Há muitos estudos sobre plantas medicinais brasileiras, porém a carência de pesquisas relacionadas à segurança alimentar de plantas brasileiras usadas como alimento ainda é consideravelmente grande. A maioria dos alcaloides são substâncias tóxicas que podem ser encontrados em uma grande variedade de plantas medicinais e alimentícias, inclusive em espécies de Passiflora chamadas popularmente no Brasil de \"maracujá\". As pesquisas sobre as diversas espécies de Passiflora, em grande parte estão relacionadas com as folhas e frutos e na maioria das vezes as sementes são consideradas como resíduo. As metodologias analíticas modernas para análises na área de alimentos, utilizando SBSE com fase extratora de PDMS, tem o objetivo de facilitar o preparo de amostras complexas e diminuir os resíduos orgânicos gerados na etapa de preparo da amostra. Para isso, este estudo teve como objetivo a análise de alcaloides harmânicos pelo método SBSE/CLAE-Flu dual, adaptado para a quantificação de harmana e de harmina em sementes de maracujá azedo. O método foi especifico e linear para os alcaloides estudados (r2 = 0,996 para harmana e r2 = 0,999 para harmina). Os ensaios de repetibilidade e de precisão intermediária confirmam a precisão do método. Os testes de recuperação (entre 92,34% a 105,90% para a harmana e 83,61% a 117,76% para a harmina) indicam a exatidão do método. A amostra analisada continha (3,094x10-2 ± 5,874x10-5) µg de harmana por grama de sementes secas de maracujá azedo e (8,108x10-3±7,599x10-4) µg de harmina por grama de sementes secas de maracujá azedo. A detecção por fluorescência foi um método sensível e seletivo para a detecção da harmana (LOD = 30 ng L-1 e LOQ = 200 ng L-1) e para a harmina (LOD = 100 ng L-1 e LOQ = 400 ng L-1). Conclui-se que o método SBSE/CLAE-Flu dual pode ser usado para a análise dos alcaloides harmânicos nas sementes de maracujá azedo. / There are many studies of Brazilian medicinal plants, but the lack of research related to food safety of Brazilian plants used as food is still high. Most of the alkaloids are toxic chemicals found in a wide variety of medicinal plants and edible plants, including Passiflora species popularly known in Brazil as \"maracujá\". Researches about most of the Passiflora species are largely related about the leaves and the fruits and in most of the cases, the seeds are considered as waste. Modern analytical methodologies for food analysis, using SBSE with PDMS as the extraction phase, have the purpose of make easier the preparation of complex samples and to reduce the organic waste generated in the sample preparation step. For that, this study had the aim to analyze harman alkaloids by a dual SBSE/HPLC-Flu method, adapted to the quantification of harmane and harmine in sour passion fruit seeds. The method was specific and linear for the studied of the alkaloids (r2 = 0.996 for harmane and r2 = 0.999 for harmine). The repeatability and accuracy intermediate assays confirm the precision of the method. The recovery assay (between 92.34% and 105.90% to harmane and between 83.61% and 117.76% to harmine) indicate the accuracy of the method. The sample analyzed contained (3.094x10-2 ± 5.874x10-5) µg of the harmane in 1.0 g of dried seeds of sour passion fruit and (8.108x10-3 ± 7.599x10-4) µg of the harmine in 1.0 g of dried seeds of sour passion fruit. The fluorescence detection was a sensitive and selective method for detection of harmane (LOD = 30 ng L-1 and LOQ = 200 ng L-1) and harmine (LOD = 100 ng L-1 and LOQ = 400 ng -1). It was concluded that the dual SBSE/HPLC-Flu method can be used for the analysis of the harman alkaloids in dried seeds of sour passion fruit.

