Master of Landscape Architecture / Department of Landscape Architecture/Regional and Community Planning / Timothy Keane / Kansas City’s outdated sewer system is presently incapable of capturing and treating the increased runoff volumes in Turkey Creek Basin during rainstorm events. As a result, 2.66 billion gallons of untreated sewer system overflow is released annually into the Kansas River and nearby properties. In 2002, the Environmental Protection Agency issued a civil action requiring the City of Kansas City, Missouri, to take appropriate and necessary actions needed to prevent or minimize the discharge of untreated sewage. In response, the City of Kansas City adopted a comprehensive Overflow Control Plan intended to reduce sewer system overflow volumes in Turkey Creek Basin by 85% at a cost of approximately $244 million. Initially, the City of Kansas City seriously considered implementing stormwater best management practices (BMPs) in place of sewer system improvements. Stormwater BMPs infiltrate, filter, store, and evaporate stormwater runoff close to its source, preventing stormwater runoff from reaching the sewer system. Subsequently, many BMPs were eliminated from the Overflow Control Plan and replaced with conventional sewer system technologies because of performance concerns. However, the Overflow Control Plan acknowledged that BMPs located on private property would indirectly benefit Kansas City’s stormwater management strategy. Using geographic information system (GIS) analysis, suitability maps were generated for twelve different BMPs to determine suitable locations in Turkey Creek Basin for reducing stormwater runoff. Analysis concluded that the most effective strategy for sustainable stormwater management would be to locate BMPs at higher elevations within the watershed to prevent upland runoff from flooding sewer system pipes at lower elevations. Areas having the highest suitability are located primarily on residential land, implying that Kansas City could benefit most from encouraging its residents to equip their properties with site-appropriate BMPs. This can be achieved through educational initiatives, policy adoption, and homeowner incentives. Therefore, policies and incentives targeting Kansas City’s residents should be implemented to reduce sewer overflow volumes and prevent future costly improvements to Kansas City’s sewer system.

Antibiotic resistance is a major threat to human health. Clinical situations are the main focus for antibiotic resistance research, but understanding the spread of resistance in the environment is also vital. A major contributor to this spread is wastewater from combined sewer overflow (CSO) events. The effect of CSO events on antibiotic resistance in the James River near Richmond, Virginia was studied using genomic and microbiological approaches. The abundance of genes associated with resistance to quinolones (qnrA) and tetracycline (tetW) was strongly correlated with the presence of fecal indicator bacteria (E. coli abundance) as well as total nitrogen and phosphorus loads, which suggests an anthropogenic source of these genes. Abundance of the blaTEM gene, which confers resistance to β-lactam antibiotics, was elevated during CSO events and increased with precipitation and river discharge. Bacteria isolated during a CSO event were resistant to more antibiotics and had higher multi-drug resistance when compared to isolates from a non-event. This study demonstrated that CSO events are contributing to the spread of antibiotic resistance.

This study showed that hydrodynamic particle breakage had potential as a method to help improve the disinfection of wastewater effluents. The physicochemical properties of flocs from four distinct effluents sources (combined sewer overflow, settled combined sewer overflow, primary effluent, and final effluent) were compared before and after hydrodynamic treatment. The use of hydrodynamic force to cause floc breakage was shown to be effective, though variable, across all source types. This variation in floc breakage did not have a significant impact on the UV disinfection achieved, as the UV dose kinetics were similar across samples from the same source type. The results of this study demonstrate how the physicochemical properties of floc are affected when exposed to shear force. These observations further the understanding of floc composition and behaviour when shear forces are applied while also providing evidence to indicate this process improves the performance of UV disinfection technology.

