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The reduction of information contained in model time series through the use of aggregating statistical performance measures is very high compared to the amount of information that one would like to draw from it for model identification and calibration purposes. It is readily known that this loss imposes important limitations on model identification and -diagnostics and thus constitutes an element of the overall model uncertainty as essentially different model realizations with almost identical performance measures (e.g. r-² or RMSE) can be generated. In three consecutive studies the present work proposes an alternative approach towards hydrological model evaluation based on the application of Self-Organizing Maps (SOM; Kohonen, 2001). The Self-Organizing Map is a type of artificial neural network and unsupervised learning algorithm that is used for clustering, visualization and abstraction of multidimensional data. It maps vectorial input data items with similar patterns onto contiguous locations of a discrete low-dimensional grid of neurons. The iterative training of the SOM causes the neurons to form a discrete, data-compressed representation of the high-dimensional input data. Using appropriate visualization techniques, information on distributions, patterns and relationships in complex data sets can be extracted. Irrespective of their potential, SOM applications have earned very little attention in hydrological modelling compared to other artificial neural network techniques. Therefore, the aim of the present work is to demonstrate that the application of Self-Organizing Maps has very high potential to address fundamental issues of model evaluation: It is shown that the clustering and classification of model time series by means of SOM can provide useful insights into model behaviour. In combination with the diagnostic properties of Signature Indices (Gupta et al., 2008; Yilmaz et al., 2008) SOM provides a novel tool for interpreting the model parameters in the hydrological context and identifying parameter sets that simultaneously meet multiple objectives, even if the corresponding model realizations belong to different models. Moreover, the presented studies and reviews also encourage further studies on the application of SOM in hydrological modelling.
This thesis presents a study of tsunami deposits created by the 2004 Indian Ocean tsunami at the Thai Andaman coast. The outcomes of a study are the characteristics of tsunami deposit for paleo-tsunami database, the identification of major sediment layers in tsunami deposit and the reconstructing tsunami run-ups from the characteristics of tsunami deposit for a coastal development program. The investigations of tsunami deposit are made almost 3 years after the event. Field investigations characterize the tsunami deposit as a distinct sediment layer variable in thickness of gray sand deposited with an erosional basis on a pre-existing soil. The best location for the observation of recent tsunami deposit is the area located about 50-200 m inland from the coastline. In most cases, the deposit layer is normally graded. In some cases, the deposit contains rip-up clasts of muddy soils and/or organic matters. The tsunami deposits are compared with three deposits from coastal sub-environments. The mean grain-size and standard deviation of deposits show that the shoreface deposits are fine to very fine sand, poorly to moderately well sorted; the swash zone deposits are coarse to fine sand, poorly to well sorted; the berm/dune deposits are medium to fine sand, poorly to well sorted; and the tsunami deposits are coarse to very fine sand, poorly to moderately well sorted. The plots of deposit mean grain-size versus sorting indicate that the tsunami deposits are composed of shoreface deposits, swash zone deposits and berm/dune deposits as well. The vertical variation of the texture of tsunami deposit shows that the mean grain-size fines upward and fining landward. The analysis and interpretation of the run-up numbers from the characteristics of tsunami deposits get three run-ups for the 2004 Indian Ocean tsunami at the Thai Andaman coast. It corresponds to field observations from the eye-witness reports and local people- affirmations. The total deposition is a major transportation pattern of onshore tsunami sediments. The sediments must fine in the direction of transport. In general, the major origins of the sediment are the swash zone and berm/dune zone where coarse to medium sand is a significant material, the minor origin of tsunami sediment is a shoreface where a significant material is fine to very fine sand. Only at an area with flat slope shorface, the major origin of tsunami sediment is the shoreface. The thicknesses, the mean grain-sizes, and the standard deviations of tsunami deposits are used to evaluate the influences of coastal morphology on the sediment characteristics. The evaluations show that the tsunami affected areas were attacked by the variable energy waves. Wave energies at the direct tsunami wave affected areas are higher than at the indirect tsunami wave affected areas. Tsunami wave energy is highly dissipated at an area with steep slope shoreface. In the same way, tsunami run-up energy is highly dissipated at an area with steep slope onshore. A channel paralleled to the coastline decreases the run-up velocity, slightly dissipates run-up energy. The road and pond highly influence the characteristics of tsunami deposit and tsunami run-up. A road obstructs the run-up velocity, dissipates run-up energy. A pond decreases run-up velocity, dissipates run-up energy. The characteristics of tsunami deposit can be interpreted for reconstructing the characteristics of tsunami run-up such as a run-up height and a flow velocity. Soulsby et al.(2007)- model is applied for reconstructing tsunami run-up at the study areas. The input parameters are sediment grain-size and sediment inundation distance. Ao Kheuy beach and Khuk Khak beach, Phang Nga province, Thailand are the areas listed for reconstructing tsunami run-up. The evaluated run-up heights are 4.2-4.9 m at Ao Kheuy beach, and 5.4-9.4 m at Khuk Khak beach. The evaluated run-up velocities are 12.8-19.2 m/s (maximum) and 0.2-1.9 m/s (mean) at the coastline and onshore, respectively. Hence, a reasonably good agreement between the evaluated and observed run-up is found. Tsunami run-up height and velocity can be used for coastal development and risk management in the tsunami affected areas. The case studies from the Thai Andaman coast suggest that in the area from coastline to about 70-140 m inland was flooded by the high velocity (high energy) run-ups, and those run-up energies were dissipated there. That area ought to be a non-residential area or a physical protection construction area (flood barrier, forest planting, etc.).
