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Avaliação analítica integrada de PM1 e nanopartículas atmosféricasSchneider, Ismael Luís January 2016 (has links)
As nanopartículas atmosféricas podem causar diversos efeitos à saúde já que durante a inalação podem penetrar mais profundamente no sistema respiratório humano, atingindo inclusive os alvéolos e a corrente sanguínea, e, desta forma, aumentar os efeitos toxicológicos prejudiciais. Estas partículas podem ser emitidas diretamente para a atmosfera (partículas primárias) ou ser formadas por conversão gás-partícula (partículas secundárias). Durante a última década, estudos têm sido realizados para uma melhor compreensão sobre as concentrações de nanopartículas, avaliando sua emissão, formação, dispersão, exposição e os efeitos à saúde. No entanto, poucos estudos foram realizados no Brasil e desta forma ainda há pouca informação em relação a esta temática. A Região Metropolitana de Porto Alegre - RMPA (Brasil) possui forte tráfego veicular, cerca de 1,8 milhões de veículos e, apesar das fontes móveis impactarem fortemente a qualidade do ar nesta região e ser a principal fonte de nanopartículas, não há estudos que avaliem em profundidade os níveis deste importante poluente ambiental. Desta forma, no presente estudo foram avaliadas as concentrações de nanopartículas em diferentes pontos da RMPA. Além disso, foram avaliadas as concentrações ambientais de partículas <1 μm (MP1) e metais associados, bem como foram identificadas e quantificadas as contribuições de fontes utilizando o modelo receptor Positive Matrix Factorization (PMF). Em adição, foi realizada uma análise das assinaturas espectrais no infravermelho de nitro-hidrocarbonetos policíclicos aromáticos (nitro-HPAs) associados ao MP1. Este grupo de compostos orgânicos possui uma ampla distribuição no ambiente e, por possuírem propriedades mutagênicas e carcinogênicas, seus riscos ambientais foram calculados. Desta forma, considerando os resultados obtidos, verificou-se uma forte contribuição antropogênica dos poluentes avaliados na RMPA. Esta degradação da qualidade do ar está em geral associada ao tráfego veicular, especialmente no que se refere aos níveis de nanopartículas e nitro-HPAs associados ao MP1. Além disso, as emissões industriais, a queima de carvão, biomassa e óleo combustível, bem como a queima de combustíveis veiculares apresentaram forte influência nos níveis de metais observados em MP1. / Atmospheric nanoparticles can cause health effects, as during inhalation penetrate more deeply into the human respiratory system, including the alveoli and reaching the bloodstream, and thereby increase the damaging toxicological effects. These particles can be emitted directly to the atmosphere (primary particles) or be formed by gas-to-particle conversion (secondary particle). Over the last decade, studies have been conducted for a better understanding of the nanoparticles, assessing the emission, formation, dispersion, exposure and health effects. However, few studies have been conducted in Brazil and thus there is little information regarding this subject. The Metropolitan Area of Porto Alegre - MAPA (Brazil) has a strong vehicular traffic, about 1.8 million vehicles and, despite mobile sources strongly impact the air quality in this region and be the main source of nanoparticles, there are no studies that evaluate in depth the levels of this important environmental pollutant. Thus, considering this knowledge gap, the present study evaluated the nanoparticle concentrations in different sites of the MAPA. In addition, environmental concentrations of particles <1 μm (MP1) and associated metals were evaluated and the sources were identified and quantified using the receptor model Positive Matrix Factorization (PMF). In addition, an analysis of the infrared spectral signatures of nitro polycyclic aromatic hydrocarbons (nitro-PAHs) associated with PM1 was performed. This group of organic compounds has a wide distribution in the environment, and because they have mutagenic and carcinogenic properties, their environmental risks were calculated. Thus, considering the obtained results, a strong anthropogenic contribution of the evaluated pollutants in the MAPA was observed. This degradation of the air quality in general is associated with vehicular traffic, especially regarding the levels of nanoparticles and nitro-PAHs associated with PM1. In addition, industrial emissions, coal, biomass and fuel oil combustion, as well as vehicle fuels combustion had a strong influence on the metal levels observed in PM1.
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Elimination des micropolluants aromatiques et persistants de boues de station d'épuration au cours de la digestion anaérobie assistée par électrolyse microbienne et matériaux conducteurs / Removal of persistent aromatic micropolluants from municipal sewage sludge in anaerobic digesters assisted by microbial electrolysis and conductive materialsKronenberg, Izabel 29 March 2018 (has links)
L’élimination des micropolluants organiques est devenue aujourd’hui un objectif de santé publique car leur toxicité et bioaccumulation au travers de la chaine trophique sont incontestables. Les hydrocarbures aromatiques polycycliques (HAP) et le nonylphénol (NP) présents en faible concentration dans l’eau usée sont peu éliminés par le traitement. Ces composés hydrophobes se retrouvent fortement sorbés à la matière organique des boues de station d’épuration. Les procédés de traitement de ces boues, comme la digestion anaérobie (DA) jouent un rôle central car ils constituent une des dernières barrières avant rejet vers l’environnement par épandage. La DA élimine les HAP et le NP mais les performances restent insatisfaisantes. L’objectif de cette thèse est d’améliorer les performances d’élimination des HAP et NP par la DA en utilisant l’électrolyse microbienne et l’ajout des matériaux conducteurs. Les résultats montrent que le graphite poreux permet de lever deux limites à la bioremédiation des HAP: le manque d'accepteurs d'électrons terminaux et la biodisponibilité limitée des HAP. En effet, le graphite poreux semble faciliter l'échange direct d'électrons avec la communauté syntrophique anaérobie ce qui améliore les performances d’hydrolyse de la matière et des contaminants associés, particulièrement, l’élimination de 12 HAP est accrue de 21 à 33 %. Pour le NP, les processus impliqués semblent être différents car aucune amélioration n’est observée quelles que soient les conditions.. L’addition du graphite non-poreux (avec une surface spécifique moindre) et du platine (avec une conductivité plus élevée) n’éliminait que deux HAP de faible poids moléculaire suggérant que la conductivité ne constitue pas un facteur majeur dans la dissipation des HAP. L’ajout du graphite poreux en plus grande quantité, par contre, n’a pas confirmé l’hypothèse qu’une augmentation de surface spécifique du matériau conducteur accroit également l’élimination des HAP. Lors de la DA assistée par différents matériaux, un enrichissement de méthanogènes hydrogénotrophs a été constaté qui pourra être à la racine des performances observées. Dans ce contexte, la composition microbienne de l’inoculum joue un rôle majeur dans l’ampleur d’une biodégradation des HAP. L'utilisation de matériaux conducteurs abordables tels que le graphite poreux et non-poreux pourrait présenter une stratégie de biodégradation alternative pour éliminer les HAP des boues, du sol ou des sédiments. / The elimination of organic micropollutants from the environment has become a public health goal today because of their toxicity and bioaccumulation through the trophic chain. Polycyclic aromatic hydrocarbons (PAHs) and nonylphenol (NP) are found at low concentrations in wastewaters. They are removed from wastewaters by sorption onto sewage sludge due to their hydrophobicity. Anaerobic digesiton (AD) that is used for sewage sludge treatmentThe treatments of these sludge, like anaerobic digestion (AD), therefore, plays a central role in reducing the micropollutant load before dissemination to the environment via sludge spreading. Removal performances of PAHs and NP are removed byunder AD are, yet, but limitedwith low performances. The aim of this PhD work is to enhance removalthese performances thanks to the use of two emerging techniques, microbial electrolysis and the addition of conductive materials. The results demonstrate that independent of the application of a potential porous graphite