A large database now exists for tardigrades in the Great Smoky Mountains National Park (GSMNP) consisting of 780 samples, 15,618 specimens, and 80 species including 14 new to science. We found 43 species of tardigrades in 150 soil/leaf litter samples. We calculated the Chao 1 species richness estimate with the species accumulation curve for the GSMNP and confirmed that our species list is virtually complete. Compared with soil data from mt. Fuji, Japan, estimated species richness in GSMNP was significantly higher. In our comparison of previous studies of soil/leaf litter tardigrades in other geographic areas, only the Kanagawa prefecture of Japan reported a higher number of species (47) than the GSMNP. Species richness estimators are valuable tools for comparing diversity in different habitats, even when sampling effort varies between studies.

Changes to vegetation architecture within a natural habitat can have profound impacts upon ecological community function, but the relative influence of vegetation architecture itself and potential indirect influences of associated food resources are often difficult to disentangle. I present the results of a three-year study designed to address the community level impacts of changes in big sagebrush (Artemisia tridentata) architecture and associated prey availability upon the ground-dwelling arthropod community. Three experimental sagebrush architecture treatments (low, intermediate, and high foliage density) and two levels of prey insect availability treatment (natural and baited/increased prey availability) were imposed in a sage-steppe ecosystem. The ground-dwelling spider (top predators) and insect (prey) response to all combinations of the six treatments were assessed through abundances in pitfall traps. Chapter 2 demonstrates that changes in a single shrub’s architecture did not markedly impact prey arthropod availability on the ground, though an overall increase in arthropod abundance was detected within baited shrubs. This indicates changes to vegetation architecture do not impact prey insect availability for ground-dwelling spiders. Thus, changes in shrub architecture and not the associated prey base directly alter the ground-dwelling spider community. Chapter 3 shows that architectural manipulation of a single sagebrush directly influences the ability of certain ground-dwelling spider guilds to persist beneath the shrub. Data within Chapters 2 and 3 support the conclusion that vegetation architecture directly affects ground-dwelling spider community structure and composition. Finally, Chapter 4 explores how the relative impacts of vegetation architecture and prey availability upon ecological communities can shift depending upon the spatial context in which the manipulations take place, given the differences in mobility of species within the grounddwelling spider and insect prey communities. Within this study, Diurnal and Nocturnal Wanderers responded to manipulations in prey availability at a larger spatial context (patches of 15 shrubs) while more stationary Trappers and Ambushers did not. Through simultaneous manipulation of vegetation architecture (Artemisia tridentata, big sagebrush) and associated insect prey base in different spatial contexts, this dissertation demonstrates that vegetation architecture directly affects ground-dwelling spider community organization at the species and guild levels of diversity. Shifts in spider (predator) functional diversity can ultimately impact arthropod decomposer and herbivore populations, influencing fundamental ecosystem processes such as decomposition and productivity.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第20437号 / 農博第2222号 / 新制||農||1049(附属図書館) / 学位論文||H29||N5058(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 北山 兼弘, 教授 神﨑 護, 教授 北島 薫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