O maracujá, nome popular atribuído ao fruto das diversas espécies do gênero Passiflora, da família Passifloraceae, é amplamente comercializado e consumido no mundo, sendo o Brasil um dos maiores produtores do fruto. Alguns estudos apontam possível toxicidade relacionada às espécies de Passiflora, principalmente P. incarnata. No entanto, há pouco conhecimento acerca das espécies P. edulis e P. alata, sobretudo em relação à polpa e sementes. Os extratos da polpa e das sementes dos frutos dessas duas espécies de \"maracujá\", Passiflora alata e Passiflora edulis, foram estudados com o objetivo de identificar alcaloides harmânicos, pelo preparo das amostras por extração por sorção em barra magnética recoberta com polidimetilsiloxano (SBSE-PDMS) e SBSE recoberta com polietilenoglicol silicone (SBSE-EG Silicone) e análise por cromatografia líquida de alta eficiência com detector por fluorescência (CLAE-Flu) e cromatografia líquida de ultra eficiência acoplada à espectrometria de massas sequencial (CLUE-EM/EM). A análise dos alcaloides harmana e harmina nos extratos da polpa de P. alata foi feita por meio do método de adição de padrão e mostrou menor quantidade destes alcaloides, em comparação com os resultados da análise dos extratos da polpa dos frutos de P. edulis, no trabalho de Pereira e colaboradores. As análises CLUE-EM e CLUE-EM/EM possibilitaram a identificação dos alcaloides nos extratos: nas sementes de P. alata, os alcaloides harmana, harmina, harmol, harmalol e harmalina foram identificados, enquanto que na polpa, harmana e harmina tiveram a confirmação da sua presença. Nos extratos da polpa dos frutos de P. edulis observou-se os alcaloides harmana, harmina e harmalina. E nas sementes de P. edulis harmina foi encontrada, porém há indícios da presença de harmana. A literatura sobre as barras de SBSE-EG Silicone Twister&®; não relata nenhum estudo relacionado ao seu uso para extração e concentração de alcaloides harmânicos nos extratos de P. alata e P. edulis. Por isso foi proposto inicialmente a aplicação do planejamento fatorial fracionário para otimização do método de extração, utilizando os padrões comerciais dos alcaloides harmana e harmina. O planejamento experimental revelou as variáveis principais e seus níveis de importância, e a partir destes resultados foi realizado o estudo cinético dos tempos de extração e de dessorção das barras de SBSE-EG Silicone. Porém, os resultados mostraram que as barras de SBSE-EG Silicone não são adequadas para a extração dos alcaloides harmana e harmina, uma vez que a recuperação obtida foi baixa, na ordem de 30%. / \"Maracujá\" is the popular name given to the fruit of several species of Passiflora genus, from Passifloraceae family, it is widely commercialized and consumed around the world, and Brazil is one of the largest producers of this fruit. Some studies pointed out the possible toxicity related to Passiflora species, mainly P. incarnata. Although, there is a lack of knowledge about the P. edulis and P. alata species, especially with regards to the pulp and seeds. The extracts of pulp and seeds from the \"maracujá\" species Passiflora alata and Passiflora edulis, were studied in order to identify harman alkaloids. The samples were prepared by extraction with sorptive stir bar coated with polydimethylsiloxane (SBSE-PDMS) and SBSE coated with polyethylene glycol silicon (SBSE-EG Silicone). The samples were analyzed by high performance liquid chromatography with fluorescence detector (HPLC-Flu), and ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Harmane and harmine alkaloids in P. alata pulp extracts were analyzed using the standard addition method and the results showed a lower amount of these alkaloids, compared with the test results for the extracts from the P. edulis pulp in the work of Pereira et al. UHPLC-MS and UHPLC-MS/MS analysis enabled to identify the alkaloids amount present in the extracts. In the P. alata seeds extract the following alkaloids were identified harmane, harmine, harmol, harmalol and harmaline, while in the pulp extract, harmane and harmine were confirmed. In the extracts of P. edulis pulp the alkaloids identified were harmane, harmine and harmaline. And in the P. edulis seeds extract the harmine alkaloid was found, some indications of the presence of harmana were observed. The literature about SBSE-EG Silicone Twister® bars reports no study related to their use for extraction and concentration of harman alkaloids in P. alata and P. edulis. Thus, it was initially proposed the application of fractional factorial design to optimize the extraction method using commercial standards of harmane and harmine alkaloids. The experimental design revealed the main variables and their importance levels, and from these results kinetic studies were performed for the extraction and desorption times of SBSE-EG Silicone bars. However, the results showed SBSE-EG Silicone bars are not suitable for the extraction of harmane and harmine alkaloids, since the recovery obtained was low, on the order of 30%.

