Le développement des activités industrielles et agricoles du siècle dernier a eu pour conséquence le rejet de substances chimiques dans l’environnement, qui contaminent les écosystèmes, et peuvent s’accumuler dans les plantes, les sédiments, et le long des chaînes alimentaires. Il apparait primordial d’effectuer une évaluation du risque écologique associé à la présence de ces composés dans les milieux naturels et les espèces sentinelles sont des outils judicieux pour déterminer l’exposition des individus et évaluer les conséquences sur les organismes. Les amphibiens et les reptiles possèdent des caractéristiques écologiques et biologiques qui les rendent potentiellement intéressants en tant qu’espèces sentinelles. L’objectif des études présentées dans cette thèse est de déterminer la disponibilité de certaines substances dans l’environnement, et d’évaluer les effets de la contamination environnementale sur les organismes en utilisant plusieurs espèces de reptiles et d’amphibiens, (i) en mesurant les niveaux de polluants accumulés dans les tissus d’espèces de reptiles ; (ii) en évaluant l’influence d’un habitat contaminé sur la morphologie d’adultes de crapaud commun (Bufo bufo) ; (iii) en mesurant de manière expérimentale l’impact d’une molécule fréquemment retrouvée dans l’environnement sur le développement de têtards de crapaud commun. Nos résultats montrent d’une part que les reptiles accumulent efficacement les polluants organiques et les éléments traces métalliques dans leurs tissus, et permettent d’évaluer efficacement la contamination globale d’une zone, mais aussi de mettre en évidence des pollutions ponctuelles localisées. D’autre part, nos travaux montrent une influence de l’habitat agricole sur la morphologie du crapaud commun, potentiellement causé par la présence de contaminants dans ces milieux. Enfin, nous avons pu mettre en évidence un effet de concentrations faibles d’AMPA, métabolite principal du glyphosate, sur le développement de têtards de crapaud commun. Les amphibiens constituent donc des organismes efficaces pour étudier l’impact de la contamination de l’environnement sur les caractéristiques individuelles. Nos travaux montrent ainsi l’utilité de certaines espèces de reptiles et d’amphibiens comme espèces sentinelles de la présence de contaminants dans l’environnement et de leur impact potentiel sur les organismes. Les travaux en écotoxicologie qui portent sur ces organismes restent trop rares, et il est impératif de développer la recherche dans ce domaine, pour participer de manière efficace à l’évaluation du risque environnemental. / The development of industrial and agricultural activities over the last century has resulted in the release of chemicals into the environment, which contaminate ecosystems and can accumulate in plants, sediments and across food webs. As such, it is essential to conduct an assessment of the ecological risk associated with the presence of these compounds in natural environments. Achieving this can be aided through the study of sentinel species, which can be used to determine exposure rates of individuals and assess how these subsequently impact wildlife. Amphibians and reptiles display ecological and biological characteristics that may make them particularly attractive as sentinel species. The collection of studies presented in this thesis aims to determine the availability of contaminants in the environment and estimate the effects of this on individual traits of living organisms. Across several species of reptiles and amphibians we, (i) measured levels of pollutants accumulated in the tissues of several reptile species ; (ii) assessed how contaminated habitats impact the morphology of adult common toads (Bufo bufo) ; and (iii) experimentally measured the influence of a molecule frequently detected in the environment on common toad tadpoles’ growth. Results show that reptiles efficiently accumulate organic pollutants and metal trace elements in their tissues, and thereby allow efficient evaluation of the global contamination of an area, as well as the occurrence of localized pollution events. In addition, our work shows an influence of the agricultural habitat on the morphology of adult common toads, which is potentially a consequence of the presence of chemicals in these environments. Finally, we were able to demonstrate an effect of low concentrations of AMPA, the main metabolite of glyphosate, on the development of common toad tadpoles. Amphibians are therefore efficient organisms to study the impact of environmental contamination on individual characteristics. Our work shows the usefulness of certain species of reptiles and amphibians as sentinel species for the presence of contaminants in the environment, and their potential impact on organisms. Ecotoxicological studies on these organisms remain too scarce, and it is imperative to develop research in this field, to participate effectively in the assessment of environmental risk.

