Diese Dissertation beschäftigt sich mit der passiven Probenahme von hydrophoben organischen Schadstoffen in Oberflächengewässern: Polyaromatische Kohlenwasserstoffe (PAK), polychlorierte Biphenyle (PCB), polybromierte Biphenylether (PBDE), Organochlorpestizide (u. a. HCH, DDX) und weitere hydrophobe Pestizide. Die Zielstellung dieser Arbeit lag bei der Validierung des Silikonstabs als Alternativmethode im Gewässermonitoring zu konventionellen Probenahmetechniken wie Schöpf- und Wochenmischproben der Wasserphase sowie Schwebstoffanalysen. Die Probenahme mit dem Silikonstab erfolgte durch dessen Exposition im Gewässer für einen Zeitraum zwischen einer Woche und mehreren Monaten. Nach Einholung wurden die im Silikonstab akkumulierten Schadstoffe (Analyten) mittels instrumenteller Analytik quantifiziert. Die Probenaufgabe erfolgte ohne vorherige Lösungsmittelextraktion durch direktes Erhitzen des Silikonstabs, wodurch die Analyten vom Polymer desorbieren (Thermodesorption). Die durch Hitze freigesetzten Analyten wurden direkt auf eine chromatographische Trennsäule gegeben und massenspektroskopisch quantifiziert. Nach Erhalt der Ergebnisse der Silikonstab-Analytik gibt es verschiedene Herangehensweisen für die Berechnung der zeitgemittelten Analytkonzentrationen im Gewässer, die in dieser Arbeit vorgestellt und diskutiert werden. Dazu gehören die Verwendung von experimentellen Daten aus Kalibrierversuchen und Berechnungen auf Grundlage von physikochemischen Eigenschaften der Analyten wie dem Sammler-Wasser-Verteilungskoeffizienten. Im Zuge dieser Arbeit wurde die Aufnahmekinetik des Silikonstabs bei verschiedenen Temperaturen und Fließgeschwindigkeiten mit Hilfe von Kalibrierversuchen untersucht. Die gewonnenen experimentellen Daten wurden für die Entwicklung von Rechenmodellen herangezogen, mit denen das Aufnahmeverhalten vorgesagt werden soll. Es wurden Sammler-Wasser-Verteilungskoeffizienten für den Silikonstab u. a. mit der Kosolvenzmethode bestimmt und als Parameter für die Berechnung von zeitgemittelten Analytkonzentrationen des Gewässers verwendet. Für die Validierung wurde der Silikonstab in zwei Gewässergütemessstationen der Fließgewässer Mulde (Dessau) und Elbe (Magdeburg) in Durchflussbehältern exponiert und die zeitgemittelten Analytkonzentrationen mit verschiedenen Rechenmodellen bestimmt. Die erhaltenen Werte werden mit gleichzeitig entnommenen Wochenmischproben der Wasserphase sowie monatlichen Schwebstoffproben verglichen und die Eignung des Silikonstabs als alternative Probenahmemethode für das Umweltmonitoring von Oberflächengewässern diskutiert.

Nenhuma / Valores-guia de qualidade de sedimentos (VGQS) têm sido usados para avaliar o risco ecológico associado à contaminação de sedimentos para organismos bentônicos. O principal objetivo deste trabalho foi o desenvolvimento de metodologias e levantamento de dados que permitissem a aplicação de VGQS para os metais Cd, Cu, Pb, Ni e Zn. O local escolhido foi a sub-bacia do ribeirão Espírito Santo, que integra a Unidade de Planejamento e Gestão de Recursos Hídricos UPGRH SF4 da bacia do rio São Francisco, localizada junto ao núcleo urbano da cidade de Três Marias. A vida na sub-bacia é afetada, principalmente, pela instalação de uma barragem de rejeitos oriundos do beneficiamento do minério de zinco na região. Os estudos foram direcionados à caracterização biogeoquímica dos sedimentos por meio da determinação das concentrações de sulfetos volatilizáveis por acidificação (SVA), da determinação da fração de metais associada aos SVA nos sedimentos totais, denominada metais extraídos simultaneamente (MES) e do estudo da partição de metais entre os sedimentos totais e águas intersticiais. Os dados levantados foram interpretados em associação aos resultados da estrutura da comunidade bentônica e testes de ecotoxicidade. Os estudos incluíram ainda, a realização de análises das variáveis físico-químicas e da concentração de metais nas amostras de águas superficiais; análises mineralógicas, granulométricas, teor de matéria orgânica e concentração de contaminantes inorgânicos em amostras de sedimentos. Elevados valores de condutividade elétrica e sólidos totais dissolvidos foram obtidos em apenas uma estação de amostragem de água superficial. Os resultados das análises por ativação neutrônica mostraram que os elementos químicos predominantes nas amostras de sedimentos são o Ti, Al, V, Mn, e Fe. Os resultados das análises de metais totais nas amostras de sedimentos demonstraram que apenas o ponto localizado no córrego Lavagem, a montante de sua foz no ribeirão Espírito Santo, apresentou valores de concentração dos metais avaliados (Cd, Cu, Pb, Ni e Zn) acima dos valores obtidos no ponto de background. Os limites de TEL (Threshold Effect Level) e PEL (Probable Effect Level) não foram ultrapassados em nenhuma das campanhas de coleta. O Valor-Guia de Unidades Tóxicas de Águas Intersticiais (VGUTAI), preconizado pela USEPA, aplicado aos sedimentos foi violado em todos os pontos de amostragem. A aplicação do VGQS, baseado na teoria do equilíbrio de partição, mostrou que os metais são controlados pela fase sulfídrica presente nos sedimentos, indicando que os metais presentes nos sedimentos não estão biodisponíveis. Os resultados de avaliação da estrutura da comunidade bentônica indicaram uma possível influência da barragem de rejeitos sobre a estrutura da comunidade bentônica nos trechos avaliados do córrego Lavagem e no ribeirão Espírito Santo, a jusante da confluência com o córrego Lavagem. Os resultados dos testes de ecotoxicidade demonstraram a ocorrência apenas de efeitos tóxicos crônicos. / Sediment Quality Values Guidelines (SQVG) have been used for evaluating ecological risk associated with the sediment contamination for benthonic organisms. The main objective of this work was to develop methodologies and to collect data that allowed the application of SQVG for the following metals: Cd, Cu, Pb, Ni and Zn. The location chosen for the case study was the sub-watershed of the Espírito Santo Stream, which is part of the Unit Planning and Hydrologic Resources Management UPGRH SF4, of the São Francisco river watershed, located in the area of the city of Três Marias. The life in the sub-watershed is significantly affected by the installation of a waste dam that controls effluents coming from the zinc-ore beneficiation plant. Our studies addressed the biogeochemical characterization of the sediments of those environments through the determination of the concentrations of acid volatile sulfide (AVS), the study of the partitioning of metals among the total sediments and interstitial waters, and the determination