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Ett allt vanligare uppdrag inom hydraulisk modellering är att undersöka hur nya bostads- och handelsområden påverkar statusen för äldre befintliga ledningsnät. I nya områden anläggs nästan uteslutande separatsystem och eftersom nya områden oftast uppförs utanför den befintliga bebyggelsen måste spillvattnet från dessa områden transporteras genom det befintliga avloppsnätet, vilket kan vara helt eller delvis kombinerat. I stadsdelen Hjorthagen i Stockholm planeras 5000 nya lägenheter att byggas och spillvattenflödet från dessa skall anslutas uppströms det befintliga kombinerade ledningsnätet i området. Examensarbetet har gått ut på att undersöka hur det kombinerade ledningssystemet påverkas av den tillkommande spillvattenbelastningen. Syftet var att undersöka hur den dimensionerande spillvattenavrinningen påverkar de åtgärder och kostnader som behövs för att få det utökade ledningsnätet att fungera, samt att utreda behovet av ytterligare anpassningar av ledningsnätet inför framtida klimatförändringar. Vid beräkning av bräddvolymer simuleras vanligen spillvattenavrinningen från medeldygnsförbrukning av vatten. I studien har det även ingått att undersöka om detta sätt ger lägre bräddvolymer jämfört med att variera vattenförbrukningen med ett veckomönster. Med modelleringsprogrammet MIKE Urban undersöktes tre fall av dimensionerande spillvattenflöde från det tillkommande området i Hjorthagen. Olika stor säkerhetsmarginal användes vid dimensioneringen. I alla tre fallen fick det tillkommande spillvattenflödet kapaciteten i det kombinerade ledningssystemet att överskridas redan vid torrväder, med bräddning som följd. Undersökningen visade att ju högre säkerhetsmarginal dimensioneringen innebar desto mer omfattande blev de åtgärder som krävdes för att få bräddningen att upphöra. Fallet med störst säkerhetsmarginal krävde ett utjämningsmagasin på 142 m3 och att dimensionen på 670 m ledning ökades från 400 mm till 600 mm med en uppskattad åtgärdskostnad på omkring nio miljoner kronor. De åtgärder som krävdes för att undvika bräddning vid torrväder var i alla tre fallen tillräckliga för att både minska bräddningen vid regnväder och för att klimatanpassa ledningsnätet. I de fall som undersöktes kunde ingen trend ses som visade att simulering av dag- och spillvattenavrinning, utan veckovariationer i vattenförbrukningen, skulle ge för låga bräddvolymer. / An increasingly common assignment in hydraulic modeling is to investigate how sewage from new residential and commercial areas affect the status of existing sewer systems. In new areas the sewer systems are constructed to operate separately from storm drains and since new areas often are founded in the outskirt of existing areas the sanitary sewage from these areas must be transported through the older existing sewer system, which can be completely or partly combined. In the district of Hjorthagen in Stockholm 5,000 new apartments are planned to be built and sewage from these must be connected upstream of existing combined sewer system in the area. The purpose of this master thesis was to investigate how combined sewer system is affected by additional sewage load. The aim was to investigate how the choice of design sewage flow affects the measures and costs necessary to get the extended sewer system to function, and to consider the need for further adjustments to future climate change. When calculating volumes of combined sewer overflow the sewage flow usually is simulated as discharge from average water use. One objective with this thesis was to investigate whether this method gave lower volumes of combined sewer overflow as compared to using a weekly pattern. By using the modeling program MIKE Urban three choices of design sewage flow, from the new area in Hjorthagen, were investigated. The margin of safety when designing the sewage flow was increased in each case. The results showed that the capacity of the combined sewer system was exceeded during dry weather, with overflow as a result. The investigation showed that the higher margin of safety used the more extensive measures was needed. The greatest margin of safety required a storage basin of 142 m3 and that the dimension of 670 m of pipes was increased. To stop the overflow at dry weather the measures required in each case were sufficient to both reduce overflow during wet weather and to adjust the sewer system to future climate. Simulating the rainfall runoff and sewage flow, without weekly variations in water consumption did not underestimate the volumes of combined sewer overflow compared to varying the water use.

