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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Isolation and identification of marine bacteria from marine mud in Vietnam with antimicrobial activity / Phân lập và nhận dạng các chủng vi sinh vật biển từ mẫu bùn biển ven bờ Việt Nam và hoạt tính kháng khuẩn của chúng

Thi, Tuyen Do, Dinh, Quyen Le, Dinh, Thi Quyen, Van, Cuong Pham 15 July 2013 (has links) (PDF)
Seventeen bacterial strains were isolated from 9 marine mud samples from the inshore environments of the East Sea. Four bacterial strains showed an inhibition against all tested microorganisms Staphylococcus aureus ATCC10832, Escherichia coli JM109, and Fusarium oxysporum. 16S rRNA sequences of four bacterial strains were obtained by PCR using specific primers. PCR products were cloned into E. coli DH5a using pJET1.2 blunt vector. The recombinant plasmids were sequenced and the lengths of these 16S rRNA sequences were ~930bp. The 16S rRNA sequence from the four bacterial DB1.2, DB1.2.3, DB4.2 and DB5.2 strain showed a high identity of 97 to 99% with the 16S rRNA sequence from Photobacterium sp., Oceanisphaera sp., Shigella sp., Stenotrophomonas sp, respectively. / Mười bảy chủng vi khuẩn đã được phân lập từ 9 mẫu bùn biển từ các vùng ven bờ biển Việt Nam. Bốn chủng vi khuẩn được ghi nhận có khả năng ức chế mạnh sự sinh trưởng và phát triển của các chủng vi khuẩn Staphylococcus aureus ATCC10832, Escherichia coli JM109, và thậm chí cả nấm Fusarium oxysporum. Trình tự gene 16S rRNA của bốn chủng vi khuẩn này đã được khuếch đại bằng PCR sử dụng cặp mồi đặc hiệu. Sản phẩm PCR được nối ghép vào vector pJET1.2 blunt sử dụng T4 ligase, hình thành plasmid tái tổ hợp và biến nạp vào E. coli DH5α. Khuẩn lạc có plasmid mang phân đoạn DNA chèn được nuôi cấy và tách plasmid. Trình tự 16S rRNA từ 4 chủng DB1.2, DB1.2.3, DB4.2 and DB5.2 chỉ ra có sự tương đồng 97 ÷ 99% so với trình tự 16S rRNA tương ứng của các chủng vi sinh vật biển trên ngân hàng gene thế giới là Photobacterium sp., Oceanisphaera sp., Shigella sp., và Stenotrophomonas sp.
2

Gefährliche Stoffe in Kläranlagen

Schneider, Petra, Günther, Thomas, Gottschalk, Nicole, Zänder, Daniel 22 April 2010 (has links) (PDF)
Gefährliche Stoffe gelangen durch Abwässer aus Haushalten, Industrie und Landwirtschaft in Kläranlagen. Während der mechanisch-biologischen Abwasserbehandlung in konventionellen Kläranlagen werden nur wenige Stoffe mit toxischen, persistenten und bioakkumulierenden Eigenschaften eliminiert. Der Bericht beschreibt die Eigenschaften gefährlicher Stoffe - darunter Industriechemikalien, Pflanzenschutzmittel, Arzneistoffe, Hormone und Metalle, ermittelt Quellen des Stoffeintrages und leitet Möglichkeiten zur Verringerung der Stoffeinträge ab. In Laborkläranlagen wurde eine Methodik entwickelt, mit der bei Anlagen mit biologischer Grundreinigung und weitergehender Reinigung (Stickstoffeliminierung) die Eliminierungsrate für ausgewählte gefährliche Stoffe ermittelt werden kann. Die Eliminationsraten der untersuchten gefährlichen Stoffe sind sehr unterschiedlich. Während ausgewählte Pestizide nur in geringem Umfang eliminiert werden (Bentazon < 40 Prozent, Dichlorprop < 30 Prozent, Dibutylzinn < 50 Prozent), werden für Nonylphenolethoxylate, Phthalate und Trichlormethan hohe Eliminationsraten mit bis zu 99 Prozent ermittelt.
3

Experimental Host-Parasite Co-Evolution in a Changing Environment / Experimentelle Parasit-Wirt Co-Evolution in einer sich verändernden Umwelt

