Orientador: Alexandre Nunes Ponezi / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-21T07:25:38Z (GMT). No. of bitstreams: 1 Torres_DaniladeLeoneFrancaeFreitas_M.pdf: 2036503 bytes, checksum: 068ab775d9c675592e34d0bfdd2ca1e9 (MD5) Previous issue date: 2012 / Resumo: O uso de efluente tratado na agricultura irrigada constitui uma alternativa adequada para a preservação de mananciais, além de ser uma importante fonte de nutrientes. No entanto, os riscos de toxicidade devem ser avaliados. Testes ecotoxicológicos apresentam-se como uma ferramenta apropriada, devido à sensibilidade dos organismos e sua ampla aceitação por órgãos governamentais e científicos. O objetivo do presente estudo foi avaliar a toxicidade aguda na fotobactéria marinha Vibrio fischeri e no microcrustáceo Daphnia similis (CE50) e do potencial citotóxico, genotóxico e mutagênico em Allium cepa (cebola) de um efluente sanitário tratado usado para irrigação. Os ensaios foram realizados no efluente bruto e nos tratados utilizados para irrigação: anaeróbio e pos-tratamento anaeróbio (nitrificado). Os resultados evidenciaram que o efluente bruto, apresentou uma maior variação dos valores de CE50 para ambos os organismos. O efluente anaeróbio apresentou um efeito tóxico agudo maior para V. fischeri que para D. similis. Já o nitrificado (pos-tratamento anaeróbio) apresentou uma toxicidade aguda menor para ambos os organismos testados. Na avaliação das aberrações cromossômicas em A. cepa, os efluentes tratados apresentaram uma redução do potencial citotóxico (p<0,05) em relação ao efluente bruto. Pode-se concluir que os testes escolhidos foram adequados para o monitoramento do efluente. Recomenda-se que o uso de efluente tratado na agricultura seja realizado de forma regrada, atentando para os padrões de Boas Práticas Agrícolas (GAP) e de toxicidade. Além disso, em escala real é necessário um estudo de monitoramento periódico que avalie impactos a longo prazo, incluindo testes de toxicidade crônica / Abstract: The use of the treated effluent in agriculture has proven to be an important alternative, since its preservation of water and its potential as source of nutrients, with low potential for negative impact on the environment. However, toxicity remains an important issue. Ecotoxicological tests are appropriate tools to evaluate the toxicity of effluents. The aim of this study was to evaluate acute toxicity on marine bacteria Vibrio fischeri, on micro crustacean Daphnia similis (EC50) and on Allium cepa (onion) (assessment of cytotoxic, mutagenic and genotoxic potencial) in treated sanitary effluent used for irrigation. We tested the raw effluent, and in its treatments, used for irrigation: anaerobic and anaerobic posttreatment (nitrified). Our results showed that the raw effluent had a greater variation of EC50 for both organisms, due to an intrinsic variability of its composition. The anaerobic effluent had a greater acute toxic effect on V. fischeri than for D. similis. Yet, the nitrified (anaerobic post-treatment stage) showed a lower toxicity for both tested organisms. We found a reduction in the citotoxical potential in A. cepa as compared to the raw efluente. We concluded that the tests chosen were adequate for monitoring the effluent since the organisms showed adequate sensitivity. It is therefore suggested that the use of treated wastewater in agriculture should be standardized, noting Good Agricultural Practices (GAP) and toxicity stardards. Furthermore for larger scale assumption it is necessary a periodic monitoring study to assess long term impacts, including chronic toxicity tests / Mestrado / Saneamento e Ambiente / Mestra em Engenharia Civil

Orientador: Alexandre Nunes Ponezi / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-21T10:44:37Z (GMT). No. of bitstreams: 1 Brota_GiovaniArchanjo_M.pdf: 4346412 bytes, checksum: b44166c5b1e0b8b8123c49c267aa9a87 (MD5) Previous issue date: 2012 / Resumo: O tratamento de esgoto doméstico é de fundamental importância para a manutenção da qualidade dos corpos hídricos e da vida aquática assim como para a conservação dos recursos naturais. Neste foco, os sistemas de tratamento como lagoas de estabilização são largamente utilizados no Brasil para tratamento de efluentes sanitários. O uso de efluente sanitário proveniente do pós-tratamento de lagoa de estabilização utilizado na irrigação de culturas agrícolas é recomendado como prática complementar ao tratamento do efluente, fornecendo assim um aporte de nutrientes necessários ao desenvolvimentos de diversas culturas agrícolas. No entanto essa prática deve ser avaliada quanto aos impactos