O maracujá azedo, Passiflora edulis, e o maracujá doce, Passiflora alata, são as duas espécies pertencentes à família Passifloraceae de maior importância econômica para o Brasil, sendo seus frutos amplamente comercializados como alimentos. Um grupo de substâncias conhecido como alcaloides β-carbolínicos são componentes minoritários desses frutos, no entanto, a literatura indica que substâncias pertencentes a esse grupo podem ter atividade tóxica. Este trabalho apresentou três objetivos específicos, com o intuito de ampliar o conhecimento sobre a fitoquímica dos frutos de P. edulis e de P. alata e da aplicação da SBSE como método de preparo de amostra: quantificação de harmana e de harmina na polpa e nas sementes de P. alata utilizando SBSE-PDMS e análise por HPLC/Flu; identificação de alcaloides β-carbolínicos nas cascas de P. edulis através de análises por HPLC/Flu e por UHPLC/MS; estudo do processo de extração de norharmana por SBSE com duas fases extratores diferentes (PDMS e EG-Silicone) por HPLC/Flu. A quantificação de harmana e harmina na polpa e nas sementes de P. alata foi realizada após preparo de amostra por SBSE-PDMS e análises por HPLC/Flu. A concentração de harmana encontrada foi 1,0328 . 10-1 ± 3,1217 . 10-3 μg L-1 na polpa, enquanto que nas sementes foi encontrada a concentração de 7,4391 . 10-5 ± 2,5501 . 10-6 μg g-1. As concentrações de harmina ficaram abaixo do LOD e do LOQ na polpa e nas sementes de P. alata, respectivamente. Na identificação dos alcaloides β-carbolínicos nas cascas de P. edulis, os extratos para análises por HPLC/Flu e por UHPLC/MS foram preparados pelo método clássico, sendo identificados os alcaloides norharmana, harmana, harmina, harmol e harmalol. Nas análises por HPLC/Flu de extrato das cascas de P. edulis preparado utilizando SBSE-PDMS, o alcaloide norharmana foi identificado como majoritário e por esse motivo, foi realizado o estudo do processo de extração de norharmana por SBSE com duas fases extratoras diferentes, PDMS e EG-Silicone, utilizando planejamento fatorial fracionário, tendo como objetivo obter a melhor recuperação percentual possível. Com as melhores condições para extração definidas, os ensaios utilizando PDMS e EG-Silicone apresentaram recuperação percentual de cerca de 50 % e 80 %, respectivamente. A quantificação de harmana na polpa e nas sementes do maracujá doce, P. alata, indicou quantidades muito pequenas desse alcaloide nessas partes dos frutos, levando à conclusão que um fruto de P. alata, apresenta cerca de 140 vezes menos alcaloides β-carbolínicos do que o fruto de P. edulis, levando em consideração as quantidades de harmana e harmina no maracujá azedo relatadas na literatura. A identificação dos alcaloides β-carbolínicos nas cascas de P. edulis é um importante passo para a compreensão sobre a fitoquímica dessa parte do fruto, que, nos últimos anos, deixou de ser apenas resíduo industrial e passou a ser produto alimentício com valor agregado, comercializadas como farinhas das cascas de maracujá. O estudo sobre a utilização da SBSE como técnica de extração de norharmana mostrou-se promissor, atingindo recuperação percentual aceitável para o futuro desenvolvimento de um método de quantificação de norharmana nas cascas de maracujá. / Sour passion fruit, Passiflora edulis, and sweet passion fruit, Passiflora alata, are the two species of Passifloraceae of greater economic importance to Brazil, and their fruits are widely marketed as food. A class of compounds known as β-carboline alkaloids is a minor component of these fruits, however, the literature indicates that substances belonging to this group may have toxic activity. This work presents three specific objectives, in order to increase knowledge about the phytochemistry of P. edulis and P. alata and the application of SBSE as a sample preparation method: quantification of harmane and harmine in the pulp and in the seeds of P. alata using SBSE-PDMS and HPLC/Flu analysis; identification of β-carboline alkaloids in P. edulis peels using HPLC/Flu and UHPLC/MS analysis; study of the SBSE extraction process of norharmane using two different phases (PDMS and EG-Silicone), by HPLC/Flu analysis. Quantification of harmane and harmine in the pulp and the seeds of P. alata was performed after sample preparation by SBSE-PDMS and analysis by HPLC/Flu. The concentration of harmane was 1,0328 . 10-1 ± 3,1217 . 10-3 μg L-1 in the pulp while the in the seeds it was found 7.4391 . 10-5 ± 2.5501 . 10-6 μg g-1 harmane. The concentrations of harmine were below the LOD and LOQ in the pulp and in the seeds of P. alata, respectively. For the identification of the β-carboline alkaloids in the peels of P. edulis, the extracts were prepared by using the classic method and analyzed by HPLC/Flu and UHPLC/MS, and the alkaloids norharmane, harmane, harmine, harmol and harmalol were identified. In the HPLC/Flu analysis of the extract of P. edulis peels prepared using SBSE-PDMS, the alkaloid norharmane was identified as a major constituent and, for this reason, it was performed a study about the extraction process of norharmane by SBSE, evaluating two different phases, PDMS and EG-Silicone, by using fractional factorial design, aiming to get the best percentage recovery. Defined the best conditions for extraction, the experiments performed by using PDMS and EG-Silicone bars showed percentage recovery about 50% and 80%, respectively. Quantification of harmane in the pulp and seeds of sweet passionfruit, P. alata, indicated very small amounts of this alkaloid in these parts of the fruits, leading to the conclusion that the fruits of P. alata, has around 140 times less β-carboline alkaloid than the fruits of P. edulis, taking into account the amounts of harmana and harmine on sour passionfruit reported in the literature. Identification of β-carboline alkaloids in P. edulis peels is an important step for understanding the phytochemistry of this part of the fruits, which, in recent years, is no longer just an industrial waste and became a food product with added value, marketed as passion fruit peels flour. The study about the utilization of SBSE as extraction technique of norharmane showed to be promising, reaching acceptable recovery percentage, for the future development of a method for quantification of norharmane in passion fruit peels.