Antibiotics have been one of the greatest scientific discoveries for humanity. Their great success has been hindered by the increasing of antibiotic resistance events. Conventional wisdom considers antibiotic resistance as a strictly clinical issue. In reality, more and more studies have proven that non-clinical environments, especially aquatic environments, are critical factors for the spread of antibiotic resistance. Antibiotic resistance genes (ARGs) are usually located on mobile genetic elements, which can disseminate among taxonomically unconnected species. Therefore, environmental bacteria can serve as hotspots for the development and dissemination of antibiotic resistance. The aim of this thesis is to determine how human originated pollution affects the antibiotic resistance abundance in the surrounding water environments. The case studies were three example environments, wastewater treatment plants (WWTPs), receiving river bodies and combined sewer overflows (CSOs). Molecular microbiology methods have been applied to analyse the resistance levels of bacterial communities from WWTP, CSOs and wastewater affected environments at different European geographical locations. Additionally, a novel approach consisting in an amplicon sequencing is described, in order to be able to investigate and asses the composition of a significant amount of integron gene cassettes. Findings consistently indicate that effluents originated from WWTPs and CSOs have a significant impact on the levels of ARGs of the receiving water bodies. Moreover, this thesis suggests that gene blaOXA-58, could be utilized as a proxy to investigate the spread of ARGs. Its occurrence has been reported to show, consistently throughout the chapters, lower concentrations upstream, but at higher concentrations in the WWTP effluent, CSOs and downstream of the effluent. In conclusion, although antibiotic resistance genes and integrons are part of the environmental resistome, water environments that are affected by anthropogenic wastewater display high levels of the above-mentioned genetic elements. These findings clearly suggest the need to limit pollution levels, as well as the need to establish a more responsible policy in antibiotic prescriptions. This must take place in order to be able to perform efficient risk assessments and to establish acceptable levels of antibiotic and ARGs pollution.:List of Figures VII List of Tables IX List of Abbreviations X 1. Introduction 11 1.1 Antibiotic resistance: an increasing problem 11 1.2 Intrinsic antibiotic resistance and horizontal gene transfer 13 1.2.1 Transformation 13 1.2.2 Transduction 13 1.2.3 Conjugation 14 1.3 Gene transfer elements 14 1.4 Antibiotic resistance and anthropogenic impact on surrounding environments 16 1.5 Study goals 17 2. Genetic variations of the resistome of wastewater treatment plants driven by the seasonality of antibiotic prescription 18 2.1 Introduction 18 2.2 Materials and methods 19 2.3 Results and discussion 20 3. Assessment of inter-laboratory variations of quantitative analyses of antibiotic resistance genes 27 3.1 Introduction 27 3.2 Materials and methods 27 3.3 Results and discussion 30 3.4 Conclusions 36 4. Antibiotic resistance genes in treated wastewater and in the receiving water bodies: A pan-European survey of urban settings 37 4.1 Introduction 37 4.2 Materials and methods 38 4.3 Results 44 4.4 Discussion 49 4.5 Conclusions 53 5. Analysis of integron gene cassettes composition in treated wastewater 55 5.1 Introduction 55 5.2 Materials and methods 55 VI 5.3 Results 56 5.4 Discussion 57 6. Role of combined sewer overflows in the dissemination of antibiotic resistance in surrounding environments 59 6.1 Introduction 59 6.2 Materials and methods 60 6.3 Results 61 6.4 Discussion 66 7. Closing Conclusions 68 References 70 List of publications 88 Note on the commencement of the doctoral procedure 89 Acknowledgments 90

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L’optimisation du système d’assainissement et la réduction des rejets d’eaux résiduaires urbaines non traitées est devenue un enjeu majeur pour de nombreuses collectivités dans le but d’atteindre les objectifs de qualité des milieux aquatiques fixés par la Directive Cadre européenne sur l’Eau (DCE 2000/60/CE). Pour cela, une parfaite connaissance du système d’assainissement est nécessaire. L’objectif de cette thèse, financée par la CDAPP et l’Agence de l’Eau Adour Garonne, était l’étude de la dynamique du système d’assainissement de la CDAPP et de sa contribution sur les flux de polluants dans le milieu récepteur (le Gave de Pau). La première étape du travail a été consacrée à la caractérisation hydraulique et physicochimique du système d’assainissement par temps sec et par temps de pluie. Une étude hydraulique a été tout d’abord réalisée et a permis de mieux appréhender la dynamique des déversements via les déversoirs d’orage (DO) en fonction de la nature des évènements pluvieux. D’autre part, la caractérisation physico chimique des eaux usées (matières en suspension ou MES, demande chimique en oxygène ou DCO, métaux, hydrocarbures aromatiques polycycliques) a mis en évidence que pour l’ensemble de ces paramètres, une nette augmentation des flux par temps de pluie était observée en particulier en début d’événement (augmentation d’un facteur de 2 à 10). Ce phénomène s’explique par l’apport de polluant par les eaux de ruissellement et l’érosion des dépôts accumulés par temps sec dans les réseaux. Seul l’azote total se comporte différemment puisqu’il est majoritairement présent sous forme dissoute. Afin de mieux appréhender la dynamique des paramètres polluants réglementaires (MES, DCO et azote), un suivi haute fréquence (au pas de temps de cinq minutes) a été mis en place durant un an à l’aide de sondes