of the fraction of metals associated with AVS in the total sediments. The data obtained were analyzed in association with those related to the analysis of the structure of the benthonic community and eco-toxicity tests. The studies also included analysis of the physico-chemical variables and concentration of metals in the samples of the surface water; mineralogical and granulometric analyses, quantity of organic matter and concentration of inorganic contaminants in the samples of sediments. High levels of electrical conductivity and total solids diluted were found in one surface water sample site. The results of a Neutron Activation Analysis (NAA) showed that the chemical elements predominant in the samples were Ti, Al, V, Mn and Fe. The results of the total metal analysis in sediment samples showed that only one point located in the Lavagem stream, situated immediately downstream the dam and before flowing into the Espirito Santo stream, presented concentration values of the evaluated metals (Cd, Cu, Pb, Ni and Zn) above those of the background value. The TEL limits (Threshold Effect Level) and PEL (Probable Effect Level) were not exceeded in any of the sampling campaigns. The Guidelines Interstitial Water Toxic Units (IWTU), established by the USEPA, was violated for all samples. The application of SQVG, based on the equilibrium partition theory, showed that metals are controlled by the sulfide-phase present in sediments. This indicates that the metals presented in the sediments may have low bio-availability. The results of the evaluation of the benthonic community structure indicated a possible influence of the waste dam in the Lavagem stream and in the Espírito Santo stream, downstream the confluence with the Lavagem stream. The results of eco-toxicity tests showed that contaminated sediment can cause only chronic effects.

As águas superficiais da Amazônia, que englobam, dentre outros, rios das mais variadas ordens, desde pequenos igarapés, até o rio Amazonas, um dos maiores do mundo, são um componente importante do funcionamento deste ecossistema de escala continental. Um paradigma acerca dos sistemas fluviais é que estes integram os processos que ocorrem em suas bacias de drenagem e, portanto, alterações na cobertura vegetal e nos usos da terra, podem interferir diretamente nos parâmetros físicos e químicos dos compartimentos aquáticos. Algumas áreas da Amazônia, como o estado de Rondônia, por exemplo, têm sofrido alterações substanciais nas suas bacias de drenagem e as conseqüências destas nos sistemas fluviais ainda são pouco compreendidas. Sabe-se que a conversão de florestas em pastagens, o tipo de alteração mais comum nos ecossistema de Rondônia, resulta na compactação dos solos e no aumento da erosão. Ao mesmo tempo, diversos estudos têm demonstrado que os sedimentos em suspensão, carreados nos rios, podem ser importantes fontes de carbono e outros nutrientes limitantes para o metabolismo aquático. Com base nestas premissas, este estudo visou avaliar a importância dos sedimentos em suspensão grossos (maiores que 63 µm) e finos (menores que 63 µm e maiores 0,1 µm) no metabolismo de alguns rios de Rondônia, sob diversas coberturas e usos da terra. Para tal, as concentrações destes sedimentos foram artificialmente aumentadas em amostras incubadas no escuro, nas quais os consumos de oxigênio indicaram as taxas respiratórias. No caso dos sedimentos em suspensão finos, observaram-se aumentos nas taxas respiratórias em praticamente todos os casos nos quais suas concentrações foram aumentadas. Os sedimentos em suspensão grossos, por outro lado, somente favorecem aumentos nas taxas respiratórias nos períodos de maiores precipitações. Estes resultados relacionam-se com a composição dos sedimentos. Sedimentos em suspensão finos, por sua maior capacidade de adsorver substâncias orgânicas e nutrientes, constituem uma fonte permanente de substrato para o metabolismo aquático. Sedimentos em suspensão grossos, por outro lado, somente carreiam fração substancial de material orgânico durante o período de maiores precipitações, quando os solos são “lavados" pelas enxurradas. Apesar destas diferenças no papel relativo destas duas frações do material particulado no metabolismo destes rios, fica evidente que o aumento da erosão pode acarretar mudanças significativas no metabolismo dos sistemas fluviais da Amazônia. / Amazonian surface waters, encompassing rivers of distinct orders, from small streams to one of the largest of the world, the Amazon, are important components of the functioning of this continental-scale ecosystem. A paradigm about fluvial systems is that they integrate the processes that occur at their watersheds and, therefore, changes in land use/cover may directly interfere on physical and chemical parameters of the aquatic compartments. In some areas of the Amazon, such as in the state of Rondonia, for example, the landscape has been significantly altered, and the consequences of these changes are still poorly understood. It is well know that the conversion of forests into pastures, a common type of land use change in Rondonia, can cause the compactation of soils and increased erosion rates. At the same time, several studies have demonstrated that suspended sediments can be important sources of carbon and other nutrients to river metabolism. Based on those assumptions, this study aimed the evaluation of the importance of coarse (larger than 63 µm) and fine (smaller than 63 µm and larger than 0,1 µm) suspended sediments on the metabolism of some rivers of Rondonia. This assessment was made through incubations in the dark of samples enriched in sediments, in which the consumption of oxygen indicated respiration rates. In the case of fine suspended sediments, most incubated samples showed increased respiration rates as a result of the concentration of these particles. Coarse suspended sediments, on another side, only favor the increase in respiration rates during the high water period. These results are related to the composition of these sediments. Fine suspended sediments constitute a constant substract for the aquatic metabolism, as a result of their larger surface area and capacity for absorving organic substances and nutrients. Coarse sediments, however, only favor respiration during the high water period, when overland flow carries organic debris from land into the water systems. Although these two fractions of suspended sediments have different relative roles in the metabolism of rivers, it becomes evident from this data that increasing the erosion in the Amazon may affect significantly the metabolism of fluvial systems in this region.