Há muitos estudos sobre plantas medicinais brasileiras, porém a carência de pesquisas relacionadas à segurança alimentar de plantas brasileiras usadas como alimento ainda é consideravelmente grande. A maioria dos alcaloides são substâncias tóxicas que podem ser encontrados em uma grande variedade de plantas medicinais e alimentícias, inclusive em espécies de Passiflora chamadas popularmente no Brasil de \"maracujá\". As pesquisas sobre as diversas espécies de Passiflora, em grande parte estão relacionadas com as folhas e frutos e na maioria das vezes as sementes são consideradas como resíduo. As metodologias analíticas modernas para análises na área de alimentos, utilizando SBSE com fase extratora de PDMS, tem o objetivo de facilitar o preparo de amostras complexas e diminuir os resíduos orgânicos gerados na etapa de preparo da amostra. Para isso, este estudo teve como objetivo a análise de alcaloides harmânicos pelo método SBSE/CLAE-Flu dual, adaptado para a quantificação de harmana e de harmina em sementes de maracujá azedo. O método foi especifico e linear para os alcaloides estudados (r2 = 0,996 para harmana e r2 = 0,999 para harmina). Os ensaios de repetibilidade e de precisão intermediária confirmam a precisão do método. Os testes de recuperação (entre 92,34% a 105,90% para a harmana e 83,61% a 117,76% para a harmina) indicam a exatidão do método. A amostra analisada continha (3,094x10-2 ± 5,874x10-5) µg de harmana por grama de sementes secas de maracujá azedo e (8,108x10-3±7,599x10-4) µg de harmina por grama de sementes secas de maracujá azedo. A detecção por fluorescência foi um método sensível e seletivo para a detecção da harmana (LOD = 30 ng L-1 e LOQ = 200 ng L-1) e para a harmina (LOD = 100 ng L-1 e LOQ = 400 ng L-1). Conclui-se que o método SBSE/CLAE-Flu dual pode ser usado para a análise dos alcaloides harmânicos nas sementes de maracujá azedo. / There are many studies of Brazilian medicinal plants, but the lack of research related to food safety of Brazilian plants used as food is still high. Most of the alkaloids are toxic chemicals found in a wide variety of medicinal plants and edible plants, including Passiflora species popularly known in Brazil as \"maracujá\". Researches about most of the Passiflora species are largely related about the leaves and the fruits and in most of the cases, the seeds are considered as waste. Modern analytical methodologies for food analysis, using SBSE with PDMS as the extraction phase, have the purpose of make easier the preparation of complex samples and to reduce the organic waste generated in the sample preparation step. For that, this study had the aim to analyze harman alkaloids by a dual SBSE/HPLC-Flu method, adapted to the quantification of harmane and harmine in sour passion fruit seeds. The method was specific and linear for the studied of the alkaloids (r2 = 0.996 for harmane and r2 = 0.999 for harmine). The repeatability and accuracy intermediate assays confirm the precision of the method. The recovery assay (between 92.34% and 105.90% to harmane and between 83.61% and 117.76% to harmine) indicate the accuracy of the method. The sample analyzed contained (3.094x10-2 ± 5.874x10-5) µg of the harmane in 1.0 g of dried seeds of sour passion fruit and (8.108x10-3 ± 7.599x10-4) µg of the harmine in 1.0 g of dried seeds of sour passion fruit. The fluorescence detection was a sensitive and selective method for detection of harmane (LOD = 30 ng L-1 and LOQ = 200 ng L-1) and harmine (LOD = 100 ng L-1 and LOQ = 400 ng -1). It was concluded that the dual SBSE/HPLC-Flu method can be used for the analysis of the harman alkaloids in dried seeds of sour passion fruit.

