Atualmente no cenário mundial a qualidade da água tem gerado muitas preocupações, pois milhares de produtos sintéticos são produzidos para facilitar muitas práticas industriais, domésticas e pessoais e com isso diversas substâncias químicas utilizadas para esses fins são introduzidas no meio ambiente. Os parabenos são substâncias químicas utilizadas pelas indústrias farmacêuticas, de alimentos e cosméticos e que cuja função é a conservação, sendo que há muitos questionamentos em relação a sua segurança, pois alguns relatórios têm mostrado que a exposição a esses parabenos pode modular ou perturbar o sistema endócrino e com isso gerar consequências prejudiciais à saúde humana e aos ambientes aquáticos. Esse estudo teve como objetivo avaliar a presença dos Parabenos Metilparabeno e Propilparabeno no ambiente aquático e os seus potenciais estrogênicos e a toxicidade aguda. A metodologia se desenvolveu a partir do ensaio in vitro YES para a determinação da atividade estrogênica, ensaios de toxicidade aguda em Daphnia similis e em Aliivibrio fischeri e a quantificação dos parabenos na água do Rio Maracanã-RJ através da cromatografia líquida acoplada ao espectrômetro de massa. Os resultados obtidos para a CE50 dos MP e PP, a partir do ensaio in vitro YES, foi de 18,91 mgL-1 e 7 mgL-1 e a magnitude da resposta foi de 10-5 e 10-4 menos potente que 17ß-estradiol para o MP e o PP, respectivamente. A partir do ensaio de toxicidade aguda, os valores de CE50 obtidos em Daphnia similis, foi de 29,42 mgL-1 e 9,94 mgL-1e em Aliivibrio fischeri foi de 3,047 mgL-1 e 1,946 mgL-1, respectivamente, com isso observou-se que o PP é mais tóxicos em todos os organismos testados, sendo mais tóxicos para um e menos para outros. A água do Rio Maracanã não foi tóxica para a Daphnia similis em nenhum dos dois pontos, já para o Aliivibrio fischeri foi tóxica em apenas um ponto. As concentrações encontradas de MP e PP foram maiores no ponto onde, de acordo com os parâmetros físico-químicos, a qualidade da água não está dentro dos padrões exigidos pela legislação, sendo quantificados na ordem de ngL-1, contudo é válido ressaltar que os DEs não são encontrados no meio ambiente separados, eles interagem entre si e provocam efeitos aditivos ou sinérgicos, sendo muito difícil de prever qual o efeito, por isso é importante o conhecimento do potencial estrogênico das substâncias simples, pois em um estudo com uma matriz ambiental, pode-se observar se houve algum efeito aditivo ou sinérgico de outras substâncias. / Currently on the world stage water quality has generated many concerns, because thousands of synthetic products are produced to facilitate many industrial, domestic and personal practices and thereby various chemicals used for these purposes are introduced into the environment. Parabens are chemicals used by the pharmaceutical, food and cosmetic and whose function is to conserve, and there are many questions regarding its safety , as some reports have shown that exposure to these parabens can modulate or disrupt endocrine system and thereby generate harmful consequences for human health and aquatic environments . The aim of this study was to evaluate the presence of Methylparaben and Propylparaben in the aquatic environment and their potential estrogenic and acute toxicity. The methodology developed from the in vitro YES assay for the determination of estrogenic activity, acute toxicity tests with Daphnia similis and Aliivibrio fischeri and quantification of parabens in water of Maracanã River RJ by liquid chromatography coupled with mass spectrometry. The EC50 results for the PP and MP, from the in vitro YES, was 18,91 mgL -1 and 7 mgL-1 and the magnitude of response was less potent 10-5 and 10-4 that 17ß - estradiol for MP and PP , respectively. From the acute toxicity test, the EC50 values obtained in Daphnia similis was 29,42 mgL-1 and 9,94 mgL-1, and Aliivibrio fischeri was 3,047 mgL-1 and 1,946 mgL-1, respectively, with it was observed that PP is more toxic in all organisms tested, and more toxic to one and less to others . The water of the Maracanã River was not toxic to Daphnia similis in either points, since for Aliivibrio fischeri was toxic at only one point. Found concentrations of MP and PP were higher at the point where, according to the physico-chemical parameters, water quality is not within the standards required by legislation and quantified in order ngL-1, however its worth to note that the DEs are not found in the middle separate environment, they interact with each other and cause additive or synergistic effects , its very difficult to predict what effect , so its important to know the estrogenic potential of simple substances , since in a study with an environmental matrix, one can observe if there were any additive or synergistic effect of other substances .