As águas superficiais da Amazônia, que englobam, dentre outros, rios das mais variadas ordens, desde pequenos igarapés, até o rio Amazonas, um dos maiores do mundo, são um componente importante do funcionamento deste ecossistema de escala continental. Um paradigma acerca dos sistemas fluviais é que estes integram os processos que ocorrem em suas bacias de drenagem e, portanto, alterações na cobertura vegetal e nos usos da terra, podem interferir diretamente nos parâmetros físicos e químicos dos compartimentos aquáticos. Algumas áreas da Amazônia, como o estado de Rondônia, por exemplo, têm sofrido alterações substanciais nas suas bacias de drenagem e as conseqüências destas nos sistemas fluviais ainda são pouco compreendidas. Sabe-se que a conversão de florestas em pastagens, o tipo de alteração mais comum nos ecossistema de Rondônia, resulta na compactação dos solos e no aumento da erosão. Ao mesmo tempo, diversos estudos têm demonstrado que os sedimentos em suspensão, carreados nos rios, podem ser importantes fontes de carbono e outros nutrientes limitantes para o metabolismo aquático. Com base nestas premissas, este estudo visou avaliar a importância dos sedimentos em suspensão grossos (maiores que 63 µm) e finos (menores que 63 µm e maiores 0,1 µm) no metabolismo de alguns rios de Rondônia, sob diversas coberturas e usos da terra. Para tal, as concentrações destes sedimentos foram artificialmente aumentadas em amostras incubadas no escuro, nas quais os consumos de oxigênio indicaram as taxas respiratórias. No caso dos sedimentos em suspensão finos, observaram-se aumentos nas taxas respiratórias em praticamente todos os casos nos quais suas concentrações foram aumentadas. Os sedimentos em suspensão grossos, por outro lado, somente favorecem aumentos nas taxas respiratórias nos períodos de maiores precipitações. Estes resultados relacionam-se com a composição dos sedimentos. Sedimentos em suspensão finos, por sua maior capacidade de adsorver substâncias orgânicas e nutrientes, constituem uma fonte permanente de substrato para o metabolismo aquático. Sedimentos em suspensão grossos, por outro lado, somente carreiam fração substancial de material orgânico durante o período de maiores precipitações, quando os solos são “lavados” pelas enxurradas. Apesar destas diferenças no papel relativo destas duas frações do material particulado no metabolismo destes rios, fica evidente que o aumento da erosão pode acarretar mudanças significativas no metabolismo dos sistemas fluviais da Amazônia. / Amazonian surface waters, encompassing rivers of distinct orders, from small streams to one of the largest of the world, the Amazon, are important components of the functioning of this continental-scale ecosystem. A paradigm about fluvial systems is that they integrate the processes that occur at their watersheds and, therefore, changes in land use/cover may directly interfere on physical and chemical parameters of the aquatic compartments. In some areas of the Amazon, such as in the state of Rondonia, for example, the landscape has been significantly altered, and the consequences of these changes are still poorly understood. It is well know that the conversion of forests into pastures, a common type of land use change in Rondonia, can cause the compactation of soils and increased erosion rates. At the same time, several studies have demonstrated that suspended sediments can be important sources of carbon and other nutrients to river metabolism. Based on those assumptions, this study aimed the evaluation of the importance of coarse (larger than 63 µm) and fine (smaller than 63 µm and larger than 0,1 µm) suspended sediments on the metabolism of some rivers of Rondonia. This assessment was made through incubations in the dark of samples enriched in sediments, in which the consumption of oxygen indicated respiration rates. In the case of fine suspended sediments, most incubated samples showed increased respiration rates as a result of the concentration of these particles. Coarse suspended sediments, on another side, only favor the increase in respiration rates during the high water period. These results are related to the composition of these sediments. Fine suspended sediments constitute a constant substract for the aquatic metabolism, as a result of their larger surface area and capacity for absorving organic substances and nutrients. Coarse sediments, however, only favor respiration during the high water period, when overland flow carries organic debris from land into the water systems. Although these two fractions of suspended sediments have different relative roles in the metabolism of rivers, it becomes evident from this data that increasing the erosion in the Amazon may affect significantly the metabolism of fluvial systems in this region.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-07-01T13:22:09Z No. of bitstreams: 1 thiagopereiradasilva.pdf: 8248708 bytes, checksum: 846bfbc45d8748f458221b2646af213c (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-07-13T16:00:55Z (GMT) No. of bitstreams: 1 thiagopereiradasilva.pdf: 8248708 bytes, checksum: 846bfbc45d8748f458221b2646af213c (MD5) / Made available in DSpace on 2016-07-13T16:00:55Z (GMT). No. of bitstreams: 1 thiagopereiradasilva.pdf: 8248708 bytes, checksum: 846bfbc45d8748f458221b2646af213c (MD5) Previous issue date: 2012-03-29 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O bacterioplâncton é um dos componentes da cadeia trófica em ecossistemas aquáticos, sendo regulado por fatores como disponibilidade de nutrientes, temperatura, predação e infecção viral. A morte de bactérias é caracterizada como o processo de perda funcional e morfológica da célula e tem função de controlar a abundância e produção bacteriana, com significado importante na ciclagem de carbono e nutrientes nos ecossistemas aquáticos. No entanto, o processo de morte celular em bactérias aquáticas é desconhecido. Ainda não se sabe se a morte celular programada (MPC), processo regulado e bem conhecido em organismos eucariotos, ocorre em bactérias aquáticas. Este