed for fecal coliforms, E. coli, and resistance to 7 antibiotics of varying concentrations. A total of 486 water samples (9,907 isolates) were analyzed during the study. The objectives of this study were verify that each sampling site exceeded state bacterial count standards (using fecal coliform data), to compare the Discriminate Analysis and Logistic Regression statistical models for use in the classification of isolates, and finally to determine the source of contamination at each site. The fecal coliform and E.coli data was used to determine if each site exceeded state standards during the assessment period. Thirty-eight of the sites exceeded the fecal coliform standard at least 10% of the time, and thirty-three exceeded the E.coli standard at least 10% of the time. Discriminate Analysis (DA) is typically used to classify isolates, but the results obtained from the DA model were unrealistic based on the watershed land uses. By statistically analyzing the original 1,280 E.coli isolates six different ways, a more appropriate classification of isolates was determined. The six analyzing methods were Regular DA and Logistic Regression (LR); DA and LR where each isolate whose probability fell below 80% was deleted; DA and LR where each isolate whose probability fell below 80% was used to create an Unknown category. The Logistic Regression model with an Unknown category proved to be the most appropriate. By using the Logistic Regression model, with Unknown category, to classify isolates, twenty five of the forty sites were discovered to be contaminated predominately with Livestock and fourteen of the sites predominately by Wildlife. One site was equally divided between these two categories. Human and Pet contamination were not dominant at any of the forty sites. This comparison of the DA and LR statistical methods could change the analysis standard for Bacterial Source Tracking and suggests that the model required to classify isolates depends on the watershed characteristics.
Master of Science

Antibiotic resistance analysis (ARA) of Enterococci was used to determine sources of fecal contamination in three District of Columbia waterways: Rock Creek, the Anacostia River, and the Potomac River. These three waterways were identified as exceeding water quality standards set for fecal coliform levels and were designated by the District of Columbia to the Environmental Protection Agency's 303 (d) impaired waters list. A library profile of 1,806 enterococcus isolates from known sources was built based on antibiotic resistance patterns from thirty concentrations of nine antibiotics. These sources included human, cattle, chicken, horse, goat, sheep, deer, raccoon, muskrat, goose, seagull, coyote, duck, wild turkey, dog, and cat. Antibiotic profiles were characterized for 24 unknown enterococci isolates on each of 198 samples (38 samples from the Potomac River, 79 samples from the Anacostia River, and 81 samples from Rock Creek) collected periodically from July 2002 through April 2003. Two major storm events were also sampled during this period. These isolate profiles were compared to the known source library using logistic regression. Three dominant sources of fecal pollution were detected in the Potomac River: livestock (30%), human (29%), and wildlife (22%). Three dominant signatures were also detected in Rock Creek: horse (26%), human (26%), and wildlife (24%). Human was the only dominant source detected in the Anacostia River, averaging 43% over the sampling period. The results of this study indicate that human is a substantial contributor to the fecal contamination problems, especially in the Anacostia River, but there are significant agricultural and wildlife contributions as well. Significant and predictable seasonal variations were also detected, indicating the influence of precipitation on source distributions. The results of this study will aid the Metropolitan Washington D.C. Council of Governments in making important management decisions to help improve the water quality in and around the Washington D.C. area. Expanding the limits of ARA was also an integral part of this research. Three new and even controversial analytical techniques were run on the data collected from this project in an attempt to improve confidence and provide direction to the results of this study. The first was a comparison of the more commonly used statistical analysis model discriminate analysis (DA) with logistic regression (LR). No significant difference was found between the output of the two models for the known source libraries, therefore no suggestion could be made in favor of one model over the other. Another analytical test of the data was the introduction of a standard requiring isolates to meet a minimum of 80% similarity to the known source profiles where it was classified. With the 80% cutoff, between 41% and 44% of the isolates could not be classified to any source and were placed in an unknown category. Based on the remaining isolates, source distributions were recalculated and were not statistically different than those calculated with no restriction for isolate similarity for matching. The last major test of the data was the analysis of the library for representativeness via pulled sample cross validation and the exclusion of all duplicate patterns from the known source library. These analyses did not confirm the representativeness of the databases, but results were further analyzed based on the implications these analyses have on library based methods.
Master of Science

This project involves developing and applying bacterial source tracking (BST) methodology to determine sources of fecal pollution in water for a rural community (Millwood, VA). Antibiotic resistance analysis (ARA) is the primary BST method for fecal source identification, followed by randomly amplified polymorphic DNA (RAPD) analysis for confirmation. Millwood consists of 66 homes, all served by individual septic systems, and a stream (Spout Run) passes through the center of the community. Spout Run drains a 5,800 ha karst topography watershed that includes large populations of livestock and wildlife. Stream and well samples were collected monthly and analyzed for fecal coliforms and fecal streptococci, starting in 5/99 and ending in 5/00. Twelve percent of the well samples and 92% of the stream samples were positive for fecal coliforms, and 26% of the stream samples exceeded the recreational water standard (1,000 fecal coliforms/100 ml). ARA of fecal streptococci recovered from the stream samples indicated that isolates of human origin appeared throughout the stream as the stream passed through Millwood with a yearly average of (approx. 10% human, 30% wildlife, and 63% livestock), and the percent human origin isolates declined downstream from Millwood. These results were obtained by comparing the antibiotic resistance profiles of stream isolates against a library of 1,174 known source isolates with correct classification rates of 94.6% for human isolates, 93.7% for livestock isolates, and 87.8% for wildlife isolates. There is a human signature in Spout Run, but it is small compared to the proportion of isolates from livestock and wildlife. The sporadic instances where well water samples were positive appeared primarily during very dry periods. Restricting livestock access to streams can dramatically lower fecal coliform counts during the unusually hot and dry periods. Reducing FC counts to below recreational water standards for Virginia (1000 per 100ml for any one sample) may be achievable, however to maintain streams below standards may prove to be difficult, as Spout Run is in an area where there are large populations of Canada geese, deer, and other wildlife, and will be hard to restrict these animals.
Master of Science

Antibiotic resistance analysis (ARA) was used to determine sources of fecal pollution in the Blackwater River in South-central Virginia. The Department of Environmental Quality designated six segments as impaired due to high fecal coliform concentrations with non-point source (NPS) agriculture the suspected source of impairment. The Blackwater River watershed encompasses 72,000 ha of dairy, beef, and intensive production agriculture, abundant wildlife populations and many homes with onsite septic systems. A library of antibiotic resistance profiles based on 30 concentrations of 9 antibiotics was developed for 1,451 enterococci isolates from human, cattle, chicken, horse, goat, sheep, deer, raccoon, muskrat, goose, duck, coyote, and wild turkey fecal samples. Each isolate was classified as human, wildlife or livestock. Correct classification rates were 82.3% for human, 86.2% for livestock and 87.4% for wildlife isolates when profiles were analyzed with discriminant analysis. Profiles were also determined for 48 isolates from 128 stream samples collected periodically from August 1999 thru April 2001 and compared to the known sources using discriminate analysis. A human signature was found at each site at least once during the year, ranging from 0.0% to 85.0% of the sample isolates. The livestock signature varied from 2.3% to 100% over sites and months, and the wildlife signature varied from 0.0% to 79.5%. The results indicate that both humans and wildlife contribute to fecal pollution in addition to the suspected source, livestock, and reducing fecal pollution will require consideration of all three sources. The results from this research are being used to develop a total maximum daily load (TMDL) project allocations for fecal coliforms in the Blackwater River. Isolates identified by ARA were also profiled using the Biolog metabolic identification system. A library of metabolic profiles was constructed from known source isolates. Stream isolates were identified by Biolog and the metabolic profile was compared to the Biolog library. Of ten stream isolates identified by ARA as human, the Biolog library identified one as human, four as livestock, and five as wildlife. Of ten isolates identified by ARA as livestock, the Biolog library identified seven as livestock and three as wildlife. Of ten isolates identified by ARA as wildlife, one was identified as human, three as livestock and six as wildlife. The overall correct classification of Blackwater isolates in the Biolog library was 14 of 30 isolates, or 47%. Although the Biolog library was constructed with some isolates from the Blackwater basin, there may not be enough isolates in the Biolog library to adequately represent the variability shown by the Blackwater isolates, resulting in lower than expected correct classifications. In spite of these results, Biolog remains promising as one of several tools with potential as a bacterial source tracking method.
Master of Science

The research used dendro-chemical analysis of ash tree rings and current year leaf litter to track Cd, Pb, Zn, Cu, Mn, Cr, and Sn spread and cycling from a closed garbage dump-toxic waste site. This technique allowed for determination of areal extent, contaminant levels and time period of initial contaminant contact. Only Zn, Sn, and Cu were found in elevated quantities in the xylem wood and Pb in the leaf litter. Elemental concentrations of Pb, Sn and Cd in xylem wood and leaves of ash were positively correlated. Tin was the only element to demonstrate a clear initial contact period and elemental accumulation with age. Significant levels of Cu accumulated in the heartwood while Zn revealed significant but inconsistent accumulated patterns. Expected attenuation zones associated with municipal solid waste landfill leachate dispersion were not found; thus the pathway for contaminant dispersion was likely through groundwater flow.
An elemental index was developed to facilitate the use of dendro-chemical analysis in periods of suppressed tree growth resulting from environmental pollution.