This study aims to estimate the cotton yield at the field and regional level via the APSIM/OZCOT crop model, using an optimization-based recalibration approach based on the state variable of the cotton canopy - the leaf area index (LAI), derived from atmospherically corrected Landsat-8 OLI remote sensing images in 2014. First, a local sensitivity and global analysis approach was employed to test the sensitivity of cultivar, soil and agronomic parameters to the dynamics of the LAI. After sensitivity analyses, a series of sensitive parameters were obtained. Then, the APSIM/OZCOT crop model was calibrated by observations over a two-year span (2006-2007) at the Aksu station, combined with these sensitive cultivar parameters and the current understanding of cotton cultivar parameters. Third, the relationship between the observed in-situ LAI and synchronous perpendicular vegetation indices derived from six Landsat-8 OLI images covering the entire growth stage was modelled to generate LAI maps in time and space. Finally, the Particle Swarm Optimization (PSO) and general-purpose optimization approach (based on Nelder-Mead algorithm) were used to recalibrate four sensitive agronomic parameters (row spacing, sowing density per row, irrigation amount and total fertilization) according to the minimization of the root-mean-square deviation (RMSE) between the simulated LAI from the APSIM/OZCOT model and retrieved LAI from Landsat-8 OLI remote sensing images. After the recalibration, the best simulated results compared with observed cotton yield were obtained. The results showed that: (1) FRUDD, FLAI and DDISQ were the major cultivar parameters suitable for calibrating the cotton cultivar. (2) After the calibration, the simulated LAI performed well with an RMSE and mean absolute error (MAE) of 0.45 and 0.33, respectively, in 2006 and 0.46 and 0.41, respectively, in 2007. The coefficient of determination between the observed and simulated LAI was 0.83 and 0.97, respectively, in 2006 and 2007. The Pearson- correlation coefficient was 0.913 and 0.988 in 2006 and 2007, respectively, with a significant positive correlation between the simulated and observed LAI. The difference between the observed and simulated yield was 776.72 kg/ha and 259.98 kg/ha in 2006 and 2007, respectively. (3) Cotton cultivation in 2014 was obtained using three Landsat-8 OLI images - DOY136 (May), DOY 168 (June) and DOY 200 (July) - based on the phenological differences in cotton and other vegetation types. (4) The yield estimation after the assimilation closely approximated the field-observed values, and the coefficient of determination was as high as 0.82, after recalibration of the APSIM/OZCOT model for ten cotton fields. The difference between the observed and assimilated yields for the ten fields ranged from 18.2 to 939.7 kg/ha. The RMSE and MAE between the assimilated and observed yield was 417.5 and 303.1 kg/ha, respectively. These findings provide scientific evidence for the feasibility of coupled remote sensing and APSIM/OZCOT model at the field level. (5) Upscaling from field level to regional level, the assimilation algorithm and scheme are both especially important. Although the PSO method is very efficient, the computational efficiency is also the shortcoming of the assimilation strategy on a regional scale. Comparisons between the PSO and general-purpose optimization method (based on the Nelder-Mead algorithm) were implemented from the RSME, LAI curve and computational time. The general-purpose optimization method (based on the Nelder-Mead algorithm) was used for the regional assimilation between remote sensing and the APSIM/OZCOT model. Meanwhile, the basic unit for regional assimilation was also determined as cotton field rather than pixel. Moreover, the crop growth simulation was also divided into two phases (vegetative growth and reproductive growth) for regional assimilation. (6) The regional assimilation at the vegetative growth stage between the remote sensing derived and APSIM/OZCOT model-simulated LAI was implemented by adjusting two parameters: row spacing and sowing density per row. The results showed that the sowing density of cotton was higher in the southern part than in the northern part of the study area. The spatial pattern of cotton density was also consistent with the reclamation from 2001 to 2013. Cotton fields after early reclamation were mainly located in the southern part while the recent reclamation was located in the northern part. Poor soil quality, lack of irrigation facilities and woodland belts of cotton fields in the northern part caused the low density of cotton. Regarding the row spacing, the northern part was larger than the southern part due to the variation of two agronomic modes from military and private companies. (7) The irrigation and fertilization amount were both used as key parameters to be adjusted for regional assimilation during the reproductive growth period. The result showed that the irrigation per time ranged from 58.14 to 89.99 mm in the study area. The spatial distribution of the irrigation amount is higher in the northern part while lower in southern study area. The application of urea fertilization ranged from 500.35 to 1598.59 kg/ha in the study area. The spatial distribution of fertilization was lower in the northern part and higher in the southern part. More fertilization applied in the southern study area aims to increase the boll weight and number for pursuing higher yields of cotton. The frequency of the RSME during the second assimilation was mainly located in the range of 0.4-0.6 m2/m2. The estimated cotton yield ranged from 1489 to 8895 kg/ha. The spatial distribution of the estimated yield is also higher in the southern part than the northern study area.
The fragmentation of landscapes has an important impact on the conservation of biodiversity. The genetic diversity is an important factor for a population- viability, influenced by the landscape structure. However, different species with differing ecological demands react rather differently on the same landscape pattern. To address this feature, we studied ten xerothermophilous butterfly species with differing habitat requirements (habitat specialists with low dispersal power in contrast to habitat generalists with low dispersal power and habitat generalists with higher dispersal power). We analysed allozyme loci for about 10 populations (Ã 40 individuals) of each species in a western German study region with adjoining areas in Luxemburg and north-eastern France. The genetic diversity and genetic differentiation between local populations was discussed under conservation genetic aspects. For generalists we detected a more or less panmictic structure and for species with lower abundance and sedentarily behaviour the effect of isolation by distance. On the other hand, the isolation of specialists was mostly reflected by strong genetic differentiation patterns between the investigated populations. Parameters of genetic diversity were mostly significantly higher in generalists, compared to specialists. Substructures within populations as an answer of low intrapatch migration, low population densities and high population fluctuations could be shown as well. Aspects of landscape history (the historical distribution of habitats resulting of the presence of limestone areas) and the changes of extensive sheep pasturing and the loss of potential habitats in the last few decades (recent fragmentation) are discussed against the gained genetic data-set of the ten butterflies.
As an interface between an individual and its environment, the skin is a major site of direct exposure to exogenous substances. Once absorbed, these substances may interact with different biomolecules within the skin. The aryl hydrocarbon receptor (AhR) signaling pathway is one mechanism whereby the skin responds to exposures, predominantly through the induction or upregulation of metabolizing enzymes. One known physiological role of the AhR in many tissues is its involvement in the control of cell cycle progression. In skin, almost nothing is known about this physiological function. Moreover, the question whether frequently used naturally occurring phenolic derivatives like eugenol and isoeugenol impact on the AhR within the skin has rarely been studied so far. Eugenol and isoeugenol are due to their odour referred to as fragrances. The ubiquitous distribution of eugenol and isoeugenol results in an almost unavoidable contact with these substances in our daily lives. Despite this fact, their molecular mechanisms of action in skin are poorly understood. There is evidence supporting the hypothesis that these substances may impact on the AhR. On the one hand, eugenol is shown to induce cytochrome P450 1A1 (CYP1A1), a well-known target gene of the AhR. On the other hand, their known anti-proliferative properties might also be mediated by the AhR, based on its physiological function. In order to proof this hypothesis, it was investigated whether eugenol and isoeugenol impact on the AhR signaling pathway in skin cells. Results revealed that eugenol as well as isoeugenol impact on the AhR signaling pathway in skin cells. Both substances caused the translocation of the AhR into the nucleus, induced the expression of the well-known AhR target genes CYP1A1 and AhR repressor (AhRR) and exhibited impact on cell cycle progression. Both substances caused an AhR-dependent cell cycle arrest in skin cells, modulated protein levels of several cell cycle regulatory proteins, inhibited DNA synthesis and thereby reduced cell numbers. The comparison of wildtype cells to AhR knockdown cells revealed an influence of the AhR on cell cycle progression in skin cells in the absence of exogenous ligands. AhR knockdown cells exhibited a slower progression through the cell cycle caused by an accumulation of cells in the G0/G1 phase of the cell cycle and a decreased DNA synthesis rate. Modulation of cell cycle regulatory proteins involved in the transition from the G0/G1 to the S phase of the cell cycle was altered in AhR knockdown cells as well. To conclude, eugenol as well as isoeugenol were able to impact on the AhR signaling pathway in skin cells. Their molecular mechanisms of action are similar to those of classical AhR ligands, although their structural characteristics strongly differ from that of these ligands. In the absence of exogenous ligands the AhR promotes cell cycle progression in many tissues and this knowledge could be expanded on skin-derived cells within the scope of this thesis.
