The aim of this thesis was to test and evaluate whether parameter regionalization of a hydrological model can be used to model the impact of forest clearcutting on streamflow in Sweden. This is an important task to be able to perform water management and impact assessments adequately. The HBV conceptual rainfall-runoff model was applied for 218 Swedish catchments of different sizes that were spread across the country and covered a wide range of different forest cover percentages. The modelling approach included calibration of the model for each catchment using a genetic algorithm and then associating the resulting optimal parameter values with the percentage of forest cover. The obtained relationship between different model parameters and forest cover was validated with help of a paired catchment study site in northern Sweden where a clear cut was done in 2006: calibrated optimal parameter sets of pre- and post-clearcutting conditions were compared to parameter sets obtained from the Sweden-wide analysis. Correlations were found for about half of the fifteen hydrological model parameters, but the validation with the paired catchment study site could only partially confirm these obtained relationships. The results suggest that the adopted parameter regionalization approach is too basic. However, some of the results seem promising and emphasize the need for further research and development of the approach to provide a more reasonable method to model the impact of forest clearcutting on streamflow. / Det huvudsakliga målet med detta examensarbete var att testa och utvärdera om parameterregionalisering av en hydrologisk modell kan vara en lämplig metod för att modellera och kvantifiera påverkan från skogsavverkning på vattenbalansen i Sverige. Detta är en viktig uppgift för att kunna hantera våra vattenresurser och utföra konsekvensanalyser på ett tillfredsställande sätt. En konceptuell hydrologisk modell tillämpades på 218 avrinningsområden av olika storlekar och som var geografiskt utspridda i hela Sverige där även andelen skog i avrinningsområdena hade ett brett spektrum. Den använda modelleringsmetoden innefattade kalibrering av varje avrinningsområde genom att använda en genetisk algoritm, varefter de optimala parametervärdeana korrelerades mot andelen skog i avrinningsområdet. Idén med denna metod är att använda dessa potentiella samband för att justera modellparametrarna och därmed simulera en skogsavverkning. De erhållna sambanden mellan modellparametrarna och skogstäcket validerades med hjälp av data från en försöksstudie i norra Sverige där en skogsavverkning gjordes under 2006. Skillnaden mellan de bäst fungerande parametervärdena före och efter skogsavverkningen jämfördes med de tidigare sambanden från andra avrinningsområden i Sverige. Signifikant korrelation hittades för ungefär hälften av de 15 hydrologiska modellparametrarna, men valideringen mot den riktiga skogsavverkningen kunde bara delvis bekräfta de erhållna sambanden. Resultaten visar att detta sätt att använda parameterregionalisering antagligen är för grundläggande. Vissa resultat är ändå lovande och fortsatt forskning och utvidgning av metoden är nödvändig för att kunna tillhandahålla en rimlig metod för att kvantifiera en skogsavverknings effekter på vattenbalansen.

Eine Monotherapie mit Tenofovir disoproxil fumarate (TDF) stellt eine hoch effiziente Therapie-option für multipel vorbehandelte Patienten mit chronischer Hepatitis-B-Virusinfektion (HBV) dar. Eine Resistenz gegen TDF wurde bislang nicht beschrieben, jedoch wird ein möglicher negativer Einfluss von Adefovir dipivoxil (ADV)-Resistenzvarianten auf die TDF-Ansprechrate diskutiert. Diese retrospektive Kohortenstudie untersucht die Dynamik von Nukleos(t)id-Analoga (NA)-Resistenzvarianten im HBV-Polymerasegen mit Fokus auf ADV-Resistenzvarianten bei 18 chronisch HBV-infizierten Patienten mit Therapieversagen auf eine vorangegangene Lamivudin (LAM)- und ADV-Therapie, sowie nur partiellem Therapieansprechen auf eine TDF-Monotherapie. Zur Detektion von NA-Resistenzvarianten wird eine HBV-Genomsequenzierung mit Tiefenpyrosequenzierung (Genome Sequencer FLX, Roche Diagnostics, Germany) (UDPS), direkte Sequenzierung (TRUGENETM HBV Genotyping Kit, OpenGeneTM DNA Sequencing Sys-tem, Siemens Healthcare Diagnostic, USA) (TG) und Line Probe Assay (INNO-LiPa DRv2 und v3, Innogenetics, Belgium) (INNO-LiPA) durchgeführt. Unter TDF kommt es zu einer quantitati-ven Shift zugunsten der ADV-Resistenzvarianten mit konstant bleibendem Anteil und deutlich höher persistierender Virämie zu Monat 12 im Vergleich zu Patienten ohne ADV-Resistenzvarianten. Vor allem werden die Varianten rtA181V und rtN236T selektiert, jedoch nicht die Variante rtA181T. Die absolute Anzahl der LAM-Resistenzvarianten