Swimming in ocean water, including beaches not impacted by known point sources of pollution, continues to pose a rising health concern. Studies which evaluate the relationship between indicator microbes, pathogens, and human health at recreational beaches are rare, especially at non-point beaches, although such studies are necessary for establishing criteria which would protect public health while minimizing economic burdens. The objective of this dissertation was to better understand the non-point source beach system, and hence determine possible directions that should be taken from a beach regulatory perspective in order to protect bather health and minimize economic burdens. This objective was accomplished through two studies. The first study included an initial evaluation of the presence of and associations among indicator microbes, pathogens and environmental conditions at a subtropical, recreational marine beach in South Florida (USA) impacted by non-point sources of pollution. Twelve water and eight sand samples were collected during four sampling events conducted at high or low tide and during elevated or reduced solar insolation conditions. Analyses included fecal indicator bacteria (FIB) (fecal coliform, Escherichia coli, enterococci, Clostridium perfringens), human-associated microbial source tracking (MST) markers (human polyomaviruses [HPyVs] and Enterococcus faecium esp gene), and pathogens (Vibrio vulnificus, Staphylococcus aureus, enterovirus, norovirus, hepatitis A virus, Cryptosporidium spp. and Giardia spp.). The objective of the second study was to capture a series of "snapshots" of the study beach with respect to pathogens and indicator microbe levels, environmental conditions, and cumulative human health effects. Two sets of daily composite samples were collected during the BEACHES epidemiology study conducted in Miami (FL) at the same non-point source subtropical marine beach. These samples were analyzed for several pathogens, microbial source tracking markers, indicator microbes, and environmental parameters similar to those in the first study. Enterococci concentrations in water and sand assessed by qPCR were greater than membrane filtration measurements in both studies. FIB concentrations in water were below recreational water quality standards for three of four sampling events; pathogens and MST markers were also generally undetectable during the first study. FIB levels exceeded regulatory guidelines during one event, accompanied by the detection of HPyVs and pathogens, including the autochthonous bacterium Vibrio vulnificus in both sand and water, the allochthonous protozoan Giardia spp. in water, and Cryptosporidium spp. in sand samples. These elevated microbial levels were detected at high tide, low solar insolation conditions, and after elevated 2 day antecedent rainfall. Analyses of the second study demonstrated that rainfall and tide were also significant factors in determining the presence of both indicator microbes and pathogens. Additional potential associations between environmental factors, microbes and human health effects were also identified. However, more analyses should be conducted as the limited number of samples renders it difficult to make conclusive statements about the relationship between these different factors. The results of this research demonstrate the complexity of beach systems characterized by non-point sources, and how more novel and comprehensive approaches will be needed to assess beach water quality for the purpose of protecting bather health. A recommended approach "The comprehensive tool box with approval process approach" to address the beach complexity is presented and applied to the study beach.

Les poblacions humanes i animals excreten una gran diversitat de virus patògens en les seves femtes i orina, de manera que l’aigua residual que generen representa un dels principals vehicles de la disseminació de patògens a les aigües superficials, subterrànies o costaneres i conseqüentment, als aliments. La contaminació del medi ambient suposa un risc greu de salut pública. S’ha estimat que en el conjunt del planeta, aproximadament 3.000 milions de persones no disposen d’aigua potable i que el 95% de l’aigua residual domèstica és abocada al medi ambient sense tractar. Aquesta tesi doctoral es basa en dues àrees d’estudi de la virologia ambiental. Per una banda s’ha avaluat l’estabilitat de virus contaminants en condicions naturals (Capítol I) i davant processos de desinfecció (Capítol II). En aquests estudis, els indicadors virals/patògens HAdV i JCPyV han estat analitzats. D’altra banda, s’ha proposat una nova eina per a la identificació de l’origen de la contaminació fecal al medi ambient (Capítol III). Al Capítol I, que inclou els estudis 1, 2 i 3, s’ha caracteritzat els factors ambientals i els mecanismes que intervenen en la inactivació natural dels HAdV a diferents ambients aquàtics i aliments, considerant la temperatura i la llum solar com els agents potencialment més rellevants, així com l’estandardització entre diferents laboratoris d’un mètode de qPCR per a la quantificació dels HAdV en mostres d’aliments (fruits rojos). Al Capítol II (estudis 4, 5 i 6) s’ha estudiat l’efectivitat de tractaments químics (clor) i físics (llum ultraviolada) en la inactivació de virus contaminants. Finalment, al Capítol III (estudi 7), s’ha considerat que era interessant completar les eines moleculars existents per a identificar les fonts de contaminació fecal en aigua i aliments, i s’ha desenvolupat una nova tècnica molecular per a traçar la contaminació fecal d’aviram al medi ambient, mitjançant la detecció i quantificació dels parvovirus de pollastre i gall d’indi. És evident que la contaminació viral en el medi representa un risc d’infecció important en països en vies de desenvolupament però també en zones industrialitzades on els tractaments d’aigua no aconsegueixen eliminar la totalitat dels virus presents. Durant el desenvolupament d’aquesta tesi doctoral s’ha demostrat que, en molts casos, els mètodes de desinfecció més habituals en la inactivació de virus contaminants no són completament eficients i sovint es detecten virus infecciosos després de tractaments a les dosis o concentracions habitualment utilitzades. Aquesta observació demostra la importància de seguir treballant per a caracteritzar els diversos factors i mecanismes particulars que condicionen la inactivació natural dels virus patògens al medi ambient, i de desenvolupar i optimitzar nous mètodes de desinfecció per assolir la inactivació d’un rang de microorganismes més ampli, més enllà dels indicadors bacterians tradicionals. Els avanços en la virologia ambiental han d’anar forçosament acompanyats d’avenços en els tècniques i mètodes utilitzats, que han de permetre detectar, quantificar i caracteritzar els virus presents al medi de manera rutinària. En aquest sentit, els mètodes moleculars representen eines valuoses que faciliten l’estudi de diversos aspectes de la disseminació de virus al medi ambient, com per exemple identificar l’origen de la contaminació fecal per a implementar mesures de remediació a les fonts de contaminació més rellevants en cada localitat. Els virus patògens contaminants representen un problema de salut pública que cal resoldre imperativament millorant els processos de control microbiològica de recursos tant imprescindibles com són l’aigua i els aliments, introduint paràmetres virals que complementin als indicadors fecals bacterians estàndards.

Sources of fecal pollution were evaluated as a function of sampling frequency with stream samples from Mill Creek, Montgomery County, VA. Samples were collected monthly for one year, plus weekly for four consecutive weeks during seasonal high flows (March), and seasonal low flows (September-October), plus daily for seven consecutive days within the weekly schedules. Thirty stream samples were collected from each of two sites (60 total) in Mill Creek, and 48 isolates of E. coli per sample (total of 2,880 stream isolates) were classified by source using antibiotic resistance analysis (ARA) and comparing the resulting patterns against a known-source E. coli library (1,158 isolates). The same process was performed with enterococci isolates against an enterococci library (1,182 isolates). The average rate of correct classification (ARCC) for the E. coli library with a three-way split (human, livestock, and wildlife) was 89.0%, and the ARCC of the species-specific E. coli library (cattle, deer, goose, human, misc. wildlife) was 88.9%. The ARCC of the enterococci library with a three-way split was 85.3%, and the ARCC of the species-specific enterococci library was 88.1%. The results did not justify the need for daily or weekly sampling, but indicated that monthly was adequate (quarterly and every-other-month were not). There was a seasonal effect as the human signature was highest during high flow while the livestock signature dominated during low flow. The results also indicated that sampling should be done over a period of time that includes both seasonal wettest and driest periods (at least 8 months).
Ph. D.

Metabolic were used to evaluate Enterococcus as an indicator of faecal pollution. Enterococci were isolated using m-Enterococcus agar and speciated using conventional biochemical tests. Forty percent of the isolates were identified and metabolically characterized by the automated Biolog system. The biochemical test scheme recognized 16 enterococcal species, while Biolog recognized nine. Both methods identified E. faecalis at the greatest frequency. Overall species frequencies varied between the two methods. Biolog was unable to identify 31% of the isolates; 7% of the isolates were unidentified by the biochemical test scheme. Of the identified isolates, metabolic profiling with Biolog achieved speciation with 60 substrates. Unique profiles were obtained for 89% of the isolates. Isolates also demonstrated inter-trial differntial metabolism of substrates. This and the large number of unidentified isolates suggest great diversity among enterococci. Diversity and inter-trial metabolic inconsistencies will complicate use of enterococcal metabolic profiles as a source-tracking tool.
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Epidemiological studies routinely demonstrate a positive correlation between ambient air pollution and respiratory aggravation. Potential benefits from federal regulations to economic and physical health of individuals, however, are not well understood by either law makers or voters. As the EPA implements new regulations targeting SO ₂ emissions, robust empirical analysis can frame the policy issue as one of statistical and economic analysis. The Asthma Call Back Survey extension of the Behavioral Risk Factor Surveillance System collects detailed data on symptoms, intervention methods, and demographics of individuals with asthma as well as associated ZIP-level spatial identification. The EPA's Acid Rain emissions database records detailed emission data for every power plant in the country. Together, these data can be used to link detailed asthma information with nearby plant emission levels, providing the foundation for an in-depth enviro-economic analysis of SO &#8322 effects on pediatric asthma aggravation and potential long term human capital investment. Using a zero-inflated negative binomial regression technique, the study estimates the effect that pollution has on days of missed school the provides insight into educational effects of emissions and presents the issue as one important to child development. The results and policy-oriented sensitivity analysis demonstrate the clear benefits of coal power plant pollution reduction, and suggest policy and education solutions which can mitigate negative asthma outcomes and promote educational attainments.