The skin is continuously challenged by environmental antigens that may penetrate and elicit a skin sensitization, which can develop into allergic contact dermatitis. Medical treatment for allergic contact dermatitis is limited - in fact only acute symptoms can be cured and for secondary prevention of the disease a lifelong avoidance of the allergen(s) is necessary. Therefore, the screening of the sensitization potential of substance used in commercially available products is indispensable to prevent such diseases. Hence, risk assessment is deduced from data obtained by murine local lymph node assay predominantly, but there exists a need to develop methods capable of providing the same information that do not require the use of animals in view of legislative initiatives such as REACH (registration, evaluation, authorization of chemicals) as well as the 7th Amendment to the Cosmetics Directive (2003/15/EC). Therefore, a number of promising in silico and in vitro approaches are being developed to address this need. In vitro test systems using the response of dendritic cells, which are the key player in the elicitation process of contact dermatitis, are established, but, although these novel methods for hazard identification might find application in the context of screening, it is not clear whether these approaches are useful for the purposes of risk assessment and risk management to predict allergic potency. Therefore, it was investigated whether on the one hand in vitro generated dendritic cells from primary blood monocytes (MoDC) and on the other hand a continuous monocytic cell line, the THP-1 cells, suggested as dendritic cell surrogate, react to a presumably weak allergen. Ascaridol, predicted as one of the possible causes for tea tree oil contact dermatitis, was studied and its effects in these two in vitro skin sensitization models were explored. Thus, the surface expression of CD86, HLADR, CD54, and CD40, which are known as activation markers in both in vitro models, were measured via flow cytometry. For MoDC, an augmented CD86 and HLADR surface expression in comparison to untreated cells were determined after 24 h exposure with ascaridol. An increased CD54 and CD40 surface expression were found only in some donors. After long term incubation of 96 h, ascaridol-treated MoDC still up-regulated CD86 and additionally an augmented CD40 expression was measured in all studied donors. An enhanced CD54 expression was determined for 50 percentage of all investigated donors. Furthermore, CD80, CD83 and CD209 protein expression were up-regulated in MoDC after 96 h of ascaridol incubation. In addition, it was determined that after 24 h ascaridol-treated MoDC showed an increased capacity to uptake antigens, whereas after 96 h this capacity got lost and antigen-capturing devices were reduced in comparison to non-treated MoDC. Moreover, the cytokine release of ascaridol-treated MoDC were measured after 24 h. Tumor necrosis factor (TNF)alpha, interleukin (IL)-1beta and IL 6 secretion were determined in some donors. Furthermore, IL-8 release was clearly increased after 24 h ascaridol treatment. By the same token, THP-1 cells were analyzed after ascaridol treatment for several activation markers. We found a similar response pattern as measured in MoDC. Ascaridol induced CD86 expression as well as CD54 after 24 h incubation. Additionally, the impact of ascaridol on phosphorylation of p38 mitogen-activated protein kinase, which had been shown to be involved in increased expression of activation markers like CD86 by others, were studied via Western blot analysis. A phosphorylation of p38 was determined after 15 min of ascaridol stimulation. Moreover, an augmented CD40 and HLADR surface expression were measured in a dose-response manner after 24 h ascaridol treatment. Also similar to MoDC an enhanced IL-8 secretion after ascaridol stimulation was observed in THP-1 cells. Hence, for the first time it was shown that ascaridol has immuno-modulating effects. The obtained data from both in vitro systems, MoDC and THP-1 cells, identified ascaridol as a sensitizer. Although for both systems there remain significant challenges to overcome for potency assessment, ascaridol is presumed to be a weak sensitizer probably. Interestingly, ascaridol treatment of THP-1 cells resulted also in an increased augmentation of CD184 and CCR2, two chemokine receptors expressed on monocyte. Therefore, these data encouraged the exploration of chemokine receptors as tools in skin sensitization prediction. Consequently, the combination of chemical assays with in vitro techniques may provide a useful surrogate to animal testing for skin sensitization. Due to the continuously changing environmental conditions, it is necessary to regularly monitor and update the spectrum of sensitizers that elicit contact dermatitis. Therefore, both debated in vitro test systems will become indispensable tools.
Evapotranspiration (ET) is one of the most important variables in hydrological studies. In the ET process, energy exchange and water transfer are involved. ET consists of transpiration and evaporation. The amount of plants transpiration dominates in ET. Especially in the forest regions, the ratio of transpiration to ET is in general 80-90 %. Meteorological variables, vegetation properties, precipitation and soil moisture are critical influence factors for ET generation. The study area is located in the forest area of Nahe catchment (Rhineland-Palatinate, Germany). The Nahe catchment is highly wooded. About 54.6 % of this area is covered by forest, with deciduous forest and coniferous forest are two primary types. A hydrological model, WaSiM-ETH, was employed for a long-term simulation from 1971-2003 in the Nahe catchment. In WaSiM-ETH, the potential evapotranspiration (ETP) was firstly calculated by the Penman-Monteith equation, and subsequently reduced according to the soil water content to obtain the actual evapotranspiration (ETA). The Penman-Monteith equation has been widely used and recommended for ETP estimation. The difficulties in applying this equation are the high demand of ground-measured meteorological data and the determination of surface resistance. A method combined remote sensing images with ground-measured meteorological data was also used to retrieve the ETA. This method is based on the surface properties such as surface albedo, fractional vegetation cover (FVC) and land surface temperature (LST) to obtain the latent heat flux (LE, corresponding to ETA) through the surface energy balance equation. LST is a critical variable for surface energy components estimation. It was retrieved from the TM/ETM+ thermal infrared (TIR) band. Due to the high-quality and cloudy-free requirements for TM/ETM+ data selection as well as the overlapping cycle of TM/ETM+ sensor is 16 days, images on only five dates are available during 1971-2003 (model ran) " May 15, 2000, July 05, 2001, July 19, August 04 and September 21 in 2003. It is found that the climate conditions of 2000, 2001 and 2003 are wet, medium wet and dry, respectively. Therefore, the remote sensing-retrieved observations are noncontinuous in a limited number over time but contain multiple climate conditions. Aerodynamic resistance and surface resistance are two most important parameters in the Penman-Monteith equation. However, for forest area, the aerodynamic resistance is calculated by a function of wind speed in the model. Since transpiration and evaporation are separately calculated by the Penman-Monteith equation in the model, the surface resistance was divided into canopy surface resistance rsc and soil surface resistance rse. rsc is related to the plants transpiration and rse is related to the bare soil evaporation. The interception evaporation was not taken into account due to its negligible contribution to ET rate under a dry-canopy (no rainfall) condition. Based on the remote sensing-retrieved observations, rsc and rse were calibrated in the WaSiM-ETH model for both forest types: for deciduous forest, rsc = 150 sm−1, rse = 250 sm−1; for coniferous forest, rsc = 300 sm−1, rse = 650 sm−1. We also carried out sensitivity analysis on rsc and rse. The appropriate value ranges of rsc and rse were determined as (annual maximum): for deciduous forest, [100,225] sm−1 for rsc and [50,450] sm−1 for rse; for coniferous forest, [225,375] sm−1 for rsc and [350,1200] sm−1 for rse. Due to the features of the observations that are in a limited number but contain multiple climate conditions, the statistical indices for model performance evaluation are required to be sensitive to extreme values. In this study, boxplots were found to well exhibit the model performance at both spatial and temporal scale. Nush-Sutcliffe efficiency (NSE), RMSE-observations standard deviation ratio (RSR), percent bias (PBIAS), mean bias error (MBE), mean variance of error distribution (S2d), index of agreement (d), root mean square error (RMSE) were found as appropriate statistical indices to provide additional evaluation information to the boxplots. The model performance can be judged as satisfactory if NSE > 0.5, RSR ≤ 0.7, PBIAS < -±12, MBE < -±0.45, S2d < 1.11, d > 0.79, RMSE < 0.97. rsc played a more important role than rse in ETP and ETA estimation by the Penman-Monteith equation, which is attributed to the fact that transpiration dominates in ET. The ETP estimation was found the most correlated to the relative humidity (RH), followed by air temperature (T), relative sunshine duration (SSD) and wind speed (WS). Under wet or medium wet climate conditions, ETA estimation was found the most correlated to T, followed by RH, SSD and WS. Under a water-stress condition, there were very small correlations between ETA and each meteorological variable.