Dusi, Eike 31 March 2016 (has links) (PDF)
Parasites with exclusive vertical transmission from host parent to offspring are an evolutionary puzzle. Any fitness costs for infected hosts risk the selective elimination of these parasites because their fitness is linked to host reproduction. One of the main evolutionary transitions from parasitism towards beneficial or mutualistic associations may therefore encompass a change from horizontal transmission to vertical transmission. In this thesis, the experimental evolution study on Paramecium and Holospora supports this hypothesis. The parasite nearly entirely lost horizontal transmission capacity in a treatment favouring vertical transmission and low virulence. However, many vertically transmitted parasites e.g. Caedibacter taeniospiralis impose detectable costs to their hosts. This endosymbiont imposes context-dependent costs to its host Paramecium tetraurelia. Fitness of infected paramecia was reduced in resource-limited conditions at all experimentally tested temperatures (16-32°C). These universal fitness costs along the temperature gradient necessitate universal cost compensation that can be the ‘killer trait’ that eliminates uninfected competitors. At acute heat stress the loss of infection indicates that cost compensation is impossible, thereby restricting conditions for parasite persistence. Surprisingly, the parasite persists in permanent stress and optimal temperature conditions. Caedibacter was able to adapt to high temperature conditions by increasing its number in the populations but without reducing virulence in high temperature conditions. Acute and intense stress harms the parasite and causes its extinction but the parasite was able to evolve and adapt to stress conditions. Moreover, the parasite reacts exactly in the opposite direction as it was expected. They do not suffer from stressful conditions, they benefit.
4

Investigation of a polyether trisiloxane surfactant

Michel, Amandine 09 May 2016 (has links) (PDF)
Thanks to their adaptability and high efficiency compared to traditional carbon based surfactants, silicone surfactants are a success in many different applications, from pesticides to cosmetics, polyurethane foam, textile and car care products. In spite of those numerous applications, no analytical method existed for their trace determination in environmental samples and no data have been available regarding their environmental occurrence and fate. An analytical method for the trace analysis of trisiloxane surfactants in the aqueous environment was developed and validated. The method, based on liquid-liquid extraction and HPLC-MS/MS, reaches limits of quantification in the ng L-1 range and allows an individual quantification of every homologue of the targeted trisiloxane surfactant. The newly developed analytical method was applied to analyze 40 river water samples. The targeted trisiloxane surfactant was detected in 14 samples, between 1 ng L-1 and 100 ng L-1. The results showed that the studied trisiloxane surfactant does not ubiquitously occur in the aquatic environment in measurable concentrations, but can reach surface waters on a local scale. In order to assess the persistence of the trisiloxane surfactant in surface waters, its hydrolysis was studied in the lab, under various conditions (temperature, pH, and concentration). The half-lifes at pH 7 and 2 mg L-1 were found to be between 29 days and 55 days at 25°C and between 151 days and 289 days at 12°C. Taking only into account the hydrolysis, these results indicate that the trisiloxane surfactant could persist several weeks in surface waters. A degradation product of the trisiloxane surfactant was tentatively identified by high resolution mass spectrometry. When used as agricultural adjuvants, trisiloxane surfactants may reach the soil compartment and might further leach to ground water. The behavior of the trisiloxane surfactant on soil was therefore investigated to assess the possibility to reach ground water. With a sorption batch equilibrium method, distribution coefficients between water and soil (Kd, Koc, and Kclay) were estimated for two standard soils (loam and sandy loam) and for every homologue of the trisiloxane surfactant. The obtained values for Kd were between 15 L kg-1 and 135 L kg-1, indicating that the trisiloxane surfactant is only slightly mobile in soil. To further investigate the possibility of leaching to ground water after application on agricultural fields, the leaching in soil was simulated in the lab in a soil column. The experimental settings were designed to simulate a worst case scenario where the application of the trisiloxane surfactant is done on quartz sand and is immediately followed by a heavy rainfall. Even in these conditions, less than 0.01 % of the initially applied trisiloxane surfactant leached through 20 cm of quartz sand. Based on the Kd values and the results of the leaching in soil column, the studied trisiloxane surfactant is considered to be unlikely to leach to ground water after application as an agricultural adjuvant.
5

Life cycles and secondary production of Ephemeroptera, Plecoptera, and Trichoptera (Insecta) under an extreme continental climate (River Kharaa, Mongolia)