hingegen halbiert sich. Varianten mit einem initial per UDPS detektierten Anteil von >20% der patientenspezifi-schen HBV-Population werden meist selektiert und nehmen im Verlauf den Hauptanteil der Quasispezies ein. UDPS stellte ein potentes Medium der Detektion, Identifikation und Quantifi-zierung von HBV-Varianten dar und ist INNO-LiPa und TG überlegen. Es ergibt sich kein Hin-weis auf TDF-Resistenzvarianten, jedoch zeigt das Vorliegen von ADV-Resistenzvarianten ei-nen tendentiell negativen Einfluss auf die virale Kinetik. Weitere größere Langzeitstudien sind zur Bestätigung dieser Beobachtung notwendig.:INHALTSVERZEICHNIS 2 ABBILDUNGSVERZEICHNIS 5 TABELLENVERZEICHNIS 6 1 BIBLIOGRAPHISCHE ZUSAMMENFASSUNG 8 ABKÜRZUNGSVERZEICHNIS 9 2 EINFÜHRUNG 10 2.1 Epidemiologie der chronischen Hepatitis-B-Virusinfektion 10 2.2 Aufbau, Replikation und Resistenzentwicklung des Hepatitis-B-Virusgenoms 10 2.3 Antivirale Therapie 13 2.4 Sequenziermethoden 14 3 AUFGABENSTELLUNG 15 4 MATERIAL UND METHODE 16 4.1 Studiendesign und Beschreibung der Kohorte 16 4.2 Evaluation der TDF-Monotherapie und Resistenzanalyse 17 4.3 Statistische Auswertung 18 4.4 Verbrauchsmaterialien und Reagenzien 18 4.5 Puffer 22 4.6 Geräte 22 4.7 Durchführung der Laborarbeiten 24 4.8 HBV-DNA Quantifizierung und Bestimmung biochemischer Parameter 24 4.9 Extraktion von Nukleinsäuren aus Serumproben 24 4.10 Tiefenpyrosequenzierung mittels Genome Sequencer FLX System (454 Life Science, Roche Diagnostic, Branford, CT) 25 4.10.1 GS FLX HBV-DNA-Library 25 4.10.2 GS FLX Emulsions-PCR 31 4.10.3 GS FLX Sequenzierung 37 4.10.4 GS FLX Datenauswertung 41 4.11 Direkte Sequenzierung mittels TRUGENETM HBV Genotyping Kit (OpenGeneTM DNA Sequencing System, Siemens Healthcare Diagnostic, USA) 42 4.11.1 PCR-Amplifikation 42 4.11.2 CLIP-Amplifikations-Reaktion 42 4.11.3 Genotyp-und Variantenanalyse 44 4.12 Sequenzierung mittels Line Probe Assay INNO-LiPa HBV DRv2 und v3 (Innogenetics, Belgium) 44 4.12.1 HBV-DNA Amplifikation 45 4.12.2 Denaturierung und Hybridisierung 46 4.12.3 Farbentwicklung 46 5 ERGEBNISSE 47 5.1 Patientencharakteristika zur Baseline 47 5.2 Virologisches Ansprechen 48 5.3 Biochemisches Ansprechen 49 5.4 Serologisches Ansprechen 50 5.5 Therapie-Adhärenz und medikamentöse Verträglichkeit 50 5.6 Ergebnisse der Tiefenpyrosequenzierung mit Genome Sequencer FLX System (454 Life Science, Roche Diagnostic, Branford, CT) 50 5.6.1 Verschiedene Einzelverläufe der NA-Resistenzvarianten 55 5.6.2 Kombiniert oder isoliert auftretenden NA-Resistenzvarianten 57 5.6.3 Entwicklung der ADV-Resistenzvarianten 61 5.6.4 Entwicklung der LAM-Resistenzvarianten 66 5.6.5 Entwicklung der ETV-Resistenzvarianten 68 5.6.6 Shift der NA-Resistenzvarianten und Auswirkung auf die Dynamik der HBV-DNA 70 5.6.7 Entwicklung der potentiellen NA-Resistenzvarianten 72 5.6.8 Entwicklung der HBsAg-Varianten 75 5.6.9 Entwicklung der HBV-Quasispezies 77 5.7 Ergebnisse der direkten Sequenzierung mit TRUGENETM HBV Genotyping Kit (OpenGeneTM DNA Sequencing System, Siemens Healthcare Diagnostic, USA) 80 5.8 Ergebnisse des Line Probe Assays mit INNO-LiPa HBV DRv2 und v3 (Innogenetics, Belgium) 81 5.9 Vergleich der Sequenziermethoden 81 6 DISKUSSION 83 6.1 Patientenkohorte und Ansprechen auf die TDF-Monotherapie 83 6.2 Entwicklung und Einfluss der ADV-Resistenzvarianten unter TDF-Monotherapie 84 6.3 Entwicklung und Einfluss der LAM-Resistenzvarianten unter TDF-Monotherapie 90 6.4 Entwicklung und Einfluss der ETV-Resistenzvarianten unter TDF-Monotherapie 92 6.5 Entwicklung und Einfluss der HBV-Wildtyp-Varianten unter TDF-Monotherapie 93 6.6 Entwicklung und Einfluss der potentiellen NA-Resistenzvarianten 94 6.7 Entwicklung und Einfluss der HBsAg-Varianten 96 6.8 Einfluss von multiplen Vortherapien unter TDF-Monotherapie 98 6.9 Entwicklung und Einfluss der HBV-DNA-Serumkonzentration 98 6.10 Entwicklung und Einfluss von HBV-Quasispezies unter TDF-Monotherapie 100 6.11 Vergleich der Sequenziermethoden 101 7 ZUSAMMENFASSUNG DER ARBEIT 106 8 LITERATURVERZEICHNIS 109 9 EIDESSTATTLICHE VERSICHERUNG 114 10 CURRICULUM VITAE 115 11 ANTEILSERKLÄRUNG AN ERFOLGTEN PUBLIKATIONEN 116 12 DANKSAGUNG 117 / Tenofovir disoproxil fumarate (TDF) is a highly efficient treatment option for nucleos(t)ide analogue (NA) pre-treated patients with chronic hepatitis B virus (HBV) infection. Little is known about the reasons for persistent virus replication in some rare cases. As of today, no TDF resistance variants have been identified, but a possible linkage to Adefovir dipivoxil (ADV) resistance associated variants negatively influencing HBV-DNA suppression by TDF has been suspected, based on the similarity of the chemical structure. In this retrospective cohort study the dynamics of NA resistance variants in