Veterinary antibiotics are released to arable agricultural soil together with manure, including nutrients, organic matter, and microorganisms. Previously, the effects of antibiotic-contaminated manure on soil microbial community activity, function, structure, and resistance have been reported under controlled experimental conditions. This thesis further evaluated the antimicrobial effects as influenced by different manure compositions, soil microhabitats and moisture regimes, plants, and different distances to roots. Microbial community responses were determined by phenotypic phospholipid fatty acid (PLFA) and genotypic 16S rRNA gene fragment analyses. (Chapter 3) demonstrates that medication of pigs with difloxacin (DIF) and sulfadiazine (SDZ) alters the molecular-chemical pattern of slurries, confounding the detection of a consistent antibiotic effect in bulk and respective rhizosphere soil. This was evaluated in a 63-day mesocosm experiment considering typical agricultural manure applications to maize planted soil. Fecal bacteria were detected even 14 days after manure amendment. Manure of DIF- and SDZ-medicated pigs clearly affected the microbial community in mesocosm bulk and rhizosphere soil, temporarily matching antibiotic effects reported in previous studies. (Chapter 4) discusses the influences of different soil microhabitats on antibiotic fate and the effects on soil microflora. Total extractable SDZ was more than two-fold larger in earthworm burrows and soil macroaggregate surfaces compared to bulk soil or the interior fraction of aggregates. Furthermore, soil microbial communities were affected by a combination of soil microhabitat and treatment, which was reflected by different structural and functional community responses to SDZ in laboratory and under field conditions. (Chapter 5) evaluates if SDZ effects on microbial communities are more pronounced in soils which undergo periodic changes in soil moisture by drying-rewetting dynamics compared to soils without such moisture fluctuations. This was tested in a 49-day climate chamber soil pot experiment grown with grass. Manure-amended pots without or with SDZ contamination were incubated under a dynamic moisture regime with repeated drying and rewetting changes of more than twenty percent maximum water holding capacity compared to the control moisture regime. The microbial biomass, but less pronouncedly the community structure, showed an increased responsiveness to the combined stress of SDZ and dynamic moisture changes in the laboratory. Similar responses were documented under field conditions. (Chapter 6) indicated adverse effects of SDZ on root geotropism, number of lateral roots, and water uptake by plants in a 40-day greenhouse experiment with willow and maize grown in soil with environmentally relevant and worst-case antibiotic contamination. (Chapter 7) showed that the associated microbial community responded to a combination of plant species, distance to the root, and antibiotic spiking concentration. In highly antibiotic-contaminated soils, the structural and functional responses of the microbial community were dominated by indirect antibiotic effects on plants and roots.
The collapse of the tailings pond of the Aznalcállar open pit mine (West of Seville, Spain) in April 1998 left more than 4000 ha of arable land and floodplains contaminated with heavy metal containing pyrite sludge. After a first remediation campaign a considerable contamination remained in the soil. The present study evaluates the possibilities of reflectance spectroscopy and airborne hyperspectral remote sensing for the qualitative and quantitative assessment of heavy metal contamination and the acidification risk related to the mining accident. Based on an extensive data set consisting of geochemical analyses and reflectance measurements of more than 300 soil samples different chemometrics methods (multiple linear regression, partial least squares and artificial neural networks) are tested for computation of concentrations of soil constituents on the basis of the spectral reflectance. Spectral mixture analysis is applied for the analysis of the spatial distribution of the contamination. The abundance information derived from spectral mixture analysis is turned into quantitative information incorporating an artificial mixture experiment. The results of this experiment provide a link between sludge abundance and sludge weight, allowing as a consequence calculation of the amount of residual sludge per pixel, the acidification potential and other parameters important for remediation planning. The application of laboratory, field and imaging spectroscopy for providing quantitative information about the contamination levels in their spatial context is a good complement to conventional methods. The advantage is the reduction of the time and labour-intensive geochemical analysis, because after the model calibration, further samples can be analysed directly with the chemometric models. Furthermore, the spatial distribution can be mapped with imaging spectroscopy data helping in a more precise remediation planning.
Since November 1997, we started to focus on the population ecology of two sympatric Sinonatrix snakes in the Chutzuhu swamp, northern Taiwan. At the same time we also examined some specimens from Senckenberg Natural History Museum, Frankfurt am Main and accumulated field data of some observation made on S. percarinata suriki from Fushan botanical garden, Sanping and Gaoshu, Taiwan. According to the specimens examined, we suspect that the close phylogeny of S. percarinata suriki may come from two ancestors, northeast Taiwan population closest to Fujien or Zehjiang and the southwest population closest to Guandong or Vietnam. This pattern was also represented in some molecular phylogeny studies of freshwater fish in Taiwan. There were 22,462 trap-nights, taken from the Chutzuhu swamp, during the period November 1999 to September 2001 and 361 snakes were collected, comprising five species and 617 snake-times. The population sizes were based on the Lincoln-Peterson index and were estimated to be 988-±326 in S. annularis and 129-±78 in S. percarinata suriki. Movement and home range data showed S. annularis is a restricted activity water snake and S. percarinata suriki possesses great mobility in spatial patterns, but movement ability seems to be influenced by the size of the aquatic environment. S. annularis is live-bearing, on average 8.19 neonates and this principally occurs in September; S. percarinata suriki lays 6-24 eggs, but due to insufficient observations no conclusions can be drawn. It must be noted that oviposition was also noted in September. The reproductive mode may reflect on thermal requirement differences of the two sympatric snakes. S. annularis tended to be a fish (98%) eater and S. percarinata suriki take 50% fish and 50% frogs in their diet. Middle to high ground cover marshland appears to be the favorite microhabitat of S. annularis, and S. percarinata suriki seems prefer open creeks and ditches. The population condition of S. annularis in the Chutzuhu swamp seems to be rapidly deteriorating and this trend is also reflected in the BCI declines, low proportion stomach contents and diseases of S. annularis. Water seems to be the major influencing factor and strongly correlates with the conservation strategy. Conservation proposals for S. annularis in the Chutzuhu swamp will be formulated. During this study period we also developed an efficient technique for snake morphological data accumulation and image database, with the aid of the following devices, PC notebook and scanner, which is adapted for practical field studies. We also want to propose a component system for the establishment of a fundamental snake population databases (FPDS) for long-term snake ecological studies and monitoring herein.