trabalho teve por objetivo investigar a ocorrência de MCP em bactérias aquáticas utilizando dois modelos de ecossistemas: lago Batata e lago dos Manacás. O lago Batata é um ecossistema amazônico de inundação e encontra-se divido em duas áreas: impactada e natural. O lago dos Manacás é um ecossistema artificial localizado em Minas Gerais. Três grupos de bactérias, provenientes do lago dos Manacás, lago Batata – área natural e lago Batata – área impactada foram estudados. As amostras de água foram coletadas na sub-superfície dos lagos e processadas para análises da viabilidade celular por microscopia de fluorescência e da fragmentação de DNA por citometria de fluxo. Em paralelo, amostras do lago Batata – área impactada foram processadas para microscopia eletrônica de transmissão (MET) para caracterizar a diversidade ultraestrutural da comunidade bacteriana e investigar a ocorrência de alterações celulares bacterianas. Os resultados da viabilidade celular utilizando marcadores específicos para integridade de membrana (LIVE/DEAD BacLight) mostraram diretamente a presença de bactérias vivas/viáveis e mortas/inviáveis . O lago Batata apresentou maior proporção de morte celular bacteriana (36,20%) em comparação com o lago dos Manacás (19,66%). As análises de fragmentação de DNA (ensaio TUNEL) mostraram que a MCP constitui um fenômeno presente nos ecossistemas aquáticos estudados, com maior ocorrência na área impactada do lago Batata. A MET revelou a presença de bactérias aquáticas com grande diversidade ultraestrutural, representada por diferenças morfológicas quanto ao envoltório celular, cápsula, mesossomos, vesículas membranosas, partículas aderidas e tilacóides. Além disso, 34,28% das bactérias apresentavam vírus no citoplasma. A diversidade ultraestrutural pode representar a ampla diversidade metabólica e adaptativa bacteriana, enquanto a presença de vírus parece relacionada com a morte bacteriana. As análises de integridade ultraestrutural mostraram que (i) a maior proporção de bactérias encontrava-se com alterações ultraestruturais indicativas de processo de morte, (ii) a frequência de ocorrência de cápsula foi menor nas bactérias vazias, (iii) bactérias com alterações apresentaram maior frequência de partículas aderidas, e (v) o número médio de fagos por secção foi significativamente maior nas bactérias alteradas em comparação com as bactérias intactas. A MET mostrou a ocorrência de alterações ultraestruturais típicas de MCP, como retração e condensação citoplasmáticas. Em conjunto, nossos dados ressaltam morte bacteriana como um evento importante atuando na regulação da comunidade bacteriana e demonstra, pela primeira vez, a ocorrência de MCP em bactérias aquáticas. A MCP pode ter significado funcional como um dos mecanismos desenvolvidos para sobrevivência da comunidade bacteriana. / The bacterioplankton is an important component of the food web structure in aquatic ecosystems and it is regulated by many factors such as nutrient supply, temperature, predation and virus infection. Bacterial death is characterized by functional and morphological loss of the bacterial cell with roles in the control process of abundance and bacterial production of aquatic ecosystems and functional meaning in the carbon and nutrient cycles. However, the cell death process in aquatic bacteria remains to be defined. Programmed cell death (PCD) is a regulated process largely known in eukaryotic organisms. There are not studies dealing with PCD occurrence in aquatic bacteria. This study aimed to investigate the occurrence of PCD in free-living aquatic bacteria in two models of aquatic ecosystems: Batata lake and Manacás lake. Batata Lake is located on floodplain Trombetas river in northern of Brazilian Amazon. This ecosystem has been impacted for bauxite tailings over teen years and it is presently divided in impacted and natural stations. Manacás Lake is an artificial system located in Minas Gerais. Three groups of bacteria from Manacás Lake, Batata lake – impacted area - and Batata lake – natural area -were studied. Water samples were collected from the subsurface of these lakes and processed for analyses of bacterial viability by fluorescence microscopy and DNA fragmentation by flow citometry. In parallel, samples from Batata lake – impacted area - were processed for transmission electron microscopy (TEM) for characterization of the bacterial community ultrastructural diversity and investigation of bacteria alterations. Our cell viability results using specific markers for membrane integrity (LIVE/DEAD BacLight) showed directly the presence of live/viable and dead/ not viable bacteria. Batata lake presented a higher proportion of bacterial death (36,20%) compared to Manacás lake (19,66%). DNA fragmentation analyses (TUNEL assay) showed that PCD is a phenomenon occurring in all aquatic ecosystems investigated, with higher frequency in the Batata lakeimpacted area compared to the other ecosystems. Our TEM analyses revealed a great bacterial ultrastructural diversity represented by morphological differences in the cellular envelope, capsule, mesosomes, membrane vesicles, attached particles and thylacoids. Moreover, around 34,28% of the bacteria showed virus in their cytoplasm. The ultrastructural diversity may represent the large metabolic and adaptative diversity of aquatic bacteria while the presence of virus may be related to bacterial death. Our ultrastructural integrity analyses showed that (i) the higher proportion of bacteria was in death process or dead (damaged and empty), (ii) the capsule frequency is lower in empty bacteria, (iii) the higher bacteria-particle association was found in altered bacteria, (iv) the mean number of viruses per cell section was higher in altered compared to intact bacteria. TEM also showed the presence occurrence of typical ultrastructural changes indicative of MPC, such as cell retraction and condensation. Altogether, our data demonstrate that PCD occur in aquatic bacteria, and that this event may be a survival mechanism for bacterial communities in these ecosystems.