de turbidité et de conductivité. Ce suivi en continu constitue la troisième partie de ce travail. Des corrélations (r² ≈ 0,9) ont été établies entre d’une part, les paramètres polluants DCO et MES, et la turbidité et d’autre part, entre la conductivité et l’azote total. Ces enregistrements ont permis une meilleure compréhension du fonctionnement du système d’assainissement : mise en évidence du phénomène de first flush, estimation des flux polluants déversés via les DO, étude des phénomènes de stockage dans les réseaux. La dernière partie de cette thèse vise à l’étude de la contribution des rejets d’assainissement dans le milieu récepteur. Elle a permis de démontrer la contribution modérée du rejet de sortie de STEP (entre 1 et 15 %) par temps sec. Par temps de pluie, la contribution du système d’assainissement via les DO est extrêmement variable suivant les conditions hydro-climatiques (de < 1 % à plus de 50 %). Ce travail a permis d’une part d’apporter des résultats utilisables par le gestionnaire de l’assainissement pour optimiser la gestion des eaux usées de la CDAPP. D’autre part, ce travail apporte des résultats plus fondamentaux relatifs à une meilleure connaissance de la dynamique hydrologique et physicochimique des eaux résiduaires urbaines et des polluants associés tels que la mise en évidence, à l’aide d’outils statistiques, des paramètres influençant les déversements et les concentrations en polluants par temps de pluie. / Optimization of sewer system and reduction of untreated wastewater discharges has become a key issue for many communities in order to achieve the good quality of aquatic environments set by the European Framework Directive (WFD 2000/60/EC). For this, a perfect knowledge of sanitation is required. The objective of this thesis, funded by CDAPP and Adour Garonne Water Agency, was the study of the dynamics of the CDAPP sanitation and its contribution to the pollutants fluxes in the receiving environment (the river Gave de Pau). The first step of the work was devoted to the hydraulic and physicochemical wastewater characterization during dry and wet weather. A hydraulic study was first carried and helped to better understand the dynamics of discharges through the combined sewer overflow (CSO) according to the rainfall events characteristics. On the other hand, the physico-chemical characterization of wastewater (suspended solids or TSS, chemical oxygen demand or COD, metals, polycyclic aromatic hydrocarbons) showed that for all these parameters, a clear increase of pollutant fluxes was observed at the beginning of the rainfall event (increase by a factor of 2 to 10). This phenomenon is explained by the runoff contribution and erosion of sediments accumulated in the networks during dry weather periods. Only total nitrogen behaves differently because it’s mostly dissolved. To better understand the dynamics of pollutants parameters (TSS, COD and nitrogen), high frequency monitoring (every five minutes) has been established for one year with turbidity and conductivity sensors. This continuous monitoring is the third part of this work. Correlation functions (r² ≈ 0.9) were found between, the pollutant parameters COD and TSS, and turbidity, and secondly, between conductivity and total nitrogen. These records allowed a better understanding of sanitation system: highlighting the first flush phenomenon, estimation of pollutant loads discharged by CSO, study of storage networks phenomenon. The last part of this thesis aims to study the contribution of wastewater discharges to the receiving environment. It demonstrated the moderate contribution of rejection output STEP (between 1% and 15%) in dry weather. In rainy weather, the contribution of sanitation through CSO is extremely variable depending on the hydro-climatic conditions (<1% to over than 50%). This work has led to provide usable results for the sanitation manager to optimize CDAPP wastewater treatment. Moreover, this work provides most fundamental results for a better understanding of the hydrological and physicochemical dynamics of urban wastewater and associated pollutants such as highlighting, using statistical tools, the parameters influencing pollutant concentrations during rainfall events.

Urban waste water collecting systems are designed to convey domestic, industrial and storm water. When sizing sewer network, heavy rainfall must be considered to provide the needed hydraulic capacity for collection. Maintenance is also required in order to avoid anomalies such as inflow, infiltration and unusual polluted discharges from Combined Sewer Overflows (CSOs). Inflow and infiltration decrease the treatment yield at the Waste water Treatment Plant (WWTP) and participate in hydraulic overloads and overflows. CSOs have a direct impact on the pollution of water bodies and must be strictly sized and monitored. Detecting sizing and maintenance anomalies is crucial to ensure public health and a good status of our natural environment. Today, numerical hydraulic models support consulting engineers in assessing overflows then in choosing the best technical and cost-effective scenario. The objectives of this paper are to review the dysfunctions of collecting system and to understand how a numerical hydraulic model is constructed, calibrated and then used to establish a Corrective Action Plan (CAP). My master thesis is based on a working project achieved at SAFEGE (Group SUEZ Environnement) in the Urban Hydraulics Department in Lyon (Rhône-Alpes, France) between September 2013 and February 2014. The paper will present some outcomes obtained from an Urban Hydraulic Project at Chazelles-sur-Lyon (Rhône-Alpes, France) in the South West of Lyon. Mike Urban is the software - developed by the company DHI Water- used for the numerical hydraulic modeling.