During and after application, pesticides enter the atmosphere by volatilisation and by wind erosion of particles on which the pesticide is sorbed. Measurements at application sites revealed that sometimes more than half of the amount applied is lost into the atmosphere within a few days. The atmosphere is an important part of the hydrologic cycle that can transport pesticides from their point of application and deposit them into aquatic and terrestrial ecosystems far from their point of use. In the region of Trier pesticides are widely used. In order to protect crops from pests and increase crop yields in the viniculture, six to eight pesticide applications take place between May and August. The impact that these applications have on the environmental pollution of the region is not yet well understood. The present study was developed to characterize the atmospheric presence, temporal patterns, transport and deposition of a variety of pesticides in the atmosphere of the area of Trier. To this purpose, rain samples were weekly collected at eight sites during the growing seasons 2000, 2001 and 2002, and air samples (gas and particle phases) were collected during the growing season 2002. Multiresidue analysis methods were developed to determine multiple classes of pesticides in rain water, particle- and gas-phase samples. Altogether 24 active ingredients and 3 metabolites were chosen as representative substances, focussing mainly on fungicides. Twenty-four of the 27 measured pesticides were detected in the rain samples; seventeen pesticides were detected in the air samples. The most frequently detected pesticides and at the highest concentrations, both in rain and air, were compounds belonging to the class of fungicides. The insecticide methyl parathion was also detected in several rain samples as well as two substances that are banned in Germany, such as the herbicides atrazine and simazine. Concentration levels varied during the growing season with the highest concentrations being measured in the late spring and summer months, coinciding with application times and warmer months. Concentration levels measured in the rain samples were, generally, in the order of rnng l-1. Though average concentrations for single substances were less than 100 ng l-1, total concentrations were considerable and in some instances well above the EU drinking water quality standard of 500 ng l-1 for total pesticides. Compared to the amounts applied for pest control, the amounts deposited by rain resulted between 0,004% and 0,10% of the maximum application rates. These low pesticide inputs from precipitation to surface-water bodies is not of concern in vinicultural areas where the impact of other sources, such as superficial runoff inputs from the treated areas and cleaning of field crop sprayers, is more important. However, the potential impacts of these aerial pesticide inputs to non-target sites, such as organic crops, and sensitive ecosystems are as yet not known. Concentration levels in the air samples were in the order of ng m-3 at sites close to the fields were pesticides were applied, while lower values, in the order of pg m-3, were detected at the site located further away from fields where applications were performed. The measured air concentration levels found in this study do not represent a concern for human health in terms of acute risk. Inhalation toxicity studies have shown that an acute potential risk only arises at air concentrations in the range of g m-3. Finally, it must be kept in mind that only a small number of chemicals that were applied in the area were analysed for in this study. In order to gain a better evaluation of the local atmospheric load of pesticides, a wider spectrum of applied substances (including metabolites) needs to be investigated.
Today obesity has been recognized as a disease. Evidence suggests that obesity often has Genetic, environmental, psychological and other factors. Growing evidence points to heredity as a strong determining factor of obesity. The characterization of uncoupling proteins (UCP) represents a major breakthrough of genetic factors towards understanding the molecular basis for energy expenditure and therefore likely to have important implication for the cause and treatment of human obesity. UCPs as mitochondrial anion carriers which creates a pathway that allows dissipation of the proton electrochemical gradient therefore which when deregulated are key risk factors in the development of obesity and other eating disorders. In order to better understand the roles of both UCP2 and UCP3 which considered as prime candidate genes involved in the pathogenesis of obesity, this study elucidate (1) Genomic organization: The human UCP2 (3) gene spans over 8.7 kb (7.5 kb) distributed on 8 (7) exons. Three UCP genes may have evolved from a common ancestor or are the result from gene duplication events. Two mRNA transcripts are generated from hUCP3 gene, the long and short form of hUCP3 is differing by the presence or absence of 37 amino acid residues at the C-terminus. (2) Mutational analysis revealed a mutation in exon 4 of hUCP2 resulting in the substitution of an alanine by a valine at codon 55 and an insertion polymorphism in exon 8 consisted of a 45 bp repeat located 150 bp downstream of the stop codon in the 3'-UTR. The allele frequencies of both polymorphisms were not significantly elevated in a subgroup of children characterized by low Resting Metabolic Rates (RMR). (3) Promoter Analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box. Functional characterization of hUCP2 promoter showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site. 75 bp further upstream a strong cis-acting regulatory element was identified which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements. the 5"-flanking region of the hUCP3 gene were characterized in which contains both TATA and CAAT boxes as well as consensus motifs for PPRE, TRE, CRE and muscle-specific MyoD and MEF2 sites. Functional characterization identified a cis-acting negative regulatory element between - 2983 and -982 while the region between -982 and -284 showed greatly increased basal promoter activity suggesting the presence of a strong enhancer element. Promoter activity was particularly enhanced in the murine skeletal muscle cell line C2C12 reflecting the tissue-selective expression pattern of UCP3.
In recent decades, the Arctic has been undergoing a wide range of fast environmental changes. The sea ice covering the Arctic Ocean not only reacts rapidly to these changes, but also influences and alters the physical properties of the atmospheric boundary layer and the underlying ocean on various scales. In that regard, polynyas, i.e. regions of open water and thin ice within thernclosed pack ice, play a key role as being regions of enhanced atmosphere-ice-ocean interactions and extensive new ice formation during winter. A precise long-term monitoring and increased efforts to employ long-term and high-resolution satellite data is therefore of high interest for the polar scientific community. The retrieval of thin-ice thickness (TIT) fields from thermal infrared satellite data and atmospheric reanalysis, utilizing a one-dimensional energy balance model, allows for the estimation of the heat loss to the atmosphere and hence, ice-production rates. However, an extended application of this approach is inherently connected with severe challenges that originate predominantly from the disturbing influence of clouds and necessary simplifications in the model set-up, which all need to be carefully considered and compensated for. The presented thesis addresses these challenges and demonstrates the applicability of thermal infrared TIT distributions for a long-term polynya monitoring, as well as an accurate estimation of ice production in Arctic polynyas at a relatively high spatial resolution. Being written in a cumulative style, the thesis is subdivided into three parts that show the consequent evolution and improvement of the TIT retrieval, based on two regional studies (Storfjorden and North Water (NOW) polynya) and a final large-scale, pan-Arctic study. The first study on the Storfjorden polynya, situated in the Svalbard archipelago, represents the first long-term investigation on spatial and temporal polynya characteristics that is solely based on daily TIT fields derived from MODIS thermal infrared satellite data and ECMWF ERA-Interim atmospheric reanalysis data. Typical quantities such as polynya area (POLA), the TIT distribution, frequencies of polynya events as well as the total ice production are derived and compared to previous remote sensing and modeling studies. The study includes a first basic approach that aims for a compensation of cloud-induced gaps in daily TIT composites. This coverage-correction (CC) is a mathematically simple upscaling procedure that depends solely on the daily percentage of available MODIS coverage and yields daily POLA with an error-margin of 5 to 6 %. The NOW polynya in northern Baffin Bay is the main focus region of the second study, which follows two main goals. First, a new statistics-based cloud interpolation scheme (Spatial Feature Reconstruction - SFR) as well as additional cloud-screening procedures are successfully adapted and implemented in the TIT retrieval for usage in Arctic polynya regions. For a 13-yr period, results on polynya characteristics are compared to the CC approach. Furthermore, an investigation on highly variable ice-bridge dynamics in Nares Strait is presented. Second, an analysis of decadal changes of the NOW polynya is carried out, as the additional use of a suite of passive microwave sensors leads to an extended record of 37 consecutive winter seasons, thereby enabling detailed inter-sensor comparisons. In the final study, the SFR-interpolated daily TIT composites are used to infer spatial and temporal characteristics of 17 circumpolar polynya regions in the Arctic for 2002/2003 to 2014/2015. All polynya regions combined cover an average thin-ice area of 226.6 -± 36.1 x 10-³ km-² during winter (November to March) and yield an average total wintertime accumulated ice production of about 1811 -± 293 km-³. Regional differences in derived ice production trends are noticeable. The Laptev Sea on the Siberian shelf is presented as a focus region, as frequently appearing polynyas along the fast-ice edge promote high rates of new ice production. New affirming results on a distinct relation to sea-ice area export rates and hence, the Transpolar Drift, are shown. This new high-resolution pan-Arctic data set can be further utilized and build upon in a variety of atmospheric and oceanographic applications, while still offering room for further improvements such as incorporating high-resolution atmospheric data sets and an optimized lead-detection.
