O araçá é um fruto, pertencente à espécie Psidium grandifolium Mart. ex DC, nativo da mata atlântica brasileira pouco caracterizado pela literatura ciêntífica. Esta pesquisa visou estudar o rendimento de extração e avaliar as atividades antioxidante e antimicrobiana do óleo essencial de araçá obtido por extração com dióxido de carbono supercrítico (SFE) e dos diferentes extratos obtidos por extração com etanol pressurizado (PLE) seguindo um delineamento composto central rotacional com temperatura e tempo estático como variáveis independentes estudadas. Avaliou-se também o perfil de voláteis do óleo essencial, o teor de compostos fenólicos totais dos extratos etanólicos e as alterações causadas na matriz vegetal por diferentes processos de extração. Com a SFE conseguiu-se extrair toda a fração lipídica do fruto (2,33% ± 0,04.10-2%), a análise dos compostos voláteis do óleo essencial obtido permitiu inferir que este possui diversos compostos com potencial bioativo tais como E-cariofileno (17.56%), α-humuleno (16,26%) e β-sesquifelandreno (14.27%); porém este óleo essencial não apresentou atividade antioxidante relevante. O extrato etanólico com maior rendimento (17, 49%) foi obtido a 74 °C por 6 min, mesma condição em que obteve-se a maior atividade antioxidante (EC50 = 6,37 mg/mL), porém o maior teor de compostos fenólicos totais (136,95 mg EGA/100 g de massa seca) foi obtido quando aplicou-se 60°C por 9 min. Em relação à atividade antimicrobiana, tanto os extratos obtidos por PLE quanto por SFE apresentaram atividade contra cepas de P. aeruginosa, S. aureus e B. cereus, somente extratos etanólicos apresentaram atividade contra cepas de L. monocytogenes e nenhum dos extratos apresentaram atividade contra cepas de B. subtilis, E. coli e Salmonella. Em ambos os extratos foi possível identificar compostos com potencial aplicação farmacológica. O estudo de processos de extração que envolvam inovação tecnológica na obtenção extratos com atividade biológica visa contribuir com o extrativismo consciente ou estimular o plantio em maior escala de mais um fruto tropical empregado na medicina popular brasileira. / The araçá is a fruit, belonging to the species Psidium grandifolium Mart. ex DC, native of the Brazilian Atlantic Forest, is not largely characterized by the scientific literature. This research work aimed to study the extraction yield and to evaluate the antioxidant and antimicrobial activities of the araçá essential oil obtained by supercritical carbon dioxide extraction (SFE) and the different extracts obtained by pressurized ethanol extraction (PLE) following a central composite design with temperature and static time as independent variables studied. It was also evaluated the volatile profile of the essential oil, the content of total phenolic compounds of the ethanolic extracts and the changes caused in the vegetal matrix by different extraction processes. SFE was able to extract the whole lipid fraction of the fruit (2.33% ± 0.04.10-2%), the analysis of the volatile compounds of the essential oil obtained allowed to infer that it has several compounds with bioactive potential such as E-caryophyllene (17.56%), α-humulene (16.26%) and β-sesquipelandene (14.27%); but this essential oil had no relevant antioxidant activity. The highest ethanolic extract (17, 49%) was obtained at 74 °C for 6 min, the same condition in which the highest antioxidant activity was obtained (EC50 = 6.37 mg/mL), but the higher content of total phenolics contente (136.95 mg EGA / 100 g dry mass) was obtained when 60 °C was applied for 9 min. In relation to the antimicrobial activity, both extracts obtained by PLE and SFE showed activity against P. aeruginosa, S. aureus and B. cereus strains, only ethanolic extracts showed activity against L. monocytogenes strains and none of the extracts presented activity against B. subtilis, E. coli and Salmonella strains. In both extracts it was possible to identify compounds with potential pharmacological application. The study of extraction processes that involve technological innovation in obtaining extracts with biological activity aims to contribute to the conscious extraction or stimulate the larger scale cultivation of another tropical fruit used in Brazilian popular medicine.

