Legionella pneumophila and other opportunistic pathogens of drinking water pose important problems at the interface of biology, environmental engineering, public health, and governance. In this thesis, I explore the molecular mechanisms permitting survival of L. pneumophila in built water systems is the nature of its physiology under different conditions and different life-phases. In the first chapter, I discuss how various physiological states of L. pneumophila affect the propensity for survival and virulence in relation to drinking water environments. This literature review should provide a perspective important for designing controlled laboratory experiments rooted in a robust understanding of how phenotype dictates experimental results. In the second chapter, I describe sequence and phylogenetic analyses performed to investigate the presence of a type 1 secretion system and virulence factor throughout the Legionella genus. While this system was previously believed to be conserved to L. pneumophila, our analysis indicates that this system is well-distributed throughout the Legionella genus, blurring the lines between "pathogenic" and "non-pathogenic" species. In the third chapter, I report the secretome of endemic Flint, Michigan L. pneumophila in corrosive water, simulating the environmental impact of the Flint Water Crisis on local L. pneumophila populations. Our results from this study have expanded the secretome of L. pneumophila, provided insight on mechanisms it may employ to resist stress in water, and created several novel lines of inquiry at the merging frontier of biochemistry and environmental engineering. / Master of Science in Life Sciences

Like many other urban areas around the world, Cape Town, South Africa relies on governmental management authorities to ensure water supply. Recently, a three year drought from 2015 to 2018 caused a major water shortage, threatening water supply to the city. In response, the City engaged in multiple mitigation efforts, amongst which was a major communications campaign to inform the public and encourage conservation behaviour. Drawing on literature on water crisis management and framing theory, this thesis analyses how the water crisis was framed in communications made available online by the City of Cape Town (CCT) to the public between March 2017 and March 2018. To answer this question, the project adopted a frames study approach to determine the types and characteristics of communication items released by the City of Cape Town, as the water managing authority, during the recent water crisis. The study established that a range of frames were employed by the CCT in communicating the drought. Through an analysis of the trends in the framing of the water crisis messages the study further identified the shifts in framing and messaging throughout the water crisis response period. Six key frames were identified, namely ‘the City success story'; ‘obscurity and ambiguity'; ‘consumption is key'; ‘the situation is controllable'; ‘together we can beat the drought'; and ‘us versus them'. It was found that while there may have been a lack of strategic planning regarding public communications which resulted in conflated messages, the City's communications campaign was nonetheless effective in that it correlated with a significant drop in private – individual and household – water consumption which delayed Day Zero (when water supply would be cut-off and daily water rations would only be available at collection points for the public). The key implication of this study is that despite contradictions, idiosyncrasies and lack of planning, a heterogenous range of messages in communicating a crisis can reach and evoke appropriate responses from multiple audiences of the public.

Orientador: Angélica Góis Morales / Coorientador: Fernando Ferrari Putti / Coorientador: Sandra Cristina de Oliveira / Resumo: O objetivo geral desta pesquisa é analisar a relação entre a gestão dos recursos hídricos, praticada em duas fecularias de mandioca do Escritório de Desenvolvimento Rural de Assis, e a Pegada Hídrica do produto final. Portanto, neste estudo multicasos, foi mensurada a Pegada Hídrica da mandioca, do processamento da mandioca para a produção de fécula e da fécula, e comparadas, por meio do coeficiente de correlação por postos de Spearman, as práticas de gerenciamento da água consideradas ideais em cada uma das fecularias estudadas. Ademais, foram identificadas as práticas efetivamente empreendidas. A coleta de dados ocorreu por meio da aplicação de formulários semiestruturados e entrevistas. Como resultado, na Fecularia A, o total da Pegada Hídrica do produto final foi de 2.607,25 m³.ton-1 de fécula e na Fecularia B de 1.764,13 m³.ton-1 de fécula, enquanto as práticas consideradas ideais, segundo o Coeficiente de Spearman de 0,5857, possuem correlação positiva de moderada a forte. Igualmente, as práticas efetivamente realizadas são semelhantes e