Abstract Freshwater ecosystems are severely threatened by a variety of anthropogenic stressors. In order to keep track with at least part of the changes, it is important to efficiently assess and monitor freshwater biological diversity. Biological assessment programs are developed to detect human-induced changes in the ecological state of aquatic systems. These programs typically rely on the assumption that environmental conditions are the sole drivers of biological communities occupying a site and, thus, these local communities would correctly inform about environmental conditions. Recently, this background principle of current bioassessment methods has faced some criticism, stemming from the idea that community structuring is a more complex process than just a mere result of local environmental conditions. In this thesis, I studied the natural and anthropogenic drivers of freshwater biodiversity. I was particularly interested if the various biodiversity metrics studied showed any spatial patterns and if so, for which reasons these patterns might occur. To obtain a comprehensive picture of spatial patterns in biodiversity, I studied multiple spatial contexts, biological groups and indices. I found that environmental conditions were not the only drivers of freshwater biodiversity. Instead, different spatial patterns, likely stemming from dispersal processes, were surprisingly powerful drivers of aquatic communities and index values derived from them. The spatial context (i.e. spatial extent and connectivity) of the aquatic study systems likely played a major role in structuring biodiversity. I also found that the distinct biological groups and indices studied were partly related to different predictor variables. The findings of this thesis are of importance to the development of new bioassessment methods. The results of this thesis also suggest that the spatial context of the study setting should be acknowledged when interpreting results based on current bioassessment methods. / Tiivistelmä Makeanveden ekosysteemit ovat hyvin alttiita ihmistoiminnalle. Ekosysteemissä mahdollisesti tapahtuvien muutosten havaitseminen vaatii tehokkaita vesistöjen ekologisen tilan ja luonnon monimuotoisuuden arviointi- ja seurantamenetelmiä. Näiden menetelmien toimintaperiaatteen yleisenä tausta-ajatuksena on, että biologiset yhteisöt määräytyvät paikallisten ympäristöolojen mukaan. Tietyn paikan yhteisön oletetaan siis heijastavan kyseisen paikan ympäristön tilaa. Viime aikoina tausta-ajatus paikallisten ympäristöolojen merkityksestä ainoana eliöyhteisöjä muovaavana tekijänä on kuitenkin kohdannut kritiikkiä. Kriitikot painottavat, että biologisten yhteisöjen rakenteeseen vaikuttavat monet muutkin asiat kuin paikalliset ympäristöolosuhteet ja niissä tapahtuvat ihmisperäiset muutokset. Väitöskirjassani tutkin sisävesien luonnon monimuotoisuuteen vaikuttavia tekijöitä. Olin erityisen kiinnostunut siitä, näkyykö tutkituissa biologisissa parametreissa maantieteellisessä tilassa ilmeneviä spatiaalisia säännönmukaisuuksia. Saadakseni mahdollisimman laaja-alaisen käsityksen luonnon monimuotoisuudessa esiintyvistä spatiaalisista säännönmukaisuuksista, tutkin useaa spatiaalista kontekstia, eliöryhmää ja indeksiä. Tutkimuksessa selvisi, että paikalliset ympäristöolosuhteet eivät ole ainoita luonnon monimuotoisuuteen vaikuttavia tekijöitä. Erilaiset spatiaaliset säännönmukaisuudet, todennäköisesti eliöiden levittäytymiseen liittyvien seikkojen aiheuttamina, olivat yllättävän yleisiä makeiden vesien eliöyhteisöjen rakenteessa ja niihin perustuvien indeksien arvoissa. Tutkimussysteemien spatiaalinen konteksti (alueen laajuus ja paikkojen väliset spatiaaliset suhteet) selvästi vaikutti luonnon monimuotoisuutta kuvastavien indeksien arvojen vaihteluun. Lisäksi selvisi, että eri eliöryhmät ja indeksit olivat useimmiten liitoksissa hyvin erilaisiin selittäviin muuttujiin, osoittaen, että nämä mittarit kuvastavat eri asioita. Väitöskirjassa esitetyt havainnot on tärkeää huomioida vesistöjen ekologisen tilan ja luonnon monimuotoisuuden arviointi- ja seurantamenetelmiä kehitettäessä. Spatiaalisen kontekstin merkitys olisi hyvä huomioida myös nykyisten arviointi- ja seurantamenetelmien tuottamien tulosten tulkinnassa.