the HBV polymerase gene with focus on ADV resistance variants were assessed. For this, we have chosen a cohort including patients with multiple failures to treatment with different NAs. Thus, data of 18 patients with previous treatment failure to LAM and ADV was analysed, showing a persistent viremia (HBV-DNA >35 copies/mL) despite switch to TDF monotherapy (median HBV-DNA at month 12 3,5±0,8 (2,1-4,9) log10 copies/mL). Sequencing analysis was performed with ultra-deep pyrosequencing (UDPS) (Genome Sequencer FLX, 454 Life Science, Roche Diagnostic, Branford, CT), direct sequencing (TG) (TRUGENETM HBV Genotyping Kit, OpenGeneTM DNA Sequencing System, Siemens Healthcare Diagnostic, USA) and line probe assay (INNO-LiPA) (INNO-LiPa DRv2/v3, Innogenetics, Belgium). Using TDF monotherapy, a quantitative shift in favour to ADV resistance variants was observed in this cohort. The percentage of substitutions conferring resistance to ADV at baseline (BL) and at the time of the last sequencing endpoint (EP) of the HBV genome remained constant (BL 35%, 13/37, EP 36%, 9/25). The variants rtA181V and rtN236T were mostly selected, whereas rtA181T was not selected. The total amount of substitutions conferring resistance to Lamivudin (LAM) showed a strong decline, however remained the majority part of all NA resistance variants (BL 51% (19/37), EP 40% (10/25)). The percentage of ETV resistance variants increased slightly (BL 14% (5/37), EP 24% (6/25)). Known ADV, Lam and ETV resistance variants emerged in variable abundance (1,0-99,6%) of quasispecies during TDF therapy. A homogenization of HBV quasispecies took place. Especially mutations occurring in higher abundance (>20% of viral population) were mostly selected (BL 51% (19/37), EP 80% (20/25)). No new HBV variants with possible association to resistance against TDF were identified, but patients with ADV resistance variants showed the highest HBV-DNA level at month 12 of TDF therapy (median HBV-DNA 3,57±0,72 (2,14-3,96) log10 copies/mL, not significant). A negative influence of ADV resistance variants on viral suppression with TDF monotherapy may be assumed, however more long-term studies are needed to confirm the role of ADV resistance variants in TDF therapy. UDPS is a potent medium for detection, identification and quantification of dominant to low level variants in HBV-DNA. It is superior to direct sequencing and line probe assay in the detection of variants.:INHALTSVERZEICHNIS 2 ABBILDUNGSVERZEICHNIS 5 TABELLENVERZEICHNIS 6 1 BIBLIOGRAPHISCHE ZUSAMMENFASSUNG 8 ABKÜRZUNGSVERZEICHNIS 9 2 EINFÜHRUNG 10 2.1 Epidemiologie der chronischen Hepatitis-B-Virusinfektion 10 2.2 Aufbau, Replikation und Resistenzentwicklung des Hepatitis-B-Virusgenoms 10 2.3 Antivirale Therapie 13 2.4 Sequenziermethoden 14 3 AUFGABENSTELLUNG 15 4 MATERIAL UND METHODE 16 4.1 Studiendesign und Beschreibung der Kohorte 16 4.2 Evaluation der TDF-Monotherapie und Resistenzanalyse 17 4.3 Statistische Auswertung 18 4.4 Verbrauchsmaterialien und Reagenzien 18 4.5 Puffer 22 4.6 Geräte 22 4.7 Durchführung der Laborarbeiten 24 4.8 HBV-DNA Quantifizierung und Bestimmung biochemischer Parameter 24 4.9 Extraktion von Nukleinsäuren aus Serumproben 24 4.10 Tiefenpyrosequenzierung mittels Genome Sequencer FLX System (454 Life Science, Roche Diagnostic, Branford, CT) 25 4.10.1 GS FLX HBV-DNA-Library 25 4.10.2 GS FLX Emulsions-PCR 31 4.10.3 GS FLX Sequenzierung 37 4.10.4 GS FLX Datenauswertung 41 4.11 Direkte Sequenzierung mittels TRUGENETM HBV Genotyping Kit (OpenGeneTM DNA Sequencing System, Siemens Healthcare Diagnostic, USA) 42 4.11.1 PCR-Amplifikation 42 4.11.2 CLIP-Amplifikations-Reaktion 42 4.11.3 Genotyp-und Variantenanalyse 44 4.12 Sequenzierung mittels Line Probe Assay INNO-LiPa HBV DRv2 und v3 (Innogenetics, Belgium) 44 4.12.1 HBV-DNA Amplifikation 45 4.12.2 Denaturierung und Hybridisierung 46 4.12.3 Farbentwicklung 46 5 ERGEBNISSE 47 5.1 Patientencharakteristika zur Baseline 47 5.2 Virologisches Ansprechen 48 5.3 Biochemisches Ansprechen 49 5.4 Serologisches Ansprechen 50 5.5 Therapie-Adhärenz und medikamentöse Verträglichkeit 50 5.6 Ergebnisse der Tiefenpyrosequenzierung mit Genome Sequencer FLX System (454 Life Science, Roche Diagnostic, Branford, CT) 50 5.6.1 Verschiedene Einzelverläufe der NA-Resistenzvarianten 55 5.6.2 Kombiniert oder isoliert auftretenden NA-Resistenzvarianten 57 5.6.3 Entwicklung der ADV-Resistenzvarianten 61 5.6.4 Entwicklung der LAM-Resistenzvarianten 66 5.6.5 Entwicklung der ETV-Resistenzvarianten 68 5.6.6 Shift der NA-Resistenzvarianten und Auswirkung auf die Dynamik der HBV-DNA 70 5.6.7 Entwicklung der potentiellen NA-Resistenzvarianten 72 5.6.8 Entwicklung der HBsAg-Varianten 75 5.6.9 Entwicklung der HBV-Quasispezies 77 5.7 Ergebnisse der direkten Sequenzierung