Avlyush, Saulyegul 27 February 2014 (has links) (PDF)
Since the 1990s water quality monitoring projects using aquatic insects or macroinvertebrates as bioindication in Mongolia has mostly occurred in rivers drainage to the Arctic Ocean. They have been conducted to identify different anthropogentic stressors and impacts upon these running water ecosystems. However, there are still knowledge gaps and uncertainties concerning the research of these macroinvertebrates, in particular, a life cycle study of representative species are one such section of information missing. The specific aim for the study was to determine their life cycles and secondary production of selected species in the Kharaa River Basin, Mongolia, where these animals are exposed to harsh environment conditions. The main challenges for the research project were selecting the most suitable methods for use in the field sampling campaigns as well as establishing biomonitoring criteria for the target species under the extreme harsh climatic conditions. The research also sorts to address the pre-existing taxonomical identification problems. Consequently, a multi-habitat quantitative sampling method, and emergence traps type ‘Model week’ were selected. Five specific traits were chosen as selection criteria from the literature, where the life cycles of numerous species were investigated under comparable conditions to this study. Based on those five distinct criteria, a total of 18 species from EPT group (Ephemeroptera, Plecoptera, and Trichoptera) were selected for deeper analysis. This thesis provided the first quantitative results on the life cycle, production, growth rate and emergence of aquatic insects from Mongolia, to allow comparisons with studies in other regions using the same methods. However, it still needs more quantitative research of population dynamics for a wider range of species including fecundity, accurate development rates, mortality losses (e.g., due to predation), and food availability across environmental gradients of hydraulic conditions and substrate types. In conclusion, last not least it is essential to obtain knowledge especially about life cycle strategies of macroinvertebrates to identify the indicator-properties of single species and to predict re-colonisation potential of disturbed habitats and to evaluate the efficiency of management measures.
6

Phytoplankton dynamics in two large rivers: / Phytoplanktondynamik in zwei großen Flüssen: Langzeittrends, longitudinale Dynamik und mögliche Auswirkungen des Klimawandels