ambientais. Com esse intuito, foi desenvolvida uma pesquisa para avaliar os potenciais efeitos tóxicos proveniente da aplicação de efluente tratado por lagoas facultativas na cultura de Eucalyptus urograndis. O local escolhido para a pesquisa foi uma área cedida pela ETE Prof. Carmelino Correa Junior (Colégio Agrícola), no município de Franca-SP. A área foi dividida em 8 tratamentos e irrigada de acordo com a dose de "água'' recomendada para a cultura de eucalipto da seguinte forma: T1 dose de Água, sem adubação química, T2 dose de Água, com adubação química, T3 Efluente - 1/3 da dose, com adubação química, T4 Efluente - 1/2 dose com, adubação química, T5 Efluente - 1 dose, com adubação química, T6 Efluente - 1 dose, sem adubação, T7 Efluente - 1,5 dose, sem adubação química e T8 - sem irrigação e sem adubação química (testemunha). A pesquisa avaliou a presença de elementos e compostos com potencial tóxico em detrimento da aplicação de efluente tratado, por meio de analises físicoquímicas e ecotoxicológicas, correlacionando a concentração de metais, sais e o potencial efeito tóxico causado por compostos alelopáticos liberados pelo eucalipto. Assim foi possível inferir a partir dos resultados obtidos nos testes com Daphnia similis e Vibrio fischeri, onde foram obtidos valores de CE50 superiores a 75% para a maior parte dos tratamentos, indicando um baixa toxicidade ou em alguns caso sua ausência / Abstract: The domestic wastewater treatment is critical for maintaining the quality of water bodies and aquatic life as well as for the conservation of natural resources. In this focus, treatment systems and stabilization ponds are widely used in Brazil for the treatment of wastewater. The use of sanitary effluent from the post-treatment stabilization pond used for irrigation of agricultural crops is recommended as a complementary practice to the treatment of the effluent, thus providing a supply of nutrients necessary for the development of several crops. However, this practice should be evaluated for environmental impacts. For this purpose, we developed a survey to assess the potential toxic effects from the application of effluent ponds for the culture of Eucalyptus optional urograndis, the site chosen for the research was an area ceded by Prof. ETE. Carmelino Correa Junior (Agricultural College) in the city of Franca-SP. The area was divided into eight treatments and irrigated according to the dose of "water'' recommended for the cultivation of eucalyptus as follows: T1 dose of water, without chemical fertilizer, T2 dose of water with chemical fertilizer, effluent T3 - third dose, with chemical fertilizer, effluent T4 - 1/2 dose, chemical fertilizer, effluent T5 - 1 dose, with chemical fertilizer, effluent T6 - 1 dose, without fertilization, T7 Effluent - 1.5 dose without chemical fertilizer and T8 - without irrigation and without chemical fertilizer. (control). This study evaluated the presence of elements and compounds with toxic potential detriment of the application of treated wastewater through physical-chemical analyzes and ecotoxicological correlating the concentration of metals, salts and the potential toxic effect caused by allelopathic compounds released from eucalyptus. Thus it was possible to conclude from the results obtained in tests on Daphnia similis and Vibrio fischeri, which were obtained EC50 values higher than 75% for most treatments, indicating a low toxicity and in some cases failing / Mestrado / Saneamento e Ambiente / Mestre em Engenharia Civil

Orientador: Gisela de Aragão Umbuzeiro / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Tecnologia / Made available in DSpace on 2018-08-21T02:00:16Z (GMT). No. of bitstreams: 1 Ribeiro_AlysonRogerio_M.pdf: 3701819 bytes, checksum: b2f0a69f435398e5de38f031d75649e3 (MD5) Previous issue date: 2012 / Resumo: As planarias de água doce possuem vasta distribuição geográfica, plasticidade biologica, facilidade de cultivo em laboratório, sensibilidade a contaminantes diversos e capacidade regenerativa, por isso vem sendo utilizadas como organismos-teste em ensaios ecotoxicologicos. Porem, ainda não ha normatização internacional ou nacional que padronize esses ensaios. O objetivo deste trabalho foi verificar o potencial do uso de planarias de agua doce (Girardia tigrina) na avaliação de alguns contaminantes e amostras ambientais utilizando protocolos padronizados avaliando-se mortalidade e atraso na regeneração encefálica. Foram empregados 15 organismos por concentração-teste nos ensaios de mortalidade de filhotes (96h) e 20 planarias adultas regenerantes por concentração-teste nos ensaios de regeneração encefálica (120h). Avaliou-se cromo e zinco como substancias de referencia e empregou-se cobre, azo corante têxtil Disperse Red 1, herbicida atrazina, nanotubos de carbono de parede múltipla, alem de amostras da ETE Carioba e do Ribeirão dos Cristais. Carta controle com substancia de referencia foi realizada, sendo estabelecido um protocolo para o monitoramento da sensibilidade das culturas de G. trigrina. O zinco foi a substancia mais apropriada que o cromo, com a vantagem do ensaio de sensibilidade poder ser finalizado em 24h. Os métodos utilizados para realização do ensaio de regeneração encefálica e de mortalidade de filhotes foram eficazes para a observação de efeitos adversos das substancias analisadas. Todas as amostras testadas apresentaram toxicidade as planarias, exceto os nanotubos de carbono. Atraso na regeneração encefálica foi o endpoint mais sensível para os contaminantes avaliados. Os protocolos utilizados neste estudo podem ser utilizados como subsídios na elaboração futura de normas técnicas de testes ecotoxicologicos com planarias de agua doce / Abstract: Freshwater planarians are worldwide distributed, have biological plasticity and a unique body regenerative capacity. They are easily maintained under laboratorial conditions and are sensitive for several compounds. For these reasons they have been used in ecotoxicological testing although the lack of national or international guidelines. This research aimed to verify the potential use of freshwater planarians (Girardia tigrina) in mortality and regeneration delay endpoints to assess the toxicity of environmental contaminants using standardized protocols. The culture sensitivity of G. tigrina was monitored with zinc, which was the most suitable substance for this purpose, providing responses in only 24h. The adopted protocols for regeneration and mortality tests were efficient to assess the adverse effects of the chemical agents (copper, an azo textile dye, atrazine), carbon nanotubes and wastewater and water samples from Carioba WWTP and Cristais River. The regeneration test was the most sensitive endpoint and all analyzed substances showed toxicity for planarians, except carbon nanotubes. The methods adopted in this research can be used as basis for the development of a testing guideline using freshwater planarians / Mestrado / Tecnologia e Inovação / Mestre em Tecnologia

Traditional wastewater treatment is known not to be specifically designed to eliminate the new generation of chemical residues that ends up in the sewage system. Polluted wastewatereffluent therefore reaches the aquatic environment possibly causing adverse effects in aquatic wildlife. The effects of effluent water from five Swedish sewage treatment plants sampled on 6 occasions 2017, were studied in developing zebrafish (Danio rerio). The study included morphological, physiological and behavioural endpoints. Overall there were few effectswhere deviations from control animals could be seen in the exposed zebrafish. The overall outcome of this assessment was that the wastewater effluent had no consistent effects on the early development of zebrafish. The consequences of continuous low-level exposure duringthe whole life-cycle of wild fish are presently unknown and further studies are needed to evaluate potential risks.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The indiscriminate use of agricultural supplies can cause soil pollution, affecting the flora, fauna and ecological relationships. Among these chemical compounds used in agriculture, there is the Bordeaux mixture. The intense use of this fungicide results in a considerable copper addition in the soil, may cause damage to exposed organisms. This study aimed to evaluate the toxicological effects of different copper doses, (0, 35, 70, 105, 140 e 175 mg.Kg-1) applied in soil, at earthworms Eisenia andrei biomass and reproduction, through ecotoxicological tests as well as the effects of exposure to antioxidant enzyme activity, by the enzymes catalase and glutathione S-transferase (GST and CAT), on neurotransmission biomarkers such as acetylcholinesterase (AChE) and cell membrane lipid peroxidation through the levels of malondialdehyde (MDA). The ecotoxicological results showed that the largest copper doses tested, (140 e 175 mg.Kg-1) were toxic, because the worms had a mean biomass reduction and cocoons. In the enzyme assessments, an increase in the glutathione-S-transferase activity was observed as the antioxidant defense mechanism. The significant MDA levels increase and the acetylcholinesterase activity inhibition, indicating the cell membrane lipid peroxidation occurrence