sugerem exercer influência sobre o resultado da Pegada Hídrica. Este trabalho possibilitou o conhecimento do consumo hídrico na produção da fécula de mandioca, de forma a evidenciar fatores, de ordem interna e externa às fecularias, influentes na apropriação do recurso hídrico nas mesmas. / Abstract: The general objective of this research is to analyze the relationship between the management of water resources, practiced in two cassava starch of the Office of Rural Development of Assis, and the Water Footprint of the final product. Therefore, in this multi-level study, was measured the Water Footprint of cassava, cassava processing for starch production, and starch, and were compared, by means of the Spearman's Rank Correlation Coefficient, water management practices considered ideal in each of the farms studied. In addition, the practices actually undertaken were identified. Data collection took place through the application of semi-structured forms and interviews. As a result, in Cassava Starch A, the total Water Footprint of the final product was 2,607.25 m³.ton-1 starch and in Cassava Starch B, 1,764.13 m³.ton-1 starch, while the practices considered ideal, according to the Spearman Coefficient of 0.5857, have a moderate to strong positive correlation. Equally, the practices actually carried out are similar and suggest influence on the result of the Water Footprint. This work made possible the knowledge of water consumption in the production of cassava starch, in order to show factors, internal and external to the cassava starch, influential in the appropriation of water resources in them. / Mestre

Submitted by Luana Ferreira Pires (luanaf.p@hotmail.com) on 2018-06-07T00:57:44Z No. of bitstreams: 1 LuanaFerreiraPires_PGAD_TUPA_032018.pdf.pdf: 3166778 bytes, checksum: a7e4eb1956fbb1924354e5dc15cef232 (MD5) / Approved for entry into archive by Eliana Katia Pupim (katiapupim@tupa.unesp.br) on 2018-06-11T14:44:19Z (GMT) No. of bitstreams: 1 pires_lf_me_tupa.pdf: 2538547 bytes, checksum: 29f57bded09910f88f2924c686829242 (MD5) / Made available in DSpace on 2018-06-11T14:44:19Z (GMT). No. of bitstreams: 1 pires_lf_me_tupa.pdf: 2538547 bytes, checksum: 29f57bded09910f88f2924c686829242 (MD5) Previous issue date: 2018-03-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo geral desta pesquisa é analisar a relação entre a gestão dos recursos hídricos, praticada em duas fecularias de mandioca do Escritório de Desenvolvimento Rural de Assis, e a Pegada Hídrica do produto final. Portanto, neste estudo multicasos, foi mensurada a Pegada Hídrica da mandioca, do processamento da mandioca para a produção de fécula e da fécula, e comparadas, por meio do coeficiente de correlação por postos de Spearman, as práticas de gerenciamento da água consideradas ideais em cada uma das fecularias estudadas. Ademais, foram identificadas as práticas efetivamente empreendidas. A coleta de dados ocorreu por meio da aplicação de formulários semiestruturados e entrevistas. Como resultado, na Fecularia A, o total da Pegada Hídrica do produto final foi de 2.607,25 m³.ton-1 de fécula e na Fecularia B de 1.764,13 m³.ton-1 de fécula, enquanto as práticas consideradas ideais, segundo o Coeficiente de Spearman de 0,5857, possuem correlação positiva de moderada a forte. Igualmente, as práticas efetivamente realizadas são semelhantes e sugerem exercer influência sobre o resultado da Pegada Hídrica. Este trabalho possibilitou o conhecimento do consumo hídrico na produção da fécula de mandioca, de forma a evidenciar fatores, de ordem interna e externa às fecularias, influentes na apropriação do recurso hídrico nas mesmas. / The general objective of this research is to analyze the relationship between the management of water resources, practiced in two cassava starch of the Office of Rural Development of Assis, and the Water Footprint of the final product. Therefore, in this multi-level study, was measured the Water Footprint of cassava, cassava processing for starch production, and starch, and were compared, by means of the Spearman's Rank Correlation Coefficient, water management practices considered ideal in each of the farms studied. In addition, the practices actually undertaken were identified. Data collection took place through the application of semi-structured forms and interviews. As a result, in Cassava Starch A, the total Water Footprint of the final product was 2,607.25 m³.ton-1 starch and in Cassava Starch B, 1,764.13 m³.ton-1 starch, while the practices considered ideal, according to the Spearman Coefficient of 0.5857, have a moderate to strong positive correlation. Equally, the practices actually carried out are similar and suggest influence on the result of the Water Footprint. This work made possible the knowledge of water consumption in the production of cassava starch, in order to show factors, internal and external to the cassava starch, influential in the appropriation of water resources in them.