As estações de tratamento de água (ETA) operam mundialmente com o intuito de tratar e disponibilizar água potável a população, a partir das águas brutas dos mananciais. Durante as fases do tratamento da água é gerada uma grande quantidade de lodo, em torno de 1 a 3% do total de água tratada pela ETA, o que pode representar uma tonelada diária de lodo gerado. Esse resíduo vem sendo descartado nos mesmos mananciais da captação das águas, porém com as novas legislações ambientais, visando à proteção e a preservação dos recursos hídricos, pesquisas estão sendo desenvolvidos com o intuito de encontrar uma alternativa para o descarte desse resíduo. O presente estudo tem como objetivo principal avaliar e relacionar a ecotoxicidade do lodo da ETA, com as características físicas e químicas dos mesmos. As quantificações dos elementos químicos presentes: nas amostras de água e nos filtros, quando foram realizadas as filtrações das águas foram realizadas pelas técnicas de fluorescência de raios X dispersiva em energia por reflexão total (TXRF), para as amostras líquidas e por fluorescência de raios X dispersiva em energia (EDXRF) para as amostras sólidas. As determinações dos elementos químicos presentes nas amostras de lodo foram realizadas por espectrometria de massa com plasma indutivamente acoplado (ICP MS) e por espectrometria de emissão óptica com plasma indutivamente acoplado (ICP OES). Os testes ecotoxicológicos foram realizados com as espécies Hydra attenuata, Daphnia similis, Danio rerio, Pseudokirchneriella subcapitata, Chironomus xanthus, Folsomia candida e Zea mays. Além das exposições para avaliação da mortalidade e da inibição do crescimento, também foram realizados testes crônicos de reprodução com as D. similis e F. candida. Os testes de toxicidade mostraram que as amostras de elutriato e principalmente as amostras do lodo filtrado foram altamente tóxicas aos organismos aquáticos e terrestres. As amostras do floculador apresentaram menor qualidade ambiental, com menores concentrações de OD, maior condutividade elétrica e concentrações de elementos químicos mais elevados. As análises de redundância (RDA), realizadas com as plântulas de Z. mays irrigadas com o elutriato, indicaram alta correlação negativa com os elementos químicos quantificados nas raízes. Entretanto a incorporação do lodo ao substrato foi benéfica, incentivando o crescimento e desenvolvimento das plântulas. Concluiu-se que a incorporação ao solo, pode ser uma alternativa de destino do lodo da ETA / The water treatment plants (WTP) operate worldwide in order to treat and deliver drinking water to the population, from the untreated waters of the springs. During the water treatment stage is generated a large amount of sludge, about 1 to 3% of total treated water, which thus can arrive at a daily ton of sludge generated by the ETA. This waste is being disposed of in the same watershed catchment of water, but with the new laws aimed at environmental protection and preservation of water resources, research is being developed in order to find an alternative for the disposal of this waste. This study aims to evaluate and relate the ecotoxicity of sludge from ETA, with the physical and chemical characteristics of the same. The quantification of chemical elements: the water samples and filters when filtrations were performed water were conducted by the techniques of fluorescence energy dispersive X-ray total reflection (TXRF) for the liquid samples and ray fluorescence energy dispersive X (EDXRF) for solid samples. The determinations of chemical elements in samples of sludge were performed by mass spectrometry with inductively coupled plasma (ICP MS) and optical emission spectrometry with inductively coupled plasma (ICP OES). The ecotoxicological tests were performed with the species Hydra attenuata, Daphnia similis, Danio rerio, Pseudokirchneriella subcapitata, Chironomus xanthus, Folsomia candida and Zea mays. In addition exposures to assess mortality and growth inhibition tests were also performed with chronic reproduction D. similis and F. candida. Toxicity tests showed that samples of elutriate and mostly filtered sludge samples were highly toxic to aquatic and terrestrial organisms. However for Z. mays incorporate the sludge was beneficial to the substrate, promoting inclusive growth and seedling development, concluding that the incorporation into the soil can be an alternative destination sludge ETA