and neurotransmission changes as a result of oxidative stress caused by the metal. / O uso indiscriminado de insumos agrícolas pode causar a poluição dos solos, comprometendo a flora, fauna e as relações ecológicas. Dentre os insumos químicos utilizados na agricultura, destaca-se a calda bordalesa. Sua intensa utilização resulta em uma adição considerável de cobre ao solo, podendo causar danos aos organismos expostos. O presente estudo teve como objetivo determinar os efeitos toxicológicos de diferentes doses de cobre: 0, 35, 70, 105, 140 e 175 mg.Kg-1 aplicado ao solo, na biomassa e reprodução das minhocas Eisenia andrei, através dos testes ecotoxicológicos assim como os efeitos da exposição a nível enzimático antioxidante, através das enzimas catalase e glutationa S-transferase (CAT e GST), sobre biomarcadores de neurotransmissão como a acetil-colinesterase (AChE) e na peroxidação lipídica da membrana celular através dos níveis de malondialdeído (MDA). Os resultados ecotoxicológicos, evidenciaram que as maiores doses de cobre testadas, (140 e 175 mg.Kg-1) foram tóxicas, pois as minhocas apresentaram redução média na biomassa e no número de casulos. Em relação aos níveis enzimáticos, foi observado um incremento na atividade das enzimas glutationa-S-transferase, como mecanismo de defesa antioxidante. O aumento significativo nos níveis de MDA observados e a inibição da atividade da acetil-colinesterase, indicam a ocorrência da peroxidação lipídica da membrana celular e alterações na neurotransmissão em decorrência do estresse oxidativo ocasionado pela presença do metal.

L’extension des activités humaines est responsable du rejet de molécules et de perturbations climatiques pouvant affecter la physiologie des organismes aquatiques. La moule zébrée possède des caractéristiques biologiques faisant d’elle une espèce intéressante en surveillance environnementale. Chez cet organisme, les hémocytes constituent une cible privilégiée pour la mise en place d’une approche multi-biomarqueurs. En effet, ces cellules à fonctionnalités multiples sont impliquées dans les grandes fonctions physiologiques de l’espèce et la régulation de l’homéostasie des individus. L’objectif de ce travail est de développer les outils analytiques permettant d’étudier les réponses hémocytaires de la moule zébrée. Les expérimentations menées ont permis de caractériser la structure des populations hémocytaires ainsi que leurs fonctionnalités propres en lien avec le processus de phagocytose. L’utilisation de ces biomarqueurs dans divers contextes indique une forte adaptabilité de l’espèce aux conditions environnementales. Les résultats montrent l’intérêt d’analyser les activités hémocytaires à l’échelle des sous populations comparativement à l’approche globale ne tenant pas compte de la diversité cellulaire. Il a été observé que certains facteurs comme le statut reproducteur ou l’espèce échantillonnée (D. polymorpha vs D. bugensis) constituent des facteurs de confusion importants. Il ressort également un positionnement fort du test de phagocytose en tant que marqueur de sensibilité aux contaminants. Ce travail constitue un ensemble de données voué à être utilisé dans des contextes multiples aussi bien en écotoxicologie qu’en écophysiologie. / Extension of human activities is responsible of molecule releases and climate changes that may affect physiology of aquatic organisms. The zebra mussel has biological traits making it an interesting species for environmental monitoring. In this organism, hemocyte cells constitute an interesting target to develop a multi-biomarker approach. These cells possess multiple functionalities and are involved in all major physiological functions of the species and in homeostasis regulation. The objective of this work was to develop analytical tools to study hemocyte responses of zebra mussels. Experiments allowed characterizing structure of hemocyte populations and their functionalities linked with phagocytosis process. Use of these biomarkers in various contexts indicated an important adaptation capacity of the species to environmental conditions. Results highlighted interest to analyze hemocyte activities at sub-population scale comparatively to global approach that does not consider hemocyte diversity. It was demonstrated that factor such as reproductive status or sampled species (D. polymorpha vs D. bugensis) constitute important confounding factors. Studies also demonstrated a strong positioning of phagocytosis assay as a sensitive marker to contaminants. This work constitutes a data set destined to be used in multiple contexts such as ecotoxicology or ecophysiology.