O araçá é um fruto, pertencente à espécie Psidium grandifolium Mart. ex DC, nativo da mata atlântica brasileira pouco caracterizado pela literatura ciêntífica. Esta pesquisa visou estudar o rendimento de extração e avaliar as atividades antioxidante e antimicrobiana do óleo essencial de araçá obtido por extração com dióxido de carbono supercrítico (SFE) e dos diferentes extratos obtidos por extração com etanol pressurizado (PLE) seguindo um delineamento composto central rotacional com temperatura e tempo estático como variáveis independentes estudadas. Avaliou-se também o perfil de voláteis do óleo essencial, o teor de compostos fenólicos totais dos extratos etanólicos e as alterações causadas na matriz vegetal por diferentes processos de extração. Com a SFE conseguiu-se extrair toda a fração lipídica do fruto (2,33% ± 0,04.10-2%), a análise dos compostos voláteis do óleo essencial obtido permitiu inferir que este possui diversos compostos com potencial bioativo tais como E-cariofileno (17.56%), α-humuleno (16,26%) e β-sesquifelandreno (14.27%); porém este óleo essencial não apresentou atividade antioxidante relevante. O extrato etanólico com maior rendimento (17, 49%) foi obtido a 74 °C por 6 min, mesma condição em que obteve-se a maior atividade antioxidante (EC50 = 6,37 mg/mL), porém o maior teor de compostos fenólicos totais (136,95 mg EGA/100 g de massa seca) foi obtido quando aplicou-se 60°C por 9 min. Em relação à atividade antimicrobiana, tanto os extratos obtidos por PLE quanto por SFE apresentaram atividade contra cepas de P. aeruginosa, S. aureus e B. cereus, somente extratos etanólicos apresentaram atividade contra cepas de L. monocytogenes e nenhum dos extratos apresentaram atividade contra cepas de B. subtilis, E. coli e Salmonella. Em ambos os extratos foi possível identificar compostos com potencial aplicação farmacológica. O estudo de processos de extração que envolvam inovação tecnológica na obtenção extratos com atividade biológica visa contribuir com o extrativismo consciente ou estimular o plantio em maior escala de mais um fruto tropical empregado na medicina popular brasileira. / The araçá is a fruit, belonging to the species Psidium grandifolium Mart. ex DC, native of the Brazilian Atlantic Forest, is not largely characterized by the scientific literature. This research work aimed to study the extraction yield and to evaluate the antioxidant and antimicrobial activities of the araçá essential oil obtained by supercritical carbon dioxide extraction (SFE) and the different extracts obtained by pressurized ethanol extraction (PLE) following a central composite design with temperature and static time as independent variables studied. It was also evaluated the volatile profile of the essential oil, the content of total phenolic compounds of the ethanolic extracts and the changes caused in the vegetal matrix by different extraction processes. SFE was able to extract the whole lipid fraction of the fruit (2.33% ± 0.04.10-2%), the analysis of the volatile compounds of the essential oil obtained allowed to infer that it has several compounds with bioactive potential such as E-caryophyllene (17.56%), α-humulene (16.26%) and β-sesquipelandene (14.27%); but this essential oil had no relevant antioxidant activity. The highest ethanolic extract (17, 49%) was obtained at 74 °C for 6 min, the same condition in which the highest antioxidant activity was obtained (EC50 = 6.37 mg/mL), but the higher content of total phenolics contente (136.95 mg EGA / 100 g dry mass) was obtained when 60 °C was applied for 9 min. In relation to the antimicrobial activity, both extracts obtained by PLE and SFE showed activity against P. aeruginosa, S. aureus and B. cereus strains, only ethanolic extracts showed activity against L. monocytogenes strains and none of the extracts presented activity against B. subtilis, E. coli and Salmonella strains. In both extracts it was possible to identify compounds with potential pharmacological application. The study of extraction processes that involve technological innovation in obtaining extracts with biological activity aims to contribute to the conscious extraction or stimulate the larger scale cultivation of another tropical fruit used in Brazilian popular medicine.

Les furanocoumarines (FCs) sont des métabolites secondaires principalement synthétisés chez quatre familles botaniques et dérivent de la voie de biosynthèse des phénylpropanoïdes. Ces phytoalexines interviennent dans les processus de défense de la plante et présentent un fort potentiel thérapeutique. Des travaux réalisés dans les années 1960 sur des cultures cellulaires en parallèle de l’utilisation de précurseurs radiomarqués ont permis de démontrer que de nombreuses enzymes impliquées dans cette voie appartenaient à la famille des cytochromes P450 (P450s). Seules deux d’entre elles avaient pu être identifiées d’un point de vue moléculaire au début de ce travail de thèse. Afin de générer des informations concernant le génome de plantes productrices de FCs, nous avons fait séquencer les ARNm extraits de feuilles de Pastinaca sativa, de Ruta graveolens et de Cullen cinereum. L’analyse in silico de ces trois banques de données a permis d’identifier près de 800 fragments d’ADNc codants pour des P450s. Des travaux antérieurs réalisés au laboratoire et l’analyse comparative des transcriptomes de ces 3 plantes nous ont amenés à nous focaliser sur la sous-famille CYP71AZ au travers d’une étude fine de CYP71AZ3 et CYP71AZ4. La caractérisation fonctionnelle de ces enzymes a été réalisée dans un système d’expression hétérologue eucaryote : Saccharomyces cerevisiae. Les résultats obtenus ont permis de montrer que CYP71AZ4 avait une spécificité de substrat assez large puisqu’elle pouvait métaboliser au moins une FC et 4 coumarines. L’analyse et la comparaison des constantes cinétiques pour chacun de ces substrats indiquent néanmoins que le psoralène est le substrat préférentiel. La caractérisation fonctionnelle de CYP71AZ3 a mis en évidence que cette enzyme pouvait hydroxyler l’esculétine, une coumarine, mais ne jouait aucun rôle dans la synthèse