A toxicidade da formulação comercial contendo o praguicida piretróide deltametrina foi testada nas espécies de peixe Danio rerio, Hyphessobrycon bifasciatus, Geophagus brasiliensis e Oreochromis niloticus por meio de testes de toxicidade aguda, avaliação de alterações comportamentais e análises anatomopatológicas do fígado e das brânquias. A espécie D. rerio apresentou CL50;48h igual a 0,078µg L-1, a espécie H. bifasciatus apresentou CL50;48h igual a 0,082µg L-1, a espécie G. brasiliensis apresentou CL50;48h igual a 0,594µg L-1 e O. niloticus apresentou CL50;48h igual a 0,954µg L-1. As análises de comportamento realizadas com as espécies D. rerio e H. bifasciatus mostrou ser a segunda mais sensível ao praguicida, revelando alterações significativas nas menores doses de exposição. As análises anatomopatológicas realizadas com a espécie G. brasiliensis quando exposta à concentrações subagudas do praguicida mostrou vacuolização e degeneração nuclear hepática e nas brânquias a ocorrência de hiperplasia lamelar e fusão nas lamelas secundárias que aumentaram em função da concentração do praguicida e do tempo de exposição. Os resultados demonstraram elevada toxicidade do praguicida para todas as espécies testadas. / The toxicity of the commercial formulation containing the pesticide pirethroid deltamethrin was tested in the species of fish Danio rerio, Hyphessobrycon bifasciatus, Geophagus brasiliensis and Oreochromis niloticus through tests of acute toxicity, evaluation of behavioral alterations and anatomopathological analysis of the liver and of the gills. The species D. rerio presented LC50;48h equals to 0,078µg L-1, the species H. bifasciatus presented LC50;48h equals to 0,082µg L-1, the species G. brasiliensis presented LC50;48h equals to 0,594µg L- 1 and O. niloticus presented LC50;48h equals to 0,954µg L-1. The analysis of behavior accomplished with the species D. rerio and H. bifasciatus showed that the second one was more sensitive to the pesticide, and revealed significant alterations in the exhibition of fewer doses. The anatomopathological analysis accomplished with the species G. brasiliensis, when exposed to underacute concentrations of the pesticide, showed vacuolization and hepatical nuclear degeneration and the occurrence of lamellar hyperplasia in the gills plus fusion in the secondary lamellas which were increased due to the concentration of the pesticide and of the time of exposition. The results demonstrated high toxicity of the pesticide for all of the tested species.

Os ecossistemas aquáticos têm sido afetados pelo uso intensivo de agrotóxicos e pela introdução de espécies exóticas. Para melhor avaliar os efeitos diretos e indiretos dessas atividades sobre as comunidades aquáticas, é essencial a utilização de experimentos em escala mais próxima a real. O objetivo desta pesquisa foi analisar os efeitos do agrotóxico Vertimec® 18 CE, cujo princípio ativo é a abamectina, sobre a comunidade planctônica através de um experimento em mesocosmos com duração de 20 dias. Para reproduzir relações biológicas mais complexas e analisar o impacto da introdução de uma espécie exótica sobre o plâncton, inseriram-se girinos de Lithobates catesbeianus em alguns tratamentos. Foram consideradas duas concentrações do agrotóxico (2 μL L-1 em dose múltipla e 8 μL L-1 em dose única) na ausência e presença de girinos, além de dois tratamentos não contaminados (controle com e sem girinos). Variáveis físicas, químicas e biológicas (clorofila a, aspectos qualitativos e quantitativos do fitoplâncton e do zooplâncton) da água e do sedimento foram monitoradas. A partir dos resultados obtidos, verificou-se que os girinos causaram um aumento na concentração de nutrientes na água e uma diminuição do oxigênio dissolvido e da densidade planctônica. Nos tratamentos contaminados, como um efeito indireto do incremento da concentração de nutrientes e da mortalidade das espécies de Cladocera e Copepoda, a densidade do fitoplâncton e de algumas espécies tolerantes de Rotifera aumentou. No tratamento com a concentração mais elevada