Legionella pneumophila is a pathogen that can proliferate in premise (i.e., building) plumbing and, when aerosolized during water use, infect the lungs of exposed individuals and cause a deadly form of pneumonia known as Legionnaires' disease. Given that it is one of the primary sources of tap-water associated disease throughout much of the world, this organism has been the subject of intense research, ranging from aiming to understand key aspects of its physiology that allow it to proliferate in premise plumbing, to the specific virulence factors that make it so infectious to humans. The work presented here starts with a comprehensive review of published studies related to the L. pneumophila proteome, i.e., the set of expressed proteins associated with a given strain under a given set of environmental conditions, showing how the field has progressed in parallel to improvements in mass spectrometry technologies and how proteomics can be used as a tool to understand this unique and important organism. Copper is a natural antimicrobial that can be present in drinking water due to passive release from copper pipes or intentionally dosed (e.g., copper-silver ionization systems) for microbial control. However, some L. pneumophila strains have recently been found to exhibit copper resistance, an adaptive process that is not fully understood at the physiological level. Chapter Two describes the copper survivability of three outbreak-associated strains of L. pneumophila and examines the copper-induced proteome of QC1, a strain found to display high resistance to copper. Pairwise comparisons of the proteomes of copper-resistant and copper sensitive strains indicated that L. pneumophila QC1 adapts to copper exposure via the induction of redox and metal homeostasis proteins, while concomitantly inducing motility and pathogenesis related proteins, suggestive that copper induces a search for a host protozoan strain for protection. In 2014 and 2015, Flint, Michigan experienced the largest per capita community-wide Legionnaires' Disease outbreak in US history. The outbreak was associated with a change in the source of the municipal drinking water supply from Detroit water, which was sourced from the Great Lakes and subject to appropriate corrosion control, to the Flint River, which was not appropriately controlled for corrosivity. The underlying drivers of this outbreak have been debated and include: elevated iron in the water serving as a nutrient for L. pneumophila, diminished chlorine in the water due to reactions with iron, reduced copper in the water due to shifts in pH influencing release from copper pipes, and shifts in potentially key components of the microbial community. In Chapter Three of this dissertation, we employ controlled microcosm studies to establish a fundamental understanding of interactive effects of pipe material and water of varying iron bioavailability (ferric chloride, ferrous chloride and ferric pyrophosphate) on the microbial community and its relationship with L. pneumophila numbers. The combination of copper pipes and Flint River water decreased the diversity of the microbial community to a larger degree than copper pipes with Detroit water, implying greater copper bioavailability in the former condition. Several Order were found to be significantly associated with high or low numbers of culturable L. pneumophila recovered from the microcosms. Most notably, the Order Pseudomonadales was significantly associated to the reactors with low culturable L. pneumophila. This order contains Pseudomonas species known to inhibit the growth of L. pneumophila. The findings reported in this dissertation can be used to develop more informed management practices for drinking water systems to reduce the risk of Legionnaires' Disease outbreaks associated with premise plumbing. Specifically, 1) copper might be inducing a more pathogenic form of copper resistant L. pneumophila, 2) the use of corrosive control in municipal water systems goes beyond the influence on lead and copper pipes, but also on the microbial community, which in part influences L. pneumophila, and 3) there are organisms, such as Pseudomonadales species, associated with environments with low culturable L. pneumophila which might be introduced to the drinking water systems as probiotics. / Doctor of Philosophy / Legionella pneumophila is a microbe found in drinking water plumbing systems. This organism causes Legionnaires' Disease, a severe form of pneumonia that particularly affects immunocompromised individuals. Due to its health and economic impact, there are worldwide efforts to understand the biology of this organism, from the conditions that allows it to grow in the drinking water plumbing, to the specific components that allows it to infect humans. In this dissertation, we first review the published studies related to the L. pneumophila proteome, a powerful tool used to functionally describe biological organisms. This first chapter showed how proteomics can be used to understand this unique and important organism. In the next chapter we studied how copper metals may influence the proteome of L. pneumophila. Copper pipes have been extensively used to control the growth of microorganisms in drinking water systems, however some studies have reported that copper may promote the growth of L. pneumophila. In this chapter, we showed that a copper resistant strain of L. pneumophila adapts to copper exposure by inducing motility and pathogenesis related proteins, suggesting that it might be more infectious. In the last chapter of this dissertation, we investigated the combined effect of pipe material and water chemistry, on the microbial community and its relationship with L. pneumophila. The combination of copper pipes and a more corrosive water decreased the diversity to a larger degree, in comparison to the other evaluated conditions. Several organisms were also identified to be significantly associated with the high or low culturable L. pneumophila. This is of particular interest because they might be used as potential probiotics to control the growth of L. pneumophila. The findings reported in this dissertation can help to better understand the significance of water chemistry and pipe material, particularly copper pipes, for the purpose of reducing risk of Legionnaires' Disease outbreaks associated with drinking water systems.