Hardenbicker, Paulin 16 March 2018 (has links) (PDF)
This study addresses the regulation of large river phytoplankton by climate-related drivers with the help of three different approaches, i.e. analyses of long-term data and spatial dynamics (longitudinal samplings) as well as mathematical modeling. The central hypothesis is that discharge has a dominant role among climate-related variables which strongly alters phytoplankton biomass development. A multi-factorial statistical analysis on the basis of long-term data (1990 – 2009; 1994 – 2009) from two measuring stations of the rivers Rhine and Elbe revealed that discharge conditions and light availability were the main driving forces regulating phytoplankton spring bloom dynamics. For the Rhine, a trend towards an earlier occurrence of the spring bloom event and a decrease in seasonal mean phytoplankton biomass could be detected, whereas for the Elbe no shift in the timing of the spring bloom and a tendency towards increasing seasonal mean phytoplankton biomass was found. Longitudinal sampling campaigns served to analyze the spatial plankton development on a short-term scale. River-internal growth and loss processes, as well as import mediated by tributaries were examined. Four longitudinal profiles were realized at different seasons in recent years (2009 – 2011) and it was revealed that tributaries mainly had a diluting impact on plankton densities in the Elbe and provided an additional import of phyto- and zooplankton densities in the Rhine. In the present study, high bivalve abundances were detected in the Rhine, probably leading to river-internal losses of phytoplankton which could compensate phytoplankton production resulting in low phytoplankton concentrations. In the Elbe, low abundances of bivalves and a low benthic grazing pressure prevailed. On the other hand, an unusually low discharge event in spring 2011 in the Rhine demonstrated that loss processes can at times be superimposed by strong phytoplankton production leading to extremely high phytoplankton biomasses and chlorophyll a values. Hence, despite the observed long-term trend of decreasing chlorophyll values in the last two decades, extreme environmental conditions can provoke regime shifts with exceptional phytoplankton mass developments. To assess the potential impacts of future climate change on water quality, the water quality simulation model QSim was used to establish a model for the free-flowing part of the Rhine. The modeling approach was implemented by changing the hydrological and climatologic input data according to different climate projections for the near (2021 – 2050) and the far future (2071 – 2100). The model results indicated a weak response of phytoplankton biomass in the Rhine towards altered climatic conditions, including discharge reductions and water temperature increases. The study suggests that changes in discharge rather than water temperature mediate climate change effects on large river phytoplankton. However, the effects are river specific as a consequence of system specific differences in main control mechanisms (e.g. ‘bottom-up’ versus ‘top-down’). / Anhand von drei verschiedenen Ansätzen analysiert die vorliegende Arbeit die Regulierung von Phytoplankton in großen Flüssen durch klimabedingte Faktoren: Auswertung von Langzeitdaten, räumliche Dynamik (fließzeitkonforme, longitudinale