circumvents two limits of PAH bioremediation: the lack of terminal electron acceptors and PAHs’ limited bioavailability. Mediatorless electron exchange between the anaerobic syntrophic community and the conductive material presumably facilitates sludge hydrolysis which, in turn, leads to the enhanced bioavailability of PAHs and their subsequent biodegradation. The reductions of 12 PAHs were improved by 21 to 33% while NP was eliminated to the same extent in the digesters with and without conductive material. The addition of non-porous graphite (with less specific surface) and platinum (exhibiting higher conductivity) eliminated only two low molecular weight PAHs suggesting that conductivity is not a major factor in the dissipation of PAHs. The addition of porous graphite in larger quantities, however, did not support the hypothesis that an increase in specific surface area of the conductive material also enhances the removal of PAHs. During conductive material supplemented AD, an enrichment of hydrogenotrophic methanogens was ascertained which could be at the root of the removal performance. In this context, the microbial composition of the inoculum plays a major role in the extent of PAH biodegradation. The use of affordable conductive materials such as porous and non-porous graphite may present an alternative biodegradation strategy for removing PAHs from sludge, soil or sediments.
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Mesure et modélisation de la contamination du sol dans les ouvrages de gestion à la source du ruissellement urbain / Assessment of soil contamination in Sustainable Urban Drainage Systems (SUDS) through experimental and modelling approachesTedoldi, Damien 03 July 2017 (has links)
L'essor que connaît aujourd'hui la gestion intégrée des eaux pluviales s'accompagne d'un recours croissant à des techniques favorisant l'infiltration de l'eau “à la source”, disséminées sur des bassins versants urbains. Néanmoins, de telles pratiques suscitent des interrogations sur la capacité du sol à jouer le rôle de “filtre” vis-à-vis des polluants présents dans le ruissellement. Ces travaux de thèse s'intéressent à la contamination du sol par les métaux et les hydrocarbures aromatiques polycycliques (HAP) dans ces dispositifs, avec trois objectifs : (i) évaluer les niveaux et l'étendue spatiale de la contamination, (ii) mieux comprendre les mécanismes qui gouvernent le devenir de ces contaminants dans le sol des ouvrages, et (iii) proposer des recommandations sur la conception et la maintenance de ces installations, afin de leur assurer un fonctionnement épuratoire durable.La première partie du travail consiste en une série d'investigations expérimentales sur dix sites d'étude aux caractéristiques contrastées, en service depuis plus de dix ans. Cette phase donne lieu à des cartographies de la contamination superficielle en éléments traces métalliques, puis à des profils verticaux de métaux et HAP, accompagnés de différentes variables explicatives. La distribution spatiale des métaux en surface, qui est toujours structurée autour de la zone d'arrivée de l'eau, porte la signature du fonctionnement hydraulique réel des dispositifs, et révèle le caractère non-uniforme de l'infiltration lors des événements pluvieux courants, qui représentent la plus grande partie du flux polluant à l'échelle annuelle. Dans la zone la plus contaminée des ouvrages, métaux et HAP présentent un enrichissement significatif sur 10 à 40 centimètres de profondeur. La rétention des contaminants résulte de la combinaison de processus physico-chimiques (adsorption) et mécaniques (sédimentation et filtration) ; leur importance relative peut être quantifiée grâce au déficit de zirconium dans le sédiment urbain par rapport au bruit de fond géochimique local. La variabilité inter-sites des niveaux de contamination s'explique à la fois par les capacités de rétention du sol et par les flux de polluants issus du bassin versant d'apport. Bien que les teneurs de surface excèdent, sur certains sites, les seuils internationaux de remédiation du sol pour des espaces “multifonctionnels”, la zone concernée représente — latéralement et verticalement — une région assez limitée dans les ouvrages.La seconde partie de la thèse consiste en une approche de modélisation, dont la finalité est de décrire l'évolution à long terme de la contamination du sol, et d'évaluer l'incidence de différentes pratiques de conception ou de gestion. Une analyse de sensibilité est d'abord mise en œuvre afin d'identifier les “éléments-clés” dans la description du système, et d'orienter le choix du modèle. Les résultats montrent qu'une mauvaise estimation de la dispersivité ou de l'isotherme d'adsorption du sol est susceptible de biaiser significativement la prévision des profils de contamination. Dans la suite du travail, on propose une méthode visant à décrire l'infiltration non-uniforme et la filtration des polluants particulaires. Le modèle ainsi construit est validé via une comparaison entre mesures et simulations sur l'un des sites d'étude faisant l'objet d'une nouvelle caractérisation approfondie. Enfin, une analyse de scénarios illustre les bénéfices d'un amendement de l'horizon superficiel par un matériau adsorbant, ainsi que d'une répartition homogène des flux d'eau en surface, en matière de durée de vie des installations et de fréquence d'intervention. / Sustainable Urban Drainage Systems (SUDS) are increasingly used for stormwater management. However, the generalization of runoff infiltration in urban watersheds raises some concerns regarding the soil's ability to retain ubiquitous micropollutants. The present work addresses soil contamination by trace metals and polycyclic aromatic hydrocarbons (PAHs) in such infiltration devices, with the aims of: (i) appraising the levels and spatial extent of soil contamination, (ii) better understanding the mechanisms which govern the fate of contaminants in these systems, and (iii) identifying design and maintenance guidelines which may enhance long-term pollutant control through SUDS.The first part of the study consists in a series of experimental investigations in ten contrasting study sites, which have been in operation for more than ten years. The two-stage methodology successively leads to cartographies of metal contamination in the surface soil, and vertical profiles of metal and PAH concentrations, along with different explanatory variables. The spatial distribution of trace metals in the upper horizon displays a systematic structure with respect to the inflow area, and bears the time-integrated signature of the infiltration fluxes and flow pathways at the surface. In the most contaminated zone of the facilities, a significant enrichment of metals and PAHs is detectable until 10 to 40 cm depth. Contaminant retention results from the combination of different physicochemical and mechanical processes (resp. sorption and sedimentation/filtration), the contribution of which can be assessed via the zirconium deficit in urban sediment in comparison to the soil's geochemical background. The inter-site variability of contamination levels is attributable to (i) differences in the soil's retention capacities, and (ii) differences in pollutant loads from the watershed. Although surface contents may exceed intervention thresholds for “multi-functional” spaces in several study sites, the area which would require soil remediation is laterally and vertically limited.These experimental assessments are complemented by a modelling approach, to describe the long-term evolution of soil contamination, and to evaluate the effect of various SUDS designs and maintenance operations. A sensitivity analysis is first carried out so as to identify the “key elements” in the system description: the results show that a wrong estimation of the soil's dispersivity or sorption isotherm is likely to induce significant biases in the predicted contamination profiles. A method is then proposed to describe non-uniform water infiltration fluxes, and the filtration of particle-bound contaminants. The model is validated via a comparison between measured and predicted metal profiles in one of the study sites, where a comprehensive soil characterization is undertaken. Finally, a “scenario analysis” illustrates the benefits of (i) using soil enrichment products with enhanced sorption capacities, and (ii) facilitating water spreading at the surface, in terms of “lifespan” of the devices and maintenance requirements.