mit TRUGENETM HBV Genotyping Kit (OpenGeneTM DNA Sequencing System, Siemens Healthcare Diagnostic, USA) 80 5.8 Ergebnisse des Line Probe Assays mit INNO-LiPa HBV DRv2 und v3 (Innogenetics, Belgium) 81 5.9 Vergleich der Sequenziermethoden 81 6 DISKUSSION 83 6.1 Patientenkohorte und Ansprechen auf die TDF-Monotherapie 83 6.2 Entwicklung und Einfluss der ADV-Resistenzvarianten unter TDF-Monotherapie 84 6.3 Entwicklung und Einfluss der LAM-Resistenzvarianten unter TDF-Monotherapie 90 6.4 Entwicklung und Einfluss der ETV-Resistenzvarianten unter TDF-Monotherapie 92 6.5 Entwicklung und Einfluss der HBV-Wildtyp-Varianten unter TDF-Monotherapie 93 6.6 Entwicklung und Einfluss der potentiellen NA-Resistenzvarianten 94 6.7 Entwicklung und Einfluss der HBsAg-Varianten 96 6.8 Einfluss von multiplen Vortherapien unter TDF-Monotherapie 98 6.9 Entwicklung und Einfluss der HBV-DNA-Serumkonzentration 98 6.10 Entwicklung und Einfluss von HBV-Quasispezies unter TDF-Monotherapie 100 6.11 Vergleich der Sequenziermethoden 101 7 ZUSAMMENFASSUNG DER ARBEIT 106 8 LITERATURVERZEICHNIS 109 9 EIDESSTATTLICHE VERSICHERUNG 114 10 CURRICULUM VITAE 115 11 ANTEILSERKLÄRUNG AN ERFOLGTEN PUBLIKATIONEN 116 12 DANKSAGUNG 117

The Håga river, located west of the Swedish city Uppsala, is currently without a proper gauging station. Knowing the streamflow is nonetheless important to, for example, be able to calculate the nutrient transport in the river. This project aimed to evaluate different indirect methods of streamflow estimation to investigate how they perform, in particular in relation to SMHI’s S-HYPE model. Two of the methods used were based on transferring streamflow of nearby catchments to Håga, either by using relationships between the mean and standard deviation of the streamflow time series (MOVE), or by simply scaling relative to catchment size (DAR). Furthermore, a hydrological model, HBV, was calibrated for Håga using different amounts and types of calibration data. All the methods were then compared to streamflow data from a previously active gauging station in Håga.  It was found that the overall best method to estimate the streamflow in Håga was using the MOVE method with one particular donor catchment. However, the performance of the simpler MOVE and DAR methods varied a lot from catchment to catchment. HBV was found to be able to produce better performing simulations than S-HYPE, despite being a simpler model. Even HBV-calibrations using alternative or limited data could perform rather well, although rarely at the level of a calibration utilising all available streamflow data. A big uncertainty of the study was the fact that the most recent available validation data for the Håga catchment was from two decades ago, when the old gauging station was decommissioned. Most likely the methods that worked well during the 90s would work well today as well, but this is a matter that could be studied further. / Hågaån, ett vattendrag som ligger väster om Uppsala, saknar i nuläget en mätstation för vattenföring. Att känna till flödet är dock ändå intressant, bland annat för att kunna beräkna näringstransporten i ån. Syftet med detta projekt var därmed att utvärdera och jämföra olika metoder för att uppskatta vattenflödet i Hågaån, särskilt för att undersöka hur de presterade i jämförelse med SMHI:s S-HYPE-modell. Två av metoderna som användes för detta baserades på att överföra flöden från närliggande vattendrag till Håga, antingen genom att använda förhållanden mellan medelvärde och standardavvikelse för flödes-datan (MOVE), eller genom att bara utgå från skillnader i områdenas storlek (DAR). Utöver det kalibrerades även den hydrologiska modellen HBV för Håga med olika typer och mängder av kalibreringsdata. Alla metoderna jämfördes sedan med data från en mätstation som tidigare funnits i Hågaån. Resultaten visade att den över lag bästa metoden för att uppskatta flödet i Håga var MOVE-metoden i kombination med ett av de närliggande vattendragen. Hur väl dessa simplare MOVE- och DAR-metoder presterade varierade dock mycket beroende på vilket vattendrag som användes som donator. Det visade sig även att det gick att erhålla bättre resultat med HBV än de som gavs av S-HYPE, trots att HBV är en enklare modell. Även HBV-kalibreringar baserade på alternativ eller begränsad data kunde producera välpresterande simulationer, dock sällan på samma nivå som den kalibrering som använt all tillgänglig flödesdata. En stor osäkerhet i projektet kretsar kring att den nyaste tillgängliga valideringsdatan från Hågaån var över två decennier gammal, då den mätstation som funnits stängdes ner. Med stor sannolikhet kommer metoderna som fungerade väl under 90-talet även fungera bra i modern tid, men detta är något som kräver vidare studier.