Beprobungen) und mathematische Modellierung. Die zentrale Hypothese ist, dass Abfluss eine dominante Rolle unter den klimabedingten Faktoren spielt und die Phytoplanktonbiomasse stark beeinflusst. Eine multifaktorielle statistische Analyse basierend auf Langzeitdaten (1990 – 2009; 1994 –2009) von zwei Stationen der Flüsse Rhein und Elbe zeigten, dass hauptsächlich Abflussbedingungen und Lichtverfügbarkeit die Antriebskräfte bei der Regulierung der Phytoplanktonfrühjahrsblüte darstellten. Während sich am Rhein ein Trend hin zu einem früheren Auftreten der Frühjahrsblüte und einer Abnahme der mittleren Phytoplankton-biomasse während der Vegetationsperiode zeigte, konnte für die Elbe keine zeitliche Verschiebung der Frühjahrsblüte festgestellt werden und mittlere Phytoplanktonbiomassen zeigten hier eine steigende Tendenz. Longitudinale Fließzeitbeprobungen dienten zur Analyse der kurzfristigen räumlichen Entwicklung des Planktons. Interne Produktions- und Verlustprozesse, sowie der Eintrag durch Zuflüsse wurden untersucht. Vier longitudinale Profile wurden zu unterschiedlichen Jahreszeiten der letzten Jahre (2009 – 2011) realisiert und es wurde gezeigt, dass die Nebenflüsse der Elbe vorwiegend einen Verdünnungseffekt auf die Planktondichten des Hauptstromes hatten, während sie für den Rhein einen wichtigen zusätzlichen Eintrag von Phyto- und Zooplankton darstellten. In der vorliegenden Arbeit wurden hohe Muscheldichten im Rhein gefunden, die möglicherweise zu hohen internen Planktonverlusten durch Fraß beigetragen haben, und somit zu niedrigen Phytoplankton-konzentrationen geführt haben könnten. In der Elbe waren die Muscheldichten dagegen gering und somit war auch der benthische Fraßdruck niedriger. Auf der anderen Seite zeigte ein ungewöhnlich niedriges Abflussereignis im Frühjahr 2011 am Rhein, dass diese Verlustprozesse zeitweise von starker Phytoplanktonproduktion überlagert werden können und dadurch extrem hohe Phytoplanktonbiomassen und Chlorophyllwerte entstehen können. Demzufolge können trotz der Beobachtung eines langfristigen abnehmenden Trends in den Chlorophyllgehalten während der letzen zwei Jahrzehnte extreme Umweltbedingungen einen Regime-shift mit außergewöhnlichen Massenentwicklungen des Phytoplanktons hervorrufen. Um den möglichen Einfluss des zukünftigen Klimawandels auf die Gewässergüte abzuschätzen, wurde mithilfe des Gewässergütesimulationsmodells QSim ein Modell für den frei fließenden Abschnitt des Rheins erstellt. Für den Modellierungsansatz wurden die hydrologischen und klimatologischen Eingangsdaten entsprechend der verschiedenen Klimaprojektionen für die nahe (2021 – 2050) und ferne Zukunft (2071 – 2100) verändert. Die Modellergebnisse zeigten, dass sich Änderungen in den klimatischen Bedingungen, einschließlich Abflussreduktion und Wassertemperaturanstieg, nur geringfügig auf die Phytoplankton¬biomasse des Rheins auswirkten. Die vorliegende Arbeit deutet darauf hin, dass Klimawandeleffekte eher durch Änderungen der Abflussverhältnisse auf das Phytoplankton in großen Flüssen wirken als durch Änderungen der Wassertemperatur. Der Effekt ist jedoch flusssystemspezifisch, da die Auswirkungen von systemspezifischen Unterschieden in den Hauptkontrollmechanismen gesteuert werden (z.B. ‚Bottom-up’ versus ‚Top-down’).
7

Effects of Single and Combined Pesticide Exposure on the Grazer–Aufwuchs–Interaction –