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Natural and synthetic estrogens are naturally excreted by humans and some vertebrates. These emerging contaminants classified as endocrine disruptors (EDs) present, even in small quantities, in water and sediment are able to affect the endocrine system and the stages of growth and reproduction of aquatic organisms and humans. In this context, this study aimed to perform a physical-chemical and microbiological characterization of reservoir water from Santo AnastÃcio, located in CearÃ, Brazil, and the develop and validate an analytical method using High-performance liquid chromatography with detector diode array (HPLC-DAD) for the determination of caffeine (CAF) and estrone hormones, 17β-estradiol, estriol (natural) and 17α-ethinylestradiol (synthetic) for water and sediment phase of reservoir water studied. The physicochemical parameters evaluated were phosphorus, chlorophyll A, pheophytin, nitrogen, chloride, sulfide, hardness, calcium, magnesium, pH, alkalinity, turbidity and total and dissolved solids. Microbiological characterization evaluated the presence of total and fecal coliforms. For the determination of EDs by HPLC-DAD was used to extract solid phase techniques (SPE) for water sample and QuEChERS for the sediment sample. The results of physicochemical analyzes indicated above the allowed values by CONAMA 357/05 for class 2, only to phosphorus and sulfide. Microbiological characterization indicated the presence of coliforms far above the values stipulated by CONAMA. In determining the DEs concentrations from 0.54 to 0.93 μg L-1 were detected caffeine in water, and 0.76 to 4.94 mg kg-1 17β estradiol and estriol in the sediment. Therefore, the water reservoir is unsuitable for direct consumption and recreation within the law. The validated analytical method was efficient for the determination of analytes can be used for routine analysis since there is no legislation to caffeine and EDs. / EstrÃgenos naturais e sintÃticos sÃo naturalmente excretados por seres humanos e alguns vertebrados. Estes contaminantes emergentes classificados como desreguladores endÃcrinos (DEs) presentes, mesmo em pequenas quantidades, na Ãgua e no sedimento sÃo capazes de afetar o sistema endÃcrino e as fases de crescimento e reproduÃÃo de organismos aquÃticos e humanos. Nesse contexto, este trabalho teve como objetivos efetuar uma caracterizaÃÃo fÃsico-quÃmica e microbiolÃgica da Ãgua do aÃude Santo AnastÃcio, localizado no CearÃ, Brazil, alÃm de desenvolver e validar um mÃtodo analÃtico empregando a cromatografia lÃquida de alta eficiÃncia com detector de arranjos de diodos (HPLC-DAD) para determinaÃÃo da cafeÃna e dos hormÃnios estrona, 17β-estradiol, estriol (naturais) e 17α-etinilestradiol (sintÃtico) na Ãgua e no sedimento do aÃude mencionado. Os parÃmetros fÃsico-quÃmicos avaliados foram fÃsforo, clorofila A, feofitina, nitrogÃnio, cloreto, sulfeto, dureza, cÃlcio, magnÃsio, pH, alcalinidade, turbidez e sÃlidos totais e dissolvidos. A caracterizaÃÃo microbiolÃgica avaliou a presenÃa de coliformes totais e termotolerantes. Para a determinaÃÃo dos DEs por HPLC-DAD utilizou-se as tÃcnicas de extraÃÃo em fase sÃlida (SPE) para a amostra de Ãgua e QuEChERS para a amostra de sedimento. Os resultados das anÃlises fÃsico-quÃmicas indicaram valores acima do permitido pelo CONAMA 357/05, para classe 2, apenas para fÃsforo e sulfeto. A caracterizaÃÃo microbiolÃgica indicou a presenÃa de coliformes muito acima dos valores estipulados pelo CONAMA. Na determinaÃÃo dos DEs foram detectados as concentraÃÃes de 0,54 a 0,93 μg L-1 de cafeÃna na Ãgua, e, 0,76 a 4,94 mg kg-1 do 17β estradiol e estriol no sedimento. Diante disso, a Ãgua do aÃude encontra-se inadequada para consumo direto e recreaÃÃo dentro da legislaÃÃo vigente. O mÃtodo analÃtico validado mostrou-se eficiente para determinaÃÃo dos analitos podendo ser utilizado para anÃlises de rotina uma vez que nÃo existe legislaÃÃo para a cafeÃna e os DEs.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / One of the major environmental problems under discussion is related to the fact that natural and synthetic hormones have been frequently found in effluent from Wastewater Treatment Sewage (TSE) and natural waters, as well as the fact that conventional treatments are not effective in removing these substances. These substances are one recent class of environmental contaminants that interfere with the functioning of the human endocrine system and are related to occurrence of several types of cancer. In the attemp to solve this problem, we propose a study, from a theoretical approach, with cyclodextrins (CDs) molecules, that can be able to form inclusion complexes with many organic substances, poorly soluble in water, thus we believe that CDs are complexing agents promising for this task. In this work we studied the complexation of four steroid drugs commonly found in Wastewaters, with fourteen βCDs chosen, under different criterion to determine one CD more promising for the removal these steroids from the environment. Initially, we propose us to identify a semiempirical method can be better in describe the crystal structure of many compounds on the inclusion of CDs. On a second step applying a study about the formation of inclusion complexes with steroids, investigating two inclusion modes: in and out. The initial calculations indicated the PM3 method as the best method in the structural description of this type of system. Since the results of the complexation with steroids suggest that, in general, the structural orientation energetically more favorable corresponds to out arrangement. An analysis about the interactions more significant to the stability of the complex shows that the LH and the interactions dipole-dipole has a small contribution to the stabilization of