Les estuaires sont des zones de transition entre les eaux fluviales et les océans soumis à de fortes contraintes d’origine anthropique, telles que la pollution chimique. Pourtant, les estuaires sont d’une importance écologique primordiale, et sont notamment des zones de nourricerie ou de reproduction pour de nombreuses espèces. Le Flet européen Platichthys flesus est un modèle souvent utilisé comme espèce sentinelle dans les estuaires. Le but de ces travaux est d’explorer les mécanismes de réponse du Flet européen à la contamination chimique, afin éventuellement d’identifier de nouveaux biomarqueurs. Il a donc été choisi de mener cette étude par une approche de protéomique par électrophorèse en deux dimensions. Les protéines différentiellement accumulées en réponse à la pollution ont été identifiées par MALDI TOF-TOF. Ces études par protéomique ont été complétées par des mesures ciblées de transcription de gènes. Tout d’abord, des flets juvéniles issus d’un élevage ont été contaminés expérimentalement avec deux doses d’un cocktail de HAP et PCB (concentration retrouvée dans la Seine, et 10 fois celle-ci). Après 29 jours de contamination, le métabolisme énergétique paraît dérégulé. Des protéines impliquées dans les défenses anti-oxydantes et la détoxification sont accumulées dans le foie des poissons contaminés. Les résultats nous ont conduit à poser l’hypothèse de l’implication de la BHMT dans un cycle aboutissant à la production de GSH, et qui pourrait donc participer à la détoxification ou aux défenses anti-oxydantes. Ensuite, les profils protéiques de foies de poissons issus de 3 estuaires de la Manche ont été comparés. Les profils protéiques des foies de poissons de la Seine et de la Tamar, 2 estuaires présentant des profils de contamination différents, se sont révélés différents des profils protéiques des foies des poissons de la Canche. Des différences moins marquées entre les profils protéiques des foies de poissons de la Seine et de la Tamar pourraient refléter les différences entre les contaminants présents dans ces estuaires. / Estuaries are important areas highly susceptible to anthropogenic degradations such as pollution. Estuarine species have to cope not only with environmental constraints inherent to estuarine habitats, but also with the presence of contaminants, the occurrence of hypoxic events or with waters warming. The European flounder Platichthys flesus is considered as a sentinel species in estuarine water quality monitoring. This study investigates the molecular mechanisms allowing the European flounder to cope with these different stress factors, both in environmental and experimental conditions, in order to eventually identify potential biomarkers of the response to contamination. A proteomic approach using 2-dimensional electrophoresis followed by MALDI TOF-TOF mass spectrometry allowed us to identify differentially expressed proteins in flounder livers, and then to better understand the mechanisms and a pathway implied in the response of flounder to environmental constraints. These observations were completed by targeted markers analyses by qPCR and enzymatic activity measurements. First, farm flounders were experimentally contaminated with two concentrations of PAH/PCB cocktails (concentrations found in the Seine and ten times this concentration). After 29 days of contamination, energetic metabolism was deregulated in contaminated flounder livers. Proteins involved in anti-oxidative defenses and detoxification were also accumulated. We suggest that BHMT could be implied in a pathway leading to the production of GSH allowing detoxification and anti-oxidative defense. Then, Flounders were fished in contrasted estuaries along the French Atlantic coast. The liver proteomic patterns of Flounders from the Seine (France) and the Tamar (UK), two estuaries displaying different contamination patterns, and from the Canche (France), were compared to characterize the proteins differentially expressed between these sites. Proteome profiles reflected the contamination patterns in each estuary.