do agrotóxico, registrou-se a morte de todos os girinos, proporcionando um aumento na concentração de nutrientes na água e na densidade de algumas espécies fitoplanctônicas, que alcançaram uma densidade total de 96.919 ind mL-1. Estes resultados indicam que a inserção dos girinos e a adição do agrotóxico causaram efeitos diretos e indiretos sobre a comunidade planctônica e a qualidade da água. Conclui-se que os mesocosmos podem contribuir para a compreensão de tais efeitos e oferecer subsídios ao manejo de ecossistemas de água doce impactados por atividades antrópicas. / Aquatic ecosystems have been impacted due to the intensive use of pesticides and the introduction of exotic species. It is essential to use more realistic experiments for assessing direct and indirect effects of these activities on aquatic communities. This thesis aimed to analyze the effects of the pesticide Vertimec® 18 EC, whose active ingredient is abamectin, on plankton community through a mesocosm experiment during 20 days. Tadpoles of the species Lithobates catesbeianus were introduced in some treatments to simulate complex biological relationships and to evaluate the impact of the introduction of such exotic species on plankton. Different treatments were analyzed, including one case of no contamination (control) and two cases of contamination: 2 μL L-1 (multiple dose) and 8 μL L-1 (single dose). Such conditions were considered both with absence and presence of tadpoles. Physical, chemical and biological (chlorophyll a, qualitative and quantitative aspects of phytoplankton and zooplankton) variables of water and sediment were also monitored. The tadpoles caused an increase in the concentration of nutrients and a decrease of dissolved oxygen concentration and plankton density. In the contaminated treatments, as an indirect effect of the nutrient increase and the mortality of species of Cladocera and Copepoda, the density of phytoplankton and some tolerant species of rotifers increased. In the treatment with the highest concentration of the pesticide, all the tadpoles died. As a result of the death of the tadpoles, there was nutrient enrichment in the water, with an associated increase in the density of some phytoplankton species, reaching a total density of 96,919 ind mL-1. These results suggested that direct and indirect effects occurred on plankton community and water quality through the addition of the contaminant and the tadpoles. We conclude that the mesocosms might contribute to the understanding of these effects and provide subsidies for management of freshwater ecosystems impacted by human activities.

O Brasil ocupa a quarta posição entre os maiores produtores mundiais de carne suína. Destaque nacional deve ser dado ao estado de Santa Catarina, que detém 25% da produção do Brasil. Em Santa Catarina, a intensificação da produção em pequena área territorial gera um grande aporte anual de dejetos de suínos por unidade de área agrícola. Isso se deve ao fato de que a alternativa mais utilizada para descarte desses resíduos é a sua aplicação como fertilizante agrícola, prática que tem gerado um dos maiores problemas de poluição ambiental, especialmente na região Oeste Catarinense. Poucos trabalhos com intuito de avaliar a toxicidade desse material aos organismos edáficos e sua interação com diferentes classes texturais de solos podem ser encontrados. Para avaliação desta toxicidade, uma ferramenta utilizada internacionalmente, e cada vez mais frequentemente no Brasil, é a ecotoxicologia terrestre. Esta técnica é utilizada para avaliar efeitos de substâncias que, quando adicionadas aos solos, causam impactos em organismos, mensurando as respostas de alterações na taxa de letalidade, reprodução, desenvolvimento e comportamento de organismos edáficos padronizados. Desta maneira, este estudo objetivou avaliar por meio de testes ecotoxicológicos os efeitos de doses crescentes de dejeto de suínos (D0: 0 m3 ha-1; D1: 25 m3 ha-1; D2: 50 m3 ha-1; D3: 75 m3 ha-1 e 100 m3 ha-1), em diferentes tipos de solo (Argissolo Vermelho Eutrófico S2; Latossolo Vermelho Distrófico S3 e Neossolo Quartzarênico S4 e Solo Artificial Tropical S1) na sobrevivência, reprodução e comportamento de minhocas (Eisenia andrei) e colêmbolos (Folsomia