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Dynamique des flux de micropolluants et mécanismes de dépollution des rejets pluviaux urbains stricts à travers une technique alternative : approche multi-échelles par des méthodes expérimentale et numérique / Dynamic of micropollutants fluxes and treatment mechanisms in an urban stormwater pond-filter constructed wetlandWalaszek, Milena 19 January 2018 (has links)
Afin de limiter les impacts négatifs des rejets pluviaux urbains stricts (RPUS) sur l’hydraulique et la qualité des milieux naturels, des techniques alternatives sont mises en place. A Strasbourg (France,67), une mare de sédimentation suivie d’un filtre planté de roseaux à écoulement vertical collecte, stocke et traite les eaux de ruissellement d’un bassin versant résidentiel (2,7ha) depuis 5 ans. L’objectif de cette thèse est de (i) déterminer l’efficacité hydraulique et de traitement de la filière, (ii) évaluer les dynamiques de stockage des métaux et des hydrocarbures aromatiques polycycliques en temps de pluie et en temps sec, (iii) anticiper l’impact des variations climatiques sur l’efficacité de la filière, (iv) évaluer le comportement de sorption du cuivre, plomb et zinc dans le filtre et (v) modéliser les flux en entrée et en sortie de la filière. Trois années de suivi hydraulique, hydrologique et physico-chimique sur le site et 13 campagnes de prélèvement en temps pluie ont permis de montrer que les efficacités de la filière sont constantes au cours de l’année. Les RPUS sont caractérisés par de fortes concentrations en zinc, et les concentrations en micropolluants sont dépendantes de l’importance de la pluie. De plus, des expériences réalisées sur le sable du filtre (analyses chimiques, extractions séquentielles, expériences batch) ont montré que la rétention des métaux dans le filtre dépend du pH et de la concentration en métaux. Un modèle basé sur l’équation de Richards a été développé et permet de reproduire avec succès les écoulements verticaux 1D dans le filtre. / Stormwaters are identified as a major source of pollution for waterbodies. Particularly, metals and Polycyclic Aromatic Hydrocarbons (PAHs) in stormwater are known as the most toxic compounds for the fauna and flora. To limit the impact of these micropollutants on hydrosystems quality, stormwater constructed wetlands (SCW) have been built worldwide. This study aims i) to assess the efficiency of a 5-years-old SCW that combined a sedimentation pond followed by a vertical flow sand filter in urban area (Strasbourg 67, France), ii) to determine micropollutants storage in water and soils during dry period, (iii) to investigate the seasonality impacts on SCW performances, (iv) to evaluate copper, lead and zinc sorption behavior on the filter sand and (v) to model micropollutants and rain water fluxes from the catchment to the filter output. 3 years of on-site monitoring and 13 rain event sampling campaigns have shown that SCW hydraulic and treatment performances are constant over the year. Rain water is characterized by high loads of zinc and its micropollutants loads depend on the rain event depth. Then, chemical analyses, sequential extractions and batch experiments performed on the SCW substrate highlight that metals mobility in the filter is affected by pH and metal concentration. A model based on the Richard’s equation has been developed and predicts with success the 1-D vertical flow in the filter.
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Avaliação da biodisponibilidade dos HPAs em Mugil curema do Estuário de Santos e de Cananéia através da análise de metabólitos de HPAs em bile de peixes / PAHs bioavailability evaluation in Mugil curema from Santos and Cananeia Estuaries through the analysis of PAHs metabolites in fish bilePatire, Vinicius Faria 25 November 2010 (has links)
Os estuários são o receptáculo final de muitos contaminantes antrópicos, como os hidrocarbonetos policíclicos aromáticos (HPAs), que podem ser tóxicos aos organismos. A biodisponibilidade dos HPAs pode ser avaliada através de biomarcadores de exposição, como os metabólitos biliares. Este trabalho teve como objetivo avaliar a biodisponibilidade do HPAs em peixes da espécie Mugil curema do Estuário de Santos e de Cananéia, através da análise de metabólitos de HPAs em bile de peixes. As coletas no Estuário de Cananéia foram realizadas nos meses de junho e novembro de 2008 e as coletas no Estuário de Santos foram realizadas entre os meses de março e maio de 2009. Os metabólitos foram analisados através de cromatografia líquida com detector de fluorescência (HPLC/F). A concentração de metabólitos biliares totais para o Estuário de Cananéia variou entre 0,91 a 89,97 ?g g-1 de bile, e para o Estuário de Santos variou de 4,68 a 528,43 ?g g-1 de bile. Houve diferença significativa entre os locais estudados. Observou-se também que não houve diferença significativa entre as amostras de machos e de fêmeas. A biodisponibilidade de HPAs foi considerada como baixa para o Estuário de Cananéia e alta para o Estuário de Santos. Valores de referência foram propostos para analise ambiental de metabólitos biliares de HPAs, sendo estipulada uma concentração de 2,22 ?g g-1 de bile para locais não contaminados. / Estuaries are the final receptacle to many anthropic contaminants, such as polycyclic aromatic hydrocarbons (PAHs), who can be toxic to organisms. The PAH bioavailability can be evaluated through biomarkers, such as biliary metabolites. This work had as objective evaluate the PAHs bioavailability in fishes of Mugil curema from Santos and Cananéia Estuaries, through the analysis of PAHs metabolites in fish bile. The Cananéia Estuary sampling was made in June and November from 2008 and the Santos Estuary sampling was made between the months of March and May from 2009. The metabolites were analyzed through a high performance liquid chromatograph with fluorescence detector (HPLC/F). The biliary metabolites concentrations from the Cananéia Estuary varied between 0,91 a 89,97 ?g g-1 of bile, and from Santos Estuary varied between 4,68 a 528,43 ?g g-1 of bile. There were significant differences between the sampling sites. There were no significant differences between the male and female samples. The PAHs bioavailability was considered low to Cananéia Estuary and high to Santos Estuary. Reference values were proposed to PAHs biliary metabolites environmental analysis, been stipulated a concentration of 2,22 ?g g-1 of bile to uncontaminated sites.