The microbial source tracking (MST) methods of antibiotic resistance analysis (ARA) and fluorometry (to detect optical brighteners in detergents) were used in the summers of 2004 and 2005 to determine the origins of fecal pollution at beaches with a past history of, or the potential for, high enterococci counts and posted advisories. At Hilton and Anderson beaches, ARA and fluorometry in the summer of 2004 detected substantial human-origin pollution in locations producing consistently high counts of Enterococcus spp. Investigations by municipal officials led to the fluorometric detection and subsequent repair of sewage infrastructure problems at both beaches. The success of these mitigation efforts was confirmed during the summer of 2005 using ARA and fluorometry, with the results cross-validated by pulsed-field gel electrophoresis (PFGE). Results at other beaches indicated that birds and/or wildlife were largely responsible for elevated enterococci levels during 2004 and 2005. The application of fluorometry proved difficult in opens waters due to high levels of dilution, but showed potential for use in storm drains. An additional study developed and tested a new library-based MST approach based on the pattern of DNA band lengths produced by the amplification of the 16S-23S rDNA intergenic spacer region, and subsequent digestion using the restriction endonuclease MboI. Initial results from small known-source libraries yielded high average rates of correct classification (ARCC). However, an increase in the library size was accompanied by a reduction in the ARCC of the library and the method was deemed unsuccessful, and unsuitable for field application. A final study focused on the potential for classification bias with disproportionate source category sizes using discriminant analysis (DA), logistic regression (LR), and k-nearest neighbor (K-NN) statistical classification algorithms. Findings indicated that DA was the most robust algorithm for use with source category imbalance when measuring both correct and incorrect classification rates. Conversely k-NN was identified as the most sensitive algorithm to imbalances with the greatest levels of distortion obtained from the highest k values. Conclusions of this project include: 1) application of a validation set, as well as a minimum detectable percentage to known-source libraries aids in accurately assessing the classification power of the library and reducing the false positive identification of contributing fecal sources; 2) the validation of MST results using multiple methods is recommended for field applications; 3) fluorometry displayed potential for detecting optical brighteners as indicators of sewage leaks in storm drains; 4) the digestion of the 16S-23S rDNA intergenic spacer region of Enterococcus spp. using MboI does not provided suitable discriminatory power for use as an MST method; and 5) DA was the least, and k-NN the most, sensitive algorithm to imbalances in the size of source categories in a known-source library.
Ph. D.

The field of molecular fecal source tracking in the water environment has developed rapidly since the first PCR assays for general and host-­‐specific Bacteroides 16s rRNA markers were published. Numerous host-­‐specific molecular markers and PCR assays have been developed, adding greater specificity, sensitivity and quantitative methods to the array of options. The public demand for readying methods for transfer to the commercial lab, so that they may be used to generate data for public utilities, citizen action groups and regulatory agencies, has fueled the development of an entire new research community. These methods, however plentiful, have not found community agreement and there is no consensus concerning the appropriate implementation of molecular fecal source tracking in the field. Some issues plaguing the implementation include imperfect marker specificity, environmental variability, DNA extraction variability, PCR inhibition and high cost of molecular analysis. This thesis presents an approach for locating hot spots of human fecal pollution in an urban watershed by using published methodologies for the collection of molecular fecal source tracking data along with a tiered watershed screening tool for cost reduction and two data normalization techniques which ameliorate several known sources of error and strengthen the efficacy of watershed application.

Bacterial water pollution is a significant problem because it is associated with reduction in the ‘quality’ of water systems with a potential impact on human health. Faecal indicator bacteria (FIB) are usually used to monitor the quality of water, and to indicate the presence of pathogens in water bodies. However, enumeration alone does not enable identification of the precise origin of these pathogens. This study aimed to monitor the quality of bathing water and associated fresh water in and out of the ‘bathing season’ in the UK, and to evaluate the use of microbial source tracking (MST) such as the host-specific based polymerase chain reaction (PCR) and quantitative PCR (qPCR) to recognize human and other animal sources of faecal pollution. The culture-dependent EU method of estimating FIB in water and sediment samples was performed on beach in the South Sands, Kingsbridge estuary, Devon, UK- a previously ‘problematic’ site. FIB were present at significant levels in the sediments, especially mud, as well as fresh water from the stream and pond flowing onto South Sands beach. However, the quality of bathing water was deemed to be ‘good’ and met with the EU bathing water directive 2006. Using MST it was possible to successfully classify the nature of the source from which the bacteria came. PCR was applied to detect the Bacteroides species 16S rRNA genetic markers from human sewage and animal faeces. All water and sediment samples displayed positive results with a general Bacteroides marker indicating the presence of Bacteroides species. Host-specific PCR showed the human Bacteroides genetic marker only in the sediment of the stream. However, limitations in the ‘types’ of probes available and in the persistence of these markers were identified. Thus, novel dog-specific Bacteroides conventional PCR and qPCR primer sets were developed to amplify a section of the 16S rRNA gene unique to the Bacteroides genetic marker from domestic dog faeces, and these were successfully used to quantify those markers in water samples at a ‘dog permitted’ and ‘dog banned’ beach (Bigbury-on-Sea, Devon, UK). Generic, human and dog Bacteroides PCR primer sets were also used to evaluate the persistence of Bacteroides genetic markers in controlled microcosms of water and sediment at differing salinities (< 0.5 and 34 psu) and temperature (10 and 17 ºC). The rates of decline were found did not differ significantly over 14 and 16 days for the water and sediment microcosms, respectively. Beach sediments which were studied in this project may act as a reservoir for adhesive FIB, and this was confirmed using fluorescence in situ hybridisation (FISH). The similarity in the persistence of these Bacteroides 16S rRNA genetic markers in environmental water and sediment suggests that viable but non-culturable (VBNC) Bacteroides spp. do not persist in the natural environment for long. Therefore, 16S rRNA genetic markers can be of value as additional faecal indicators of bathing water pollution and in source tracking. Thus, in this study MST methods were successfully used and in future applications, dog-specific primer sets can be added to the suite of host-specific Bacteroides genetic markers available to identify the source(s) of problem bacteria found on failing beaches.

Thesis (MScEng (Electrical and Electronic Engineering))--University of Stellenbosch, 2009.
ENGLISH ABSTRACT: Limited research results are available on the aging impacts of surface discharges for High Voltage Direct Current (HVDC) excitation on practical insulators using the Tracking Wheel Tester (TWT) methodology. This thesis gives details of an experimental investigation to compare the aging performance of insulator samples using the TWT for High Voltage Alternating Current (HVAC) and positive and negative polarity HVDC excitation. Two series of tests were performed. The first series of tests evaluated the aging of six insulator rods, namely three Room Temperature Vulcanized Silicone Rubber (RTV SR) coated glass samples and three uncoated glass samples. Three creepage distances were used for each of the test materials, namely 277 mm, 346 mm and 433 mm. The tests were conducted with HVAC excitation and the test methodology described in the IEC 61302 standard. The second series of tests involved the testing of actual insulator samples. The test voltage and conductivity of the salt water solution were also adapted. Six insulators from different manufacturers, representing different materials and specific creepage distances, were tested with HVAC and positive and negative polarity HVDC excitation. The test samples consisted of Ethylene Propylene Diene Monomer (EPDM), High Temperature Vulcanized Silicone Rubber (HTV SR), porcelain and SR coated porcelain insulators. The aging performance of the different test samples for the three types of excitation are compared in terms of peak leakage current, visual observations of surface degradation and hydrophobicity properties. The leakage current data for HVAC excitation shows that the insulators from the different manufacturers perform differently, even for the same type of material. The results also indicate differences in the way the surfaces degrade, as well as the rate of degradation. For insulators representing the same specific creepage distance, but different materials, it has been shown that the material does influence the aging performance. Comparison of the leakage current data for the HTV SR insulators from the same manufacturer, but with different specific creepage distances, shows that the specific creepage distance affect the aging performance significantly. In general, the test insulators showed higher peak leakage currents with HVDC excitation compared to HVAC excitation. The results for positive polarity HVDC excitation show that the dry band arcing, as well as the discharges, has the same form for all six insulators. The colour of the dry band discharges ranged from a blue-ish orange to a dark yellow, depending on the intensity of the leakage currents. The hydrophobic insulators, namely the HTV SR, EPDM and RTV SR coated porcelain insulators, had lost hydrophobicity within the first week of testing. The results for negative polarity HVDC excitation show severe surface degradation compared to the results for HVAC excitation. The effect of positive polarity HVDC excitation, however, seems to be more severe in terms of leakage currents and aging compared to negative polarity HVDC excitation.