the complexes, on the other hand, the hydrophobicity of the substituents on steroids rings, has been pointed as a crucial force to decide how s more stable the steroid enters towards the cavity of the CD. And finally, the results show that in general, all the CDs investigated, theoretically, have similar behavior among themselves. / Um dos grandes problemas ambientais em discussão está relacionado ao fato de hormônios naturais e sintéticos estarem sendo freqüentemente encontrados em efluentes de Estações de Tratamento de Esgoto (ETEs) e águas naturais, bem como o fato de os tratamentos convencionais não serem eficientes em remover tais substâncias. Estas substâncias são uma categoria recente de poluentes ambientais que interferem no funcionamento do sistema endócrino humano e de animais, estando relacionadas com o aparecimento de diversos tipos de câncer e disfunções nestes organismos vivos. Na tentativa de contribuir para resolver este problema, propomos um estudo, a partir de uma metodologia teórica, com ciclodextrinas (CDs), moléculas hábeis para formar complexos de inclusão com muitas substâncias orgânicas pouco solúveis em meio aquoso, pois, acreditamos que as CDs são agentes complexantes promissores para tal tarefa. Assim, neste trabalho estudamos a complexação dos 4 esteróides mais comuns encontrados nas ETEs, com 14 βCDs diferentes escolhidas a critério, a fim de determinar uma CD mais promissora para a remoção destes esteróides do meio ambiente. Inicialmente, procuramos identificar um método semiempírico capaz de melhor descrever a estrutura cristalográfica de vários compostos de inclusão a base de CDs, para numa segunda etapa aplicá-lo no estudo da formação de complexos de inclusão com os esteróides, investigando dois modos de inclusão: in e out. Os cálculos iniciais apontaram o método PM3 como o melhor método na descrição estrutural deste tipo de sistema. Já os resultados dos estudos de complexação com os esteróides sugerem que, no geral, o arranjo estrutural energeticamente mais favorável corresponde ao arranjo out. Uma análise das interações mais significativas para a estabilidade dos complexos mostra que, as LHs e as interações dipolo-dipolo têm uma contribuição pequena para a estabilização dos complexos, por outro lado, a hidrofobicidade dos substituintes nos esteróides, se mostrou fundamental para decidir o modo de inclusão mais favorável com que o esteróide entra na cavidade da CD. E finalmente, os resultados mostram que de modo geral, todas as CDs investigadas, teoricamente, possuem comportamento semelhante entre si, na inclusão dos esteróides estudados.

Estudos que avaliam a presença de interferentes endócrinos (IEs) em águas superficiais destinadas ao abastecimento público de grandes metrópoles aumentam a cada ano, devido aos efeitos que estas substâncias podem causar ao homem e animais mesmo quando em concentrações de ng L-1. Este estudo determinou 14 compostos orgânicos, entre interferentes endócrinos e marcadores de atividade antropogênica, em amostras de água e sedimento superficial da represa Guarapiranga, um dos maiores reservatórios que abastecem a Região Metropolitana de São Paulo, Brasil. O estudo indicou a presença de concentrações máximas entre 1061 ng L-1 e 12921 ng L-1 (para bisfenol A e dibutilftalato, respectivamente) na classe dos IEs e 114 ng L-1 e 12818 ng L-1 (para o colestanol e colesterol, respectivamente) na classe dos traçadores de atividade humana. Em sedimento, a concentração máxima variou de 3 ng g-1 a 872 ng g-1 (para o estradiol e nonilfenol, respectivamente). As concentrações mais elevadas foram observadas principalmente no ponto de amostragem G103-12 (23º41\'88,5\"S e 46º44\'67,3\"W), sendo a causa associada à densa urbanização próxima a este ponto. Embora, para a maioria dos compostos não haja concentrações limites estabelecidas em água superficial, abastecimento e sedimento, é muito importante que mais estudos sejam realizados a fim de conhecer o risco real para a saúde da população atendida por essas águas. A avaliação da qualidade da água quanto à presença de compostos emergentes em reservatórios é considerada um dos principais instrumentos que balizam a tomada de decisão dos gestores e fornecem subsidios para revisão da legislação. / Studies assessing the presence of endocrine disruptors compounds (EDCs) in surface water for public supply of major metropolis increase every year due to the effects that these substances can cause to human and wildlife even in concentrations of ng L-1. The present study determined 14 organic compounds, including endocrine disruptors and anthropogenic markers in surface water and sediment samples from Guarapiranga dam, one of the largest reservoirs that supply the Metropolitan Region of Sao Paulo, Brazil. The study indicated the presence of these compounds at maximum concentration ranging from 1061 ng L-1 and 12921 ng L-1 (of bisphenol A and dibutyl phthalate, respectively) for EDCs and 114 ng L-1 and 12818 ng L-1 (for cholestanol and cholesterol, respectively) for tracers of human activity. In sediment, maximum concentration ranged from 3 ng g-1 to 872 ng g-1 (for estradiol and nonylphenol, respectively). The highest concentrations were observed at the sampling point G103-12 (23º41\'88.5 \"S and 46º44\'67.3\" W), which is associated to the dense urbanization near to this point. Although, to the most of compounds studied there is no concentration limits established for surface and supply water and sediment, it is very important that more studies should be conducted to ascertain the actual risk to the health of the population served by these waters. The assessment of water quality for the presence of emerging compounds in reservoirs is considered one of the main instruments to guide the managers decision making and provide subsidies for review of the regulation.