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Isolation, characterization and exploitation of soil micro-organisms for bioremediation of benzo(a)pyrene contamination.January 2005 (has links)
by Ho, Kai-Man. / Thesis submitted in: December 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 158-179). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstracts --- p.ii / Contents --- p.vii / List of figures --- p.xiv / List of tables --- p.xvii / Abbreviations --- p.xx / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Poly cyclic Aromatic Hydrocarbons (PAHs) / Chapter 1.1.1 --- Characteristics of PAHs --- p.1 / Chapter 1.1.2 --- Sources of PAHs --- p.3 / Chapter 1.1.3 --- Environmental fates of PAHs --- p.5 / Chapter 1.1.4 --- Effects of PAHs on living organisms --- p.7 / Chapter 1.1.5 --- Summary --- p.10 / Chapter 1.2 --- Target PAH: Benzo[a]pyrene (BaP) --- p.10 / Chapter 1.3 --- PAH contamination in Hong Kong --- p.14 / Chapter 1.4 --- Remediation for PAHs contaminated soils --- p.15 / Chapter 1.4.1 --- Chemical/ Physical methods --- p.15 / Chapter 1.4.2 --- Bioremediation --- p.16 / Chapter 1.5 --- Biodegradation of PAHs by bacteria and fungi --- p.18 / Chapter 1.5.1 --- Tolerance and degradation --- p.18 / Chapter 1.5.2 --- Biodegradation of PAHs by microorganisms --- p.20 / Chapter 1.5.2.1 --- Bacteria --- p.22 / Chapter 1.5.2.2 --- Fungi --- p.23 / Chapter 1.6 --- Environmental standards --- p.26 / Chapter 1.7 --- Strategies of soil sampling and microbial isolation --- p.26 / Chapter 1.7.1 --- Contaminated soil vs. uncontaminated soil --- p.26 / Chapter 1.7.2 --- Native species vs. foreign species --- p.29 / Chapter 1.7.3 --- Background of the sampling areas --- p.30 / Chapter 1.7.3.1 --- North Tsing Yi shipyard --- p.30 / Chapter 1.7.3.2 --- "Tsam Chuk Wan, Sai Kung" --- p.33 / Chapter 1.8 --- Objectives of this study --- p.33 / Chapter 2. --- Materials and Methods --- p.35 / Chapter 2.1 --- Soil Collection --- p.35 / Chapter 2.1.1 --- Abandoned shipyard soil and its sediment soil --- p.35 / Chapter 2.1.2 --- "Tsam Chuk Wan, Sai Kung" --- p.35 / Chapter 2.2 --- Characterization of soils --- p.35 / Chapter 2.2.1 --- Sample preparation --- p.36 / Chapter 2.2.2 --- Soil pH --- p.36 / Chapter 2.2.3 --- Electrical conductivity --- p.36 / Chapter 2.2.4 --- Salinity --- p.36 / Chapter 2.2.5 --- Total organic carbon contents --- p.38 / Chapter 2.2.6 --- Metal analys --- p.is / Chapter 2.2.7. --- Oil and grease content --- p.38 / Chapter 2.2.8 --- Soil texture --- p.39 / Chapter 2.2.9 --- Available ammoniacal nitrogen and oxidized nitrogen --- p.39 / Chapter 2.2.10 --- Available Phosporus --- p.40 / Chapter 2.2.11 --- Total Nitrogen and total Phosporus --- p.40 / Chapter 2.2.12 --- Moisture / Chapter 2.2.13 --- DTPA-extractable metals --- p.41 / Chapter 2.2.14 --- Extraction of PAHs and organic pollutants --- p.41 / Chapter 2.2.14.1 --- Extraction procedures --- p.41 / Chapter 2.2.14.2 --- GC-MSD conditions --- p.41 / Chapter 2.2.14.3 --- Extraction efficiency --- p.43 / Chapter 2.2.15 --- Soil colour --- p.43 / Chapter 2.3 --- Screening and selection of microorganismms --- p.43 / Chapter 2.3.1 --- Isolation of potential BaP-degrading microorganisms --- p.44 / Chapter 2.3.1.1 --- Isolation of bacteria --- p.44 / Chapter 2.3.1.2 --- Isolation of fungi --- p.44 / Chapter 2.3.2 --- Cultures preserving microorganisms --- p.46 / Chapter 2.3.3 --- Screening and selection of microbes --- p.46 / Chapter 2.3.3.1 --- Bacteria --- p.46 / Chapter 2.3.3.2 --- Fungi --- p.46 / Chapter 2.3.4 --- Survival test --- p.47 / Chapter 2.3.5 --- Removal efficiency (RE) towards BaP by the microorganisms --- p.47 / Chapter 2.3.5.1 --- Bacteria --- p.47 / Chapter 2.3.5.2 --- Fungi --- p.48 / Chapter 2.3.6 --- Removal efficiency (RE) --- p.48 / Chapter 2.3.7 --- Relationship of absorbance of bacterial culture and bacterial biomass --- p.49 / Chapter 2.4 --- Identification of selected microorganisms --- p.49 / Chapter 2.4.1 --- Identification of bacterium --- p.49 / Chapter 2.4.1.1 --- 16S rDNA sequencing --- p.49 / Chapter 2.4.1.1.1 --- Primers --- p.49 / Chapter 2.4.1.1.2 --- DNA extraction --- p.51 / Chapter 2.4.1.1.3 --- Specific PCR --- p.51 / Chapter 2.4.1.1.4 --- Gel electrophoresis --- p.51 / Chapter 2.4.1.1.5 --- Purification of PCR products --- p.52 / Chapter 2.4.1.1.6 --- DNA sequencing --- p.52 / Chapter 2.4.1.2 --- Midi Sherlock® Microbial Identification System (MIDI) --- p.53 / Chapter 2.4.1.3 --- Biolog MicroLog´ёØ system (Biolog) --- p.55 / Chapter 2.4.2 --- Identification of fungi --- p.56 / Chapter 2.4.2.1 --- ITS DNA sequencing --- p.56 / Chapter 2.4.2.2 --- Observation under electronic microscope --- p.58 / Chapter 2.5 --- Growth curve of the microorganism --- p.58 / Chapter 2.5.1 --- Bacterium --- p.58 / Chapter 2.5.2 --- Fungi --- p.58 / Chapter 2.6 --- Preparation of Benzo[a]pyrene (BaP) stock solution --- p.58 / Chapter 2.7 --- Comparison of isolated bacterium and fungi --- p.60 / Chapter 2.8 --- Optimization of BaP degradation by selected fungus --- p.60 / Chapter 2.8.1 --- Preparation of straw compost inoculated with selected fungus --- p.60 / Chapter 2.8.2 --- Effect of incubation time --- p.61 / Chapter 2.8.3 --- Effect of initial BaP concentration --- p.61 / Chapter 2.8.4 --- Effect of inoculum size / Chapter 2.8.5 --- Effect of temperature --- p.61 / Chapter 2.8.6 --- Effect of soil pH --- p.62 / Chapter 2.8.7 --- Study of BaP degradation pathway by the microorganisms using GC-MSD --- p.62 / Chapter 2.9 --- Chitin Assay --- p.62 / Chapter 2.10 --- Enzyme assay --- p.63 / Chapter 2.10.1 --- Laccase assay --- p.63 / Chapter 2.10.2 --- Manganese peroxidase assay --- p.63 / Chapter 2.10.3 --- Lignin peroxidase assay --- p.64 / Chapter 2.11 --- Toxicity of treated soil --- p.64 / Chapter 2.12 --- Statistical analysis --- p.65 / Chapter 3. --- Results --- p.66 / Chapter 3.1 --- Soil Collection --- p.66 / Chapter 3.1.1 --- North Tsing Yi shipyard --- p.66 / Chapter 3.1.2 --- "Tsam Chuk Wan, Sai Kung" --- p.66 / Chapter 3.2 --- Characterization of soil samples --- p.71 / Chapter 3.3 --- Extraction efficiency of Benzo[a]pyrene --- p.79 / Chapter 3.4 --- Screening and selection of microorganisms --- p.79 / Chapter 3.4.1 --- Isolation ofpotential BaP-degrading microorganisms --- p.79 / Chapter 3.4.2 --- Screening and selection of microbes --- p.87 / Chapter 3.4.2.1 --- Bacteria --- p.87 / Chapter 3.4.2.2 --- Fungi --- p.93 / Chapter 3.4.3 --- Growth curve of the microorganisms --- p.95 / Chapter 3.4.3.1 --- Bacterium --- p.95 / Chapter 3.4.3.2 --- Fungi --- p.99 / Chapter 3.5 --- Comparison of isolated bacterium and fungi --- p.99 / Chapter 3.6 --- Identification of selected microorganisms --- p.102 / Chapter 3.6.1 --- Identification of bacterium --- p.103 / Chapter 3.6.1.1 --- 16S rDNA sequencing --- p.103 / Chapter 3.6.1.2 --- Midi Sherlock® Microbial Identification System (MIDI) --- p.103 / Chapter 3.6.1.3 --- Biolog MicroLog´ёØ system (Biolog) --- p.107 / Chapter 3.6.2 --- Identification of fungi --- p.107 / Chapter 3.6.2.1 --- ITS DNA sequencing --- p.107 / Chapter 3.6.2.2 --- Observation under electronic microscope --- p.113 / Chapter 3.7 --- Optimization of BaP degradation by the selected fungus: Trichoderma asperellum --- p.107 / Chapter 3.7.1 --- Effect of incubation time --- p.107 / Chapter 3.7.2 --- Effect of inoculum size --- p.113 / Chapter 3.7.3 --- Effect of initial BaP concentration --- p.113 / Chapter 3.7.4 --- Effect of soil pH --- p.113 / Chapter 3.7.5 --- Effect of temperature --- p.117 / Chapter 3.8 --- Determination of breakdown products of BaP by BaP-degrading microorganisms --- p.117 / Chapter 3.9 --- Enzyme assay --- p.117 / Chapter 3.10 --- Evaluation of toxicity by using indigenous bacteria --- p.121 / Chapter 4. --- Discussion --- p.128 / Chapter 4.1 --- Physico-chemical properties of soil samples --- p.128 / Chapter 4.2 --- Determination of BaP and other organic compounds --- p.131 / Chapter 4.3 --- Identification of the microorganisms --- p.132 / Chapter 4.3.1 --- Bacteria --- p.132 / Chapter 4.3.2 --- Fungi --- p.134 / Chapter 4.4 --- Biodegradation by BaP-degrading microorganisms --- p.135 / Chapter 4.4.1 --- Isolation and screening of BaP-degrading microorganisms --- p.135 / Chapter 4.4.2 --- Biodegradation of BaP --- p.137 / Chapter 4.4.2.1 --- Bacteria --- p.137 / Chapter 4.4.2.2 --- Fungi --- p.138 / Chapter 4.4.3 --- BaP degradation pathway --- p.140 / Chapter 4.4.3.1 --- Bacteria --- p.140 / Chapter 4.4.3.2 --- Fungi --- p.140 / Chapter 4.5 --- Optimization of PAH degradation by T. asperellum --- p.143 / Chapter 4.5.1 --- Effect of incubation time --- p.143 / Chapter 4.5.2 --- Effect of initial BaP concentration --- p.144 / Chapter 4.5.3 --- Effect of inoculum size fungus --- p.144 / Chapter 4.5.4 --- Effect of soil pH --- p.145 / Chapter 4.5.5 --- Effect of temperature --- p.146 / Chapter 4.6 --- Comparison the selected bacterium and fungi --- p.146 / Chapter 4.7 --- Evaluation of toxicity by using in indigenous bacteria --- p.148 / Chapter 4.8 --- Post treatment by crude enzyme of Pleurotus pulmonarius --- p.149 / Chapter 4.9 --- Limiting factors for BaP degradation --- p.150 / Chapter 4.10 --- Further Investigations --- p.152 / Chapter 5. --- Conclusion --- p.155 / Chapter 6. --- References --- p.158
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Génotoxicité des hydrocarbures aromatiques polycycliques en melanges, une classe majeure de polluants atmosphériques / Genotoxicity of mixtures of polycyclic aromatic hydrocarbons, a major class of atmospheric pollutantsGenies, Camille 18 November 2013 (has links)