Rybicki, Marcus 16 March 2015 (has links) (PDF)
Die Eutrophierung von Fließgewässern führt besonders im Frühjahr zur Zunahme der Biomasse des benthischen Aufwuchses. Dies kann zur reduzierter Infiltration von Wasser in die Gewässersohle führen und wichtige Funktionen von Fließgewässerökosystems stören. Verschiedene Studien konnten zeigen, dass invertebrate Weidegänger (Grazer) in der Lage sind die Biomasse des Aufwuchses in Fließgewässern effektiv zu reduzieren. Diese wichtige ökologische Funktion, das benthische Grazing, kann dazu beitragen negative Effekte der Eutrophierung in Fließgewässern zu kompensieren. Relevante invertebrate Grazer sind unter anderem die Larven der Eintagsfliege Rhithrogena semicolorata, die hohe Biomassen in Fließgewässern erreichen und eine hohe Spezialisierung auf Aufwuchs zeigen. Die Larven von R. semicolorata sind während ihres einjährigen Lebenszykluses Pestiziden ausgesetzt, welche ihre Entwicklung beeinträchtigen können. Um die Effekte von Pestiziden auf die Entwicklung dieser wichtigen Grazer zu untersuchen wurden drei Expositionsszenarien in Anlehnung an reale Bedingungen im Freiland untersucht. Das erste Szenario untersucht die Effekten einer Exposition mit dem Herbizid Terbutryn auf die Grazer-Aufwuchs-Interaktion, wie sie im Herbst und zeitigen Frühjahr anzutreffen ist. Terbutryn verursachte ab 6 µg/L eine deutliche Reduktion der Aufwuchsquantität, was indirekt das Wachstum und die Fettreserven der Grazer verringerte. Die Grazer zeigten ab 0,6 µg/L eine deutliche erhöhte Drift. Da bei dieser Konzentration noch keine Effekte auf die Aufwuchsbiomasse detektiert wurde, ist dies ein Indiz für verringerte Qualität des Aufwuchses. Insektzide werden verstärkt im Frühjahr ausgebracht und beeinflussen die Entwicklung der Grazer direkt. Das zweite Szenario untersuchte die Effekte des Insektizids lambda-Cyhalotrhin (LCH) auf die Grazer-Aufwuchs-Interaktion. LCH ist stark lipophil und adsorbiert schnell an organisches Material und Aufwüchse in Gewässer, weshalb es besonders für aufwuchsfressende Organismen bioverfügbar ist. Neben den erwarteten letalen Effekten wurden in Bereichen um 1 µg/g OC bereits subletale Effekte auf das Driftverhalten sowie das Wachstum der Grazer gefunden. Die Fettreserven waren ab ca. 0,01 µg/g OC reduziert, was auf verringerte Fraßaktivität in diesen niedrigen Konzentrationsbereichen schließen lässt. Die Analyse des Aufwuchses zeigte einen indirekt Effekt von LCH auf die Aufwuchsentwicklung, der einerseits auf die erhöhte Mortalität der Grazer aber auch auf LCH-induzierte Verhaltensänderungen zurückgeführt werden konnte. Das finale Szenario befasste sich mit den Effekten der kombinierten Exposition von Herbizid und Insektizid, wie sie im Lebenszyklus von R. semicolorata auftreten. Dabei zeigte sich entgegen der Erwartung keine erhöhte Mortalität durch die kombinierte Exposition. Die Ergebnisse weisen jedoch daraufhin, dass die Reduzierung der physiologischen Kondition der Grazer, durch die indirekten Effekte von Terbutryn, in subletalen Konzentrationsbereichen von LCH die Dauer bis zur Wiedererholung verlängert. Außerdem zeigte sich am Ende eine erhöhte Aufwuchsbiomasse, was eine reduzierte Leistung des Grazings durch die kombinierte Exposition indiziert. / The eutrophication of running water ecosystems leads especially in spring to an increase of benthic biofilms or aufwuchs, which reduces the infiltration of water into the river bed and, hence, disturb important ecological functions. Different studies already showed that invertebrate grazers of running water ecosystems are able to reduce the aufwuchs biomass effectively. This important ecological function, the benthic grazing, can therefore contribute to compensate the negative effects of eutrophication in lotic ecosystems. Relevant grazers are for instance the larvae of the mayfly Rhithrogena semicolorata, which reach high biomasses in streams and show a high feeding specialisation on aufwuchs. During their on year lasting life cycle the larvae of R. semicolorata are exposed to different pesticides, which can disturb their development. To evaluate the effects of pesticides on the development of this important grazer three exposition scenarios were performed following real exposure patterns from the environment. In the first scenarios the effects of a solely exposure with the herbicide Terbutryn on the grazer-aufwuchs-interaction was investigated, as it occurs preferably during autumn and spring. Terbutryn induced a clear reduction of aufwuchs quantity at 6 µg/L, which indirectly affected the growth and the energy storages of the grazers. Furthermore, the grazers showed an increased drift activity at 0.6 µg/L. Considering that the aufwuchs biomass was not affected at this low concentrations, the increased drift was interpreted as an indirect effect of reduced aufwuchs quantity. Inseciticides are preferably applied during spring and are able to disturb the development of grazers directly. The second scenario investigated therefore the effects of the insecticide lambda-Cyhalothrin (LCH) on the grazer-aufwuchs-interaction. LCH has a high lipophilicity and thus rapidly binds to organic matter or aufwuchs after entering aquatic ecosystems. This behaviour leads to a increased bioavailability of this substance to aufwuchs feeding organisms. Apart from the expected lethal effects, sublethal effects in form of increased drift and reduced grazer growth were found in a range of 1 µg/g OC LCH. Moreover, the energy storages of grazers were reduced at 0,01 µg/g OC, which is an indication of disturbed feeding behaviour at this low concentration. The analysis of the aufwuchs revealed additionally an indirect effect of LCH on the aufwuchs development, which was caused by increased grazer mortality and behavioural changes of the grazers induced by sublethal LCH concentrations. The final scenario aimed on the effects of a combined exposure of herbicide and insecticide, as it occurs during the life cycle of R. semicolorata. A increased mortality of grazers, due to the combinded exporsure, was not found. However, the results indicate that the reduction of the physiological condition by the indirect effect of Terbutryn extends the time until grazers recover from sublethal LCH exposure. Finally, also an increased aufwuchs biomass was found in treatments with combined exposure, indicating a reduced performance of grazers exposed to both substances.
8