O maracujá azedo, Passiflora edulis, e o maracujá doce, Passiflora alata, são as duas espécies pertencentes à família Passifloraceae de maior importância econômica para o Brasil, sendo seus frutos amplamente comercializados como alimentos. Um grupo de substâncias conhecido como alcaloides β-carbolínicos são componentes minoritários desses frutos, no entanto, a literatura indica que substâncias pertencentes a esse grupo podem ter atividade tóxica. Este trabalho apresentou três objetivos específicos, com o intuito de ampliar o conhecimento sobre a fitoquímica dos frutos de P. edulis e de P. alata e da aplicação da SBSE como método de preparo de amostra: quantificação de harmana e de harmina na polpa e nas sementes de P. alata utilizando SBSE-PDMS e análise por HPLC/Flu; identificação de alcaloides β-carbolínicos nas cascas de P. edulis através de análises por HPLC/Flu e por UHPLC/MS; estudo do processo de extração de norharmana por SBSE com duas fases extratores diferentes (PDMS e EG-Silicone) por HPLC/Flu. A quantificação de harmana e harmina na polpa e nas sementes de P. alata foi realizada após preparo de amostra por SBSE-PDMS e análises por HPLC/Flu. A concentração de harmana encontrada foi 1,0328 . 10-1 ± 3,1217 . 10-3 μg L-1 na polpa, enquanto que nas sementes foi encontrada a concentração de 7,4391 . 10-5 ± 2,5501 . 10-6 μg g-1. As concentrações de harmina ficaram abaixo do LOD e do LOQ na polpa e nas sementes de P. alata, respectivamente. Na identificação dos alcaloides β-carbolínicos nas cascas de P. edulis, os extratos para análises por HPLC/Flu e por UHPLC/MS foram preparados pelo método clássico, sendo identificados os alcaloides norharmana, harmana, harmina, harmol e harmalol. Nas análises por HPLC/Flu de extrato das cascas de P. edulis preparado utilizando SBSE-PDMS, o alcaloide norharmana foi identificado como majoritário e por esse motivo, foi realizado o estudo do processo de extração de norharmana por SBSE com duas fases extratoras diferentes, PDMS e EG-Silicone, utilizando planejamento fatorial fracionário, tendo como objetivo obter a melhor recuperação percentual possível. Com as melhores condições para extração definidas, os ensaios utilizando PDMS e EG-Silicone apresentaram recuperação percentual de cerca de 50 % e 80 %, respectivamente. A quantificação de harmana na polpa e nas sementes do maracujá doce, P. alata, indicou quantidades muito pequenas desse alcaloide nessas partes dos frutos, levando à conclusão que um fruto de P. alata, apresenta cerca de 140 vezes menos alcaloides β-carbolínicos do que o fruto de P. edulis, levando em consideração as quantidades de harmana e harmina no maracujá azedo relatadas na literatura. A identificação dos alcaloides β-carbolínicos nas cascas de P. edulis é um importante passo para a compreensão sobre a fitoquímica dessa parte do fruto, que, nos últimos anos, deixou de ser apenas resíduo industrial e passou a ser produto alimentício com valor agregado, comercializadas como farinhas das cascas de maracujá. O estudo sobre a utilização da SBSE como técnica de extração de norharmana mostrou-se promissor, atingindo recuperação percentual aceitável para o futuro desenvolvimento de um método de quantificação de norharmana nas cascas de maracujá. / Sour passion fruit, Passiflora edulis, and sweet passion fruit, Passiflora alata, are the two species of Passifloraceae of greater economic importance to Brazil, and their fruits are widely marketed as food. A class of compounds known as β-carboline alkaloids is a minor component of these fruits, however, the literature indicates that substances belonging to this group may have toxic activity. This work presents three specific objectives, in order to increase knowledge about the phytochemistry of P. edulis and P. alata and the application of SBSE as a sample preparation method: quantification of harmane and harmine in the pulp and in the seeds of P. alata using SBSE-PDMS and HPLC/Flu analysis; identification of β-carboline alkaloids in P. edulis peels using HPLC/Flu and UHPLC/MS analysis; study of the SBSE extraction process of norharmane using two different phases (PDMS and EG-Silicone), by HPLC/Flu analysis. Quantification of harmane and harmine in the pulp and the seeds of P. alata was performed after sample preparation by SBSE-PDMS and analysis by HPLC/Flu. The concentration of harmane was 1,0328 . 10-1 ± 3,1217 . 10-3 μg L-1 in the pulp while the in the seeds it was found 7.4391 . 10-5 ± 2.5501 . 