Les virus entériques sont à l’origine de pathologies liées au péril fécal et dans l’état actuel des connaissances, la recherche des indicateurs de pollution fécale conventionnels (i.e. Escherichia coli, entérocoques) peut s’avérer inefficace pour évaluer le danger viral. La définition d’autres indicateurs pour gérer le danger lié à la présence des virus entériques dans les matrices hydriques et alimentaires est aujourd’hui nécessaire. Parmi eux, les bactériophages ARN F-spécifiques (FRNAPH) présentent plusieurs intérêts. Ces virus d’origine entérique sont présents en quantité importante dans les eaux usées. Très proches des virus entériques en termes de structure, ces microorganismes présentent l’avantage d’être facilement cultivables. Ils sont enfin souvent étudiés pour déterminer l’origine d’une pollution fécale (i.e. humaine ou animale). Certaines limites leur sont cependant fréquemment associées, que ce soit en termes de corrélation avec les pathogènes entériques ou concernant leur potentiel pour discriminer l’origine d’une pollution. Dans ce contexte, l’objectif du travail présenté ici était de préciser l’intérêt des FRNAPH en tant qu’indicateurs de pollution fécale mais aussi en tant qu’indicateurs de pollution virale dans l’environnement et les coquillages. Ces travaux ont permis dans un premier temps d’améliorer la capacité des FRNAPH à identifier les contaminations d’origine humaine. Nos résultats soulignent par ailleurs la plus-value apportée par la recherche des FRNAPH en cas de pollution fécale massive, en particulier si on s’intéresse à la contamination des coquillages. En effet, contrairement aux indicateurs bactériens, l’accumulation des FRNAPH ainsi que leur persistance dans ces aliments est très comparable à celles des virus entériques (i.e. norovirus). Enfin, en utilisant des méthodes de détection comparables, une forte corrélation entre la présence des FRNAPH d’origine humaine et celle des norovirus a été observée dans les coquillages. Compte tenu de ces résultats, une méthode de détection assurant la détection sensible des FRNAPH infectieux d’origine humaine dans différents types de matrices hydriques ou alimentaires (e.g. eaux de surface, fruits de mer, fruits rouges, salades) est proposée pour améliorer la gestion du danger viral
Enteric viruses are a leading cause of fecal-oral route transmitted diseases and currently, conventional fecal indicator bacteria (i.e. Escherichia coli, enterococcus) fail to assess this kind of hazard. In this context, the use of more efficient indicators to assess the hazard linked to viruses in water or foodstuff is required. F-specific RNA bacteriophages (FRNAPH) present numerous benefits for this purpose. Of enteric origin, these viruses are found in high concentrations in wastewater. Sharing many structural similarities with pathogenic enteric viruses, FRNAPH are easily cultivable and their potential to track the origin of the pollution is also often investigated. However, some limits are still associated with these indicators, regarding to their ability to track the origin of the pollution or concerning the lack of correlation with pathogens. In this context, the aim of this work was to make clear the potential of FRNAPH as fecal and as viral indicators in environmental waters and shellfish. As a first step, their ability to track human pollution was optimized. In addition, our results underlined the gains bringing by FRNAPH detection, especially when focusing on shellfish microbiological quality management. Indeed, unlike fecal indicator bacteria, the accumulation of FRNAPH and their persistence in shellfish have been found to be close to that of enteric viruses (i.e. norovirus). Furthermore, when using comparable methods for their detection, high correlation was observed between human FRNAPH and norovirus in shellfish. Taking into account these observations, a sensitive method allowing the detection of infectious FRNAPH of human origin was developed to improve viral hazard management in water and food commodities (e.g. environmental waters, shellfish, soft fruits, leaf)

The principal aims of this thesis were to examine whether there were hydraulic links between individual on-site wastewater systems in sandy soils at Salt Ash and the Tilgerry Creek estuary near Port Stephens, New South Wales, and whether the chemical and microbiological contaminants from on-site systems could reach surface and groundwaters, and possibly lead to impacts on estuarine oyster growing waters. The research outcomes are contained within the thesis and in four refereed papers presented at conferences, and which have been subsequently published, or are in press. Copies of each of these papers are contained within the thesis Appendices. The presence of faecal contamination from domestic systems in the estuary, and surface drains in particular, has been confirmed by work contained in this thesis. The potential level of risk to human health from the consumption of contaminated oysters is, however, regarded as very low, although an assessment of health risk using established microbial assessment models has not been undertaken

Aquesta Tesi Doctoral ha estat pensada per avaluar, mitjançant l’estudi de virus contaminants, l’origen de la contaminació fecal i l’avast de la disseminació a l’aigua. Amb aquesta finalitat s’han utilitzat eines moleculars per a la detecció i quantificació de virus especifics humans, bovins i porcins i a més a més s’ha desenvolupat una nova eina per traçar la contaminació ovina a l’ambient. Els principals agents virals amb risc per als humans són d’origen humà però les femtes animals, amb potencials patògens zoonòtics, representen també un risc per a la salut humana. La intensificació de la producció animal o l’increment de plujes torrencials previst amb els escenaris proposats per al canvi climàtic presenten nous reptes per al control de la contaminació fecal a l’aigua. Bona part de la Tesi, ha anat lligada al projecte europeu VIROCLIME permetent dur a terme estudis a nivell internacional. S’ha caracteritzat la disseminació de virus humans contaminants a l’aigua d’un riu situat en una conca Mediterrània. Mitjançant el registre de variables hidro-climàtiques, la recollida de mostres d’aigua de riu, aigua de mar i aigua residual, i l’anàlisi de FIB i virus indicadors de contaminació fecal humana (HAdV i JCPyV), virus patògens (NoVGII, HEV), i un virus emergent (MCPyV), s’ha estudiat durant un any, l’impacte que l’aigua residual crua o els efluents de les depuradores tenen en la qualitat microbiològica del riu. En un segon estudi realitzat conjuntament amb laboratoris de Suècia, Grècia, Hongría i Brasil, s’ha pogut estandarditzar el mètode de floculació amb llet descremada, per a la concentració de virus, i els assajos de qPCR per identificar i traçar l’origen de la contaminació fecal a l’aigua. Durant 18 mesos de mostrejos a conques de rius situats a les regions Mediterrània, Àrtica, Continental i Tropical, s’han identificat virus humans (HAdV i JCPyV), bovins (BPyV) i porcins (PAdV), demostrant que aquestes eines virals són fiables i útils per a qualsevol àrea geogràfica o matriu d’aigua. Fins al principi d’aquesta tesi, només es disposava de marcadors virals de contaminació fecal porcina i bovina. Analitzant mostres de femta i orina d’ovella, amb un assaig de PCR d’ample espectre per a la detecció de poliomavirus, es van obtenir per primera vegada seqüències d’un putatiu nou poliomavirus oví. A partir d’aquí, s’han dissenyat dos assajos de PCR específics per poder traçar la contaminació fecal ovina a l’ambient. Arrel d’aquest treball, durant l’últim any de la tesi, es va plantejar una estada a Nova Zelanda perquè es tracta del país amb més ovelles per càpita i representa bona part de la indústria ramadera del país. Traçar i identificar l’origen de la contaminació fecal animal a l’aigua és doncs bàsic, tant per la gestió dels residus com per l’avaluació dels riscos per a la salut humana. El quart estudi inclòs a la tesi, es va dissenyar per identificar les fonts principals de contaminació fecal i avaluar, al mateix temps, les diferents eines de MST utilitzades a Nova Zelanda i al laboratori de la doctoranda. S’han recollit mostres de riu als territoris ramaders més importants de l’illa del sud, i s’han analitzat E. coli, virus específics indicadors de contaminació fecal humana (HAdV, JCPyV) bovina (BPyV) i ovina (OPyV), marcadors bacterians específics d’humans (BacH, BacHum-UCD, BiAdo) i de remugants (BacR) i esterols i estanols indicadors de contaminació fecal humana o de remugant.
This thesis was designed to evaluate, through the study of viruses, the source of the fecal contamination and its spread in water matrices. For this purpose molecular tools have been used for the detection and quantification of specific human porcine and bovine viruses, and a new tool has been developed to trace ovine pollution in the environment. Human waterborne viruses pose the main risk to humans, but animal feces, with potential zoonotic pathogens, also represent a risk to human health. The intensification of animal production and the predicted scenarios for climate change, with increase of torrential rains, present new challenges for the control of fecal contamination in water . Much of this work has been linked to the European project VIROCLIME allowing studies conducted worldwide. We characterized the spread of contaminant human viruses in a Mediterranean river basin . By recording hydro-climatic variables, collecting samples of river water, sea water and wastewater, and analysing FIB and human virus indicators of fecal pollution (HAdV and JCPyV), pathogenic viruses (NoVGII , HEV) and an emerging virus (MCPyV) we have studied, thoughouth a year, the impact of raw wastewater and efluents from the wastewater treatment plants the microbiological quality of the river. In a second study, conducted with laboratories in Sweden, Greece, Hungary and Brazil, we standardized the viral concentration method, Skimmed Milk Flocculation, and specific qPCR assays to identify and trace the origin of contamination in water. During an 18 months sampling period, in river basins located in the mediterranean region, arctic, continental and tropical regions, we identified human (HAdV and JCPyV), bovine (BPyV ) and porcine viruses (PAdV), demonstrating that these tools are reliable and useful for any geographic area or water matrix. At the beginning of this thesis, only porcine and bovine viral fecal indicators were available to trace fecal pollution. Since no ovine viral indicator was described before the present work, we designed a study to develop a new tool to trace sheep fecal contamination. Firstly we analyzed urine and stool samples from sheep, with a broad spectrum PCR assay for the detection of polyomavirus. We obtained sequences of a putative new ovine polyomavirus and designed two PCR assays specific for the new ovine virus. During the last year of the thesis raised a stay in New Zealand because it is the country with more sheep per capita and represents much of the livestock industry in the country. Trace and identify the source of animal fecal contamination in water is therefore essential for both waste management for the evaluation of risks to human health. The fourth study included in this thesis was designed to identify the main sources of fecal contamination and evaluate the same time , the different tools used MST in New Zealand and the student laboratory. River samples were collected at major livestock areas south of the island , and analyzed E.coli , viruses specific indicators of human fecal contamination (HAdV, JCPyV) bovine (BPyV) and sheep (OPyV) specific bacterial markers in humans (BacH, BacHum-UCD, BiAdo) and ruminants ( BacR) and sterols and stanols indicators of human fecal contamination or ruminants.

Thesis (M.S.)--Worcester Polytechnic Institute.
Keywords: classification; k-nearest-neighbor (k-n-n); neural networks; linear discriminant analysis (LDA); support vector machines; microbial source tracking (MST); quadratic discriminant analysis (QDA); logistic regression. Includes bibliographical references (p. 59-61).