de FCs. Ces travaux mettent en évidence la diversité fonctionnelle au sein d’une même sous-famille enzymatique et permettent d’émettre des hypothèses nouvelles quant à l’apparition de cette voie de biosynthèse chez les Apiacées d’une part, et chez les autres familles botaniques d’autre part / Furanocoumarins (FCs) are secondary metabolites mainly synthetized in four botanical families deriving from the phenylpropanoid biosynthetic pathway. These phytoalexins are involved in plant defense mechanisms and present strong therapeutic potential. Early studies in the 1960s based on cell cultures and the use of radiolabeled precursors have shown that many enzymes involved in this pathway belong to the cytochrome P450 family (P450s). Only two of them had been identified from a molecular point of view at the beginning of this thesis. In order to generate information regarding the genome of plants producing FCs, we sequenced the mRNA extracted from leaves of Pastinaca sativa, Ruta graveolens, and Cullen cinereum. In silico analysis of these three libraries identified nearly 800 cDNA fragments encoding for P450s. Previous studies in the laboratory and comparative transcriptome analysis of these three plants have led us to focus on the subfamily CYP71AZ through a detailed study of CYP71AZ3 and CYP71AZ4. Functional characterization of these enzymes was performed in an eukaryote heterologous expression system: Saccharomyces cerevisiae. The results showed that CYP71AZ4 had a broad substrate specificity enough as it could metabolize one FC and 4 coumarins. The analysis and comparison of the kinetic constants for each of these substrates indicate, however, that the preferred substrate is psoralen. The functional characterization of CYP71AZ3 showed that this enzyme could hydroxylate esculetin, a coumarin, but played no role in the synthesis of FCs. This study highlights the functional diversity within a single enzyme subfamily and allows to issue new hypotheses about the emergence of this biosynthetic pathway in Apiaceae on one hand, and among other botanical families on the other hand

The effect of cattle grazing on the abundance and distribution of vegetation, burrowing crabs (Uca rapax, Uca pugnax, and Sesarma cinereum), marsh periwinkles (Littoraria irrorata), horn snails (Cerithidea pliculosa), and salt marsh snails (Melampus bidentatus) was evaluated over four seasons (summer 2000, fall 2000, winter 2001, and spring 2001) in grazed and ungrazed treatments. A Galveston Island salt marsh adjacent to Snake Island Cove was sampled at five elevations, from the water's edge to the high tidal flats. Data were analyzed for statistical differences using a two-way ANOVA in SAS. Cattle grazing may affect the vegetation and macroinvertebrate communities in salt marshes through trampling and herbivory. Vegetation resources available to other herbivores are decreased by the direct consumption of plant material by cattle. Spartina alterniflora and Salicornia virginica heights were significantly greater in ungrazed treatments than grazed for every season in the edge, upper, and middle elevation zones. Total aerial vegetative cover was also reduced significantly in grazed treatments, with the greatest impact in the edge and upper marsh. In the ungrazed treatments, S. alterniflora stem density was significantly greater in edge elevations, while both S. virginica percent cover and stem density in the edge elevation was greater. Burrowing crab populations were greater in the upper marsh and edge habitat of ungrazed treatments, while significantly greater in most of the middle marsh habitats of the grazed treatment. Size of burrowing crabs was generally significantly greater in ungrazed treatments. Cerithidea pliculosa size decreased in grazed treatments, but population had an overall increase in grazed treatments. Littoraria irrorata had very few differences between treatments, although few specimens were found. Melampus bidentatus populations were too small to evaluate thoroughly. Macroinvertebrate populations could be used to assess the overall health of grazed salt marshes.

The effect of cattle grazing on the abundance and distribution of vegetation, burrowing crabs (Uca rapax, Uca pugnax, and Sesarma cinereum), marsh periwinkles (Littoraria irrorata), horn snails (Cerithidea pliculosa), and salt marsh snails (Melampus bidentatus) was evaluated over four seasons (summer 2000, fall 2000, winter 2001, and spring 2001) in grazed and ungrazed treatments. A Galveston Island salt marsh adjacent to Snake Island Cove was sampled at five elevations, from the water's edge to the high tidal flats. Data were analyzed for statistical differences using a two-way ANOVA in SAS. Cattle grazing may affect the vegetation and macroinvertebrate communities in salt marshes through trampling and herbivory. Vegetation resources available to other herbivores are decreased by the direct consumption of plant material by cattle. Spartina alterniflora and Salicornia virginica heights were significantly greater in ungrazed treatments than grazed for every season in the edge, upper, and middle elevation zones. Total aerial vegetative cover was also reduced significantly in grazed treatments, with the greatest impact in the edge and upper marsh. In the ungrazed treatments, S. alterniflora stem density was significantly greater in edge elevations, while both S. virginica percent cover and stem density in the edge elevation was greater. Burrowing crab populations were greater in the upper marsh and edge habitat of ungrazed treatments, while significantly greater in most of the middle marsh habitats of the grazed treatment. Size of burrowing crabs was generally significantly greater in ungrazed treatments. Cerithidea pliculosa size decreased in grazed treatments, but population had an overall increase in grazed treatments. Littoraria irrorata had very few differences between treatments, although few specimens were found. Melampus bidentatus populations were too small to evaluate thoroughly. Macroinvertebrate populations could be used to assess the overall health of grazed salt marshes.