Paramos are unique ecosystems situated in the tropical Andes at an elevation of 2800-4800 meters above sea level. They are home to a variety of endemic species and have a great water regulating capacity and supply around 80 % of the population in Colombia with freshwater. These ecosystems have been found to be particularly vulnerable towards climate change, which is a major challenge when it comes to safeguarding this water source. This study aimed to investigate paramos on three spatial scales: the large, medium and small scale. The large scale aimed at investigating the paramos of Colombia on a national level, the medium scale aimed at modeling the hydrology of a small paramo catchment, and the small scale aimed at exploring the general landscape and methods for monitoring the hydroclimate. At the national scale, the paramos were characterized based on hydrological and climatological variables, in the end focusing on precipitation and discharge, and producing descriptive statistics, in addition to finding temporal trends. The aim was to describe the paramos of Colombia hydroclimatologically on a spatial and temporal scale to get a more updated and comprehensive picture of the hydrology in these ecosystems and to investigate whether any possible relationships between the hydroclimatic variables could be detected that could affect the water availability, that is, if there were any signs of climatic forcing and hydrological response. Another aim was to investigate whether any significant trends could be seen in historical data and if there were any signs of climate change or ENSO (El Niño Southern Oscillation) events affecting the time series. The results showed that there was a large range in the amount of annual precipitation, between 500 and 5000 mm/yr, depending on the location of the precipitation station. The annual precipitation differed with up to around 2000 and 3500 mm/yr within and between different paramo regions, respectively. The annual mean daily discharge ranged between 0 and 25-30 m3/s and the total annual specific discharge ranged between 0 and 2500 mm/yr. It was not possible to detect any general, common temporal trend in neither precipitation nor discharge for all of the stations and the majority of the trends for individual stations were not significant. However, most locations that did present significant trends in precipitation showed an increase rather than a decrease, on a monthly rather than an annual scale. The trends that were significant for discharge were also increasing. No evident relationship between precipitation and discharge could be detected in neither amount nor pattern or trend over time, although some similarities and common traits could be found. Some local maxima and minima in precipitation and discharge coincided with ENSO events, but whether the significant trends are due to climate change remains unclear. The changes that were seen could have an impact on societies that are close to and rely on the paramos, but more studies on a seasonal and monthly scale are needed. At the medium scale, the HBV light model was used to simulate the hydrology in the small paramo catchment called Monterredondo. The aim was to investigate how a parsimonious model like the HBV model would perform compared to the more comprehensive SWAT model, which previously succeeded in modeling the hydrology in this watershed. Since the data is scarce in these regions, a simpler model could be more desirable. The results showed that the HBV was sufficient for the task, receiving a NSE value of 0.81 (calibration period) and 0.56 (validation period), and able to perform almost as well as the SWAT model, which received NSE values of 0.83 (calibration period) and 0.59 (validation period). Furthermore, since the paramos are vulnerable towards climate change, it was considered important to investigate how discharge might be affected by climate change. Simulations of future climate towards the end of the century, representing two emission scenarios, SSP2-4.5 and SSP5-8.5, were performed with data from the general circulation models EC-Earth3-CC (Europe) and MRI-ESM2-0 (Japan). The results from the EC-Earth3-CC model indicate a change towards drier dry seasons and wetter wet seasons. On an annual scale, it shows a decrease in discharge, but no change in annual precipitation. The MRI-ESM2-0 model was not considered reliable due to the large distance between the catchment area in question and the location where the data came from. Large uncertainties are associated with the data quality and quantity for both parts of this study. This is a well-known issue in these quite unapproachable locations, which is why the small scale investigation focused on learning more about the monitoring of these ecosystems. A horizontal precipitation station was built to capture horizontal precipitation. Even if built as a rather rough structure, it seemed to work and could be useful for future investigations. The results showed that horizontal precipitation contributed with a non-negligible amount to the total precipitation.  In summary, there is a risk that climate changes will have an impact on the water resources, due to drier dry seasons and wetter wet seasons. This could lead to water shortages in periods, but more studies are necessary to confirm the result. It is especially important to investigate more variables such as land use, soil moisture and radiation. It is also important to study more paramo areas in detail since they respond differently towards climate change. / Paramos är unika ekosystem som ligger på en höjd av 2800-4800 meter över havet i de tropiska Anderna. De är hem för en mängd olika endemiska arter och har en stor vattenreglerande förmåga och försörjer cirka 80 % av befolkningen i Colombia med dricksvatten. Dessa ekosystem har visat sig vara särskilt sårbara för klimatförändringar, vilket innebär en stor utmaning när det kommer till att skydda paramos som vattenkälla. Denna studie syftade till att undersöka paramos på tre rumsliga skalor: stor, medelstor och liten skala. Den stora skalan syftar till att undersöka paramos i Colombia på nationell nivå, den medelstora skalan syftar till att modellera hydrologin i ett litet paramoavrinningsområde, och den lilla skalan syftar till att utforska det allmänna landskapet och metoder för att övervaka hydroklimatologin. På nationell skala karaktäriserades paramos utifrån hydrologiska och klimatologiska variabler, i slutändan med fokus på nederbörd och avrinning, genom att producera beskrivande statistik, och därtill hitta trender över tid. Syftet var att beskriva paramos i Colombia hydroklimatologiskt både rumsligt och tidsmässigt för att få en mer uppdaterad och heltäckande bild av hydrologin i dessa ekosystem och för att undersöka om möjliga samband mellan de hydroklimatologiska variablerna kunde upptäckas som skulle kunna påverka vattentillgången, det vill säga om det fanns några tecken på klimatpåverkan och hydrologisk respons. Ett annat syfte var att undersöka om statistiskt signifikanta trender kunde ses i historiska data och om det