candida), através de metodologias padronizadas internacionalmente. Os resultados das avaliações com Eisenia andrei mostraram que no solo com menor teor de argila a toxicidade do dejeto causou letalidade de 100% dos indivíduos nas duas maiores doses testadas. Para os testes de toxicidade crônica, avaliando o potencial reprodutivo, as doses testadas em Neossolo Quartzarênico tiveram efeitos negativos em Eisenia andrei, mas nos outros solos não ocorreu efeito perceptível. Os resultados obtidos para testes com Folsomia candida mostraram toxicidade em todas as doses testadas, causando letalidade significativa dos indivíduos na menor dose de dejeto aplicada e, baseado nessa resposta, doses menores foram usadas para avaliar efeito crônico e no comportamento (0 m3 ha-1, 10 m3 ha-1, 15 m3 ha-1, 20 m3 ha-1 e 25 m3 ha-1). Os efeitos das doses usadas para o teste de reprodução foram significativamente diferentes dos observados no controle causando redução no número de juvenis gerados em todos os solos testados. As doses D3 e D4 no S2, assim como todas as doses no S3 causaram significativa fuga de colêmbolos para o solo não contaminado. Os resultados obtidos revelam a importância da utilização de solos naturais de diferentes texturas e diferentes organismos na avaliação de toxicidade de dejetos de suínos e também mostram que nesse caso os resultados dos testes com solo artificial tropical não podem ser extrapolados para solos naturais. / Brazil occupies the fourth place worldwide among the largest swine producers. National highlight should be given to Santa Catarina State, which stands for 25% of this production. In this state, the intensification of the production in a small area leads to a yearly huge supply of swine manure per unit of agricultural area. This is due to the fact that the most used methods to dispose swine manure is its application as a fertilizer on soils, which has been causing one of the largest environmental pollution problems, especially in the Western region of Santa Catarina State. Very few studies are available aiming to evaluate swine manure toxicity on edaphic organisms and its interaction with different textural classes of soil. To evaluate this toxicity ecotoxicology has been used around the world, and increasingly in Brazil. This technique is used to assess the effects of substances that, when applied on soil, cause impacts on the organisms, measuring their responses with regard to mortality rates, reproduction, development and behavior of soil organisms. Thus, this study aimed to evaluate, by means of ecotoxicological tests the effects of increasing swine manure doses (D0: 0 m3 ha-1; D1: 25 m3 ha-1; D2: 50 m3 ha-1; D3: 75 m3 ha-1 e 100 m3 ha-1) in different types of soil (Ultisols - S2; Oxisol S3 and Entisols S4) and in an Tropical Artificial Soil S1, on the survival, reproduction and behavior of earthworms (Eisenia andrei) and springtails (Folsomia candida), using standard international methodologies. The results of Eisenia andrei evaluations showed that, in sandy soils, swine manure toxicity caused 100% mortality of the organisms at the two largest doses tested. For chronic toxicity tests, evaluating the reproductive potential, there were negative effects of tested doses on Eisenia andrei in Entisols, but for the other soils there was no perceptible effect. The results of Folsomia candida tests showed toxicity for all the tested doses, causing significant mortality of the organisms even at the smallest dose applied. Therefore, smaller doses were used to evaluate chronic and behavioral effects (0 m3 ha-1, 10 m3 ha-1, 15 m3 ha-1, 20 m3 ha-1 e 25 m3 ha-1). The effects of applied doses for the reproduction test were highly different from those observed in the control treatment, always causing reduction in the number of produced juveniles for all types of soil tested. The D3 and D4 doses for S2, as well as, all the doses for S3, showed significant avoidance of springtails towards the uncontaminated soil. These data show the importance of the usage of natural soils with different textural classes and also different organisms in the evaluation of swine manure toxicity. Also, they show that the results for artificial tropical soils cannot be extrapolated to natural soils.