Chaos theory holds that systems act in unpredictable nonlinear ways and that their behavior can only be observed, never predicted. This is an informative model for an organization in crisis. The West Virginia water contamination crisis, which began on January 9, 2014, fits the criteria of a system in chaos. Given the lack of appropriate response from the established organizations involved, many emergent organizations formed to help fill unmet informational and physical needs of the affected population. Crisis researchers have observed these ephemeral organizations for decades, but the recent proliferation of information communication technologies (ICT’s) have made their activities more widespread and observable. In West Virginia, their activities were indispensable to the affected population and helped restore a sense of normalcy. In this chaotic system, the emergent organizations functioned as strange attractors, helping move the system away from bifurcation and towards normalcy. This dissertation uses a qualitative approach to study the emergent organizations and how their presence and efforts were the mechanism that spurred the self-organization process.

A escassez hídrica devido às baixas vazões e problemas de captação de água são objeto de debates e conflitos na bacia hidrográfica do rio Piracicaba. A restauração florestal é tida como uma alternativa para a minimização destes conflitos. No entanto, as florestas modificam significativamente os processos hidrológicos, influenciando a dinâmica da água na bacia hidrográfica. Portanto, a fim de se obter disponibilidade hídrica a longo prazo, é necessário identificar as áreas aptas para a restauração florestal, onde os efeitos sejam positivos para os recursos hídricos. Dentro deste contexto, o presente trabalho teve como objetivo utilizar um modelo hídrico climático distribuído para espacializar as características hídricas e climáticas favoráveis, ou não, à restauração florestal. Para isto, foi utilizada a bacia hidrográfica do rio Piracicaba como área de estudo (12.500 km2). Foram obtidos dados mensais normais de precipitação, temperatura, radiação solar, topografia e capacidade de armazenamento de água no solo, distribuídos espacialmente na bacia hidrográfica. A partir destas informações, foi calculado o balanço hídrico climático distribuído (BHCD) através da ferramenta \"Water Balance\", que calcula a evapotranspiração potencial e real, o déficit e o excedente hídrico em um ambiente SIG (Sistema de Informações Geográficas). Os resultados foram comparados com o balanço hídrico climático (BHC), tradicional ou pontual, e uma análise estatística de agrupamentos de áreas com características hidrológicas semelhantes foi realizada. A divisão da bacia hidrográfica em classes (ou grupos) com características semelhantes permite compreender a sensibilidade natural das áreas e possíveis efeitos na disponibilidade hídrica. A capacidade de determinar, em diferentes escalas de estudo, a dinâmica dos recursos hídricos possibilitou melhores condições de análise diante da heterogeneidade natural da bacia hidrográfica. Os dados gerados permitiram identificar áreas onde as recomendações locais podem ser feitas com base nas variáveis do balanço hídrico. Dessa forma, a espacialização das variáveis e o agrupamento por características hidrológicas são ferramentas que podem ser utilizadas em projetos de restauração florestal, uma vez que podem subsidiar decisões a respeito das áreas mais favoráveis a restauração florestal. / Water shortage due to low flows and water basin capitation problems are subject of debates and conflicts in Piracicaba river basin. Forest restoration is an alternative to minimize these conflicts. However, forests significantly modify the hydrological processes, influencing the dynamics of the water in the watershed. Therefore, in order to obtain water availability in the long term, it is necessary to identify suitable areas for forest restoration, where effects are positive for water resources. In this context, this study aimed at using a climatic model water to spatialize the water features and favorable climate, or not, to forest restoration. For this purpose, the Piracicaba river basin was used as study area (12,500 km2). Monthly data were obtained from precipitation, temperature, solar radiation, topography, and water storage capacity in the soil. Trough these data, distributed climatic water balance was calculated by the Water Balance tool, integrated in a GIS (Geographic Information System) which calculates the potential and actual evapotranspiration, deficit and the surplus water. Results were compared with traditional climatic water balance. Later statistical analyses was performed to identify areas with similar hydrological characteristics. This division of the watershed into classes (or groups) with similar characteristics allows us to understand the natural sensitivity of the areas and possible effects on water availability. The ability to determine, in different study scales, the dynamics of water resources, has enabled better analysis of conditions on the natural heterogeneity of the basin. Raised info has identified areas where local recommendations can be made based on the variables of water balance. Thus, spatial distribution of variables and grouping by hydrological characteristics are tools that can be used in forest restoration projects as it can subsidize decisions regarding the most favorable areas for forest restoration.