Arctic and Antarctic polynya systems are of high research interest since extensive new ice formation takes place in these regions. The monitoring of polynyas and the ice production is crucial with respect to the changing sea-ice regime. The thin-ice thickness (TIT) distribution within polynyas controls the amount of heat that is released to the atmosphere and has therefore an impact on the ice-production rates. This thesis presents an improved method to retrieve thermal-infrared thin-ice thickness distributions within polynyas. TIT with a spatial resolution of 1 km × 1 km is calculated using the MODIS ice-surface temperature and atmospheric model variables within the Laptev Sea polynya for the winter periods 2007/08 and 2008/09. The improvement of the algorithm is focused on the surface-energy flux parameterizations. Furthermore, a thorough sensitivity analysis is applied to quantify the uncertainty in the thin-ice thickness results. An absolute mean uncertainty of -±4.7 cm for ice below 20 cm of thickness is calculated. Furthermore, advantages and drawbacks using different atmospheric data sets are investigated. Daily MODIS TIT composites are computed to fill the data gaps arising from clouds and shortwave radiation. The resulting maps cover on average 70 % of the Laptev Sea polynya. An intercomparison of MODIS and AMSR-E polynya data indicates that the spatial resolution issue is essential for accurately deriving polynya characteristics. Monthly fast-ice masks are generated using the daily TIT composites. These fast-ice masks are implemented into the coupled sea-ice/ocean model FESOM. An evaluation of FESOM sea-ice concentrations is performed with the result that a prescribed high-resolution fast-ice mask is necessary regarding the accurate polynya location. However, for a more realistic simulation of other small-scale sea-ice features further model improvements are required. The retrieval of daily high-resolution MODIS TIT composites is an important step towards a more precise monitoring of thin sea ice and sea-ice production. Future work will address a combined remote sensing " model assimilation method to simulate fully-covered thin-ice thickness maps that enable the retrieval of accurate ice production values.
Exposure to fine and ultra-fine environmental particles is still a problem of concern in many industrialized parts of the world and the intensified use of nanotechnology may further increase exposure to small particles. Since many years air pollution is recognized as a critical problem in western countries, which led to rigorous regulation of air quality and the introduction of strict guidelines. However, the upper thresholds for particulates in ambient air recommended by the world health organization are often exceeded several times in newly industrialized countries. Such high levels of air pollution have the potential to induce adverse effects on human health. The response triggered by air pollutants is not limited to local effects of the respiratory system but is often systemic, resulting in endothelial dysfunction or atherosclerotic malady. The link between air pollution and cardiovascular disease is now accepted by the scientific community but the underlying mechanisms responsible for the pro-atherogenic potential still need to be unraveled in detail. Based on the results from in- vivo and in vitro studies the production of reactive oxygen species due to exposure to particles is the most important mechanism to explain the observed adverse effects. However, the doses that were applied in many in vivo and in vitro studies are far beyond the range of what humans are exposed to and there is the need for more realistic exposure studies. Complex in vitro coculture systems may be valuable tools to study particle-induced processes and to extrapolate effects of particles on the lung. One of the objectives of this PhD thesis was the establishment and further improvement of a complex coculture system initially described by Alfaro-Moreno et al. [1]. The system is composed of an alveolar type-II cell line (A549), differentiated macrophage-like cells (THP-1), mast cells (HMC-1) and endothelial cells (EA.hy 926), seeded in a 3D-orientation on a microporous membrane to mimic the cell response of the alveolar surface in vitro in conjunction with native aerosol exposure (VitrocellTM chamber). The tetraculture system was carefully characterized to ensure its performance and repeatability of results. The spatial distribution of the cells in the tetraculture was analyzed by confocal laser scanning microscopy (CLSM), showing a confluent layer of endothelial and epithelial cells on both sides of the Transwellâ„¢. Macrophage-like cells and mast cells can be found on top of the epithelial cells. The latter cells formed colonies under submerged conditions, which disappeared at the air-liquid-interface (ALI). The VitrocellTM aerosol exposure system was not significantly influencing the viability. Using this system, cells were exposed to an aerosol of 50 nm SiO2-Rhodamine nanoparticles (NPs) in PBS. The distribution of the NPs in the tetraculture after exposure was evaluated by CLSM. Fluorescence from internalized particles was detected in CD11b-positive THP-1 cells only. Furthermore, all cell lines were found to be able to respond to xenobiotic model compounds, such as benzo[a]pyrene (B[a]P) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with the upregulation of CYP1 mRNA. With this tetraculture system the response of the endothelial part of the alveolar barrier was studied in- vitro in a still realistic exposure scenario representing the conditions for a polluted situation without direct exposure of endothelial cells. After exposure to diesel exhaust particulate matter (DEPM) the expression of different anti-oxidant target genes and inflammatory genes such as NAD(P)H dehydrogenase quinone 1 (NQO1), superoxide dismutase 1 (SOD1) and heme oxygenase 1 (HMOX1), as well as the nuclear translocation nuclear factor erythroid-derived 2 (Nrf2) was evaluated. In addition, the potential of DEPM to induce the upregulation of CYP1A1 mRNA in the endothelium was analyzed. DEPM exposure led not to an upregulation of the anti-oxidant or inflammatory target genes, but to clear nuclear translocation of Nrf2. The endothelial cells responded to the DEPM treatment also with the upregulation of CYP1A1 mRNA and nuclear translocation of the aryl hydrocarbon receptor (AhR). Overall, DEPM triggered a response in the endothelial cells after indirect exposure of the tetraculture system to low doses of DEPM, underlining the sensitivity of ALI exposure systems. The use of the tetraculture together with the native aerosol exposure equipment may finally lead to a more realistic judgment regarding the hazard of new compounds and/or new nano-scaled materials in the future. For the first time, it was possible to study the response of the endothelial cells of the alveolar barrier in vitro in a realistic exposure scenario avoiding direct exposure of endothelial cells to high amounts of particulates.