Studies on the interaction of chemicals with cellular efflux transporter proteins Danio rerio Abcb4 and Homo sapiens ABCB1

Burkhardt-Medicke, Kathleen 06 June 2018 (has links) (PDF)
ABCB1, a member of the ATP binding cassette (ABC) transporter family, hydrolyses ATP as energy source for the translocation of substrate chemicals across the cell membrane. ABCB1-like transporters are found in all studied species. Typically, these transporters are abundant in tissues that separate compartments of the body such as the blood-brain barrier. Among the ABC transporters the ABCB1-like transporter proteins are of particular interest because they accept a broad variety of substrates and are therefore able to confer multidrug resistance (MDR) and multixenobiotic resistance (MXR) in wildlife, respectively. Inhibitors of the ABCB1-like transporter function can cause chemosensitisation, i.e. accumulation and increased sensitivity of organisms towards potentially harmful (natural/man-made) ABCB1-like substrate chemicals. In zebrafish (Danio rerio) Abcb4 was identified as functionally homologous to ABCB1. The aim of this study was to further characterise Danio rerio Abcb4 and to provide a database to approach the question to what extent ABCB1-like transporter related functions/effects are of ecotoxicological relevance. Main objectives are whether and how known ABCB1 ATPase stimulators and inhibitors interact with Abcb4 ATPase activity; to what extent ABCB1 ATPase assay data are transferable to Abcb4 ATPase assay data; and whether and how environmental chemicals interact with Danio rerio Abcb4 ATPase activity. In this study we established a test system – the ATPase assay with recombinant Danio rerio Abcb4 – to study the interaction of chemicals with the ATPase activity of the transporter protein. To relate obtained data to data for the well-known Homo sapiens ABCB1 and because available data for Homo sapiens ABCB1 were not in all cases suitable for a comparison, the ATPase assay with recombinant ABCB1 was adapted accordingly. Chemicals were tested up to concentrations in the range of their water solubilities to modulate basal and stimulator co-treated Abcb4 and/or ABCB1 ATPase activities. ATPase stimulators are often transported substrates. However, lipophilic compounds stimulating the transporter ATPase activity are not or little transported by transporter action. Therefore, experiments revealing whether compounds are translocated by transporters chemical interference with the transporter protein will not be indicated. Chemicals inhibiting the stimulator (here verapamil) co-treated ATPase activity compete with the verapamil to stimulate ATPase activity or are non-competitive inhibitors. When tested individually, these chemicals can be stimulators or inhibitors of basal ATPase activity, or do not interact with basal ATPase activity. ATPase inhibitors mitigate ATPase activity and ABCB1-like transporter mediated translocation of substrate chemicals. Obtained ATPase assay data were analysed with regard to concentrations at half-maximal effects (EC50s) and effect strengths (percent modulation). ATPase assays with recombinant Abcb4 (at 27 °C) are comparable to ABCB1 ATPase assay data obtained at 37 °C. Danio rerio Abcb4 seems less temperature-sensitive than ABCB1. Calculated activation energies for Abcb4 ATPase activities (40.75 kJ/mol for basal ATPase activity) were up to half as high as those for ABCB1 ATPase activities (81.61 kJ/mol for basal ATPase activity). Larger activation energies were previously proposed to be indicative for larger conformational rearrangements and hence possibly smaller rearrangements take place in Abcb4 compared to ABCB1. Known standard modulators of Homo sapiens ABCB1 ATPase activity interacted specifically with Danio rerio Abcb4 ATPase actitiy. The EC50s of the tested chemicals – 16 of 17 tested chemiacals interacted with the ABCB1 and the Abcb4 ATPase activity – ranged from 0.09 to 296 µM for ABCB1 and from 0.14 to 171 µM for Abcb4. Qualitative ATPase assay data for ABCB1, as interaction or not, seems transferable to Danio rerio Abcb4. Furthermore, when aligning amino acid sequences of mammalian ABCB1 transporter proteins and Danio rerio Abcb4 and comparing ABCB1 residues known to bind to (lipophilic) chemicals no obvious hints were found that chemical binding to Abcb4 is certainly different from ABCB1. Twenty-five of 33 studied environmental chemicals modulated the Abcb4 ATPase activity as stimulators and/or inhibitors. Stimulation of basal Abcb4 ATPase activity was lower for environmental chemicals than for known standard modulators. EC50s of environmental chemicals ranged from below 10 to 357 µM. Effects by environmental chemicals on Abcb4 ATPase activity with EC50s close to their water solubilities may be rather unspecific. The results of this work underline that Abcb4 function is of ecotoxicological importance as on the one hand several environmental chemicals were identified to inhibit Abcb4 ATPase activity – likely acting as chemosensitisers, while on the other hand chemicals stimulating basal ATPase activity suggest that these chemicals are possibly transported. A number of environmental chemicals also inhibited the basal Abcb4 ATPase activity. Especially non-transported inhibitors of the basal Abcb4 ATPase activity would be of ecotoxicological relevance as organisms (here Danio rerio) exposed to these chemicals would not be protected by Abcb4 mediated multixenobiotic resistance and were moreover threatened by chemosensitisation. Future studies should systematically elucidate under which circumstances chemicals are apparently net transported by ABCB1-like transporters and relate these findings to concentrations of environmental chemicals and ABCB1-like transporter protein abundance in wildlife.
9

Controlling Factors Of Life Cycle And Distribution Of Chironomid Key Species In The Mesotrophic Saidenbach Reservoir