Less than 10% of the world’s domestic wastewater is disinfected prior to discharge into surface waters; therefore, human exposure to diverse wastewater-related pathogens results in millions of cases of illness each year. Among the enteric pathogens, viruses represent an important group of emerging pathogens and are frequently the cause of food- and water-borne outbreaks of illness. Although the World Health Organization and many government agencies mandate the use of bacterial indicators to identify poor microbial water quality, it is well known that these indicators poorly correlate with fecal pollution contamination events and risk of disease. The field of public health-related environmental microbiology has made significant advances over the last twenty years; however, there is still a need for improved methods to identify poor microbial water quality and manage health risks associated with water-related activities (e.g., recreation, shellfish harvesting, irrigation). Furthermore, it is imperative to effectively detect fecal pollution in the environment as well as determine the extent of pathogen removal during (waste)water treatment to meet the Sustainable Development Goals associated with water and food security as well as the water reuse recommendations by the U.S. National Research Council. This dissertation directly addressed the need to identify an improved viral indicator by exploring the application of pepper mild mottle virus (PMMoV), a virus of dietary origin that is extremely abundant in human feces, as a surrogate for enteric viruses in diverse settings and contexts. Using a reverse transcription quantitative polymerase chain reaction (RT-qPCR) method, PMMoV was compared with other indicators, index surrogates, and reference pathogens for assessing surface water quality in a developed country (Appendix A) and developing country context (Appendix B and C). In addition, the applicability of PMMoV as a (waste)water treatment process indicator was demonstrated in natural treatment systems in developing countries (Appendix C and D) and artificial treatment systems (Appendix E). In all settings, PMMoV always co-occurred with at least one reference pathogen, index organism, and/or indicator; additionally, PMMoV was detected more frequently and in higher concentrations than other reference pathogens and indicators. Three investigations also associated PMMoV detection with predicted human health risks greater than the health benchmark for recreation and water reuse activities (Appendix A, C, and D). Additionally, PMMoV facilitated an improved understanding of virus-particle interactions in wastewater treatment pond systems (Appendix D) and allowed for an improved understanding of virus removal with respect to riverbank filtration systems and wastewater reuse in agriculture (Appendix C). PMMoV was established as a valuable component of the microbial source tracking toolbox in Costa Rica (Appendix B) and appeared to be useful in Bolivia (Appendix C). Finally, a paradigm shift in (waste)water management is occurring, in which routine monitoring is being replaced by a more holistic approach that includes sanitary surveys, targeted water quality monitoring, and exploratory quantitative microbial risk assessment. To support and complement this paradigm shift, field-based, laboratory-free methods are needed to identify and/or infer the presence of enteric viruses (Appendix F). Collectively, all the investigations presented here confirm the use of PMMoV as a surrogate for enteric viruses; however, its utility depends on the context and research question.

Water quality worldwide is assessed by enumeration of fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli, and enterococci) intended to act as surrogates for human enteric pathogens. In environmental waters, this predictive relationship is confounded by many possible sources of FIB with varying implications for human health. Many physico-chemical and biological factors influence the fate of enteric pathogens and FIB in aquatic habitats, but are poorly understood, thus limiting our understanding of the usefulness of FIB as fecal pollution indicators. These studies explored the field application of a “toolbox” approach to microbial source tracking (MST) intended to discriminate between human and non-human fecal pollution: a) in a Florida estuary used for shellfishing and recreational activities and b) at public beaches before and after remediation of wastewater infrastructure. Lastly, the effects of environmental factors (sediments, protozoa, sunlight) on survival of culturable E. coli were investigated in freshwater and seawater mesocosms simulating environmental conditions. Detection of a human- associated MST marker (the esp gene of Enterococcus faecium) at sites with suspected sewage contamination indicated that human fecal pollution is impacting water quality in Wakulla County, while Lagrangian drifters designed to follow current and tidal movement suggested that local hydrology plays an important role in bacterial transport and deposition pathways. Elevated FIB concentrations and frequent detection of human-associated MST markers (esp and human polyomaviruses) identified human sewage pollution at a public beach, facilitating remediation efforts (sewage main repair, removal of portable/abandoned restrooms), followed by significant decreases in FIB concentrations and MST marker detection. These studies show that comprehensive microbial water quality assessment can reliably identify contamination sources, thereby improving pollution mitigation and restoring recreational water quality. Protozoan predation, freshwater vs. seawater habitat and sediment vs. water column location affected the concentration of culturable E. coli in outdoor mesocosms. Sediments offered a refuge from predation where freshwater vs. seawater habitat was amore important determinant of survival. These findings provide important insight into the ecology of E. coli and their natural predators in aquatic habitats and underscore the inherent effect different habitats play in their survival.

Nearshore marine ecosystems like seagrass meadows face a wide range of anthropogenic influences, impacting the system at different spatial and temporal scales. Managing these systems in the face of these pressures requires detailed knowledge of how seagrass habitats respond to these various threats. A plethora of useful indicators have been developed to help managers and policy makers track seagrass meadow health and status, detect environmental impacts or measure the effectiveness of management interventions. However, choosing between these indicators can often be a daunting task since they vary considerably in their overall behaviour in relation to ecosystem and environmental changes. This thesis assesses the most commonly employed seagrass indicators to determine if they are adequate and appropriate to the specific needs of coastal ecosystem management. This assessment is based on evaluating three fundamental characteristics of each indicator – the robustness of its response, the specificity or generality of its response, and the time of response. We use a variety of complementary approaches to explore indicator behaviour. In Chapter 3, we use field-based studies to assess how seagrass indicators respond to the construction of a breakwater in the vicinity of a Posidonia oceanica seagrass meadow. Chapters 4 and 5 examine long-term trends in seagrass indicators to improving water quality conditions after significant regional management interventions. In addition, in Chapter 6, we comprehensively review seagrass indicator responses to multiple stressors. Chapters 3, 4 and 5 focus largely on the Catalan Coast in the Mediterranean with Posidonia oceanica as a target species. Chapter 6 in contrast is a broad review of a wide range of indicators used across several seagrass species worldwide. A central learning across these studies was that the level of biological organisation of the indicator (i.e. Physiological, biochemical, growth, morphological, structural or demographic) is critical in determining the specificity or generality of response: the lower the level (e.g. biochemical), the most specific the response, while the higher the level (e.g. population, community), the wider the response. Thus, biochemical indicators are ideal to determine the identity or even the origin of a pressure while structural indicators, in contrast, are useful as generic indicators of declining conditions. Response times are also heavily determined by the level of organisation, particularly in the detection of improving environmental quality along the Catalan coast. Biochemical indicators responded unequivocally to water quality improvements observed both in the experimental field study (Chapter 3) as well as in the analysis of the long-term data series (Chapters 4 and 5). The meta-analysis confirmed that these trends in specificity and response time were not unique to Posidonia oceanica or the Catalan coast and highlighted the critical role of plant size in determining indicator time responses. Large species take considerably longer to register a response to environmental degradation as well as improvement – a critical factor that needs to be accounted for in designing monitoring programmes and interpreting ecosystem trends. Taken together, these results suggest that differences in the behaviour of seagrass indicators require that they should be carefully selected to match the objectives of management. Based on the results reported in this thesis, where different sets of indicators have been proven successful for given management objectives, we finally develop a simple decision tree to help managers chose the most reliable sets of indicators matching their objectives. Understanding the diversity of responses that seagrass indicators display can make them a powerful set of tools in the ecosystem manager’s toolkit. Carefully employed, they can serve as bespoke solutions to a wide range of management objectives as we seek to monitor and protect these vital ecosystems and coastal water quality in the face of increasing coastal pressures.
Entendre com responen les fanerògames marines a les pressions, és clau per a poder gestionar tant els herbeis com les aigües costaneres. Actualment, disposem d’un gran nombre d’indicadors basats en fanerògames marines. Però, la manca d’informació sobre com responen als canvis ambientals, fa que no sigui fàcil escollir quins indicadors són els més adients per a cada tipus i objectiu de gestió. Aquesta tesi avalua les tres característiques bàsiques de la resposta dels indicadors més utilitzats als canvis ambientals: la robustesa de la resposta, la especificitat dels indicadors a diferents pressions i el temps de resposta. Per analitzar aquestes tres característiques, fem servir diferents aproximacions complementàries. Al capítol 3, analitzem la resposta de diferents indicadors a les obres d’ampliació del port de Blanes, situat just al costat d’un herbei de Posidonia oceanica. Als capítols 4 i 5, estudiem com responen els indicadors a la millora de la qualitat de l’aigua a la costa catalana. Finalment, al capítol 6, presentem una metaanàlisi que estudia com responen els indicadors a diferents factors d’estrès. De totes tres aproximacions, hem pogut comprovar que el nivell d’organització dels indicadors (i.e. bioquímic, estructural) és clau a l’hora de determinar el grau d’especificitat de la resposta dels indicadors a les pressions: generalment, a més baix nivell d’organització (e.g. bioquímic), més específica és la resposta i com més alt (e.g. demogràfic), més ampli és el rang de pressions que un indicador pot detectar. El temps de resposta dels indicadors varia també en funció del nivell d’organització dels indicadors, especialment, quan es tracta de la detecció de millores ambientals. A més a més, la metaanàlisi destaca la importància de la mida de les plantes per determinar el temps de resposta. Les espècies grans triguen més a detectar la degradació de les condicions ambientals i, molt més, a detectar la millora, especialment, si s’utilitzen indicadors estructurals o demogràfics. Basant-nos en els resultats d’aquesta tesi, hem elaborat un esquema per ajudar els gestors a escollir el conjunt d’indicadors que més s’ajusti a cada objectiu de gestió. Utilitzats correctament, aquests indicadors són molt útils per fer el seguiment, tant de l’estat de salut dels herbeis, com de la qualitat del medi.