Les perturbateurs endocriniens (PE) sont des polluants environnementaux naturels ou fabriqués par l'Homme capables d'interférer avec les systèmes hormonaux endogènes. Parmi ces molécules, le di(2-éthylhexyl) phtalate (DEHP) et le nonylphénol (NP) sont très répandus dans les produits du quotidien et figurent sur la liste des Substances Prioritaires de l'eau. Cependant, peu d'études s'intéressent au contrôle central de la reproduction suite à l'exposition adulte et à de faibles doses de ces PE. L'exposition chronique de souris mâles adultes à ces PE seuls ou combinés montre des altérations à des doses proches de l'exposition environnementale humaine et de la dose journalière tolérable. Nous avons constaté une diminution de l'émission des vocalisations ultrasonores (USV) et de l'attraction des femelles par les mâles exposés au DEHP. Ceci a pu être relié à une diminution de l'expression du récepteur des androgènes dans l'aire préoptique médiane, région clé dans l'expression du comportement sexuel mâle. Ces effets ont été inversés lors de l'arrêt de l'exposition au DEHP. L'exposition au NP augmente l'émission des USV et les nombres de montées, d'intromissions et de poussées pelviennes avant d'atteindre l'éjaculation. Par ailleurs, l'exposition au mélange provoque des altérations comportementales différentes de celles observées pour les molécules isolées. Ces résultats ont permis de mettre en évidence une vulnérabilité du circuit neural régulant l'expression du comportement sexuel à l'exposition à ces PE. Des cibles moléculaires ont également pu être identifiées, particulièrement pour le DEHP, aidant ainsi à la compréhension de leur mode d'action dans le système nerveux central / Endocrine disruptors are natural or man-made environmental pollutants capable of interfering with endogenous hormonal systems. Among these molecules, phthalates and particularly di(2-éthylhexyl) phtalate (DEHP) and nonylphenol are widely present in everyday products and classified as priority substances in the Water Framework Directive. These two molecules are highly studied but few publications address the neural control of reproductive behaviors following adult exposure to low doses of these molecules.Chronic exposure of adult male mice to these two molecules, alone or in combination, showed behavioral alterations at doses close to human environmental exposure and tolerable daily intake. We observed a decreased emission of ultrasonic vocalisations and attraction of receptive females by males exposed to DEHP. These behavioral alterations were related to down-regulation of the androgen receptor in the medial preoptic area, the key region involved in the expression of male sexual behavior. These behavioral and molecular effects were reversed by DEHP exposure arrest. Adult exposure to NP increased the emission of ultrasonic vocalisations and number of mounts, intromissions and pelvic thrusts, without any amelioration of sexual behavior. Interestingly, exposure to both molecules caused behavioral alterations, which are are different from those observed for each molecule alone.Altogether, these results highlight the vulnerability of the neural circuitry underlying sexual behavior to exposure to these molecules. Furthermore, molecular targets have been identified, particularly for DEHP, thereby helping to understand their mode of action in the central nervous system.

The normal operation of the endocrine (hormonal) system can be disrupted by a number of man-made and naturally-occurring chemicals, thereby affecting those physiological processes that are under hormonal control. Such substances are called endocrine disrupting compounds (EDCs). The endocrine disruption issue has alarmed the environmental authorities since the substances involved can hinder hormonal processes causing far-reaching effects on reproduction and development in current and future human and wildlife generations. Effects on some species of fish triggered worldwide concern and initiated a research scheme which is being undertaken by various organisations e.g. United States Environmental Protection Agency (USEPA), United Kingdom Environment Agency (UKEA), Oslo and Paris Commission (OSPAR), Japan Environment Agency (JEA) and World Wildlife Fund (WWF) in order to assess the effects (present and potential), point of generation, levels of contamination and exposure limits. The findings showed that most of the oestrogens are produced by humans and animals and get discharged into river streams mainly through sewage effluents. Fish in particular have been found to be affected the most even when the oestrogenic levels in water are very low. The probability of future European legislation to eliminate hormonally active compounds from wastewaters suggests that new and alternative methods should be developed for their removal. In this work, the adsorption of 17ß-oestradiol (E2) and 17a-ethinyl oestradiol (EE2) onto several granular activated carbons and Hypersol-Macronet hypercrosslinked polymers was investigated by batch experiments after a low level detection system had been developed using Gas Chromatography Mass Spectrometry (GC/MS). Equilibrium experiments were carried out for all adsorbents to quantify the sorption capacity for E2 and EE2. For better assessment of the sorbents performance, their physical properties such as surface area, average pore diameter and micropore volume and chemical structure were characterised by N2 adsorption experiments, scanning electron microscopy (SEM), FTIR spectroscopy, elemental analysis, sodium capacity determination, pH titration, proton binding curves and zeta potential measurements. Adsorption isotherm data were fitted to the Langmuir and Freundlich equations. Activated carbons were found to be preferable to Hypersol-Macronet hypercrosslinked polymers for adsorption purposes. The adsorption of oestrogens appears to be controlled by hydrophobic interactions. Kinetic experiments were performed with different size ranges of adsorbents at different concentrations and the results were analysed by a particle diffusion model. It was found that concentration did not seem to influence the kinetics of the oestrogen sorption whereas the particle size of the adsorbents influenced the adsorption rate of both molecules. The particle diffusion model seemed to fit the data collected for the adsorption rate of 17B-oestradiool onto the adsorbents but gave a poor fit for most of the data collected for 17a-ethinyl oestradiol.