La prise de conscience des problèmes d'environnement et des effets directs et indirects des activités humaines a conduit à renforcer la surveillance haute fréquence des écosystèmes marins par l'installation de stations de mesures multicapteurs autonomes. Les capteurs, installés dans des milieux hostiles, sont sujets à des périodes de calibration, d'entretien voire des pannes et sont donc susceptibles de générer des données bruitées, manquantes voire aberrantes qu'il est nécessaire de filtrer et compléter avant toute exploitation ultérieure. Dans ce contexte, l'objectif du travail est de concevoir un système numérique automatisé robuste capable de traiter de tel volume de données afin d’améliorer les connaissances sur la qualité des systèmes aquatiques, et plus particulièrement en considérant le déterminisme et la dynamique des efflorescences du phytoplancton. L'étape cruciale est le développement méthodologique de modèles de prédiction des efflorescences du phytoplancton permettant aux utilisateurs de disposer de protocoles adéquats. Nous proposons pour cela l'emploi du modèle de Markov caché hybridé pour la détection et la prédiction des états de l'environnement (caractérisation des phases clefs de la dynamique et des caractéristiques hydrologiques associées). L'originalité du travail est l'hybridation du modèle de Markov par un algorithme de classification spectrale permettant un apprentissage non supervisé conjoint de la structure, sa caractérisation et la dynamique associée. Cette approche a été appliquée sur trois bases de données réelles : la première issue de la station marine instrumentée MAREL Carnot (Ifremer) (2005-2009), la seconde d’un système de type Ferry Box mis en œuvre en Manche orientale en 2012 et la troisième d’une station de mesures fixe, installée le long de la rivière Deûle en 2009 (Agence de l’Eau Artois Picardie - AEAP). Le travail s’inscrit dans le cadre d’une collaboration étroite entre l'IFREMER, le LISIC/ULCO et l'AEAP afin de développer des systèmes optimisés pour l’étude de l’effet des activités anthropiques sur le fonctionnement des écosystèmes aquatiques et plus particulièrement dans le contexte des efflorescences de l’algue nuisible, Phaeocystis globosa. / Because of the growing interest for environmental issues and to identify direct and indirect effects of anthropogenic activities on ecosystems, environmental monitoring programs have recourse more and more frequently to high resolution, autonomous and multi-sensor instrumented stations. These systems are implemented in harsh environment and there is a need to stop measurements for calibration, service purposes or just because of sensors failure. Consequently, data could be noisy, missing or out of range and required some pre-processing or filtering steps to complete and validate raw data before any further investigations. In this context, the objective of this work is to design an automatic numeric system able to manage such amount of data in order to further knowledge on water quality and more precisely with consideration about phytoplankton determinism and dynamics. Main phase is the methodological development of phytoplankton bloom forecasting models giving the opportunity to end-user to handle well-adapted protocols. We propose to use hybrid Hidden Markov Model to detect and forecast environment states (identification of the main phytoplankton bloom steps and associated hydrological conditions). The added-value of our approach is to hybrid our model with a spectral clustering algorithm. Thus all HMM parameters (states, characterisation and dynamics of these states) are built by unsupervised learning. This approach was applied on three data bases: first one from the marine instrumented station MAREL Carnot (Ifremer) (2005-2009), second one from a Ferry Box system implemented in the eastern English Channel en 2012 and third one from a freshwater fixed station in the river Deûle in 2009 (Artois Picardie Water Agency). These works fall within the scope of a collaboration between IFREMER, LISIC/ULCO and Artois Picardie Water Agency in order to develop optimised systems to study effects of anthropogenic activities on aquatic systems functioning in a regional context of massive blooms of the harmful algae, Phaeocystis globosa.

Many polycyclic aromatic compounds (PACs) are toxic, carcinogenic, and mutagenic. As a result, their effects on aquatic biota and ecosystems are of great concern. Research on PACs in aquatic biota often overlooks the role of amphibians, alkylated PACs, and sediment as an uptake route. In order to study the accumulation and toxicokinetics of PACs following sediment and aqueous exposure, and to compare the bioaccumulation potentials of parent and alkyl PACs, two accumulation-elimination experiments using wood frog tadpoles (Lithobates sylvaticus) of Gosner stage 28-32 were conducted (one evaluating exposure to contaminated sediment and water, and the other to contaminated water alone). A complementary field study was then conducted near Fort McMurray, Alberta to assess PAC body burdens in field-collected amphibian larvae, and to determine whether PAC body burdens are related to exposure to sediment and/or water in the field. The results of our studies showed that PAC concentrations and uptake rates in wood frog tadpoles were highest when they were exposed to PAC-contaminated sediment. Consequently, we determined that the dominant route of exposure of wood frog tadpoles to PACs is sediment rather than water. This finding supports other studies that have shown dietary uptake to be an important route of PAC exposure in other aquatic organisms. In both the laboratory and field study, alkyl PAC concentrations exceeded those of parent PACs in wood frog tadpoles, which also demonstrated petrogenic PAC profiles. Interestingly, parent PACs seemed to have greater bioaccumulation potential than alkyl PACs in the laboratory-exposed wood frog tadpoles (in relation to sediment), possibly due to greater bioavailability or lower metabolism of parent PACs or alternatively, due to a saturation in uptake of alkyl PACs. Nevertheless, only a few compounds, including anthracene, fluoranthene, retene, and C1-benzofluoranthenes/benzopyrenes, were found to have higher bioaccumulation potentials. Lithobates sylvaticus tadpoles seemed to be efficient at eliminating and metabolizing both parent and alkyl PACs. However, the elimination of some compounds, such as C4-naphthalenes, was not as efficient. Furthermore, C3-fluorenes and C2-dibenzothiophenes were isolated as potential markers of amphibian larvae exposure to PAC-contaminated sediment due to their positive correlation with the wetland sediment concentrations. Additional field collections in the Athabasca oil sands are warranted to verify the utility of these markers in the natural environment. Evidently, this thesis highlights the importance of including sediment exposure and alkylated PACs in toxicological and field studies of benthic and epibenthic organisms. The results of this study are the largest, most comprehensive set of toxicokinetic and bioaccumulation information of PACs (52 analytes) in the amphibian larvae Lithobates sylvaticus obtained to date.