Les Hydrocarbures Aromatiques Polycycliques (HAP) représentent une famille de polluants atmosphériques issus de la combustion incomplète de la matière organique. Ils sont ainsi présents dans l'atmosphère polluée des villes, la fumée de cigarette et certaines industries. Une exposition aux HAP peut être à l'origine de cancers du poumon, de la peau et de la vessie. A ce titre, certains HAP sont suspectés ou reconnus cancérigènes pour l'Homme, comme le Benzo[a]pyrène (B[a]P), par leur capacité à induire des dommages à l'ADN après métabolisation. Malgré une émission systématique des HAPs en mélanges, la majorité des études s'est intéressée à l'effet génotoxique des HAP purs et principalement au B[a]P. Afin de fournir des données mécanistiques sur la génotoxicité et le mode d'action des mélanges de HAP, nous avons réalisé une étude in vitro dans des lignées cellulaires de poumons (A549), de vessie (T24) et de foie (HepG2). Les dommages à l'ADN ont été suivis par la mesure des adduits par HPLC-MS/MS et des dommages oxydatifs par la méthode des comètes, ainsi que le métabolisme par l'induction des gènes par RT-qPCR et les activités enzymatiques des CYP540 de phase I (EROD) et de phase II (GST). Dans un premier temps, l'utilisation du B[a]P, comme composé modèle a montré une absence quasi-totale de métabolisation et de génotoxicité pour T24. Par contre, la formation d'adduits et l'induction de la métabolisation a été mise en évidence pour A549, avec des effets notamment de dose-réponse « en cloche » similaires à ceux observés dans d'autres modèles de poumons. Nous avons ensuite étendu cette démarche à 12 HAP prioritaires et étudié leur métabolisation et la formation d'éventuels adduits en se focalisant sur la lignée pulmonaire A549. La combinaison de ces HAP au B[a]P dans des mélanges binaires ou dans des mélanges plus complexes mimant des mélanges environnementaux entraine une forte inhibition de la formation des adduits issus du B[a]P sans apparition d'adduits d'autres HAP. Par ailleurs, nous avons observé dans le cas des mélanges complexes une bonne corrélation entre l'activité EROD et la formation des adduits à l'ADN, alors que les gènes de phase sont eux surexprimés par l'exposition au mélange par rapport au B[a]P pur. Les mécanismes par lesquels s'exerce cette inhibition des adduits restent encore à élucider mais la métabolisation des HAPs constitue une étape clé dans la génotoxicité des mélanges à travers des phénomènes d'inhibition ou de compétition au niveau des CYP entrainant une inhibition de l'activité EROD. Il est donc clair que l'étude des HAP de façon individuelle n'est pas suffisante pour appréhender la génotoxicité des mélanges complexes. L'approche FET, couramment utilisée pour évaluer le risque lié à l'exposition aux mélanges de HAP, repose sur l'additivité des effets toxiques et néglige les interactions métaboliques entre les différents HAP. L'amélioration de cet outil de prédiction est nécessaire et passe obligatoirement par l'étude des mécanismes sous-jacents qui relient la composition des mélanges, leur métabolisation et leur génotoxicité. / Polycyclic Aromatic Hydrocarbons (PAH) represent a family of ubiquitous atmospheric pollutants produced upon incomplete combustion and thus present in polluted atmosphere of the cities, in cigarette smoke and in certain industries. Exposure to HAP can cause lung, skin and bladder cancers. For this reason, some PAHs are suspected or recognized carcinogenic to humans, especially Benzo[a]pyrene (B[a]P), through their ability at inducing the formation of DNA damage after metabolization. In spite of the systematic emission of PAHs in mixtures, the majority of the studies was interested in the genotoxic effect of pure PAH and mainly B[a]P. In order to provide mechanistic data on the genotoxicity and the mode of action of PAH mixtures, we designed an in vitro study using cell lines representative of lungs (A549), bladder (T24) and liver (HepG2). DNA damage was investigated through the quantification of adducts by HPLC-MS/MS and of oxidative damage by the Comet assay. In addition, the metabolism was studied by analyzing genes induction by RT-qPCR and enzymatic activities of phase I CYP540 (EROD) and phase II (GST). First, the use of B[a] P, as a reference compound showed a quasi-total absence of metabolization and genotoxicity for T24. In contrast, the formation of DNA adducts formation and the induction of metabolization was highlighted for A549, with a bell-shaped dose-response curve similar to those observed in other lungs models. Then we extended this approach to 12 priority PAH and analyzed their metabolization and the possible formation of adducts focusing on the pulmonary cell line A549. The combination of these HAP to B[a]P in binary mixtures or in complex mixtures representative to environmental exposures led to a strong inhibition in adducts formation induced by B[a]P without outbreak of adducts from other PAH. In addition, we observed, in the case of complex mixtures, a good correlation between the EROD activity and the formation of adducts in DNA, while phase I genes were always overexpressed after exposure to mixtures when compared to pure B[a]P. The mechanisms involved in the inhibition of DNA adducts remain to be elucidated but PAHs metabolization represents a key step in the mixtures genotoxicity through inhibition or competition of CYP resulting in an inhibition of EROD activity. It is thus clear that the study of the HAP in an individual way is not sufficient to understand the genotoxicity of complex mixtures. The TEF Approach, usually used to asses the risk related to PAH mixtures exposure, relies on toxic effects additivity and ignores metabolic interactions between the various PAH. The improvement of this prediction tool is essential and involves necessarily the study of the underlying mechanisms which connect mixtures composition, their metabolization and their genotoxicity.