O naftaleno é um dos principais constituintes do petróleo solúvel em água e esta classe de poluentes está enquadrada na lista prioritária de contaminantes orgânicos persistentes. O presente trabalho traz informações sobre o efeito do naftaleno em ensaios ecotoxicológicos com quatro diferentes grupos de organismos aquáticos. Ensaios crônicos com a microalga marinha Dunaliella tertiolecta e com ouriço-do-mar Lytechinus variegatus, ensaios crônicos e agudos com o copépodo estuarino bentônico Nitokra sp e ensaios agudos com o anfípodo estuarino Leptocherius plumulosus. Na avaliação do efeito crônico, o valor médio da CI(I)50-96h encontrada para microalga marinha foi de 29,01 mg.L-1 e o valor médio da CE(I)50-24h para ouriço-do-mar foi de 0,30 mg.L-1. Para o copépodo, os ensaios crônicos tiveram os valores de 0,19 mg.L-1 para CENO(I) e 0,40 mg.L-1 para CEO(I). Nos ensaios agudos, este mesmo organismo apresentou uma média de CL(I)50-96h em 13,4 mg.L-1 e a média da CL(I)50-96h para o anfípodo estuarino foi de 2,38 mg.L-1. Estes resultados evidenciam potenciais de efeito para diferentes classes de organismos aquáticos e poderão auxiliar nas avaliações ambientais, quando houver limites disponíveis e diretrizes que estabeleçam condições e padrões de lançamento de efluentes nos corpos de água no Brasil. / Naphthalene is a major constituent of the oil soluble in water and this class of pollutants is framed in the priority list of persistent organic pollutants. The present study provides information on the effect of naphthalene in ecotoxicological tests with four different groups of aquatic organisms. Chronic tests with the marine microalgae Dunaliella tertiolecta and sea-urchin Lytechinus variegatus, acute and chronic tests with the estuarine benthic copepod Nitokra sp and acute tests with the estuarine amphipod Leptocheirus plumulosus. In the assessment of chronic effect, the average value of IC(I)50-96h to marine microalgae was 29.01 mg.L-1 and the mean EC(I)50-24h for sea urchin 0.30 mg.L-1. In chronic tests for estuarine copepod the obtained values to NOEC and OEC was ranged from 0.19 mg L-1 to 0.40 mg L-1. In acute tests the same organism have showed an average of LC(I)50-96h in 13.4 mg.L-1 and for the estuarine amphipod the mean LC(I)50-96h was 2.38 mg.L-1. These results demonstrate the potential effect on different classes of aquatic organisms. This may assist in environmental assessments in Brazil, when limits and guidelines are available and allow establishing conditions and standards of effluent discharge into water bodies in Brazil.