A escassez hídrica devido às baixas vazões e problemas de captação de água são objeto de debates e conflitos na bacia hidrográfica do rio Piracicaba. A restauração florestal é tida como uma alternativa para a minimização destes conflitos. No entanto, as florestas modificam significativamente os processos hidrológicos, influenciando a dinâmica da água na bacia hidrográfica. Portanto, a fim de se obter disponibilidade hídrica a longo prazo, é necessário identificar as áreas aptas para a restauração florestal, onde os efeitos sejam positivos para os recursos hídricos. Dentro deste contexto, o presente trabalho teve como objetivo utilizar um modelo hídrico climático distribuído para espacializar as características hídricas e climáticas favoráveis, ou não, à restauração florestal. Para isto, foi utilizada a bacia hidrográfica do rio Piracicaba como área de estudo (12.500 km2). Foram obtidos dados mensais normais de precipitação, temperatura, radiação solar, topografia e capacidade de armazenamento de água no solo, distribuídos espacialmente na bacia hidrográfica. A partir destas informações, foi calculado o balanço hídrico climático distribuído (BHCD) através da ferramenta \"Water Balance\", que calcula a evapotranspiração potencial e real, o déficit e o excedente hídrico em um ambiente SIG (Sistema de Informações Geográficas). Os resultados foram comparados com o balanço hídrico climático (BHC), tradicional ou pontual, e uma análise estatística de agrupamentos de áreas com características hidrológicas semelhantes foi realizada. A divisão da bacia hidrográfica em classes (ou grupos) com características semelhantes permite compreender a sensibilidade natural das áreas e possíveis efeitos na disponibilidade hídrica. A capacidade de determinar, em diferentes escalas de estudo, a dinâmica dos recursos hídricos possibilitou melhores condições de análise diante da heterogeneidade natural da bacia hidrográfica. Os dados gerados permitiram identificar áreas onde as recomendações locais podem ser feitas com base nas variáveis do balanço hídrico. Dessa forma, a espacialização das variáveis e o agrupamento por características hidrológicas são ferramentas que podem ser utilizadas em projetos de restauração florestal, uma vez que podem subsidiar decisões a respeito das áreas mais favoráveis a restauração florestal. / Water shortage due to low flows and water basin capitation problems are subject of debates and conflicts in Piracicaba river basin. Forest restoration is an alternative to minimize these conflicts. However, forests significantly modify the hydrological processes, influencing the dynamics of the water in the watershed. Therefore, in order to obtain water availability in the long term, it is necessary to identify suitable areas for forest restoration, where effects are positive for water resources. In this context, this study aimed at using a climatic model water to spatialize the water features and favorable climate, or not, to forest restoration. For this purpose, the Piracicaba river basin was used as study area (12,500 km2). Monthly data were obtained from precipitation, temperature, solar radiation, topography, and water storage capacity in the soil. Trough these data, distributed climatic water balance was calculated by the Water Balance tool, integrated in a GIS (Geographic Information System) which calculates the potential and actual evapotranspiration, deficit and the surplus water. Results were compared with traditional climatic water balance. Later statistical analyses was performed to identify areas with similar hydrological characteristics. This division of the watershed into classes (or groups) with similar characteristics allows us to understand the natural sensitivity of the areas and possible effects on water availability. The ability to determine, in different study scales, the dynamics of water resources, has enabled better analysis of conditions on the natural heterogeneity of the basin. Raised info has identified areas where local recommendations can be made based on the variables of water balance. Thus, spatial distribution of variables and grouping by hydrological characteristics are tools that can be used in forest restoration projects as it can subsidize decisions regarding the most favorable areas for forest restoration.