The development of our society contributed to increased occurrence of emerging substances (pesticides, pharmaceuticals, personal care products, etc.) in wastewater. Because of their potential hazard on ecosystems and humans, Wastewater Treatment Plants (WWTPs) need to adapt to better remove these compounds. Technology or policy development should however comply with sustainable development, e.g. based on Life Cycle Assessment (LCA) metrics. Nevertheless, the reliability or consistency of LCA results can sometimes be debatable. The main objective of this work was to explore how LCA can better support the implementation of innovative wastewater treatment options, in particular including removal benefits. The method was applied to support solutions for pharmaceuticals elimination from wastewater, regarding: (i) UV technology design, (ii) choice of advanced technology and (iii) centralized or decentralized treatment policy. The assessment approach followed by previous authors based on net impacts calculation seemed very promising to consider both environmental effects induced by treatment plant operation and environmental benefits obtained from pollutants removal. It was therefore applied to compare UV configuration types. LCA outcomes were consistent with degradation kinetics analysis. For the comparison of advanced technologies and policy scenarios, the common practice (net impacts based on EDIP method) was compared to other assessments, to better consider elimination benefits. First, USEtox consensus was applied for the avoided (eco)toxicity impacts, in combination with the recent method ReCiPe for generated impacts. Then, an eco-efficiency indicator (EFI) was developed to weigh the treatment efforts (generated impacts based on EDIP and ReCiPe methods) by the average removal efficiency (overcoming (eco)toxicity uncertainty issues). In total, the four types of comparative assessment showed the same trends: (i) ozonation and activated carbon perform better than UV irradiation, and (ii) no clear advantage distinguished between policy scenarios. It cannot be however concluded that advanced treatment of pharmaceuticals is not necessary because other criteria should be considered (risk assessment, bacterial resistance, etc.) and large uncertainties were embedded in calculations. Indeed, a significant part of this work was dedicated to the discussion of uncertainty and limitations of the LCA outcomes. At the inventory level, it was difficult to model technology operation at development stage. For impact assessment, the newly developed characterization factors for pharmaceuticals (eco)toxicity showed large uncertainties, mainly due to the lack of data and quality for toxicity tests. The use of information made available under REACH framework to develop CFs for detergent ingredients tried to cope with this issue but the benefits were limited due to the mismatch of information between REACH and USEtox method. The highlighted uncertainties were treated with sensitivity analyses to understand their effects on LCA results. This research work finally presents perspectives on the use of transparently generated data (technology inventory and (eco)toxicity factors) and further development of EFI indicator. Also, an accent is made on increasing the reliability of LCA outcomes, in particular through the implementation of advanced techniques for uncertainty management. To conclude, innovative technology/product development (e.g. based on circular economy approach) needs the involvement of all types of actors and the support from sustainability metrics.
The main research question of this thesis was to set up a framework to allow for the identification of land use changes in drylands and reveal their underlying drivers. The concept of describing land cover change processes in a framework of global change syndrome was introduced by Schellnhuber et al. (1997). In a first step the syndrome approach was implemented for semi-natural areas of the Iberian Peninsula based on time series analysis of the MEDOKADS archive. In the subsequent study the approach was expanded and adapted to other land cover strata. Furthermore, results of an analysis of the relationship of annual NDVI and rainfall data were incorporated to designate areas that show a significant relationship indicating that at least a part of the variability found in NDVI time series was caused by precipitation. Additionally, a first step was taken towards the integration of socio-economic data into the analysis; population density changes between 1961 and 2008 were utilized to support the identification of processes related to land abandonment accompanied by cessation of agricultural practices on the one hand and urbanization on the other. The main findings of the studies comprise three major land cover change processes caused by human interaction: (i) shrub and woody vegetation encroachment in the wake of land abandonment of marginal areas, (ii) intensification of non-irrigated and irrigated, intensively used fertile regions and (iii) urbanization trends along the coastline caused by migration and the increase of mass tourism. Land abandonment of cultivated fields and the give-up of grazing areas in marginal mountainous areas often lead to the encroachment of shrubs and woody vegetation in the course of succession or reforestation. Whereas this cover change has positive effects concerning soil stabilization and carbon sequestration the increase of biomass involves also negative consequences for ecosystem goods and services; these include decreased water yield as a result of increased evapotranspiration, increasing fire risk, decreasing biodiversity due to landscape homogenization and loss of aesthetic value. Arable land in intensively used fertile zones of Spain was further intensified including the expansion of irrigated arable land. The intensification of agriculture has also generated land abandonment in these areas because less people are needed in the agricultural labour sector due to mechanization. Urbanization effects due to migration and the growth of the tourism sector were mapped along the eastern Mediterranean coast. Urban sprawl was only partly detectable by means of the MEDOKADS archive as the changes of urbanization are often too subtle to be detected by data with a spatial resolution of 1 km-². This is in line with a comparison of a Landsat TM time series and the NOAA AVHRR archive for a study area in the Greece that showed that small scale changes cannot be detected based on this approach, even though they might be of high relevance for local management of resources. This underlines the fact that land degradation processes are multi-scale problems and that data of several spatial and temporal scales are mandatory to build a comprehensive dryland observation system. Further land cover processes related to a decrease of greenness did not play an important role in the observation period. Thus, only few patches were identified, suggesting that no large-scale land degradation processes are taking place in the sense of decline of primary productivity after disturbances. Nevertheless, the land cover processes detected impact ecosystem functioning and using the example of shrub encroachment, bear risks for the provision of goods and services which can be valued as land degradation in the sense of a decline of important ecosystem goods and services. This risk is not only confined to the affected ecosystem itself but can also impact adjacent ecosystems due to inter-linkages. In drylands water availability is of major importance and the management of water resources is an important political issue. In view of climate change this topic will become even more important because aridity in Spain did increase within the last decades and is likely to further do so. In addition, the land cover changes detected by the syndrome approach could even augment water scarcity problems. Whereas the water yield of marginal areas, which often serve as headwaters of rivers, decreases with increasing biomass, water demand of agriculture and tourism is not expected to decline. In this context it will be of major importance to evaluate the trade-offs between different land uses and to take decisions that maintain the future functioning of the ecosystems for human well-being.
Floods are hydrological extremes that have enormous environmental, social and economic consequences.The objective of this thesis was a contribution to the implementation of a processing chain that integrates remote sensing information into hydraulic models. Specifically, the aim was to improve water elevation and discharge simulations by assimilating microwave remote sensing-derived flood information into hydraulic models. The first component of the proposed processing chain is represented by a fully automated flood mapping algorithm that enables the automated, objective, and reliable flood extent extraction from Synthetic Aperture Radar images, providing accurate results in both rural and urban regions. The method operates with minimum data requirements and is efficient in terms of computational time. The map obtained with the developed algorithm is still subject to uncertainties, both introduced by the flood mapping algorithm and inherent in the image itself. In this work, particular attention was given to image uncertainty deriving from speckle. By bootstrapping the original satellite image pixels, several synthetic images were generated and provided as input to the developed flood mapping algorithm. From the analysis performed on the mapping products, speckle uncertainty can be considered as a negligible component of the total uncertainty. In the final step of the proposed processing chain real event water elevations, obtained from satellite observations, were assimilated in a hydraulic model with an adapted version of the Particle Filter, modified to work with non-Gaussian distribution of observations. To deal with model structure error and possibly biased observations, a global and a local weight variant of the Particle Filter were tested. The variant to be preferred depends on the level of confidence that is attributed to the observations or to the model. This study also highlighted the complementarity of remote sensing derived and in-situ data sets. An accurate binary flood map represents an invaluable product for different end users. However, deriving from this binary map additional hydraulic information, such as water elevations, is a way of enhancing the value of the product itself. The derived data can be assimilated into hydraulic models that will fill the gaps where, for technical reasons, Earth Observation data cannot provide information, also enabling a more accurate and reliable prediction of flooded areas.