Les métaux traces comme le plomb, le zinc sont essentiellement émis par les activités anthropiques et se retrouvent de ce fait à des concentrations beaucoup plus élevées en milieux urbains qu’en milieux ruraux. Durant ma thèse, j’ai tout d’abord testé les effets écotoxicologiques d’une exposition chronique au plomb et/ou au zinc, deux métaux particulièrement abondants en milieu urbain, chez le pigeon biset (Columba livia). J’ai ainsi pu montrer des effets nocifs du plomb, et bénéfiques du zinc sur l’immunité, le maintien de la corpulence et plusieurs paramètres de la reproduction. Du fait de la variabilité des réponses écophysiologiques des individus, les métaux traces sont susceptibles d’exercer de nouvelles pressions de sélection sur les populations urbaines et favoriser les individus capables de se détoxifier ou de tolérer de fortes concentrations en métaux. Au cours de ma thèse, j’ai mis en évidence le rôle de la mélanine dans la fixation du zinc et du plomb au niveau des plumes. Par ailleurs la coloration mélanique du plumage semblent moduler les effets du plomb et du zinc sur certains paramètres physiologiques, et les juvéniles au plumage davantage mélanique survivent mieux dans un environnement pollué en plomb. Quels que soient les mécanismes sous-jacents (i.e. rôle détoxifiant de la mélanine ou effets pléiotropes associés à sa synthèse), mes résultats soulignent l’avantage sélectif potentiel de la mélanisation du plumage dans un environnement pollué en métaux traces, dont notamment le milieu urbain. Cette étude apporte des réponses essentielles sur l’impact écologique de l’urbanisation et les mécanismes permettant le maintien du polymorphisme de coloration mélanique du plumage, et plus largement des phanères. / Trace metals, such as lead and zinc are mainly emitted by human activities, explaining their high concentrations in urban areas in comparison with rural environments. During my PhD, I first investigated the ecotoxicological effects of a chronic exposure to lead and/or zinc, two abundant metals in urban areas, in feral pigeons (Columba livia). I stressed deleterious effects of lead, while beneficial effects of zinc on immunity, body mass index maintenance and several parameters of reproduction. Because sensitivity to trace metals differs between individuals, trace metals may exert new selective pressures on urban populations and favour individuals with higher detoxification capacities and that are more tolerant to elevated environmental trace metals concentrations. My work puts ahead the role of melanin in the storage of zinc and lead in the feathers. Moreover, melanin-based plumage colouration seems to modulate the effects of lead and zinc on some of the physiological parameters measured and darker juveniles were more prone to survive than paler ones when exposed to lead. Whatever the underlying mechanism (i.e. the detoxification role of melanin or the pleiotropic effects associated with its synthesis), my results suggest a selective advantage of plumage melanism in environments polluted with trace metals, such as urban areas. This study brings key answers on the ecological impact of urbanization and on the mechanisms explaining melanin-based plumage colouration polymorphism maintenance.