Depuis le début des années 1990, de nombreuses équipes de recherche ont consacré leurs travaux à étudier les effets des perturbateurs endocriniens (PEs) sur l'homme et les espèces animales. Plusieurs polluants libérés dans l’environnement tels que les pesticides, les herbicides, les plastifiants industriels ou les résidus de médicaments ont ainsi le potentiel de perturber les équilibres hormonaux des organismes, induisant ainsi de graves conséquences sur leur développement et leur reproduction. Pour lutter contre ces dommages, diverses réglementations spécifiques et des systèmes de criblages se mettent en place pour détecter ces molécules. Ces systèmes doivent être rapides et fiables et les poissons téléostéens comme le medaka ou le poisson zèbre constituent d'excellents modèles pour détecter la présence de ces molécules et étudier leurs effets in vivo. Durant cette thèse nous avons étudié l’impact de certains PEs sur le développement embryonnaire en utilisant le poisson zèbre et le médaka comme modèle. Nous avons pu caractériser l'action du Bisphenol A sur la formation des otholites de l’oreille interne du poisson zèbre. En alliant des approches pharmacologiques et génétiques, nous avons identifié le récepteur nucléaire orphelin ERR comme une nouvelle cible in vivo de cette molécule. En parallèle, nous avons développé des lignées transgéniques rapportrices de médaka permettant de détecter la présence de polluants à activités androgénique ou anti-androgénique ce qui accroît ainsi la gamme des outils disponibles pour évaluer la présence des PEs dans les effluents liquides. / Since the early 1990s, many research teams have devoted their work to study the effects of endocrine disruptors (EDCs) in humans and animal species. Many pollutants released into the environment such as pesticides, herbicides, industrial plasticizers or drug residues have the potential to disrupt bodies’ hormonal balances, thus inducing a severe impact on their development and reproduction. To fight against these damages, various specific regulations and screening systems are setting up to detect these molecules. These systems must be fast, reliable. Teleost fish such as medaka and zebrafish are excellent models to detect the presence of these molecules and study their effects in vivo. In this thesis, we studied the impact of certain EDCs on the embryonic development using zebrafish and medaka as models. We characterized the action of Bisphenol A on the otoliths formation in the inner ear of zebrafish. By combining genetic and pharmacological approaches, we identified the orphan nuclear receptor ERRγas a new target of this molecule in vivo. In parallel, we have developed reporter transgenic lines of medaka to detect the presence of pollutants containing androgenic or anti-androgenic activities and thereby increasing the range of tools available to assess the presence of EDCs in liquid effluents.

Les produits phytosanitaires contiennent des molécules qui peuvent interférer avec le système endocrinien et induire des effets délétères sur les êtres vivants. Ce sont des perturbateurs endocriniens. Afin d’assurer un niveau de protection satisfaisant pour les mollusques aquatiques, une bonne compréhension des perturbations du système endocrinien est nécessaire. L’organisation pour le développement et la coopération économique s’est exprimée en faveur de la mise en place de protocoles standards d’écotoxicité chez les mollusques, notamment sur la reproduction, dans le cadre des processus d’évaluation du risque écologique et des autorisations de mise sur le marché de nouvelles molécules. Cette thèse vise à proposer des tests standards et des outils d’analyse afin d’évaluer les effets de substances chimiques chez un escargot d’eau douce, Lymnaea stagnalis. Une série de tests de toxicité a été conduite dans laquelle les limnées ont été exposées à des androgènes, des oestrogènes et dessolvants. Les effets observés ont été évalués à l’aide d’approches statistiques standards et d’un modèle de toxicokinétique-toxicodynamique (TKTD). Les limnées ont montré une sensibilité uniquement au tributyltin et triphényltin à des concentrations réalistes d’un point de vue environnemental. Le triéthylène glycol et l’acétone étaient les solvants les moins nocifs. De plus, le modèle TKTD suggère un mécanisme généraliste dans l’action du composé. La manifestation des perturbations endocriniennes chez L. Stagnalis restent évasives et nécessitent d’autres recherches. Enfin, la thèse a montré que / Plant protection products and biocides may contain chemicals that interfere with the endocrine system and consequently induce adverse effects on wildlife. Such chemicals are known as endocrine disruptors (EDs). To ensure a satisfactory protection level for aquatic molluscs a comprehensive understanding of endocrine disruption is needed. The Organisation for Economic Co-operation and Development supports the standardisation of toxicity tests guideline on mollusc reproductive toxicity, which is intended for environmental risk assessments purposes. This PhD thesis aimed at improving toxicity test protocols and data analysis tools to evaluate effects of chemicals on the great pond snail Lymnaea stagnalis. A suite of toxicity tests was conducted in which snails were exposed to putative androgens (tributyltin, triphenyltin, methyltestosterone), putative oestrogens (alkylphenols and ethynyloestradiol), and solvents. Resulting effects were evaluated using standard statistical approachesand/or a toxicokinetic-toxicodynamic (TKTD) model. Snails were sensitive to tributyltin and triphenyltin, while none of the oestrogenic chemicals and methyltestosterone significantly impacted animals at environmentally relevant concentrations. Triethylene glycol and acetone were the least harmful solvents. Further, the analysis of data from the tributyltin test with the TKTD model suggested a generic mechanism of action of the compound (i.e., not specific for EDs). The demonstration of endocrine disruption in L. stagnalis remained elusive and deserves further research. Finally, the thesis highlight