abstract: Sustainability and environmental justice, two fields that developed parallel to each other, are both insufficient to deal with the challenges posed by institutional environmental violence (IEV). This thesis examines the discursive history of sustainability and critiques its focus on science-based technical solutions to large-scale global problems. It further analyzes the gaps in sustainability discourse that can be filled by environmental justice, such as the challenges posed by environmental racism. Despite this, neither field is able to contend with IEV in a meaningful way, which this thesis argues using the case study of the Flint Water Crisis (FWC). The FWC has been addressed as both an issue of sustainability and of environmental justice, yet IEV persists in the community. This is due in part to the narrative of crisis reflected by the FWC and the role that knowledge production plays in that narrative. To fill the gap left by both sustainability and environmental justice, this thesis emphasizes the need for a transformational methodology incorporating knowledge produced by communities and individuals directly impacted by sustainability problems. / Dissertation/Thesis / Masters Thesis Sustainability 2019

In 2017 and 2018, the city of Cape Town, in South Africa, suffered one of the most severe water crises ever seen, becoming the first big city to face a realistic scenario of a "Day Zero", the day in which the dams reach a water storage level unable to provide water services to other than critical services. In the wake of this emergency, several organisations and movements started to organise themselves to mitigate the effects of the drought and find a solution. The measures undertaken by the local government, which included punitive tariffs for the citizens, caused a big discontent among the population, who protested in the streets to demand a proper solution. Amid the protesters, the social movements rose to demand from the authorities democratic and reasonable management of the water in the city, putting pressure by protesting, creating petitions, mobilising people and spreading facts about the crisis and what they believed were the true problems behind it.   This qualitative research included a field study in the city of Cape Town and uses abductive research for the analysis of data. The study is exploratory, as it intends to understand and explore what happened during the crisis and the role of social movements to create a narrative. Five interviews were conducted between two different target groups: social movement actors and authorities.   This thesis focuses on the role that social movements played and their dynamics in the outcome of the actions taken by the authorities to address the water crisis in Cape Town. Using social movement theory and alliances theory, this explores what actors were involved, what actions and activities the social movements conducted, and what was the outcome of the role they played. This is done in order to create a narrative of the facts that occurred during the crisis until the Day Zero was officially called off by the local authorities, the moment in which the organisations stopped their engagement due to whether the loss of the momentum, the collapse of the alliances or the accomplishment of their minimum demands.   The study concludes that there were two moments that determined the role of social movements during the water crisis: first, with the emergence of the crisis, the movements gathered and played a communicator role, delivering information and sharing facts; secondly, after the measures taken by the authorities were announced, the movements played an instigator role as an opposition to the local government, putting pressure mainly in the streets. We conclude that the outcome delivered by the authorities, the so-called Water Strategy, was an important step but did not respond to the demands of the movement sufficiently, as it was not conducted in a participatory way, although it included some of the demands of the movement. It is not possible to conclude that the role played by the social movements was key to determine the outcome of the crisis, but they contributed to put pressure and make visible the demands for a more democratic water management.