Diese Dissertation beschäftigt sich mit der passiven Probenahme von hydrophoben organischen Schadstoffen in Oberflächengewässern: Polyaromatische Kohlenwasserstoffe (PAK), polychlorierte Biphenyle (PCB), polybromierte Biphenylether (PBDE), Organochlorpestizide (u. a. HCH, DDX) und weitere hydrophobe Pestizide. Die Zielstellung dieser Arbeit lag bei der Validierung des Silikonstabs als Alternativmethode im Gewässermonitoring zu konventionellen Probenahmetechniken wie Schöpf- und Wochenmischproben der Wasserphase sowie Schwebstoffanalysen. Die Probenahme mit dem Silikonstab erfolgte durch dessen Exposition im Gewässer für einen Zeitraum zwischen einer Woche und mehreren Monaten. Nach Einholung wurden die im Silikonstab akkumulierten Schadstoffe (Analyten) mittels instrumenteller Analytik quantifiziert. Die Probenaufgabe erfolgte ohne vorherige Lösungsmittelextraktion durch direktes Erhitzen des Silikonstabs, wodurch die Analyten vom Polymer desorbieren (Thermodesorption). Die durch Hitze freigesetzten Analyten wurden direkt auf eine chromatographische Trennsäule gegeben und massenspektroskopisch quantifiziert. Nach Erhalt der Ergebnisse der Silikonstab-Analytik gibt es verschiedene Herangehensweisen für die Berechnung der zeitgemittelten Analytkonzentrationen im Gewässer, die in dieser Arbeit vorgestellt und diskutiert werden. Dazu gehören die Verwendung von experimentellen Daten aus Kalibrierversuchen und Berechnungen auf Grundlage von physikochemischen Eigenschaften der Analyten wie dem Sammler-Wasser-Verteilungskoeffizienten. Im Zuge dieser Arbeit wurde die Aufnahmekinetik des Silikonstabs bei verschiedenen Temperaturen und Fließgeschwindigkeiten mit Hilfe von Kalibrierversuchen untersucht. Die gewonnenen experimentellen Daten wurden für die Entwicklung von Rechenmodellen herangezogen, mit denen das Aufnahmeverhalten vorgesagt werden soll. Es wurden Sammler-Wasser-Verteilungskoeffizienten für den Silikonstab u. a. mit der Kosolvenzmethode bestimmt und als Parameter für die Berechnung von zeitgemittelten Analytkonzentrationen des Gewässers verwendet. Für die Validierung wurde der Silikonstab in zwei Gewässergütemessstationen der Fließgewässer Mulde (Dessau) und Elbe (Magdeburg) in Durchflussbehältern exponiert und die zeitgemittelten Analytkonzentrationen mit verschiedenen Rechenmodellen bestimmt. Die erhaltenen Werte werden mit gleichzeitig entnommenen Wochenmischproben der Wasserphase sowie monatlichen Schwebstoffproben verglichen und die Eignung des Silikonstabs als alternative Probenahmemethode für das Umweltmonitoring von Oberflächengewässern diskutiert.:I ZUSAMMENFASSUNG ...................................................................................................... 2 II INHALTSVERZEICHNIS .................................................................................................. 3 III ABBILDUNGSVERZEICHNIS .......................................................................................... 5 IV TABELLENVERZEICHNIS ................................................................................................ 6 V GLEICHUNGSVERZEICHNIS ............................................................................................ 7 VI ABKÜRZUNGSVERZEICHNIS........................................................................................... 9 0 VIELEN DANK AN … ...................................................................................................... 11 1. EINLEITUNG ................................................................................................................ 12 1.1 Wasser, seine Nutzung und Verschmutzung ............................................................ 12 1.2 Das Wasser und seine Schadstoffe .......................................................................... 15 1.3 Monitoring von Oberflächengewässern .................................................................... 17 1.3.1 Entnahme konventioneller Schöpfproben .............................................................. 17 1.3.2 Entnahme von Mischproben (integrative oder Kompositproben) ........................... 18 1.3.3 Probenahme des Schwebstoffanteils in der Wasserphase .................................... 19 2. PASSIVSAMMLER IN DER WASSERANALYTIK ................................................................ 21 2.1 Theoretische Grundlagen ......................................................................................... 21 2.1.1 Allgemeiner Aufbau von Passivsammlern ............................................................... 23 2.1.2 Die einzelnen Schritte von der Wasser- in die Sammelphase ................................ 25 2.1.3 Adsorptive und absorptive Akkumulation des Analyten in der Sammelphase ........ 26 2.2 Passivsammlersysteme in der Wasseranalytik ......................................................... 28 2.2.1 Absorbierende Passivsammler für hydrophobe Analyten ....................................... 28 2.2.1.1 Semipermeable membrane device (SPMD) .......................................................... 28 2.2.1.2 LDPE-Streifen (LDPE strips) ................................................................................ 29 2.2.1.3 Silikonplatten (silicone sheets) ........................................................................... 30 2.2.1.4 Chemcatcher ...................................................................................................... 31 2.2.1.5 Lösungsmittelfreie Passivsammler (MESCO / Silikonstab) .................................. 32 2.2.2 Absorbierende Passivsammler für polare Analyten ............................................... 35 2.2.2.1 Polar organic integrative Sampler (POCIS) ......................................................... 35 2.2.2.2 Chemcatcher ...................................................................................................... 35 2.3 Auswertung von Passivsammlerdaten ..................................................................... 35 2.3.1 Gleichgewichtssammler ......................................................................................... 36 2.3.2 Laborkalibrierung .................................................................................................. 37 2.3.3 In-situ-Kalibrierung mit Performance Reference Compounds (PRC) ...................... 38 2.3.4 Validierung von Passivsammlern............................................................................ 39 3. LÖSLICHKEIT UND THERMODYNAMISCHES GLEICHGEWICHT ...................................... 41 3.1 Freie Enthalpie und chemisches Potential ................................................................ 41 3.2 Lineare freie Energie-Beziehungen (LFER) für die Abschätzung von KSW ................ 41 3.3 Kosolvenzmodelle für die Modellierung von KSW ...................................................... 43 3.3.1 Log-Linear-Modell von Yalkowsky .......................................................................... 43 3.3.2 Freie Enthalpie-Ansatz (Khossravi-Connors-Modell) .............................................. 44 3.3.3 Jouyban-Acree-Modell ............................................................................................ 