Orientador: Wilson de Figueiredo Jardim / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-26T13:39:08Z (GMT). No. of bitstreams: 1 Pescara_IgorCardoso_D.pdf: 5286232 bytes, checksum: 74793adc4ea590e2eb8fdeb51a796399 (MD5) Previous issue date: 2014 / Resumo: Este trabalho teve como objetivo avaliar a eficiência dos tratamentos empregados em cinco estações de tratamento de esgoto (ETE) e duas estações de tratamento de água (ETA) na remoção dos hormônios endógenos estrona (E1), 17?-estradiol (E2), estriol (E3), testosterona (TTN) e progesterona (PROG), hormônios sintéticos 17a-etinilestradiol (EE2), dietilestilbestrol (DES), norgestrel (NGT) e mestranol (MEE), produtos de uso industrial bisfenol A (BPA), fenolftaleína (PhPh), n-nonilfenol (nNP), n-octilfenol (nOP) e triclosan (TCS), o fármaco cafeína (CAF) e o pesticida atrazina (ATZ). Foi possível quantificar BPA, E1, E3, CAF e ATZ em esgoto bruto e BPA, E1, CAF e ATZ em água bruta. Para cada composto, tanto os tratamentos realizados pelas ETE quanto os realizados pelas ETA apresentaram eficiências de remoção variáveis. A cafeína foi o composto-alvo mais frequentemente encontrado nas amostras, sendo escolhida como indicador químico para avaliar a eficiência dos tratamentos. Foi encontrada uma taxa média de 99% para a remoção de CAF pelas ETE, sendo as etapas com tratamentos biológicos as mais eficientes. Os tratamentos anaeróbios apresentaram remoções médias superiores a 80% na remoção de CAF, e os tratamentos aeróbios foram capazes de remover cerca de 20% da carga do composto. Nas ETE, os tratamentos de coagulação e desinfecção foram pouco eficientes nas remoções dos compostos-alvo. Já para as ETA, foram observadas remoções de até 80% de CAF pelos tratamentos empregados, sendo a etapa de coagulação a responsável pela maior remoção de CAF da água bruta. A etapa de filtração em carvão ativado apresentou eficiências de remoção menores que 10%, enquanto a desinfecção demonstrou baixa eficiência na remoção de CAF. Na cidade de São José do Rio Preto ¿ SP foi possível quantificar, ao menos uma vez, BPA, E1, E3, PhPh, CAF, TCS e ATZ em água superficial, BPA, TCS e CAF em água subterrânea e BPA, CAF e ATZ em água de abastecimento. / Abstract: Water and wastewater conventional treatments are often considered inefficient to remove micropollutants. The aim of this work was to evaluate the removal efficiency of the endogenous hormones estrone (E1), 17? estradiol (E2), estriol (E3), testosterone (TTN), and progesterone (PROG), the synthetic hormones 17a-ethinylestradiol (EE2), diethylstilbestrol (DES), norgestrel (NGT), and mestranol (MEE), the industrial chemicals bisphenol A (BPA), phenolphthalein (PhPh), n nonylphenol (nNP), n octylphenol (nOP), and triclosan (TCS), the pharmaceutical caffeine (CAF), and the pesticide atrazine (ATZ) by five wastewater treatment plants (WWTP) and two drinking water treatment plants (DWTP). The target compounds BPA, E1, E3, CAF, and ATZ were found in raw sewage, while BPA, E1, CAF, and ATZ in raw water. Both WWTP and DWTP treatments presented variable removal efficiency. Caffeine was the target compound more often detected in samples, thus it was chosen as a chemical indicator to evaluate treatment efficiencies. The average removal of CAF by WWTP was 99%, in which the biological treatment was more efficient. The anaerobic treatment presented an average removal of CAF above 80%, and around 20% for the aerobic treatments. Coagulation and disinfection treatments of WWTP showed low efficiency to remove the target compounds. DWTP treatment efficiency reached up to 80% removal, with coagulation step presenting highest removals of CAF. The activated carbon filtration showed removal efficiency under 10%, whereas the disinfection step presented low efficiency to remove CAF. At São José do Rio Preto city were found, at least once, BPA, E1, E3, PhPh, CAF, TCS, and ATZ in surface water; BPA, TCS, and CAF in groundwater, and BPA, CAF, and ATZ in drinking water / Doutorado / Quimica Analitica / Doutor em Ciências

<p>Nespu&scaron;teni testis predstavlja odsustvo testisa u skrotumu sa jedne ili obe strane. Faktori rizika za pojavu nespu&scaron;tenog testisa obuhvataju genetsku predispoziciju, prevremeno rođenje, nisku porođajnu masu i prenatalnu izloženost endokrinim disruptorima ili duvanskom dimu. Endokrini disruptori se defini&scaron;u kao egzogene supstance koje imaju uticaj na homeostazu organizma i proizvodnju reproduktivnih hormona. U ovoj grupi nalaze se organofosforni pesticidi koji se &scaron;iroko upotrebljavaju u poljoprivredi. Većina organofosfornih pesticida ima antiandrogeni uticaj i uz činjenicu da živimo u pretežno agrarnoj sredini predmet su na&scaron;eg interesovanja. Cilj istraživanja je da se utvrdi razlika u izloženosti organofosfornim pesticidima kori&scaron;ćenjem upitnika kreiranog po modelu standardizovanog Evropskog upitnika QLK 4-1999-01422 kod