Os paradigmas ecológicos atualmente em prática apresentam dificuldades quando aplicados na avaliação, monitoração e gerenciamento dos ecossistemas altamente deteriorados. No entanto uma abordagem baseada na metáfora da saúde ambiental promete vir a ser muito útil. Pela discussão a respeito do conceito de saúde ambiental, o presente ensaio propõe algumas definições bem como um modelo e métodos preliminares de diagnóstico, baseado nos protocolos de Avaliação de Risco Ambiental da agência norte americana. A proposta de avaliação da saúde dos ecossistemas deriva de um nova ciência integradora, a ecotoxicologia, a qual estaria a desenvolver procedimentos para diagnosticar as condições de saúde dos ecossistemas de forma semelhante a como se estima a condição da saúde humana. As principais idéias a respeito de saúde dos ecossistemas estão sendo condensadas em três capítulos principais. Em primeiro lugar é apresentada uma revisão da literatura referente ao conceito de saúde dos ecossistemas e às suas diferentes formas de estimação. Uma vez definidas as bases conceituais, e a modo de exemplo, passa-se a um estudo de caso no qual procura-se estimar alguns aspectos relacionados ao estudo da saúde da Represa de Salto Grande (Americana, SP) mediante a aplicação da metodologia de Avaliação de Risco Ambiental. Finalmente, são apresentadas algumas considerações a respeito do diagnóstico da saúde de reservatórios. / The assessment, monitoring and management of highly deteriorated ecosystems are not easily served for by existing environmental paradigms. The approach to the concept of ecosystem health metaphor is seen to be very useful. This essay proposes some definitions of ecosystem health along with a preliminary diagnostic model and methods based on the USEPA\'s Environmental Risk Assessment framework. The ecosystem health assessment idea derives from the emerging integrative science of ecotoxicology, which would be to seek a procedure about diagnosis of ecosystem health condition similarly to the diagnosis of human health condition. The main ideas about ecosystem health are condensed into three main sections. First, a review of the literature regarding definitions and general approaches of biophysical landscape health is presented. This background stage is followed by a case study, as example, which details how different phases of the USEPA\'s Environmental Risk Assessment framework can be applied to accomplish some aspects of the ecosystem health status from the Salto Grande Reservoir (Americana, SP - Brazil). At the essay\'s conclusion, some remarks regarding the tropical reservoir\'s health diagnosis are presented.

Les toxines phytoplanctoniques présentent une large diversité, tant du point de vue de leurs structures chimiques que de leurs effets biologiques. La multiplicité des familles de toxines ainsi que l'observation de plus en plus fréquente d'épisodes toxiques non expliqués ont rendu l'interprétation des tests in vivo très délicate. Des méthodes complémentaires aux tests in vivo telles que les analyses chimiques ont été développées et permettent une détection spécifique des toxines connues dans un cadre réglementaire. En vigilance, il est nécessaire de pouvoir détecter de nouvelles toxines et analogues. Les lignées cellulaires font partie des méthodes complémentaires prometteuses pour ce faire. Les réponses obtenues à l'échelle cellulaire donnent des informations sur les cibles moléculaires et les mécanismes de toxicité associés aux molécules ou extraits testés. Ces connaissances sont un support pour (i) la meilleure évaluation du risque pour l'homme et (ii) l'apport de connaissances sur les épisodes toxiques. Ce travail de thèse s'inscrit dans ce contexte en se focalisant spécifiquement sur l'analyse des réponses de modèles cellulaires exposés à des neurotoxines. La lignée cellulaire Neuro-2a a été sélectionnée comme modèle cellulaire pour étudier les réponses face à différentes neurotoxines pour lesquelles les niveaux de connaissances des modes d'action sont bien ou peu décrits. L'objet de la première partie de ce travail est de caractériser les réponses de ce modèle cellulaire exposé aux saxitoxines, neurotoxines agissant sur le site 1 de la sous unité α du canal sodium voltage-dépendant. Cette démarche nous a permis de mettre en évidence, pour la première fois en France, des saxitoxines en eau douce et de discuter le risque pour l'homme. Ensuite, les réponses du modèle Neuro-2a exposé à la palytoxine et à ses analogues, neurotoxines émergentes agissant sur la pompe Na+, K+-ATPase, ont été caractérisées. Enfin, les effets à l'échelle cellulaire du 13-desméthyle-C spirolide, neurotoxine émergente sur les côtes françaises pour laquelle les études toxicologiques sont limitées, ont été investigués. La discussion générale replace l'ensemble des données acquises au cours de cette thèse dans un contexte plus général de l'utilisation des modèles cellulaires pour contribuer à une meilleure connaissance des épisodes de toxicité du phytoplancton et du risque qu'ils représentent pour l'homme.