Dans le présent travail, nous abordons une problématique essentielle liée aux accessoires des lignes électriques et plus particulièrement au connecteur à perforation d’isolant (TTDC 45501 FA), commercialisé par la société Sicame située à Arnac-Pompadour (Corrèze). Ce produit permet d’effectuer des dérivations de courant en moyenne tension gainé HTA entre 15 et 25kV. Cependant, il devient nécessaire de concevoir une nouvelle génération de connecteurs capable de fonctionner à une tension de 52 kV en raison de la constante augmentation des besoins en électricité. En réalité, le principal souci concernant ces produits réside dans l’apparition de décharges partielles dans le volume constitué par la matière isolante (polyamide 6 chargée à 50% en poids de fibres de verre). Ces décharges engendrent un vieillissement prématuré du produit sous l’action conjuguée d’une atmosphère corrosive (brouillard salin) et d’un champ électrique. Ainsi, l’objectif de la présente thèse est de faire, dans un premier temps, un choix approprié de matériaux isolants parmi des polymères thermoplastiques, thermodurcissables époxydes ou élastomères en EPDM, capables de résister à plusieurs contraintes : thermique, électrique, mécanique, chimique et climatique. La partie expérimentale concerne, tout d’abord, la mesure de la tension de contournement, de l’activité des décharges partielles se propageant à la surface des matériaux isolants et de la tension de claquage, en tension alternative (A.C.). Pour cela, des échantillons polymères qui comportent des surfaces planes ou des surfaces pourvues de protubérances ont été moulés en vue de comparer leur comportement diélectrique en présence d’atmosphères polluées de type brouillard salin; ces mesures sont nécessaires pour effectuer un choix approprié du matériau à intégrer dans le connecteur. Par la suite, l’étude et la caractérisation optique et électrique de décharges glissantes, se propageant en surface de divers matériaux polymères, en tension impulsionnelle (L.I.) pour les deux polarités (positive et négative) et pour deux types de champ électrique (normal et tangentiel) sont présentées pour le cas d’une interface solide/air. La mesure de la longueur finale des décharges glissantes et la visualisation de l’onde de courant sont les deux paramètres principaux pour différencier les caractéristiques des matériaux utilisés. L’ensemble des résultats de mesures a permis de sélectionner les résines époxydes cyclo-aliphatiques (CEP) comme candidates potentielles pour la conception d’une nouvelle génération de connecteurs. De plus, la simulation du champ électrique, à l’aide du logiciel Flux 2D/3D, sur le connecteur électrique (TTDC 45501FA) a permis d’aboutir à la proposition d’un prototype de connecteur présentant une nouvelle géométrie et pouvant fonctionner à 52kV
In the present study, we deal with an essential problem related to electrical lines accessories and more particularly to piercing connectors (TTDC 45501FA) commercialized by a private company named Sicame and which is located in Arnac-Pompadour (Corrèze, France). This product/accessory is designed to provide power diversions on medium voltage live lines HVA operating between 15 and 25kV. However, it becomes required to design a new generation of piercing connectors able to operate on 52kV live lines because of the electrical power demands that are steadily increasing. Actually, the main concerns about these products are the occurrence of partial discharges within the insulating material volume (polyamide 6 containing 50%wt of glass fibers). Most of the time, these discharges induce an early ageing process of the accessory under the combined effects of a corrosive environment (as salt fog) and an electric field threshold. Thus, the main objective of the present Ph.D. thesis is to carry out an appropriate choice of insulating materials among several tested polymers such as thermoplastics and thermosetting epoxy resins and EPDM elastomers that are capable of withstanding several constraints: thermal and electrical and mechanical and chemical conditions. The experimental section is focusing on the measurements of flashover voltage and partial discharges activity propagating on polymeric surfaces and breakdown voltage within material bulks under A.C voltage. That is why polymeric samples with plane surfaces and textured surfaces are molded to compare their resistance to partial discharges when being subjected to polluted environments such as salt fog; these measurements are necessary to choose the suitable materials for the required application to be integrated in the new generation of connectors. Then, the optical and electrical characterization results of creeping discharges propagating on several polymeric surfaces under lightning impulse (L.I.) voltage with its both polarities (positive and negative) and for two kinds of applied electric field (normal and tangential) are presented in the case of solid/air interfaces. Final length measurements of creepage discharges are the main parameter for distinguishing material properties. The whole obtained results (under AC and LI voltages) allow us to point out the cycloaliphatic epoxy resins as potential candidates to the conception of a new generation of piercing connectors. In addition, the electric field simulation and modelling of the TTDC 45501FA connector by using the Flux 2D/3D software seems to be helpful to design a connector prototype which exhibits a new geometry and is able to reach a voltage level that equals 52kV

Conventional methods for health assessment of wild-caught and stranded seals were used to describe the disease status of harbor seals in California. Clinical chemistry, infectious disease prevalence, immune function, and contaminant data were collected to evaluate harbor seal health with data collected from three groups of seals. Wild-caught seals of all ages were sampled at two locations: San Francisco Bay (a heavily urbanized estuary) and Tomales Bay (a less developed control site). Stranded seals entered rehabilitation from a more extensive portion of the California coast which included the locations where seals were caught. Hematology reference intervals were generated to provide a baseline for health assessment among the seals. Individual variability in blood variables among seals was affected by age, sex, location, and girth. Disease surveillance focused on pathogens known to cause lesions in harbor seals, zoonoses, and those with terrestrial sources. Specific pathogens of interest were E coli, Clostridium perfringens, Vibrio spp, Campylobacter spp, Salmonella, Giardia, Cryptosporidium, avian influenza virus, Brucella, Leptospira spp., Toxoplasma gondii, Sarcocystis neurona, and Neospora caninum, Leptospira spp, and phocine and canine distemper virus. There was evidence of exposure to all pathogens except for phocine distemper virus. Simple measures of immune response were used to evaluate the immune function of harbor seal pups in rehabilitation that had evidence of previous bacterial infection. The swelling response to a subcutaneous injection of phytohemagglutinin (PHA) was positively associated with growth rate, possibly illustrating the energetic trade-offs between growth and immunity. Blubber contaminant concentrations (PCBs, DDTs, PBDEs, CHLDs, and HCHs) in harbor seal pups were grouped by extent of suckling and strand location. The ratio of PCB:DDT was increased in San Francisco Bay and decreased in Monterey Bay compared with other locations along the coast. Pups that weaned in the wild, lost weight and then stranded had the highest contaminant levels, equivalent to the concentrations detected in stranded adult harbor seals. Dispersal and survival were monitored by satellite telemetry in harbor seal pups released from rehabilitation and recently weaned wild-caught pups to assess the effect of condition, health, and contaminant levels on survival probability. Increased contaminant levels and decreased thyroxine (T4) were associated with decreased survival probability. Increased mass, particularly among the rehabilitated pups, was associated with increased survival probability. This study demonstrates that health and survival of harbor seals pups along the central California coast are impacted by human activities such as contaminant disposal, pathogen pollution and boat traffic, although the variability in individual health measures requires carefully designed studies to detect these effects.

碩士
國立中興大學
通訊工程研究所
107
In recent years, outdoor air pollution is getting more and more serious, and people often close their doors and windows to block dirty air outside. But when we are in a sealed building, it does not certainly means that the indoor air quality will be better. When the indoor ventilation is insufficient, harmful substances in the air are easily to accumulated, and the indoor air quality will deteriorating. Therefore, if the window is closed for outdoor air pollution, it is possible to increase the concentration of harmful gases in the room. Therefore, we use wireless sensor networking (WSN) technology to establish indoor and outdoor sensing nodes and become a smart Internet of things (IOT) system. In this system, indoor and outdoor sensing node consolidate the sensing values, transmit them to the computer database by Lora to stored, and put the data in the deep learning module for predictive control with a set of rules. When the new data is input, the most suitable control method will be predicted according to the current learning state. Accordingly, integrating indoor and outdoor sensed pollution concentrations for analysis and regulation may improve the air quality in our room.

碩士
國立成功大學
地球科學系碩博士班
101
Taiwan is a densely populated country with lots of crowded and overused land. The rapidly developing economic growths and industrialization resulted serious heavy metal pollution on agriculture land nearby the factories. Cadmium (Cd) has eight stable isotopic at a mass range from 106 to 116 amu, where a natural isotope fractionation of -0.08~0.05‰/amu was found in various terrestrial samples. Significant isotope fractionation of Cd (-0.28~0.28‰/amu) was detected only during industrial processes at very high temperature, resulting in the lighter nuclides enriched in aerosols and ashes, and heavy nuclides remained in slag. The fractionated Cd isotopic ratios driven by the artificial activities have therefore become specific markers for distinguishing the sources from other naturally available Cd. In this study, we present a high precision Cd isotopic determination method using multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS, Neptune) with a combined standard-sample bracketing (SSB) procedures and empirical external normalization (EEN) for mass bias correction during the measurement. A two-stage chemical separation using AG 1-X8 and TRU resin was applied to purify Cd from potential interference elements (Pd, In and Sn). The long term reproducibility of replicated standard measurement was δ114/110CdJMC-SPEX = 0.34±0.08‰ (2S.D., n=18). Significant isobaric interference occurred in some rock samples that containing high amounts of Sn, however it can be corrected successfully by monitoring the 113Cd. The industrial wastes, including effluent water, irrigation water, ground water, sediment, and dust, were systematic processed in this study to further quantify the possible pollution sources and the contamination spreading pathways. Our preliminary result reveals that about 10ppt~1ppb of Cd was found in irrigation water, lower than the EPA’s standard, and about 1~11ppm of Cd in sediments was found. The δ114/110CdJMC value of -0.5‰~0.7‰ and -0.65‰~0.95‰ were determined in irrigation water and sediment, respectively. However, the enrichment factor and geo-accumulation index both shows great pollution of other heavy metals of Hg, Pb, Cr and As, etc. The result of Cd isotope reveals that the main polluted reason for the area surrounds the factory is contributed to the industrial sludge but affected by the atmospheric transport away from the factory. Due to the limited isotopic data available, industrial materials and dusts samples are being processed for clarifying the possibility of Cd contamination on crops and soils. The high precision Cd isotopic determination conducted in this study helps to clarify the extent of Cd isotopic fractionation in local high pollution industries, their possible sources and migration of Cd pollution in natural environment.