osoba koje su rodile zdravu mu&scaron;ku decu i osoba koje su rodile decu sa nespu&scaron;tenim testisom. Pored toga, cilj istraživanja je i da se odredi i uporedi vrednost metabolita organofosfornih pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat i dietilditiofosfat) u urinu majki koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima i majki koje su rodile zdravu mu&scaron;ku decu. Metodologija: Rad je radomizovano, prospektivno, kliničko istraživanje sprovedeno na Klinici za ginekologiju i aku&scaron;erstvo Kliničkog centra Vojvodine i Katedri za farmakologiju i toksikologiju Medicinskog fakulteta, Univerziteta u Novom Sadu. U kliničko istraživanje uključeno je 50 porodilja koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima (eksperimentalna grupa) i 53 porodilje koje su rodile zdravu mu&scaron;ku decu (kontrolna grupa) u periodu od oktobra 2012. godine do aprila 2018. godine. Tokom boravka u porodili&scaron;tu ispitanice su popunjavale upitnik o navikama nakon čega im je uzet uzorak urina radi analiziranja nivoa metabolita OF pesticida. Uzorci urina su pripremljeni metodom koju su opisali Wu i saradnici 2010. godine, a potom analizirani na gasnom hromatografu masenom spektrofotometaru marke Agilent 7890A. Rezultati: Ispitivane grupe se ne razlikuju po starosti ispitanica (prosečna starost kontrolne grupe 29,41 &plusmn; 5,58 godina, a eksperimentalne 30,54 &plusmn; 4,87 godina). U obe grupe prosečno je ispitanicama ovo bila druga trudnoća. Ispitanice se nisu razlikovale ni po načinu porođaja. Prosečna gestacijska nedelja trudnoće na porođaju iznosila je 39,45 &plusmn; 1,38 nedelja za kontrolnu grupu i 39,20 &plusmn; 1,38 nedelja za eksperimentalnu grupu, a porođajna masa novorođenčeta 3527,30 &plusmn; 470,16 g u kontrolnoj grupi i 3404,37 &plusmn; 508,20 g u eksperimentalnoj grupi. Statistički značajna razlika postoji u odnosu na mesto stanovanja (50,9 % ispitanica kontrolne grupe i 77,6 % ispitanica eksperimentalne grupe žive u gradu), jedinicu stanovanja (67,9 % kontrolne i 45,7 % ispitanica eksperimentalne grupe žive u kući) i načinu začeća (6 % ispitanica eksperimentalne i 1,9 % ispitanica kontrolne grupe prijavilo je IVF kao način začeća). Skoro polovina ispitanica obe grupe su pu&scaron;ači, a njih 32,7 % kontrolne grupe i 38,8 % eksperimentalne pu&scaron;ile su i tokom trudnoće. Izloženost pesticidima prijavilo je 50,9 % ispitanica kontrolne i 44 % ispitanica eksperimentalne grupe, a profesionalnu izloženost prijavilo je 3 ispitanice kontrolne i 2 ispitanice eksperimentalne grupe. Ispitanice se ne razlikuju ni po poreklu voća i povrća koje konzumiraju, kao ni po vrsti voća koje su konzumirale tokom trudnoće. Prosečne izmerene vrednosti DMF u kontrolnoj grupi iznose 5,604 &plusmn; 6,103 ug/L, a u eksperimentalnoj 4,815 &plusmn; 6,729 ug/L. Izmerene vrednosti DEF u kontrolnoj grupi su 0,408 &plusmn; 0,447 ug/L, a u eksperimentalnoj 0,461 &plusmn; 0,593 ug/L. Nivo DMDTF u kontrolnoj grupi bio je 0,431 &plusmn; 0,508 ug/L, a u eksperimentalnoj 0,547 &plusmn; 0570 ug/L, a DETF 0,403 &plusmn; 0,606 ug/L u kontrolnoj i 0,529 &plusmn; 0,725 ug/L u eksperimentalnoj grupi. Ni jedan metabolit ne pokazuje statistički značajnu razliku u ispitivanim grupama. Slične vrednosti dobijene su i za vrednosti korigovane za nivo kreatinina. Univarijantna regresiona analiza pokazala je da ispitanice koje žive u gradu imaju 3,3 puta veće &scaron;anse da rode dete sa nespu&scaron;tenim testisom, a one koje žive u stanu imaju 2,5 puta veće &scaron;anse za isti ishod. Statistički značajna razlika primećena je u nivou DEDTF u zavisnosti od starosti ispitanica i jedinici stanovanja. Vi&scaron;e vrednosti DETF dobijene su kod ispitanica koje su bile na hormonskoj terapiji tokom trudnoće. Ispitanice koje su prijavile da su bile izložene pesticidima tokom trudnoće u urinu su imale statistički značajno vi&scaron;e vrednosti DMDTF u odnosu na ispitanice koje su se izjasnile da nisu bile izložene pesticidima. Slični rezultati za vrednost DEDTF dobijeni su kod ispitanica koje su se izjasnile da poseduju kućne ljubimce. Statistički vi&scaron;e vrednosti DEF i DETF korigovano za nivo kreatinina dobijene su kod ispitanica koje nisu konzumirale jabuke, a vi&scaron;e vrednosti DEF i DEDTF dobijene su kod ispitanica koje su konzumirale maline i kupine tokom trudnoće. Ostale grupe nisu pokazale statistički značajnu razliku među ispitivanim grupama. Zaključci: Izloženosti trudnica OF pesticidima nije značajno veća u grupi majki koje su rodile decu sa nespu&scaron;tenim testisom u odnosu na izloženost OF pesticidima kod majki zdrave mu&scaron;ke dece. Vrednosti metabolita OF pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat, dietilditiofosfat) u urinu majki koje su rodile mu&scaron;ku decu sa nespu&scaron;tenim testisima nije vi&scaron;a u odnosu na vrednosti metabolita OF pesticida (dimetilfosfat, dimetilditiofosfat, dietilfosfat, dietiltiofosfat, dietilditiofosfat) izmerene u jutarnjem urinu majki koje su rodile zdravu mu&scaron;ku decu.