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Perfil dos HPA prioritários na exaustão de veículo a diesel, no combustível diesel utilizado durante os ensaios de emissão veicular e no óleo lubrificante do motor / HPA priority profile in diesel vehicle exhaust, in diesel fuel used during the emissions testing and in engine lubricant oilCarolina Vieira de Souza 04 March 2015 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As concentrações na exaustão e os fatores de emissão dos hidrocarbonetos policíclicos aromáticos (HPA) prioritários de um veículo a diesel e as suas respectivas concentrações no diesel usado durante os ensaios de emissão veicular foram determinados com a finalidade de estimar a contribuição dos HPA provenientes do combustível nas emissões. Os produtos da combustão foram coletados diretamente nas emissões brutas do escapamento, utilizando um sistema de amostragem a volume constante sem diluição dos gases da exaustão. Os HPA associados ao MP foram amostrados de forma estratificada, utilizando um impactador em cascata MOUDI e filtros de fibra de vidro como substratos, e os HPA em fase gasosa foram amostrados usando cartuchos de amberlite XAD-2. A concentração dos HPA no óleo lubrificante do motor também foi monitorada ao longo do tempo até a sua troca após 12.000 km de uso. Após a extração e tratamento das amostras, a identificação e quantificação dos HPA foram realizadas, utilizando cromatografia de fase gasosa acoplada à espectrometria de massas (CG-EM) com injetor de grande volume de vaporização com a temperatura programável (PTV-LVI). Cinco variáveis do PTV-LVI foram otimizadas, utilizando planejamento de experimentos, o que permitiu obter limites de detecção menores do que 2,0 g L-1. Somente 7 dos 16 HPA prioritários foram identificados na exaustão: NAP, ACY, ACE, FLU, FEN, FLT e PYR. Os ensaios de emissão veicular foram realizados com o veículo em modo estacionário, sem aplicação de carga e com baixa velocidade de rotação do motor (1500 rpm), utilizando um diesel com menor teor de enxofre (10 mg kg-1) e com 5% v/v de biodiesel. Esses fatores possivelmente contribuíram para reduzir as emissões dos outros 9 HPA a valores abaixo dos limites de detecção do método desenvolvido. Aproximadamente 80% da massa dos HPA totais associados ao MP estavam presentes em partículas com tamanho entre 1,0 m e 56 nm, e aproximadamente 4,5% estavam presentes em partículas menores do que 56 nm. Partículas menores que 2,5 m são facilmente inaladas e depositadas no trato respiratório e na região alveolar, justificando a preocupação com relação às emissões de HPA associados a partículas provenientes da exaustão veicular de motores a diesel. Somente 5 dos 7 HPA identificados na exaustão foram detectados no diesel: NAP, ACY, FLU, FEN e PYR. A razão entre os fatores de emissão (g L-1diesel) dos HPA na exaustão e suas respectivas concentrações do diesel (g L-1) variaram de 0,01 0,02 a 0,05 0,029, dependendo do HPA. Esses valores indicam que pelo menos 95 a 99% dos HPA identificados no diesel foram destruídos e/ou transformados em outros compostos durante a combustão, e/ou foram retidos no reservatório do óleo lubrificante. Por outro lado, os HPA que tiveram maiores concentrações no diesel também apresentaram maiores fatores de emissão, o que sugere que os HPA provenientes do diesel possuem uma contribuição significativa para as emissões dos HPA totais. O perfil dos HPA prioritários no óleo lubrificante mostrou-se semelhante ao perfil dos HPA no diesel e nas emissões totais, onde o NAP, FEN e PYR foram os HPA majoritários / A diesel vehicle exhaust concentrations and emission factors of priorities polycyclic aromatic hydrocarbons (PAHs) and their respective concentrations in the diesel fuel used during the vehicle emissions testing were determined in order to estimate the fuel PAHs contribution to vehicle emissions. Combustion products are collected directly from the raw vehicle exhaust using a constant volume sampling system that does not employ exhaust products dilution. The particle-phase PAHs were sampled in stratified way using a Micro-orifice Uniforme Deposite Impactor (MOUDI) and glass fiber filter as substrates and the gas-phase PAHs were sampled using Amberlite XAD-2 cartridge. The PAHs concentrations in the engine lubricant oil were also monitored over time until its change after 12,000 km of use. After sample extractions and treatments, the PAHs identification and quantification were performed using gas chromatography coupled with mass spectrometry (GC-MS) and programmable temperature vaporization large volume injection (PTV-LVI). Five PTV parameters were optimized using statistical design experiments, which allowed to be obtained detection limits below 2.0 g L-1. Only 7 of 16 priority PAH were identified in exhaustion: NAP, ACY, ACE, FLU, FEN, FLT and PYR. The vehicle emission tests were carried out with the vehicle in stationary mode, without load, low engine speed (1500 rpm), using diesel fuel with lower sulfur (10 mg kg-1) and 5% v/v biodiesel. These factors may have contributed to reduce the other 9 PAH emissions to values below the detection limits of the method developed. Approximately 80% of the PAHs total mass associated with the particulate matter were present in particle size between 1.0 m and 56 nm and approximately 4.5% were present in particles smaller than 56 nm. Particles smaller than 2.5 m are easily inhaled and deposited in the respiratory tract and in the alveolar region, justifying the concern about PAHs emissions associated with particles from vehicle exhaust diesel engine. Only 5 of the 7 PAHs identified in exhaust were detected in diesel fuel: NAP, ACY, FLU, PHE and PYR. The ratio between the PAHs emission factors in exhaust (g L-1 diesel) and their respective concentrations in diesel fuel (g L-1) ranged from 0.01 0.02 a 0.05 0.029, depending on the PAH. These values indicate that al least 95 to 99% of the PAH identified in diesel fuel were destroyed and/or converted into other compounds during the combustion, and/or been retained in lubricant oil reservoir. On the other hand, PAHs that had higher concentration in diesel also had higher exhaust emission factors, which suggests that the PAH from diesel have a mean contribution to total PAHs emissions. The PAH priority profile in lubricant oil was similar to PAH profile in diesel and in vehicle exhaust, where the NAP, FEN and PYR were the majorities PAHs
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Tratamento eletroqu?mico de ?gua produzida sint?tica para remo??o de hidrocarbonetos polic?clicos arom?ticosPontes, Juliana Patr?cia Souza Duarte 06 July 2010 (has links)