Contamination of recreational and shellfish waters with fecal pollution is a major water quality issue with associated economic impacts and human health risks. Reliable fecal source identification and rapid, quantitative analyses are essential components of risk assessment. Enteric bacteria that are endemic to specific hosts have a potential role as public health indicators of fecal pollution. Building on previous work to discriminate ruminant and human fecal contamination, we cloned class Bacteroidetes 16S rRNA genes from pig, elk, dog, cat, and seagull fecal DNAs. Unique restriction patterns were identified among clones from each of the host species using Terminal Restriction Fragment Length Polymorphisms (T-RFLP). Clones exhibiting unique patterns were sequenced and analyzed phylogenetically, along with human, horse, and cattle sequences recovered from previous work. The analysis revealed both endemic and cosmopolitan (global) host distributions. The sequence data were used to identify host-specific genetic markers for pig and horse feces, and to design PCR primers that identify these sources of fecal pollution in water. There was a high degree of sequence overlap among the fecal Bacteroidetes of wild and domestic ruminants, and among human, domestic pet, and seagull Bacteroidetes. We compared fecal Bacteroidetes rRNA genes from these hosts using subtractive hybridization, a method that identifies differences between closely related genomes or gene sequences. A Bacteroidetes rDNA marker that distinguishes elk and cow feces was identified, as well as a host-specific marker for dog fecal Bacteroidetes. The four newly designed PCR primers were tested for specificity and sensitivity, and the dog primer was successfully used, along with the human and ruminant-specific primers, in a collaborative study comparing fecal source tracking methods. We also developed a real time Taq nuclease assay for quantification of fecal Bacteroidetes 16S rDNA, and compared it with an EPA-approved enumeration method for the current standard public health indicator, Escherichia coli, in serial dilutions of sewage primary influent. There was a strong, positive correlation between the methods, and the Taq nuclease assay was sensitive and much more rapid than the E. coli assay. PCR source identification and enumeration of fecal Bacteroidetes 16S rDNA show promise for application in a health risk-based analysis of fecal pollution.
Graduation date: 2005

碩士
國立成功大學
電腦與通信工程研究所
98
Spilled invisible pollutions, such as oil or biochemical materials, damage not only the ocean environment but also the regional economics. To prevent such damages from expanding, a wireless sensor network deployed over the ocean, referred to as Water-surface Wireless Sensor Network (WWSN) in this study, is a suitable candidate for timely monitoring of such wide range and continuous moving objects. In WWSN, this study develops an aquatic-pollution detection and tracking scheme, designated as AquaView, based on an autonomous self-localization cluster mechanism. The proposed scheme enables each normal sensor node to automatically discover its location without maintaining the addition information (i.e., the locations of neighboring nodes, a global or a subset of network topology, and so forth) and to efficiently detect and track the boundary of the current moveable pollution in its sensing field. The numerical results obtained using a Qualnet simulator confirm the effectiveness and robustness of the proposed approach.

碩士
國立中央大學
通訊工程學系
102
Owing to the rising awareness of environment protection and the trend of global warming, more and more people increase awareness of air pollution. Because most of air quality monitoring stations set up in outdoor to monitor pollution and greenhouse gases of metropolitan area, that ignore the importance of indoor air quality (IAQ). However, damage and number of deaths of indoor air pollution are more than outdoors, but sensing device rarely for a long time monitoring. In this study, we focus on volatile organic compounds (VOC) for a long time monitoring in indoor environment. This study uses experimental data to establish indoor Gaussian diffusion model and proposes prediction method to track sources of VOC air pollution. From simulation results of indoor VOC air pollution diffusion and source tracking to verify our proposed method. This system builds on an open architecture for Arduino AVR(Microcontroller) as the main operational unit with ZigBee module (802.15.4) to transit and receive data in wireless sensor network (WSN). The sensing element uses a metal oxide semiconductor sensor element called TGS-2602 via ADC to get measured value.

Autonomous underwater vehicles (AUVs) have been increasingly applied to oceanographic missions. Despite technological advances, adaptability to environmental changes or intelligent behaviour is still at an early stage of development, especially for large survey class AUVs. This research addresses the real-world challenge of using an AUV to delineate an oil plume in sea water, in the management of an environmental spill. A review of current literature showed the most common methodology in conventional adaptive AUV missions involved gradient-following methods with point-based sensors such as fluorometers. In this work, an adaptive sampling algorithm was developed for real discontinuous oil plumes and validated through simulation, The search and detection phase of the algorithm was tested through ocean trials with an Explorer survey-class AUV. In order to develop the algorithm, I complete a series of preparatory tests in a seawater tank, the ocean and in a lake. I used a seawater tank to assess the formation of oil in seawater, waves and currents and to assess the utility of sonar as a sensor for these oil droplets. The ocean was investigated to try to find an environment to test an AUV in an area of naturally occurring oil seeps. Finally, a lake was used to assess different proxies to represent oil in the ocean in order to avoid environmental damage and to obtain sonar records for use in developing real time analysis methods. The oil formation tests showed that real oil plumes are composed of countless undissolved droplets, especially after exposure to waves and currents. This result focused attention on the critical need for a sensor that could identify oil plumes at distance and a sampling strategy that could take into account the coalescent and clustering characteristics of real oil droplets in the water column. The seawater tank experiment found that a higher frequency sonar (1.35 MHz) was more effective than a lower frequency sonar (450 kHz) in capturing an oil plume in the water column. Thereafter, a scanning sonar (750 kHz frequency) was proposed as the primary sensor on the AUV in searching for plumes. An adaptive sampling mission can be achieved by utilising real-time processing of in-situ data to optimise an AUVs capability to complete a task, Here I developed modular algorithms for acoustic detection and in-situ analysis. In accordance with the use of sonar as an oil detection sensor, I introduced a new search path inspired by bumblebee flight patterns and validated this through field trials. This search design significantly improved the more conventional survey plans based on lawnmower paths by reducing the time to seek an oil plume by approximately 75.3%. For the detection strategy, the key assumption was to approximate an oil plume in a two-dimensional sensing layer with limited vertical extent. This improved detection performance by using a higher order two-dimensional scanning method that minimised the plausible impacts of mixing energies on the plume. A ‘Measure – Analysis – Action’ schema was employed in a recursive algorithm for oil plume tracking. The tracking modules analysed the recovered sensor data in real time to produce the desired best new heading of the AUV. To produce the best tracking performance, all the relevant parameters of each component term of the algorithm (such as thresholds, coefficients and weighting factor) had to be tuned and adjusted. Thereafter, I was able to track different forms of simulated oil plumes successfully with the AUV. The search and detection algorithms, as well as the backseat driver control system of the AUV using MOOS IvP routines (some of which I developed), were tested on the Explorer AUV during in ocean trials in Holyrood Bay, Newfoundland and Labrador, Canada. The algorithm implemented on the real AUV successfully demonstrated the AUV backseat driver control. Having appropriate configuration parameters, the guided path could be followed through updating the AUV state in pose (translational position (x, y) and rotational (θ) angle). AUV heading direction was corrected by adapting to the computed desired heading, which was achieved in 2.8 seconds on average after the command was generated. Overall, the vehicle trajectory remained within 5.5m of the guided path. The field experiments verified the feasibility and utility of the designed search and detection algorithm in the ocean environment. Overall, this research has overturned the most common gradient-following methods employed in AUV-based oil spill investigation and tracking. The adaptive sampling system developed in this thesis provides an efficient tool to i) seek a realistic oil plume whose chemical characteristics may have altered over time, ii) detect patches of the plume with varying shapes and density and iii) track and map a plume without losing contact with it. The knowledge obtained through this research and the presented algorithm modules will support the next generation of researchers seeking to advance the autonomy of AUVs.