Ce travail de thèse vise à décrire l'écodynamique d'herbicides -tricétones synthétiques (sulcotrione et mésotrione) et naturel (leptospermone) et à estimer leur impact écotoxicologique sur la communauté bactérienne de sols agricoles. Les processus impliqués dans la dissipation de ces herbicides (adsorption et biodégradation) ont été étudiés dans des microcosmes de sol. Deux souches bactériennes, Bradyrhizobium sp. SRl dégradant la sulcotrione et la mésotrione, et Methylophilus sp. LS1 dégradant la leptospermone ont été isolées. Une banque de 12000 mutants de Bradyrhizobium sp. SRl a été construite et deux mutants Sul• ont été sélectionnés, mais les gènes interrompus ne codent pas pour les enzymes de dégradation de la sulcotrione. L'impact écotoxicologique des tricétones synthétique (sulcotrione) et naturelle (leptospermone) sur la communauté bactérienne du sol a été estimé à l'aide d'outils de métagénomique et de métabolomique. La leptospermone modifie de manière transitoire la diversité et la composition de la communauté bactérienne, en accord avec sa rémanence dans les sols. La sulcotrione ne modifie ni la diversité ni la composition de la communauté bactérienne. La combinaison des approches de métagénomique et de métabolomique est prometteuse pour évaluer l'impact écotoxicologique des herbicides sur les microorganismes du sol. / This work aims to describe the ecodynamics of synthetic (sulcotrione and mesotrione) and natural (leptospermone) -triketone herbicides and to estimate their ecotoxicological impact on the bacterial community in arable soils. The processes involved in the dissipation of these herbicides (adsorption and biodegradation) have been studied in soil microcosms. Two bacterial strains, Bradyrhizobium sp. SRl able to degrade sulcotrione and mesotrione, and Methylophilus sp. LS1 degrading leptospermone, have been isolated. A bank of 12 ooo mutants of Bradyrhizobium sp. SR1 was established allowing the selection of two Sul•mutants but interrupted genes didn't code for enzymes degrading sulcotrione. The ecotoxicological impact of synthetic (sulcotrione) and natural (leptospermone) triketones on soil bacterial community was estimated using metagenomics and metabolomic tools. Leptospermone transitory modified the diversity and composition of the bacterial community, in accordance with its persistence in soil. Sulcotrione did not modified neither the diversity nor the composition of the bacterial community. The combination of metagenomics and metabolomics is promising for the assessment of ecotoxicological impact of herbicides on soil microorganisms.

Common observations of reduced gonad size and spawning inhibition in wild and laboratory raised fish exposed to pulp mill effluents indicate that reproductive neuroendocrine signalling pathways may be upset. This thesis supported the neuroendocrine disruption of reproduction hypothesis by identifying potential disruptors and targets where these impacts may occur. A mechanistic study of the in vivo fathead minnow (FHM) spawning assay used by industry to assess effluent quality showed that ovulation, but not milt production, was impaired. This finding supported the hypothesis that the neuroendocrine cascade that triggers ovulation may be disrupted. I hypothesized that neuroactive constituents previously described in effluents were originating in wood feedstocks and neuroactive extracts of hardwood and conifer feedstocks were identified. Phytochemicals associated with effluents were neuroactive. Structurally similar phenolic phytochemicals showed monoamine oxidase (MAO) inhibition, and resin acid diterpenes displayed glutamic acid decarboxylase (GAD) inhibition. Inhibitors of these enzymes may have impacts on the control of reproduction since MAO metabolizes dopamine, an inhibitor of the neuroendocrine reproductive axis, while GAD synthesizes -aminobutyric acid (GABA), a stimulator of this axis. Bioassay-guided fractionations of effluents and wood feedstocks identified that medium polar extracts of primary- and secondary-treated effluents and balsam fir feedstock contained high GAD inhibitory activity. This activity was associated with chemically complex fractions rather than single active principles. Advanced metabolomic comparison of medium polar extracts of feedstock and treated effluent identified 15 common plant metabolites, demonstrating that phytochemicals entering the mill in wood are surviving pulp production and effluent treatment processes and may be responsible for observed GAD inhibition. Discriminant metabolomics analysis identified 4-acetylpyridine, a novel compound to be described in effluents, as well as two other tentatively identified compounds, as chemical markers of GAD inhibitory effluent fractions. Five tentatively identified chemical markers and (+)-lariciresinol were found in inhibitory balsam fir feedstock fractions. Neuroendocrine pathways that control reproduction in fish, such as dopamine and GABA pathways, are also important drug targets for the treatment of neurological disorders in mammals; therefore these results also have implications for the development of natural health products from phytochemicals and tree extracts common to Canadian forests. By using an interdisciplinary approach (phytochemistry, neuroendocrinology, ecotoxicology), I was able to explore the various implications of my research on the fields of natural health products chemistry and aquatic toxicology.