This thesis is focused on improving the knowledge on a group of threatened species, the European cave salamanders (genus Hydromantes). There are three main sections gathering studies dealing with different topics: Ecology (first part), Life traits (second part) and Monitoring methodologies (third part). First part starts with the study of the response of Hydromantes to the variation of climatic conditions, analysing 15 different localities throughout a full year (CHAPTER I; published in PEERJ in August 2015). After that, the focus moves on identify which is the operative temperature that these salamander experience, including how their body respond to variation of environmental temperature. This study was conducted using one of the most advanced tool, an infrared thermocamera, which gave the opportunity to perform detailed observation on salamanders body (CHAPTER II; published in JOURNAL OF THERMAL BIOLOGY in June 2016). In the next chapter we use the previous results to analyse the ecological niche of all eight Hydromantes species. The study mostly underlines the mismatch between macro- and microscale analysis of ecological niche, showing a weak conservatism of ecological niches within the evolution of species (CHAPTER III; unpublished manuscript). We then focus only on hybrids, which occur within the natural distribution of mainland species. Here, we analyse if the ecological niche of hybrids shows divergences from those of parental species, thus evaluating the power of hybrids adaptation (CHAPTER IV; unpublished manuscript). Considering that hybrids may represent a potential threat for parental species (in terms of genetic erosion and competition), we produced the first ecological study on an allochthonous mixed population of Hydromantes, analysing population structure, ecological requirements and diet. The interest on this particular population mostly comes by the fact that its members are coming from all three mainland Hydromantes species, and thus it may represent a potential source of new hybrids (CHAPTER V; accepted in AMPHIBIA-REPTILIA in October 2017). The focus than moves on how bioclimatic parameters affect species within their distributional range. Using as model species the microendemic H. flavus, we analyse the relationship between environmental suitability and local abundance of the species, also focusing on all intermediate dynamics which provide useful information on spatial variation of individual fitness (CHAPTER VI; submitted to SCIENTIFIC REPORTS in November 2017). The first part ends with an analysis of the interaction between Hydromantes and Batracobdella algira leeches, the only known ectoparasite for European cave salamanders. Considering that the effect of leeches on their hosts is potentially detrimental, we investigated if these ectoparasites may represent a further threat for Hydromantes (CHAPTER VII; submitted to INTERNATIONAL JOURNAL FOR PARASITOLOGY: PARASITES AND WILDLIFE in November 2017). The second part is related to the reproduction of Hydromantes. In the first study we perform analyses on the breeding behaviour of several females belonging to a single population, identifying differences and similarities occurring in cohorting females (CHAPTER VIII; published in NORTH-WESTERN JOURNAL OF ZOOLOGY in December 2015). In the second study we gather information from all Hydromantes species, analysing size and development of breeding females, and identifying a relationship between breeding time and climatic conditions (CHAPTER IX; submitted to SALAMANDRA in June 2017). In the last part of this thesis, we analyse two potential methods for monitoring Hydromantes populations. In the first study we evaluate the efficiency of the marking method involving Alpha tags (CHAPTER X; published in SALAMANDRA in October 2017). In the second study we focus on evaluating N-mixtures models as a methodology for estimating abundance in wild populations (CHAPTER XI; submitted to BIODIVERSITY & CONSERVATION in October 2017).
The main objective of the present thesis was to investigate whether antibody effects observed in earlier in vitro studies can translate into the protection against chemical carcinogenesis in vivo as the basis of an immunoprophylactic approach against carcinogens. As model for chemical carcinogenesis, we selected B[a]P the prototype polycyclic aromatic hydrocarbon (PAH), an environmental pollutant emanating from both natural and anthropogenic sources. Many in vivo models conveniently use high doses of carcinogens mostly given as single bolus, which provides simple surrogate readouts, but poorly reflects chronic exposure to the low concentrations found in the environment. In addition, these concentrations cannot be matched with equimolar antibody concentrations obtained by immunisation. However, low B[a]P concentrations do not permit to directly measure chemical carcinogenesis. Therefore, in the present thesis, the pharmacokinetic, metabolism and B[a]P mediated immunotoxicity were chosen as experimental read-outs. B[a]P conjugate vaccines based on ovalbumin, tetanus toxoid and diphtheria toxoid (DT) as carrier proteins were developed to actively immunise mice against B[a]P. B[a]P-DT conjugate induced the most robust immune response. The antibodies reacted not only with B[a]P but also with the proximate carcinogen 7,8-diol-B[a]P. Antibodies modulated the bioavailability of B[a]P and its metabolic activation in a dose-dependent manner by sequestration in the blood. In order to further improve the vaccination, we replaced the protein carrier by promiscuous T-helper cell epitopes to induce higher antibody titer with increased specificity for the B[a]P hapten. We hypothesised that a reduction of B cell binding sites on the carrier, compared to whole protein carrier, should favour the activation of B cells recognising the hapten instead of the carrier protein. An internal processing of the carrier and cleavage of the B[a]P-BA and subsequent presentation of the carrier peptide by MHC II molecules to T cell receptor should induce a B cell dependent immune response by activating B cells capable to recognise B[a]P. We demonstrated that a vaccination against B[a]P using promiscuous T-helper cell epitopes as a carrier is feasible and some tested peptide conjugates were more immunogenic as whole protein conjugates with increased specificity. We showed that vaccination against B[a]P reduces immunotoxicity. B[a]P suppressed the proliferative response of both T and B cells after a sub-acute administration, an effect that was completely reversed by vaccination. In immunized mice the immunotoxic effect of B[a]P on IFN-γ, Il-12, TNF-ï¡ production and B cell activation was restored. In addition, specific antibodies inhibited the induction of Cyp1a1 by B[a]P in lymphocytes and Cyp1b1 in the liver, enzymes that are known to convert the procarcinogen B[a]P to the ultimate DNA-adduct forming metabolite, a major risk factor of chemical carcinogenesis. In order to replace Freund adjuvant and to improve the immunisation strategy in terms of antibody quantity and quality, several adjuvants that are potentially compatible with their use in humans were tested. In combination with Freund adjuvant, the conjugate-vaccine induced high levels of B[a]P-specific antibodies. We showed that all adjuvants tested induced specific antibodies against B[a]P and 7,8-diol-B[a]P, its carcinogenic metabolite. The highest antibody levels were obtained with Quil A, MF-59 and Alum. Biological activity in terms of enhanced retention of B[a]P was confirmed in mice immunised with Quil A, Montanide, Alum and MF-59. Our findings demonstrate that a vaccination against B[a]P is feasible in combination with adjuvants licensed in humans. Based on these results and with the current understanding of the mechanisms of chemical carcinogenesis of the ubiquitous carcinogen B[a]P and of the effects of specific antibodies, an immunoprophylactic approach against chemical carcinogenesis is absolutely warranted. Nevertheless, the direct effects of B[a]P-specific antibodies on the different stages of carcinogenesis (e.g. adduct formation) and whether these effects may translate into long-term protective effect against tumourigenesis needs to be proven in further experiments.