10-6 μg g-1 harmane. The concentrations of harmine were below the LOD and LOQ in the pulp and in the seeds of P. alata, respectively. For the identification of the β-carboline alkaloids in the peels of P. edulis, the extracts were prepared by using the classic method and analyzed by HPLC/Flu and UHPLC/MS, and the alkaloids norharmane, harmane, harmine, harmol and harmalol were identified. In the HPLC/Flu analysis of the extract of P. edulis peels prepared using SBSE-PDMS, the alkaloid norharmane was identified as a major constituent and, for this reason, it was performed a study about the extraction process of norharmane by SBSE, evaluating two different phases, PDMS and EG-Silicone, by using fractional factorial design, aiming to get the best percentage recovery. Defined the best conditions for extraction, the experiments performed by using PDMS and EG-Silicone bars showed percentage recovery about 50% and 80%, respectively. Quantification of harmane in the pulp and seeds of sweet passionfruit, P. alata, indicated very small amounts of this alkaloid in these parts of the fruits, leading to the conclusion that the fruits of P. alata, has around 140 times less β-carboline alkaloid than the fruits of P. edulis, taking into account the amounts of harmana and harmine on sour passionfruit reported in the literature. Identification of β-carboline alkaloids in P. edulis peels is an important step for understanding the phytochemistry of this part of the fruits, which, in recent years, is no longer just an industrial waste and became a food product with added value, marketed as passion fruit peels flour. The study about the utilization of SBSE as extraction technique of norharmane showed to be promising, reaching acceptable recovery percentage, for the future development of a method for quantification of norharmane in passion fruit peels.

Malgré une sensible baisse des ventes depuis quelques années, la France reste le premier utilisateur européen de phytosanitaires. Ceci n’est pas sans conséquences sur l’environnement et les organismes non visés par ces produits. De nombreuses études ont permis d’une part de mieux comprendre les phénomènes de transfert des pesticides dans les compartiments eau-air sols et d’autre part de mettre en évidence leur contamination. Les données concernant l’eau sont plus nombreuses du fait de son statut de réceptacle ultime des pollutions.et des normes établies par les textes réglementaires. La Directive Cadre européenne sur l’Eau (DCE) adoptée en 2000 vise à atteindre le bon état chimique et écologique des eaux d’ici 2015 avec une obligation de résultats. Le bon état chimique dépend du respect des normes de qualité environnementales (NQE) établies notamment pour 24 pesticides. En 2012, 5% des points de mesure en cours d’eau et 19% des points de mesure en eaux souterraines ne respectent pas ces NQE.Le Languedoc Roussillon est depuis de nombreuses années la première région viticole de France et souffre d’une pression phytosanitaire importante. En 2014, un rapport de l’Agence Régionale de Santé (ARS) fait même état de l’utilisation encore récente de pesticides interdits (simazine, terbuthylazine…) depuis le début des années 2000. L’ARS a de plus affirmé que la plupart des cas de non-conformité avérés pouvaient être rattachés à la présence d’activité agricole dans le bassin versant du captage.Remédier à ces contaminations passe par le suivi de la qualité des eaux et la caractérisation de la pollution. Les outils actuels s’avèrent ne pas être toujours représentatifs de cette pollution ;de nouveaux outils sont donc mis au point. L’étude présentée ici porte sur l’utilisation de l’échantillonnage passif de type POCIS (Polar Organic Chemical Integrative Sampler) pour le suivi de la contamination en pesticides de trois bassins versants languedociens : Paulhan,Puissalicon, Roujan.Dans un premier temps, l’étude des limites possibles du dispositif POCIS a été faite pour évaluer son applicabilité dans les conditions d’exposition imposées par le terrain. Le comportement des molécules au sein de l’outil a été étudié pour une meilleure compréhension des phénomènes de diffusion qui s’y produisent. Ces études ont mené à la mise en place de calibrations en laboratoire en conditions contrôlées. [...] La mise en place des dispositifs POCIS sur les trois sites d’études a mis en évidence la prédominance de pesticides utilisés en viticulture et a montré tout l’intérêt de l’outil d’un point de vue qualitatif. La comparaison de l’outil avec l’échantillonnage automatisé montre des disparités entre concentration réelle et concentration estimée via le POCIS. Ce dernier reste à améliorer d’un point de vue quantitatif pour obtenir un échantillonneur fiable et robuste pour ce type de milieu d’exposition. / Despite a noticeable sales decrease since few years, France remains the first european user of phytosanitary products. This is not without consequences on the environment and the untargeted organisms. Several