fanns tecken på att klimatförändringar eller ENSO-händelser (El Niño Southern Oscillation) påverkade tidsserierna. Resultatet visade att det fanns ett stort spann i uppmätt årsnederbörd, mellan 500 och 5000 mm/år, beroende på nederbördsstationens lokalisering. Årsnederbörden skiljde sig med upp till cirka 2000 och 3500 mm/år, inom respektive mellan olika paramoregioner. Det årliga medeldygnsflödet låg mellan 0 och 25-30 m3/s och det totala årliga specifika flödet låg mellan 0 och 2500 mm/år. Det var inte möjligt att upptäcka någon generell, gemensam tidsmässig trend i varken nederbörd eller avrinning för samtliga stationer och majoriteten av trenderna för enskilda stationer var inte signifikanta. De flesta platser som faktiskt visade signifikanta trender i nederbörd visade en ökning snarare än en minskning, på månadsbasis snarare än årsbasis. Trenderna som var signifikanta för avrinning visade också en ökning. Inget uppenbart samband mellan nederbörd och avrinning kunde ses i varken kvantitet, mönster eller trend över tid, även om vissa likheter och gemensamma egenskaper kunde hittas. Vissa lokala maxima och minima i nederbörd sammanföll med ENSO-händelser, men huruvida de signifikanta trenderna beror på klimatförändringar är fortfarande oklart. De förändringar som kunde identifieras kan ha en inverkan på samhällen som är nära och förlitar sig på paramos, men fler studier på säsongs- och månadsskala behövs.  På den medelstora skalan användes HBV-lightmodellen för att simulera hydrologin i ett litet paramoavrinningsområde kallat Monterredondo. Syftet var att undersöka hur en mindre komplex modell som HBV-modellen kunde prestera jämfört med den mer omfattande SWAT-modellen, som tidigare lyckats modellera hydrologin i samma avrinningsområde. Eftersom mängden data i dessa regioner är begränsad kan en mindre komplex modell vara att föredra. Resultaten visade att HBV presterade tillräckligt bra för uppgiften då modellen erhöll ett NSE-värde på 0,81 (kalibreringsperioden) och 0,56 (valideringsperioden) och kunde prestera nästan lika bra som SWAT-modellen, som erhöll NSE-värden på 0,83 (kalibreringsperioden) och 0,59 (valideringsperioden). Då paramos är sårbara för klimatförändringar ansågs det dessutom viktigt att undersöka hur avrinningen kan påverkas av klimatförändringar. Simuleringar för framtida klimat motsvarande slutet av seklet modellerades med två utsläppsscenarier, SSP2-4.5 och SSP5-8.5. De representerades med data från de allmänna cirkulationsmodellerna EC-Earth3-CC (Europa)och MRI-ESM2-0 (Japan). Resultaten från EC-Earth3-CC-modellen indikerade en förändring mot torrare torrperioder och blötare regnperioder. På en årsskala visade modellen en minskning i avrinning, men ingen förändring i årsnederbörd. MRI-ESM2-0-modellen bedömdes inte vara tillförlitlig på grund av det stora avståndet mellan det aktuella avrinningssområdet och platsen där datan kom ifrån. Stora osäkerheter är förknippade med datakvalitet och kvantitet för båda delarna av denna studie. Detta är ett välkänt problem på dessa ganska otillgängliga platser, varför den småskaliga utredningen fokuserade på att öka förståelsen för den hydroklimatologiska övervakningen i dessa ekosystem. En horisontell nederbördsstation byggdes för att fånga horisontell nederbörd. Även om den byggdes med en ganska grov struktur, verkade den fungera och kan potentiellt vara användbar för framtida undersökningar. Resultaten visade att horisontell nederbörd bidrog med en icke-försumbar mängd till den totala nederbörden. Sammanfattningsvis finns det en risk att klimatförändringarna påverkar vattenresurserna, på grund av att torrperioderna blir torrare och regnperioderna blir blötare. Detta kan leda till en periodvis vattenbrist, men fler studier krävs för att bekräfta resultatet. Det är särskilt viktigt att undersöka fler variabler såsom markanvändning, markfuktighet och strålning. Det är också viktigt att studera fler paramoområden i detalj, eftersom de reagerar olika på klimatförändringar.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-08-22T19:48:35Z No. of bitstreams: 1 FranciscoMagalhaes.pdf: 1065814 bytes, checksum: ed97c291154490e367bbb044c576aefa (MD5) / Made available in DSpace on 2017-08-22T19:48:35Z (GMT). No. of bitstreams: 1 FranciscoMagalhaes.pdf: 1065814 bytes, checksum: ed97c291154490e367bbb044c576aefa (MD5) Previous issue date: 2015-06-24 / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) / Hepatitis B virus (HBV) chronic infection has a heterogeneous distribution around the world. Its clinical presentation depends on factors of the carrier such as the age at which he got infected, immunological status and associated diseases. Viral characteristics like the genotype can also influence. The aim of this study was to evaluate the influence of the HBV genotype in the clinical presentation of the chronic infection. This study was performed at the Clinical Research Center and the Liver Unit of the Federal University of Maranhão, with 119 chronic carriers of HBV with defined genotypes. We identified those followed for at least 12 months for their clinical definition. It was included 101 patients: 68 (68%) with genotype A1, 26 (26%) D4, 4(3%) F2a, 2 (2%) D3 and 1 (1%) D2. For the purpose of comparison, two groups were defined: A (n=68) and D (n=29). In the HBV-D group the presence of HBeAg (28% vs 10% P=0.03) and immune tolerant (21.5% vs 1.5% P=0.006) patients were more frequent when compared to carriers of genotype A. There was no age difference between groups (39±13 vs 42±11 P=0.26). In conclusion, we can suggest that among the carriers of genotype D, especially the subgenotype D4, which corresponds to 90% of this, there might be late seroconversion of HBeAg, favoring a higher risk of virus transmission. / A infecção crônica pelo vírus da hepatite B tem distribuição heterogênea em todo o mundo. Sua apresentação clínica depende de fatores do hospedeiro tais como a idade em que o indivíduo se infectou, estado imunológico e doenças associadas. Características virológicas também podem influenciar tais como o genótipo do vírus. Este estudo teve o objetivo de avaliar a influência do genótipo do HBV na apresentação clínica. A pesquisa foi conduzida no Centro de Pesquisa Clínica e no Núcleo de Estudos do Fígado da Universidade Federal do Maranhão, com 119 portadores crônicos do HBV com genótipos definidos. Foram identificados aqueles acompanhados por pelo menos 12 meses para definição clínica. Incluídos 101 pacientes: 67 (68%) com genótipo A1, 26 (26%) D4, 4 (3%) F2a, 3 (2%) D3 e 01 (1%) D2. Para fins de comparação foram definidos dois grupos: A (n=67) e D (n=30). No grupo HBV-D foram mais frequentes a presença do HBeAg (28% vs 10% P=0.03) e de imunotolerantes (21,5% vs 1.5% / P=0.006) quando comparados com portadores do genótipo A. Não houve diferença entre as idades nos dois grupos (39±13 vs 42±11 / P=0.26). Em conclusão, pode-se sugerir que entre os portadores do genótipo D, especialmente o subgenótipo D4, que corresponde a 90% do total, pode haver soroconversão tardia do HBeAg, favorecendo maior risco de transmissão do vírus.