</p> / <p>Undescended testis is the absence of testis in the scrotum on one or both sides. Risk factors for the occurrence of undescended testis include genetic predisposition, premature birth, low birth weight and prenatal exposure to endocrine disruptors or tobacco smoke. Endocrine disruptors are defined as exogenous substances that can affect homeostasis of the organism and the production of reproductive hormones. In this group are organophosphorus pesticides that are widely used in agriculture. Most of organophosphorus pesticides have anti-androgenic effect and with the fact that we live in a predominantly agricultural area, they are the focus of our interest. The aim of the research: The aim of this study is to determine the difference in exposure to organophosphorous pesticides using questionnaires created by standardized European model questionnaire QLK 4-1999-01422 in individuals who gave birth to a healthy male children and women who gave birth to children with undescended testis. In addition, the aim of this study is to determine and compare the value of metabolites of organophosphorus pesticides (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) in the urine of mothers who gave birth to male children with undescended testis and mothers who gave birth to healthy male children. Methodology: This work is randomized, prospective, clinical research conducted at the Clinic for gynaecology and obstetrics of the Clinical center of Vojvodina and the Department of Pharmacology and toxicology of the Faculty of Medicine, University of Novi Sad. This clinical research includes 50 new mothers that gave birth to male children with undescended testes (experimental group) and 53 new mothers that gave birth to healthy male children (control group) in the period from October 2012 to April 2018. During their stay at the maternity hospital the subjects were asked to fill out a questionnaire about habits and to give a urine sample for analyzing the level of metabolites of organophosphus pesticides. Urine samples were then prepared using the method described by Wu and associates 2010, and analyzed on gas chromatograph with a mass spectrophotometer Agilent 7890A brand. Results: Study groups do not differ according to the age of women (average age of control group is 29.41 &plusmn; 5.58 years, and experimental 30.54 &plusmn; 4.87 years). In both groups this was second pregnancy on average. The subjects did not distinguish either by the way of delivery. The average gestational weeks of pregnancy to childbirth was 39.45 &plusmn; 1.38 weeks for the control group and 39.20 &plusmn; 1.38 weeks for the experimental group, and birth weight of newborn was 3527.30 &plusmn; 470.16 g in control group and 3404.37 &plusmn; 508.20 g in the experimental group. There is no statistically significant difference in relation to the place of residence (50.9 % of the control group and 77.6 % of experimental live in the city), the living unit (67.9 % and 45.7 % of the control and experimental groups are living in the house) and the way of conception (6 % of experimental and 1.9 % of the control group reported IVF as a way of conception). Nearly half of both groups are smokers, and 32.7 % of women in the control group and 38.8 % in experimental smoked during pregnancy. Exposure to pesticides reported 50.9 % of mothers in control and 44 % of mothers in the experimental group. Professional exposure was reported by 3 control subjects and 2 subjects in experimental group. The subjects did not differ according to the origin of fruits and vegetables they were consuming, neither regarding the type of fruits they consumed during pregnancy. Average level of dimethylphosphate in control group was 5.604 &plusmn; 6.103 ug/L, and in experimental 4.815 &plusmn; 6.729 ug/l. Levels of diethylphosphate in control group were 0.408 &plusmn; 0.447 ug/L, and in experimental 0.461 &plusmn; 0.593 ug/l. DMDTP level in the control group was 0.431 &plusmn; 0.508 ug/L, and in experimental 0.547 &plusmn; 0570 ug/L, and the DETP was measured 0.403 &plusmn; 0.606 ug/L in control, and 0.725 &plusmn; 0.529 ug/L in the experimental group. These metabolites showed no statistically significant difference in the examined groups. Similar values are obtained for the adjusted values for creatinine level. Univariate regression analysis showed that the subjects who live in town are 3.3 times more likely to have child with undescended testis, and those who live in the apartment are 2.5 times more likely for the same outcome. Statistically significant difference was noticed in DEDTP level depending on the age of the subject and the living unit. Higher levels of DETP metabolites were detected in subjects that have been on hormonal therapy during pregnancy. The subjects who reported being exposed to pesticides during pregnancy had statistically significantly higher DMDTP values in relation to the subjects that were not exposed to pesticides. Similar results are obtained for the DEDTP level with higher levels in subjects owning pets. Statistically higher levels of DEP and DETP adjusted for creatinine were obtained in subjects that were not reporting eating apples, and higher levels of DEP and DEDTP were obtained in subjects that consumed raspberries and blackberries during pregnancy. Other groups showed no statistically significant difference between the study groups. Conclusion: Exposure of pregnant women to OP pesticides is not significantly greater in the group of mothers who gave birth to children with undescended testis in relation to exposure to OP pesticides in mothers of healthy male children. The level of OP metabolites (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) in the urine of mothers who gave birth to children with undescended testis is not higher in relation to the levels of OP metabolites (dimethylphosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate and diethyldithiophosphate) recorded in urine of mothers who gave birth to healthy male children.</p>