Hempel, Esther 24 August 2011 (has links) (PDF)
In den Jahren 2005 bis 2010 erfolgte im Rahmen der Erarbeitung der vorliegenden Dissertationsschrift eine ökologische Untersuchung der Chironomidenfauna in der Talsperre Saidenbach (Sachsen, Erzgebirge). Drei Arten mit hoher Abundanz konnten bei der umfassenden Artenanalyse im Jahr 2005 ermittelt werden: Procladius crassinervis, P. choreus und Chironomus anthracinus. Zusätzlich wurde die Art C. plumosus aufgrund ihrer engen Verwandtschaft zu C. anthracinus in die Untersuchung einbezogen. Die Arbeit ist auf vier Schwerpunkte fokussiert, wobei die Larven und Puppen der vier Arten analysiert wurden. (1) Die Erarbeitung einer zuverlässigen Methode zur Unterscheidung der Larven der beiden eng verwandten Procladius-Arten basierend auf morphologischen Kriterien (Imaginalscheiden-entwicklung, Kopfkapselgröße und Körperlänge) sowie die Tiefenverteilung der Puppen waren eine Vorbedingung für weiterführende Analysen des Lebenszyklus (LZ) dieser beiden Arten. (2) Die Untersuchung des LZ war der zweite Schwerpunkt. Der LZ wurde stark von abiotischen Faktoren wie Temperatur, Sauerstoff und Biovolumen des Phytoplanktons kontrolliert. (3) Die raum-zeitlichen Verteilungsmuster der vier Arten wurden zunächst bezüglich großräumiger Unterschiede über einen Tiefengradienten innerhalb eines Transektes analysiert. Hierbei zeigte sich bei allen vier Arten eine zeitliche Änderung im jeweiligen Hauptverbreitungsgebiet. Die jungen Larven von C. anthracinus, C. plumosus und P. choreus wanderten im Verlauf ihrer Entwicklung bis zur Verpuppung in flachere Bereiche, P. crassinervis wanderte in tiefere Bereiche. Die Analyse der Verteilungsunterschiede der Larven zwischen zwei verschieden stark eutrophierten Buchten ergab höhere Dichten der beiden Chironomus-Arten in der Bucht mit dem größeren Zulauf und der höheren Phytoplanktonkonzentration im Vergleich mit den anderen Arten. Eine Analyse des kleinräumigen Verteilungsmusters fokussierte auf der Frage, ob die Larven gleichmäßig verteilt oder aggregiert auftraten. Die Untersuchung des vertikalen Verteilungsmusters erforschte das Schwimmverhalten der Larven. (4) Der letzte Aspekt war eine experimentelle Untersuchung, die am Beispiel von C. anthracinus durchgeführt wurde mit dem Ziel, den Proximatfaktor für die beobachtete Wanderung der Larven zu ermitteln, der letztendlich die Temperatur war. Puppen bevorzugten im Experiment wärmere Temperaturen und junge Larven kühlere Temperaturen. Die fünfjährige Untersuchung der Chironomiden in der Talsperre Saidenbach beschreibt insgesamt die komplexe Verhaltensreaktion der Chironomiden, die einen wesentlichen Teil der benthischen Lebensgemeinschaft darstellen, bezüglich der Lebenszyklusmuster (Voltinismus, Verpuppung), der Abundanzänderungen (inner- und zwischenjährlich) sowie der groß- und kleinräumigen Verteilung unter der Einwirkung der wichtigsten Umweltfaktoren. / In the context of the present dissertation an ecological study was performed about chironomids in Saidenbach Reservoir in the Saxony Ore Mountains, Germany during the five years from 2005 to 2010. A preliminary overall species analysis in 2005 showed that three species were most abundant: Procladius crassinervis, P. choreus and Chironomus anthracinus. Additionally, the species C. plumosus was examined because of its close relationship to C. anthracinus. The study is focussed on four subjects, whereby larvae and pupae of the four species were analysed. (1) The elaboration of a reliable method to distinguish the larvae of the two closely related species P. crassinervis and P. choreus on the basis of morphological criteria (imaginal disc development, larval head capsule size and body length) as well as the depth distribution of their pupae was a precondition to the profound analysis of their life cycles. (2) The investigation of the life cycle pattern of the four species was the second focus. The life cycle of the four species was found to be strongly influenced by abiotic conditions such as temperature, oxygen and biovolume of the phytoplankton. (3) The spatial and temporal distribution pattern of the four species was analysed in view of large scale differences over a depth gradient in one transect. Here, in all four species a shift in the mainly settled lake bottom area occurred. The young larvae of C. anthracinus, C. plumosus and P. choreus migrated during maturing and pupation towards shallower areas; P. crassinervis migrated to deeper areas. The distribution differences between two different bays showed that the two Chironomus species had higher densities in the bay with the higher inflow which resulted in a higher phytoplankton standing stock compared to the other species. A small scale distribution pattern analysis focussed on finding out whether the larvae were aggregated or randomly distributed. The vertical distribution analysis examined the swimming behaviour of the larvae. (4) The last aspect was an experimental setup exemplarily driven with C. anthracinus which showed that the migration was stimulated by the proximate factor temperature as pupae preferred warmer temperatures and young larvae colder temperatures. Altogether, the five year study about chironomids in Saidenbach Reservoir pointed out the complex reaction in the behaviour of an important part of the benthic community concerning the life cycle pattern (voltinism, pupation pattern), the changing in abundances (inter-annual and intra-annual) and the large scale and small scale distribution pattern under the rule of the most important environmental factors.

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