L'infection chronique ou occulte par le virus de l'hépatite B (HBV) est l'un des facteurs de risque les plusimportants pour le développement de carcinome hépatocellulaire viro-induit. Malgré la petite taille dugénome et les contraintes imposées par l'organisation de ce génome, le HBV présente une grandevariabilité génétique qui est le résultat des erreurs générées lors de l'étape de reverse transcription maisaussi, lors des processus de sélection et d'adaptation. Durant l'infection, une relation étroite hôte-viruss'établit. La population virale est caractérisée par la présence de quasi-espèces pouvant influencerl'évolution de la maladie hépatique et favoriser les phénomènes d'échappement. Le but de notre travail aété 1) d'étudier la variabilité virale associée aux infections occultes et l'impact de certaines modificationsde l'AgHBs (sY100C) sur la reconnaissance des tests de détection enzymatiques, et l'impact des mutations(sK122R, sD144E) sur la reconnaissance des anticorps anti-HBs d'un patient 2) de développer un nouveloutil d'amplification de génomes entiers de HBV que nous avons appliqué à l'étude préliminaire de quasiespècesd'un mutant Core. Cet outil d'amplification est plus sensible que les techniques existantes, et ilpermet l'étude des propriétés moléculaires et fonctionnelles des isolats virales. En conclusion, nous avonspu montrer que les processus d'échappement sont complexes, et font intervenir des facteurs du virus et del'hôte. Ce travail contribue à l'amélioration de la compréhension de la biologie de l'infection par le HBV,et illustre la complexité des interactions hôte-virus.

A changing climate can severely perturb regional hydrology and thereby affect human societies and life in general. To assess and simulate such potential hydrological climate change impacts, hydrological models require reliable meteorological variables for current and future climate conditions. Global climate models (GCMs) provide such information, but their spatial scale is too coarse for regional impact studies. Thus, GCM output needs to be downscaled to a finer scale either through statistical downscaling or through dynamic regional climate models (RCMs). However, even downscaled meteorological variables are often considerably biased and therefore not directly suitable for hydrological impact modeling. This doctoral thesis discusses biases and other challenges related to incorporating climate model output into hydrological studies and evaluates possible strategies to address them. An analysis of possible sources of uncertainty stressed the need for full ensembles approaches, which should become standard practice to obtain robust and meaningful hydrological projections under changing climate conditions. Furthermore, it was shown that substantial biases in current RCM simulations exist and that correcting them is an essential prerequisite for any subsequent impact simulation. Bias correction algorithms considerably improved RCM output and subsequent streamflow simulations under current conditions. In addition, differential split-sample testing was highlighted as a powerful tool for evaluating the transferability of bias correction algorithms to changed conditions. Finally, meaningful projections of future streamflow regimes could be realized by combining a full ensemble approach with bias correction of RCM output: Current flow regimes in Sweden with a snowmelt-driven spring flood in April will likely change to rather damped flow regimes that are dominated by large winter streamflows.

Le portage chronique du virus de l'hépatite B (HBV) en Afrique est très élevé, avoisinant parfois jusqu'à 30% dans certaines régions. Selon l'OMS, sur les 400 millions de personnes souffrant d'une infection chronique HBV, 70 à 140 millions vivent en Afrique avec un taux de décès annuel d'environ 250 000 cas par an. Les données concernant l'infection concomitante par le virus de l'hépatite D (HDV) virus satellite de l'HBV, sont rares car peu d'études ont été réalisées en Afrique. Malgré ce fort taux de prévalence, les données concernant la caractérisation moléculaire des souches HBV et HDV sont limitées ou inexistantes dans la plupart des pays d'Afrique Subsaharienne et en particulier dans région du Sahara, vaste zone multiethnique, de passage et de brassage de populations. Au cours de cette étude, nous avons voulu déterminer la prévalence et l'épidémiologie moléculaire des souches HBV et HDV circulant la région du Sahara (Niger et Mauritanie). Tout d'abord, nous avons étudié une cohorte de donneurs de sang du Niger porteurs de l'AgHBs. Nous avons trouvé que 80% des souches étudiées appartenaient au génotype E. De plus, nous avons identifié et caractérisé un nouveau recombinant HBV/D-HBV/E représentant près de 20% des souches étudiées. Les points de cassure se situaient dans des " points chauds " de recombinaison, régions impliquées dans les événements d'intégration du génome de l'HBV. Des analyses phylogénétiques extensives nous ont permis de le classer comme un nouveau sous génotype. Nous avons proposé HBV/D8. Nous avons par la suite, en collaboration avec des équipes locales, étudié la diversité génétique HBV en Mauritanie, pays voisin du Niger, au sein de différents groupes représentatifs de la population : femmes enceintes (n=1020), consultants (n=954), donneurs de sang (n=11110) et patients suivis pour une infection HBV chronique (n=300). Le taux de portage de l'AgHBs, était de 11 à 18 % selon les populations étudiées, classant la Mauritanie comme pays à haute endémie pour l'HBV. L'exposition à l'HBV était associée en analyse multivariée, au niveau d'éducation, à l'ethnie, à des antécédents de transfusion et à la profession chez les femmes enceintes et, chez les consultants, au sexe masculin. Sur le plan moléculaire, 3 génotypes différents circulaient en Mauritanie (n=240) : l'HBV/D (56,3%), l'HBV/E (34,6%) et l'HBV/A (8,8%). De façon intéressante, 30% des génotypes D circulant en Mauritanie étaient l'HBV/D8. Cette diversité de l'HBV peut être expliquée par la localisation géographique du Niger et de la Mauritanie comme zones de passage entre l'Afrique du Nord et l'Afrique Sub Saharienne où l'HDV/D et l'HBV/E respectivement sont prédominants. D'autre part, 14 à 33% des patients HBV positifs étaient également infectés par l'HDV. La présence d'anticorps anti-Delta était associée en analyse multivariée chez les consultants, à l'âge et au sexe masculin, et chez les donneurs de sang, à l'âge, au nombre de mariages, à la profession (militaire), à la résidence (région du désert) et à des antécédents d'hospitalisation. Sur le plan moléculaire, le génotype HDV-1 est largement majoritaire (90%) mais l'HDV-5 a aussi été isolé (10% des cas). En conclusion, ce travail souligne encore la forte prévalence des hépatites B et Delta au Niger et en Mauritanie. Nous avons aussi mis en évidence une diversité génétique importante des souches circulant et notamment la caractérisation d'un nouveau sous-génotype HBV/D8 hautement prévalent. Il convient d'évaluer la sévérité de la maladie hépatique liée à cette diversité génétique et à ce nouveau variant, notamment son implication éventuelle dans l'oncogenèse hépatique par des événements de recombinaison génétique.