44 4. MATERIAL UND METHODEN ......................................................................................... 45 4.1 Präparation der verwendeten Passivsammler .......................................................... 45 4.2 Laborkalibrierung zur Bestimmung von Sammelraten ............................................... 45 4.2.1 Beschreibung der Versuche für die Silikonstab-Kalibrierung .................................. 45 4.3 Experimentelle Bestimmung von Sammler-Wasser-Verteilungskoeffizienten KSW ... 48 4.3.1 Zeitabhängige KSW-Bestimmung in der Wasserphase .......................................... 48 4.3.2 KSW-Bestimmung mit der Kosolvenzmethode ....................................................... 50 4.4 Validierung des Silikonstabs an limnischen Gewässergütemessstationen ............... 52 5. ERGEBNISSE UND DISKUSSION ................................................................................... 55 5.1 Sammelraten RS für den Silikonstab aus Kalibrierversuchen .................................... 55 5.1.1 Temperaturabhängigkeit ....................................................................................... 58 5.1.2 Einfluss der Hydrodynamik auf die Aufnahmekinetik von PAK ................................ 59 5.1.3 Modellierung von Sammelraten .............................................................................. 62 5.1.3.1 Polynomisches Modell nach Vrana [137] ............................................................. 62 5.1.3.2 Diffusionsmodell nach Booij [71] ......................................................................... 64 5.1.3.3 Diffusionsmodell nach Rusina [85] ...................................................................... 66 5.1.4 Wahl der geeigneten In-situ-Kalibrierung am Beispiel eines Kalibrierversuchs ..... 67 5.1.4.1 Berechnung von In-situ-Sammelraten mit RS-Modellen ...................................... 68 5.1.4.2 Berechnung von In-situ-Sammelraten über Eliminierung von PRCs .................... 69 5.1.4.3 Vergleich Modelle und PRCs mit experimentellen Sammelraten .......................... 70 5.2 Experimentelle Bestimmung des Sammler-Wasser-Verteilungskoeffizienten KSW ... 73 5.2.1 Zeitabhängige KSW-Bestimmung in der Wasserphase .......................................... 73 5.2.2 Zusammenfassung KSW(t)-Versuche in der Wasserphase .................................... 81 5.2.3 KSW-Bestimmung mit der Kosolvenzmethode ....................................................... 81 5.2.3.1 Kosolvenzmodelle ............................................................................................... 83 5.2.4 Zusammenfassung ................................................................................................ 90 5.3 Empirische Modelle zur Abschätzung von KSW-Werten ............................................ 92 5.3.1 Lineare Korrelation des KSW mit physikochemischen und Molekülparametern ...... 92 5.3.2 Berechnung mit Mehrparameter-Regression (LSER) .............................................. 95 5.3.3 Zusammenfassung Abschätzung von KSW-Werten für den Silikonstab ................. 97 5.4 Freilandvalidierung des Silikonstab-Passivsammlers ................................................ 97 5.4.1 Ausbringung an Gewässergütemessstationen....................................................... 97 5.4.1.1 Validierung des Silikonstabs mit Wasserproben ............................................... 100 5.4.1.2 Validierung des Silikonstabs mit Sedimentproben ............................................ 102 5.4.2 Validierung des Silikonstabs bei Laborvergleichsstudien ..................................... 105 6. ERGEBNISSE UND AUSBLICK ..................................................................................... 105 VII LITERATURVERZEICHNIS ......................................................................................... 107 VIII ANHANG ................................................................................................................. 116

Il est de plus en plus clair que nos écosystèmes sont liés les uns aux autres de façons multiples et parfois complexes. De plus en plus nous identifions de nouvelles façons dont deux systèmes échangent des ressources et de l’énergie. Le rôle de ces échanges et transferts est encore difficile à quantifier. Nous en savons relativement peu sur les interactions de ceux-ci avec des processus locaux et spatiaux, ainsi que sur leur importance relativement à ces processus locaux et régionaux plus connus. À travers ce projet, nous cherchons à éclaircir l’importance relative des subsides particulaires de feuilles (et particulièrement de leur stoechiométrie) sur les communautés littorales d’invertébrés benthiques. Nous avons récolté des invertébrés et des feuilles mortes sur 23 sites à travers 7 lacs relativement isolés de l’influence humaine et avons comparé l’influence de la stoechiométrie de ces subsides à l’influence de facteurs locaux de qualité de l’eau et de facteurs spatiaux. Les résultats suggèrent que l’importance de la qualité nutritive (i.e. stoechiométrie) est secondaire aux facteurs locaux de qualité de l’eau dans des milieux naturels. L’importance des subsides particulaires de feuilles semble être grandement dépendante du contexte et pourrait gagner en importance dans des contextes de fortes perturbations. Les résultats indiquent que la qualité nutritive des subsides de feuilles, leur contenu relatif en azote et phosphore, est associée avec de plus grande abondance de plusieurs taxons. La richesse spécifique n’étant pas affecté, c’est à travers l’équitabilité (i.e. une distribution des abondances plus stable) que la qualité nutritive des feuilles pourrait promouvoir une plus grande biodiversité d’invertébrés benthiques. / It is clear now, more than ever before, that our ecosystems are linked to one another in multiples and sometimes complex ways. The role of these exchanges and transfers is still hard to quantify. We know little of the interactions of these fluxes with local and spatial processes happening in ecosystems, as well as their relative importance compared to local and regional drivers of ecosystem functions and community structure. With this project, we aim to untangle the relative importance of particulate subsidies of leaves (and specifically their stoichiometry) on littoral communities of benthic invertebrates in lakes. We collected invertebrates and leaf litter on 23 sites across 7 lakes somewhat isolated from human influence and compared the influence of leaf litter stoichiometry to the influence of local water quality and spatial variables. Results suggest that nutritive quality of leaf litter (i.e. stoichiometry) is secondary to water quality. Importance of leaf subsidies appears to be context dependant and could gain in importance in systems with higher degrees of disturbances. Results that nitrogen and phosphorus content of leaf subsidies is associated with higher abundances of several taxonomic groups. Species richness being unaffected, it is through higher evenness (i.e. even distribution of abundances) that higher quality of leaves may promote higher biodiversity of benthic invertebrates.