studies allowed on one hand to better understand phenomena of pesticides transfer in water-air and soils compartments and on the other hand to highlight their contamination. Data concerning the water are more numerous cause of its status of ultimate receptacle of pollution and standards established by regulatory texts. The european Water Framework Directive (WFD) adopted in 2000 aims to achieve good chemical and ecological status of water bodies by 2015 with an obligation of results. The good chemical status depends on respect for the Environmental Quality Standards (EQS) established, for instance, for 24 pesticides. In 2012, 5% of river measurement points and 19%of groundwater measurement points do not respect these EQS. For many years, Languedoc Roussillon is the first wine-producing region of France and suffers from a significant pesticide pressure. In 2014, a report from the Regional Health Agency (RHA) has mentioned a relatively recent use of banned pesticides since the early 2000’s like simazin and terbutylazin. Moreover, RHA stated that most of recognized cases of non-compliance could be linked to the presence of agricultural activity in the catchment basin.[...] The establishment of POCIS devices on the three study sites showed the predominance ofpesticides used in viticulture and showed all the interest of the tool on a qualitative point ofview. The comparison of the tool with the automated sampling shows disparities between real concentration and concentration estimated with the POCIS. However, it remains to improvethe tool on a quantitative way to obtain a reliable and robust sampler for this type of exposure medium.

Identifying the best control method for problematic wildlife is an ever present issue in wildlife management. Popular control methods have ranged from lethal techniques, extirpating the animal, to multiple non-lethal methods focused on deterring undesired behavior. In the past, lethal methods were the preferred choice. However, with increased awareness of the need for biodiversity conservation, new management methods focus on non-lethal control, with emphasis on exploiting aspects of naturally occurring organismal behaviors and ecology. Over the past decade, technological advances in extraction method’s and equipment have also developed new techniques providing a broader range of information about species biology for management use. One of the most well documented conflicts between wildlife and humans is that of the wolf. Using advanced technology and new techniques, we investigated the implication of using chemosensory signals in canid urine to modify behavior as a possible non-lethal alternative in large predator management. Here we used the SBSE method coupled with improved GC/MS equipment to analyze the volatile organic compounds in the urine of four canid species, gray wolf (Canis lupus), red wolf (Canis rufus), wolf-dog hybrids (Canis familiaris) and the domestic dog (Canis familiaris) in order to create working urinary profiles. The extraction method identified several compounds also seen in the urinary profiles of other large predators. In addition, similarities and differences were also noted between taxa and the sexes, and these can be further explored in future studies. Two identified urinary compounds, acetophenone and methyl propyl sulfide, were selected for further behavioral evaluation. We focused on these compounds and their influence as chemosensory signals triggering urine marking events in both the gray wolf and red wolf. Behavioral observations of the effects of these two chemicals indicated they elicited responses from captive wolves. At each of the three study sites, the combination of these chemicals produced urine-marking events along the territory boundary by dominant animals. As a result, the investigation focused on what triggered the urine-marking events, the chemicals themselves, their combination, or the breakdown of the chemicals producing other odorants. It was found that there was no significant degradation of the chemicals over time and environmental conditions produced no significant breakdown of the acetophenone prior to the addition of methyl propyl sulfide. This posed a number of new questions and illustrated the need for additional behavioral studies. The results of this study analyzing chemosensory signals in canid urine, provides biologists with new information to aid in the development of new non-lethal management strategies for handling problematic wildlife as well as providing useful information for future research involving reproduction, predator/prey dynamics, territory maintenance, and a host of other studies focusing on animal ecology in association with chemosensory signaling.