Dissertação de Mestrado em Ecologia apresentada à Faculdade de Ciências e Tecnologia
A poluição química por metais é atualmente um problema nos ecossistemas marinhos e os organismos, expostos, podem acumular níveis altos de metais, provenientes de várias fontes antropogénicas. Altas concentrações induzem o stress oxidativo levando a danos fisiológicos que podem ser detetados com biomarcadores específicos. As aves marinhas são bons bioindicadores de qualidade ambiental, fornecendo informações sobre as relações tróficas e qualidade dos recursos. Os objetivos deste estudo foram avaliar a qualidade ambiental do Atlântico Norte, em particular na costa portuguesa e áreas adjacentes, com ênfase na contaminação por metais, e (1) avaliar a contaminação em três espécies de aves marinhas nas respetivas áreas de alimentação, (2) relacionar a ecologia trófica com as concentrações de metais e (3) avaliar o stress oxidativo das aves de acordo com a ecologia trófica e contaminação.Os dados foram recolhidos durante a época de reprodução de 2017 nas Berlengas (costa oeste) e Ria Formosa (costa sul), Portugal.Foram colhidas amostras de sangue e penas de: 28 cagarras Calonectris borealis em dois períodos de reprodução (pré-postura e alimentação das crias) e duas sub-colónias da Ilha da Berlenga (Furado Seco e Melreu) separadas por 800 m; 27 gaivotas-de-patas-amarelas Larus michahellis em duas colónias (Ilhas da Berlenga e Deserta); e 13 gaivotas de Audouin Larus audouinii (Ilha da Deserta). Foi efetuada quantificação de metais (Al, As, Pb, Cd, Cr, Fe, Cu, Mn, Ni, Hg, Se, Ag, Zn) por ICP-MS e avaliação do stress oxidativo usando os testes de danos no DNA, peroxidação lipídica, OXY e d-ROM. Análise de isótopos estáveis (SIA) e dispositivos GPS foram usados para relacionar a contaminação por metais e o stress oxidativo com o comportamento de alimentação das espécies (nicho trófico e uso de habitat).Os resultados revelaram uma clara segregação na contaminação entre as três espécies e entre colónias/sub-colónias. No geral, a gaivota de Audouin e a cagarra foram as espécies mais contaminadas para a maioria dos metais, provavelmente devido à sua dieta composta exclusivamente por peixe e outros recursos marinhos. Por outro lado, a gaivota-de-patas-amarelas apresentou menores níveis de contaminação para a maioria dos metais. Esta espécie alimenta-se regularmente de outras fontes de alimento terrestres para além de peixe, o que pode induzir a menores níveis de contaminação. Curiosamente, observaram-se diferenças significativas na contaminação entre duas sub-colónias de cagarras separadas por apenas 800 m, em alguns metais específicos. Os dados GPS mostram uma segregação espacial parcial entre sub-colónias, suportada por diferenças nos valores de δ13C nas células do sangue, o que pode explicar em parte as diferenças nos padrões de contaminação. No entanto, as aves do Furado Seco reproduzem-se dentro de uma caverna que pode estar exposta a níveis mais altos de alguns metais. Assim, a contaminação pode também estar relacionada com o habitat de reprodução e não apenas com a dieta. Em contraste, as gaivotas-de-patas-amarelas de colónias distantes não mostraram diferenças substanciais na maioria dos metais. No entanto, maiores níveis de Hg na Ilha da Deserta podem estar relacionados com o maior consumo de peixes demersais da pesca de arrasto, enquanto níveis mais altos de Mn e Ni na Ilha da Berlenga podem estar relacionados com o habitat de reprodução.Diferenças no stress oxidativo entre os períodos de reprodução das cagarras foram evidentes. Apesar de diferenças nos valores de δ13C e δ15N do plasma e células do sangue, não foi encontrada nenhuma relação entre os isótopos estáveis e biomarcadores. Estes resultados sugerem que níveis mais altos de stress durante a pré-postura podem estar relacionados com o papel reprodutor das aves. Nas gaivotas, registaram-se diferenças entre espécies e colónias no stress oxidativo, que se relacionaram com o uso de habitat, mas não com a contaminação. Níveis mais altos de stress nas gaivotas-de-patas-amarelas da Berlenga poderão estar relacionados com a exploração de habitats de alimentação mais diversos. Além disso, o controlo populacional de gaivotas-de-patas-amarelas na ilha da Berlenga durante o período de incubação, destruindo ovos, poderá levar a um aumento dos níveis de stress nestas aves. Este estudo sugere também que níveis mais baixos de stress oxidativo da gaivota de Audouin, em comparação com as gaivotas-de-patas-amarelas na Ilha Deserta, podem estar relacionados com a ecologia trófica, mas não com a contaminação.Este estudo permitiu avaliar a qualidade do ambiente no Atlântico Norte e na costa portuguesa. Os níveis de contaminação obtidos foram comparados com outros estudos após uma extensa revisão bibliográfica, mostrando uma maior vulnerabilidade ou exposição das espécies/colónias de estudo para alguns metais (Pb, Cr e Se). Futuros estudos podem ajudar a entender os padrões de distribuição destes contaminantes e respetivas interações tróficas na costa portuguesa
Metal pollution is currently a major issue in marine ecosystems, because organisms are exposed to increasing and accumulating levels from several anthropogenic sources. High concentrations of metals can disturb oxidative balance leading to physiological damages, which can be measured using specific biomarkers. Seabirds are suitable bioindicators of marine environmental health, providing valuable information on trophic relationships and quality of resources. This study aimed to assess environmental health status in the North Atlantic, with emphasis on metal contamination. The main objectives were to: (1) assess contamination in three seabird species within their distinct foraging areas, (2) relate their trophic ecology with respective metal concentrations, and (3) evaluate the oxidative stress of seabirds in relation to their trophic ecology and concentration of metals.Data were collected during the breeding season of 2017 in two distinct areas of Portugal (Berlengas in the west coast and Ria Formosa in the south coast). Blood and feathers were collected from: 28 Cory’s shearwaters Calonectris borealis in two breeding periods (pre-laying and chick-rearing) and two sub-colonies of Berlenga Island (Furado Seaco and Melreu) separated by only 800 m; 27 Yellow-legged gulls Larus michahellis in two colonies (Berlenga and Deserta Islands); and 13 Audouin’s gulls Larus audouinii (only in Deserta Island). Metal quantification (Al, As, Pb, Cd, Cr, Fe, Cu, Mn, Ni, Hg, Se, Ag, Zn) using ICP-MS and oxidative stress assessment using DNA damage, lipid peroxidation, OXY and d-ROM tests were performed. In addition, SIA and GPS data were used to link metal contamination and oxidative stress with the foraging behaviour of each species.Results revealed a clear segregation of metal contamination among the three species and between colonies/sub-colonies. Overall, Audouin’s gull and Cory’s shearwater were the most contaminated species for most metals, presumably because they rely mainly on fish and other marine resources. On the other hand, Yellow-legged gull showed lower contamination levels for most metals. This species feeds regularly on other food sources besides fish, including from terrestrial environments, which may buffer contamination levels. Interestingly, and unexpected, there were significant differences in metal contamination between the two adjacent sub-colonies of Cory’s shearwaters. There was a partial spatial segregation between sub-colonies, supported by differences in δ13C values in red blood cells, which may partly explain such differences in the contamination patterns. However, other potential explanations may be possible, such as the breeding habitat; birds from Furado Seco have their nests inside a long and muddy cave, which can be exposed to higher contamination levels. Thus, contamination might also be related with the breeding habitat itself and not only to foraging habitat. In contrast, Yellow-legged gulls from distant colonies did not show substantial differences in most metal concentrations. However, higher levels of Hg for birds breeding in Deserta could be related with their higher consumption of demersal fish from trawl fisheries, while higher levels of Mn and Ni in Berlenga Island individuals could be related with the respective breeding habitat.Oxidative stress parameters differed significantly between Cory’s shearwaters breeding periods. Despite differences in δ13C and δ15N of plasma and red blood cells, no relationships were found between stable isotopes and biomarkers, suggesting that higher stress levels during pre-laying could be related with intrinsic physiological factors of breeding duties, and not exclusively with birds’ foraging niche. Concerning gulls, differences in oxidative stress between species and colonies were related with birds’ foraging behaviour, but not with metal contamination. Higher stress levels in Yellow-legged gulls from Berlenga Island could probably be related with different exploited environments for foraging. Moreover, a population control of Yellow-legged gulls in Berlenga Island during the incubation period (i.e. egg destroying), may potentially lead to increased stress levels in these birds. This study also suggests that lower oxidative stress levels of Audouin’s gull in comparison with Yellow-legged gulls in Deserta Island, could be related with diet and trophic ecology, but not with metal contamination.This study allowed to assess environmental health in the North Atlantic, particularly along the Portuguese coast and adjacent areas. After comparing contamination levels with an extensive literature review, this study shows a greater vulnerability or exposure of these species/colonies for some specific metals (Pb, Cr and Se). Further studies would help to understand these contaminants’ distribution patterns in seabirds, because very few information exists for the Portuguese coast.

abstract: Air pollution has been linked to various health problems but how different air pollutants and exposure levels contribute to those diseases remain largely unknown. Researchers have mainly relied on data from government air monitoring stations to study the health effects of air pollution exposure. The limited information provided by sparse stations has low spatial and temporal resolution, which is not able to represent the actual exposure of individuals. A tool that can accurately monitor personal exposure provides valuable data for epidemiologists to understand the relationship between air pollution and certain diseases. It also allows individuals to be aware of any ambient air quality issues and prevent air pollution exposure. To build such a tool, sensors with features of fast response, small size, long lifetime, high sensitivity, high selectivity, and multi-analyte sensing are of great importance. In order to meet these requirements, three generations of novel colorimetric sensors have been developed. The first generation is mosaic colorimetric sensors based on tiny sensor blocks and by detecting absorbance change after each air sample injection, the target analyte concentration can be measured. The second generation is a gradient-based colorimetric sensor. Lateral transport of analytes across the colorimetric sensor surface creates a color gradient that shifts along the transport direction over time, and the sensor tracks the gradient shift and converts it into analyte concentration in real-time. The third generation is gradient-based colorimetric arrays fabricated by inkjet-printing method that integrates multiple sensors on a miniaturized sensor chip. Unlike traditional colorimetric sensors, such as detection tubes and optoelectronic nose, that are typically for one-time use, the presented three generations of colorimetric sensors aim to continuously monitor multiple air pollutants and the sensor lifetime and fabrication methods have been improved over each generation. Ozone, nitrogen dioxide, formaldehyde and carbon monoxide are chosen as analytes of interest. The performance of sensors has been validated in the lab and field tests, proving the capability of the sensors to be used for personal exposure monitoring.
Dissertation/Thesis
Doctoral Dissertation Chemistry 2019