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The rapid spread of antibiotic resistance is a major global health concern, jeopardizing the successful treatment of bacterial infections. Natural environments are potential hotspots for the emergence and spread of antibiotic resistance genes (ARGs). Among these potential hotspots, aquatic ecosystems are of particular concern, as they receive wastewater containing antibiotic-resistant bacteria and ARGs originating from both human and animal sources. Several key questions remain to be addressed. What is the fate of ARGs in receiving water bodies? What are implications of environmental ARGs for human health? How does wastewater discharge impacts aquatic microbial communities with regard to the overall ecosystem well-being? The objective of this work was to investigate the impact of wastewater, seasonal variations, and the riverine compartment on the prevalence of selected ARGs and the composition of natural microbial communities in a near-pristine river, and to specifically assess the effect of antibiotics on riverine microbial communities. Quantitative real-time PCR was used to monitor the abundance of three indicator ARGs (sul1 and sul2, conferring resistance against sulfonamide antibiotics, and intI1, a marker for anthropogenic pollution) upstream and downstream from a wastewater treatment plant (WWTP). Furthermore, the impact of WWTP effluent on the riverine microbial community was examined through 16S rRNA amplicon sequencing. Wastewater was the main source of all three target genes and significantly altered the microbial community in the river. The surface water compartment served as a dissemination route for ARGs, with increased prevalence even 13 km downstream of the WWTP, particularly during the summer season when the proportion of wastewater in the river was high. Notably, riverbed biofilms served as a local reservoir for ARGs only at the discharge point, with little abundance of target genes further downstream. The sulfonamide antibiotic sulfamethoxazole (SMX) was persistent in both near-pristine and wastewater-impacted river water when introduced at a concentration of 12.5 µg/L, but had neglectable effects on the microbial community diversity. Interestingly, concentrations as high as 100 µg/L SMX induced a short-term increase in microbial activity in both surface water and biofilm compartment, as revealed by bulk and nanoscale measurements. Altogether, this work underscores the fundamental role of wastewater treatment in combating the environmental dissemination of antibiotic resistance.:Summary 1 Zusammenfassung 5 1 Introduction 9 1.1 Rundown of the global antibiotic resistance crisis 9 1.1.1 History of antibiotics 9 1.1.2 Emergence of antibiotic resistance 9 1.1.3 Integrons as vehicles for antibiotic resistance 10 1.1.4 Risks related to environmental antibiotic resistance 12 1.2 Fate of antibiotic resistance genes in the aquatic environment 14 1.2.1 Genetic indicators for antibiotic resistance 14 1.2.2 River surface water compartment as dissemination route for antibiotic resistance 15 1.2.3 River biofilm compartment as reservoir for antibiotic resistance 17 1.3 Impact of antibiotics on aquatic microbial communities 18 1.4 Fate and effect of sulfamethoxazole in surface waters 20 2 Scope of the thesis 22 3 Main findings and scientific implications 24 3.1 Fate of antibiotic resistance genes after wastewater discharge into a near-pristine river 24 3.1.1 Wastewater is the primary source for aquatic antibiotic resistance 24 3.1.2 Drought increases the antibiotic resistance prevalence in surface waters 25 3.1.3 Riverbed biofilms serve as local reservoirs for antibiotic resistance genes 26 3.2 Anthropogenic pollution is the key driver for microbial community alteration 26 3.3 Sulfamethoxazole increases the microbial activity of aquatic microbial communities 27 4 Conclusions and future perspective 29 5 References 31 6 Publications 43 6.1 Publication 1 43 6.2 Publication 2 56 6.3 Publication 3 69   7 Appendix 94 7.1 Declaration of independent work 94 7.2 List of publications and conference contributions 95 7.2.1 Publications 95 7.2.2 Conference contributions 96 7.3 Contribution of Co-authors 97 7.4 Curriculum vitae 101 7.5 Acknowledgements 104 7.6 Supplementary Material 105 7.6.1 Supplementary Material for Publication 1 105 7.6.2 Supplementary Material for Publication 2 118 7.6.3 Supplementary Material for Publication 3 125

The relative importance of abiotic versus biotic top-down factors on structuring tundra plant communities is debated. With climate change already strongly affecting the tundra ecosystem, understanding which factors will prevail is vital. Tundra plant communities are presumed to be predominantly structured by their abiotic conditions yet grazing by reindeer (Rangifer tarandus) has a major effect on composition and diversity. It is increasingly recognized, however, that these factors cannot be considered in isolation. Here, I aim to test the relative and interactive effect of abiotic and top- down factors on vegetation structure in the Swedish mountain tundra. Using direct measurements of reindeer grazing via tri-axial accelerometers (from two summers, 2019 and 2020) coupled with remotely-gathered data on landscape features, I examine how species richness and coverage of vascular plants, bryophytes and lichens (sampled in 2022) are driven by grazing duration and abiotic conditions. Abiotic factors, specifically aspect and soil wetness, prevailed as the dominant drivers of local vegetation patterns. Clear interactions between factors were also observed. Reindeer grazing duration had predictable but weak effects on richness, with responses predominantly observed on south-facing slopes. Additionally, soil wetness interacted with grazing duration, with wetter areas grazed far less. These results demonstrate the importance of considering interactions between abiotic and biotic factors, providing a better understanding of how tundra plant communities in northern Sweden might change under future climate change or different grazing regimes. The observed interactions imply divergence in vulnerability between slopes and the potential for effects of herbivory to be altered under future hydrological conditions. I emphasize that future studies should continue to disentangle these relationships.

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