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Previous issue date: 2010-07-06 / The aim of this work is the treatment of produced water from oil by using electrochemical technology. Produced water is a major waste generated during the process of exploration and production in the oil industry. Several approaches are being studied aiming at the treatment of this effluent; among them can be cited the biological process and chemical treatments such as advanced oxidation process and electrochemical treatments (electrooxidation, electroflotation, electrocoagulation, electrocoagulation). This work studies the application of electrochemical technology in the treatment of the synthetic produced water effluent through the action of the electron, in order to remove or transform the toxic and harmful substances from the environment by redox reactions in less toxic substances. For this reason, we used a synthetic wastewater, containing a mixture H2SO4 0,5M and 16 HPAs, which are: naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo (a) anthracene, chrysene, benzo(b)fluoranthene, benzo(k) fluoranthene, benzo(a)pyrene, indeno(1,2,3-cd)pyrene, dibenzo(a, h)anthracene, benzo(g, h, i)perylene. Bulk electrochemical oxidation experiments were performed using a batch electrochemical reactor containing a pair of parallel electrodes, coupled with a power supply using a magnetic stirrer for favoring the transfer mass control. As anodic material was used, a Dimensionally Stable Anode (DSA) of Ti/Pt, while as cathode was used a Ti electrode. Several samples were collected at specific times and after that, the analysis of these samples were carried out by using Gas Chromatography Coupled to Mass Spectrometry (GC - MS) in order to determine the percentage of removal. The results showed that it was possible to achieve the removal of HPAs about 80% (in some cases, more than 80%). In addition, as an indicator of the economic feasibility of electrochemical treatment the energy consumption was analyzed for each hour of electrolysis, and based on the value kWh charged by ANEEL, the costs were estimated. Thus, the treatment costs of this research were quite attractive / O presente trabalho ? dirigido ao tratamento de ?gua produzida de petr?leo usando uma tecnologia eletroqu?mica. A ?gua produzida ? um dos principais res?duos gerados durante o processo de explora??o e produ??o na ind?stria petrol?fera, onde corresponde a 98 % de todos os efluentes gerados. V?rias metodologias est?o sendo estudadas visando o tratamento desse efluente, dentre elas est?o os processos biol?gicos, os processos de oxida??o avan?ados (POAs), tratamentos eletroqu?micos como eletrooxida??o, eletrocoagula??o, eletroflota??o e eletroredu??o. Esse trabalho mostra a aplica??o da tecnologia de eletrooxida??o no tratamento da ?gua produzida, pois se trata de uma t?cnica com alta compatibilidade ambiental, onde atrav?s da a??o do pr?prio el?tron, as subst?ncias t?xicas e nocivas ao meio ambiente podem ser removidas ou transformadas atrav?s de rea??es de ?xido-redu??o em subst?ncias menos t?xicas. Para isso, foi utilizado efluente sint?tico contendo uma mistura de H2SO4 0,5M e de 16 hidrocarbonetos polic?clicos arom?ticos (HPAs), que s?o: naftaleno, acenaftileno, acenafteno, fluoreno, fenantreno, antraceno, fluoranteno, pireno, benzo(a)antraceno, criseno, benzo(b)fluoranteno, benzo(k)fluoranteno, benzo(a)pireno, indeno(1,2,3-cd)pireno, dibenzo(a,h)antraceno, benzo(g,h,i)perileno. Utilizou-se o processo de oxida??o eletroqu?mica com um reator eletroqu?mico em batelada contendo um par de eletrodos paralelos, acoplados a uma fonte de corrente cont?nua e usando um agitador magn?tico para favorecer o transporte de massa na solu??o sint?tica. Como material an?dico utilizou-se um eletrodo de ?nodo dimensionalmente est?vel (DSA), constitu?do de Ti/Pt, enquanto o material cat?dico foi o Ti. Para detec??o e quantifica??o dos HPAs utilizou ? t?cnica de cromatografia gasosa acoplada a espectrometria de massa (CG - EM), onde o efluente foi analisado antes e ap?s o tratamento eletroqu?mico. Os resultados se mostraram eficazes, obtendo-se remo??es significativas dos HPAs superiores a 80%. Como indicador da viabilidade econ?mica do tratamento eletroqu?mico foram analisados os consumos energ?ticos do processo para cada hora do tratamento eletroqu?mico com base no valor kWh cobrado pela ANEEL. Os custos dos tratamentos desta pesquisa foram bastantes atrativos
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Développement et application de l’analyse dirigée par l’effet pour la recherche et l’identification de contaminants à risque présents dans les sols des sites pollués / Development and application of the effect-directed analysis for research and identification of risk contaminants in soils of polluted sitesDelafoulhouze, Maximilien 13 December 2016 (has links)
Le sol est une ressource rare remplissant de nombreuses fonctions nécessaires au bon fonctionnement des écosystèmes et de nos sociétés. Malgré tout, il subit des pressions anthropiques à l’origine de sa déterioration. Ainsi en Europe, 16% du territoire est sujet à des degradations des sols. Une des menaces affectant l’homeostasie des sols est la contamination chimique issue des sites à activités industrielles. Ces sites contaminés, estimés au nombre 2,5 millions en Europe sont à l’origine d’un risque pour la santé humaine et environnementale. Malheureusement, à l’heure actuelle, il n’existe pas de cadre réglementaire européen commun concernant la gestion de ces sites, dont les diagnostics sont réalisés uniquement sur des listes restreintes de molécules et les polluants prioritaires. Cependant, la contamination des sols est généralement issue de mélanges complexes dont la toxicité est due à un ensemble de contaminants. L’objectif de cette thèse concerne l’identification de composés à l’origine de la toxicité des sols pollués en appliquant une analyse dirigée par l’effet (EDA), une technique qui combine des analyses biologiques et chimiques à un fractionnement physico-chimique. Après le développement de l’EDA pour les matrices solides, cette approche a été utilisée sur site afin d’identifier des composés liant le recepteur des hydrocarbures aromatiques (AhR). Ceci à permis de mettre au jour des contaminants à activité biologique non pris en compte dans les diagnostics des sols pollués tels que des hydrocarbures aromatiques polycycliques non prioritaires selon l’US-EPA, leurs produits d’oxydation ainsi que des composés contenant des N-,S-,O-Heterocyles. / The soil is a scarce resource fulfilling many functions necessary for the proper functioning of ecosystems and societies. Nevertheless, it suffers human pressures causing its deterioration. In Europe, 16% of the territory is subject to soil degradations. One of the threats to the soil homeostasis is the chemical contamination from industrial activity sites. These contaminated sites, estimated to number of 2.5 million in Europe are causing a risk to human and environmental health. Unfortunately, at present there is no common European regulatory framework for the management of these sites, whose diagnoses are made only on short lists of molecules and priority pollutants. However, soil contamination generally comes from complex mixtures whose toxicity is due to a combination of contaminants. The aim of this work concerns the identification of compounds responsible for the toxicity of the contaminated soil by applying an effect-directed analysis (EDA), a technique which combines biological and chemical analyzes to a physico-chemical fractionation. After the development of EDA on the solid matrices, this approach has been used on polluted sites to identify the compounds binding to the aryl hydrocabon receptor (AhR). This allowed to highlight contaminants with a biological activity not included in the diagnoses of polluted soils such as polycyclic aromatic hydrocarbons non-priority according to US-EPA, their oxidation products and compounds containing N -, S-, O-heterocycles.
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