Made available in DSpace on 2015-05-14T12:36:52Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2070301 bytes, checksum: e15a4fc1a4f27fd39d2a55b1400d522b (MD5) Previous issue date: 2014-08-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The great difficulty of selecting software components is still an obstacle to achieving the success of Component-Based Development (CBD). With the growing market for components, the tendency is always to increase the number of options for assembling applications in different contexts, making it impractical to manual selection. Dealing with a problem of gigantic and complex search space, it is required automation performed by optimization techniques. The proposed approach aims to automate the process of selecting components using techniques of Search-Based Software Engineering (SBSE), whose optimization technique is driven by structural metrics (i.e., connections between components of a software architecture) and information reuse (i.e., aggregated values to the component itself). The metrics used in this component selection context are intended to assess the structural perspective of an architectural instance, since they predict possible integration problems between implementations of components produced by third parties. Note that other proposals ignore this perspective and focus only on the component itself. In addition, reuse information can bring an alternative to represent the perception of the developers about the quality attributes of the software components in a reuse scenario as: the degree of consumers' satisfaction who have already purchased this component and the number of downloads of it. The proposed evaluation is carried out through experiments, which are validated by applying statistical tests. / A grande dificuldade de selecionar componentes de software ainda é um obstáculo para alcançar o sucesso do Desenvolvimento Baseado em Componentes (DBC). Com o crescimento do mercado de componentes, a tendência é sempre aumentar o número de opções para montagem de aplicações em diferentes contextos, tornando impraticável a seleção manual. Tratando de um problema com gigantesco espaço de busca e complexo, é requerida a automatização efetuada por técnicas de otimização. O trabalho proposto visa automatizar o processo de seleção de componentes utilizando técnicas da Engenharia de Software Baseada em Busca (ESBB), cuja técnica de otimização é orientada por métricas estruturais (avaliam as conexões entre componentes de uma arquitetura de software) e informações de reúso (i.e., valores correspondentes ao reúso de software agregados ao próprio componente). As métricas utilizadas neste contexto de seleção de componentes têm o propósito de avaliar a perspectiva estrutural de uma instância arquitetural, já que elas preveem possíveis problemas de integração entre implementações de componentes produzidas por terceiros, além de que muitos problemas ignoram essa perspectiva e só focam no componente em si. Já as informações de reúso trazem consigo uma alternativa para representar a percepção dos desenvolvedores sobre os atributos de qualidade dos componentes de software em cenários de reúso, tais como: o grau de satisfação dos consumidores que já adquiriram tal componente e o número de downloads do mesmo. A avaliação da proposta é conduzida através de experimentos, que são validados aplicando testes estatísticos.

L’application de la Directive Cadre sur l’Eau et l’obligation de surveillance de la qualité de l'eau pour la consommation humaine et les activités industrielles entraînent une forte demande pour des systèmes d’évaluation et de suivi de la qualité de l’eau. Le projet de recherche MIGR'HYCAR (http://www.migrhycar.com) a donc été mis en place pour répondre à un besoin opérationnel et à un défaut d'outils d'aide à la décision adaptés face aux déversements d'hydrocarbures en eaux continentales (rivières, lacs et estuaires) qui représentent plus de 50% des déversements accidentels en France. Ce travail de thèse s'inscrit dans le cadre de ce projet et a pour but l’étude du comportement des nappes d'hydrocarbures en milieu contrôlé pour les eaux continentales. A cet effet, il propose une approche expérimentale à l’échelle laboratoire pour étudier les phénomènes agissant sur une nappe d'hydrocarbures déversée : dissolution, étalement, évaporation et volatilisation. Les données expérimentales réunis ont permis de constituer une base de données de référence, qui servira de support aux outils de modélisation mathématique de dérive de nappe d'hydrocarbures développés dans le cadre de ce projet. / The application of the European Water Framework Directive on water quality for human consumption and industrial activities creates a need for water quality assessment and monitoring systems. The MIGR'HYCAR research project (http://www.migrhycar.com) was initiated to provide decisional tools for risks connected to oil spills in continental waters (rivers, lakes and estuaries), which represent more than 50% of accidental spills in France. This work joins within the framework of this project and aims at the study of the behavior of hydrocarbons in a controlled environment for continental waters. For that purpose, the work proposes an experimental approach at laboratory scale to study the phenomena acting in oil spill: dissolution, spreading, evaporation and volatilization. The experimental data allowed establishing a reference database, which will serve as support in tools of oil spill modeling within the framework of this project.