Snowmelt runoff in the mountainous eastern part of Turkey is of great importance as it constitutes 60-70% in volume of the total yearly runoff during spring and early summer months. Therefore, forecasting the amount and timing of snowmelt runoff especially in the Euphrates Basin, where large dams are located, is an important task in order to use the water resources of the country in an optimum manner. The HBV model, being one of the well-known conceptual hydrological models used more than 45 countries over the world, is applied for the first time in Turkey to a small basin of 242 km2 on the headwaters of Euphrates River for 2002-2004 water years. The input data are provided from the automatic snow-meteorological stations installed at various locations and altitudes in Upper Euphrates Basin operating in real-time. Since ground based observations can only represent a small part of the region of interest, spatially and temporally distributed snow cover data are acquired through the use of MODIS optical satellite. Automatic model parameter estimation methods, GML and SCE_UA, are utilized to calibrate the HBV model parameters with a multi-objective criteria using runoff as well as snow covered area to ensure the internal validity of the model and to generate a Pareto front. Model simulations show that the choice of study years and timing of satellite images affect the results and further suggest that more study catchments and years should be included to achieve more comprehensible conclusions. In the second part of the study, the calibrated HBV model is applied to forecast runoff with a 1-day lead time using gridded input data from numerical weather prediction models of ECMWF and MM5 for the 2004 snowmelt period. Promising results indicate the possible operational use of runoff forecasting using numerical weather prediction models in order to prevent or at least take precautions before flooding ahead of time.

Introdução: A vacinação contra a hepatite B, aliada às precauções padrão, é um importante instrumento para a prevenção deste agravo em profissionais de saúde, que podem sofrer exposição a material biológico potencialmente contaminado. Objetivos: Estimar a cobertura da vacinação contra a hepatite B, a prevalência de imunizados e de exposição ocupacional a material biológico por acidentes perfurocortantes nos profissionais de saúde do município de Santa Rosa/RS/Brasil. Metodologia: Foram pesquisados, através de um estudo transversal, 322 profissionais de saúde que atuam em hospitais, no banco de sangue e nas unidades de saúde (Médicos, Enfermeiros, Técnicos e Auxiliares de Enfermagem, Odontólogos, Auxiliares de Consultório Dentário (ACD), Farmacêuticos e Auxiliares de Laboratório). A significância estatística das associações foi avaliada usando-se o teste de qui-quadrado para a comparação de variáveis categóricas, e regressão logística para a análise multivariável. Resultados: A média de idade dos entrevistados foi de 38,1 anos, com média de 13 anos de atuação na profissão. 24,8% eram do sexo masculino e 75,2 do sexo feminino. A cobertura vacinal nestes profissionais foi de 87,9% (IC 95% 83,8 a 91,2%). As coberturas de vacinação foram maiores em profissionais com nível médio e que não atuam em hospitais. A taxa de acidentabilidade com instrumento perfurocortante foi de 40,5% na amostra. A prevalência de imunizados com testagem realizada até 6 meses foi de 78,8%. Conclusão: A cobertura de vacinação e a prevalência de imunizados foram inferiores ao preconizado pelo Ministério da Saúde e Centers for Diseases Control and Prevention. Há necessidade de maiores investimentos na prevenção da infecção ocupacional pelo vírus da hepatite B para esses profissionais. / Introduction: The vaccination against hepatitis B, together with standard precautions, is an important tool for the prevention of this disease in health professionals, who can suffer exposure to biological potentially contaminated. Goals: Estimate the coverage of vaccination against hepatitis B, the prevalence of immunized and the occupational exposure to biological material from perforating injuries in health professionals located in city of Santa Rosa/RS/Brazil. Methodology: 322 health professionals who work in hospitals, in blood bank and in health services (Phisicians, Nurses, Technicians and Assistants in Nursing, Dentists, Dental Office Assistants (DOA), Pharmacists and Laboratory Assistants) were investigated, through an cross-sectional study. The statistic significance of the associations was evaluated by using the chi-square test to compare categorical variables and logistic regression for multivariable analysis. Results: The interviewed are, on average, 38,1 years old, and have been working in the profession for around 13 years. 24,8% werw male and 75,2 female. The vaccination in those professional was 87.9% (IC 95% 83.8 to 91.2%). The vaccination was larger in professionals with high school and that do not work in hospitals. The rate of injury due perforating instrument was 40.5% in the sample. The prevalence on immunized with testing performed up to 6 months was 78.8%. Conclusion: The vaccination and the prevalence of immunization were below the recommended by Brazilian Ministry of Health and Centers for Diseases Control and Prevention. More investments in the prevention of occupational infection by Hepatitis B virus for those professionals are necessary.