The extensive agricultural usage of organohalogenated pesticides has raised many concerns about their potential hazards especially in the soil environment. Environmental metabolomics is an emerging field that investigates the changes in the metabolic profile of native organisms in their environment due to the presence of an environmental stressor. Research presented here explores the potential of Nuclear Magnetic Resonance (NMR)-based metabolomics to examine the sub-lethal exposure of the earthworm, Eisenia fetida to sub-lethal concentrations of organohalogenated pesticides. Various one-dimensional (1-D) and two dimensional (2-D) NMR techniques were compared in a contact filter paper test earthworm metabolomic study using endosulfan, a prevalent pesticide in the environment. The results determined that both the 1H Presaturation Utilizing Gradients and Echos (PURGE) and the 1H-13C Heteronuclear Single Quantum Coherence (HSQC) NMR techniques were most effective in discriminating and identifying significant metabolites in earthworms due to contaminant exposure. These two NMR techniques were further explored in another metabolomic study using various sub-lethal concentrations of endosulfan and an organofluorine pesticide, trifluralin to E. fetida. Principal component analysis (PCA) tests showed increasing separation between the exposed and unexposed earthworms as the concentrations for both contaminants increased. A neurotoxic mode of action (MOA) for endosulfan and a non-polar narcotic MOA for trifluralin were delineated as many significant metabolites, arising from exposure, were identified. The earthworm tissue extract is commonly used as the biological medium for metabolomic studies. However, many overlapping resonances are apparent in an earthworm tissue extract NMR spectrum due to the abundance of metabolites present. To mitigate this spectral overlap, the earthworm’s coelomic fluid (CF) was tested as a complementary biological medium to the tissue extract in an endosulfan exposure metabolomic study to identify additional metabolites of stress. Compared to tests on the tissue extract, a plethora of different metabolites were identified in the earthworm CF using 1-D PURGE and 2-D HSQC NMR techniques. In addition to the neurotoxic MOA identified previously, an apoptotic MOA was also postulated due to endosulfan exposure. This thesis also explored the application of 1-D and 2-D NMR techniques in a soil metabolomic study to understand the exposure of E. fetida to sub-lethal concentrations of endosulfan and its main degradation product, endosulfan sulfate. The earthworm’s CF and tissue extract were both analyzed to maximize the significant metabolites identified due to contaminant exposure. The PCA results identified similar toxicity for both organochlorine contaminants as the same separation, between exposed to the unexposed earthworms, were detected at various concentrations. Both neurotoxic and apopotic MOAs were observed as identical fluctuations of significant metabolites were found. This research demonstrates the potential of NMR-based metabolomics as a powerful environmental monitoring tool to understand sub-lethal organohalogenated pesticide exposure in soil using earthworms as living probes.

Triclosan (TCS) is a personal care product with many fields of application and is of public interest for several years now. Monitoring studies showed that TCS is a ubiquitous chemical in the aquatic environment. Aquatic organisms are exposed to TCS in a broad range of concentrations, from ng L-1 up to lower μg L-1. TCS has a bactericidal effect for various types of gram-positive and gram negative bacteria. TCS targets a specific bacterial fatty acid biosynthetic enzyme, enoyl-[acyl-carrier protein] reductase (Schweizer, 2001). Therefore the terminal reaction in the fatty acid elongation cycle is inhibited (Levy et al., 1999). Although effects on non-target organisms are reported, the Mode of Action (MoA) of TCS is not well examined for those organisms. The aim of this PhD thesis was to investigate effects of TCS on non-target autotrophic organisms at different levels of biological complexity in the aquatic environment. In this thesis microalgae have been found to be very sensitive to TCS. In some cases even higher sensitivities than in bacteria were observed, which is in accordance with published effect data (Harada et al., 2008; Orvos et al., 2002). Similarly to bacteria, high species sensitivity differences were observed for algae (Franz et al., 2008). In bacteria these sensitivity differences can be ascribed to several resistance mechanisms reported in Schweizer (2001). These findings lead to the question about the reasons for species sensitivity differences in algae. A mesocosm study was performed to detect effects of TCS across levels of biological organization and to investigate the impact of sensitivity differences on complex aquatic communities. For that purpose, structural and functional effects parameters were observed.