O trabalho apresenta uma análise do desempenho de vários métodos de extração de fatores de intensidade de tensão a partir de soluções numéricas obtidas com o método dos elementos finitos generalizados (MEFG). A convergência dos fatores de intensidade de tensão é comparada com a da energia de deformação a fim de investigar a superconvergência dos métodos. Para extração dos fatores de intensidade de tensão e o cálculo da taxa de energia disponibilizada para propagação da fissura, implementam-se os métodos da integral de contorno (MIC), da função cutoff (MFC) e da integral-J no contexto do MEFG. Desenvolve-se a formulação dos métodos de extração de forma a obter uma implementação independente da malha utilizada na modelagem do problema. Aplica-se a extração dos fatores de intensidade de tensão, para modos puros e mistos, em problemas clássicos da mecânica da fratura. Verifica-se a convergência dos fatores de intensidade de tensão e da taxa de energia disponibilizada para a propagação da fissura, obtidos com cada método de extração, com o enriquecimento da ordem polinomial da solução do MEFG. Investiga-se a robustez dos métodos com relação ao tamanho dos domínios de extração / The performance of several techniques to extract stress intensity factors (SIF) from numerical solutions computed with the generalized finite element method (GFEM) is investigated. The convergence of the stress intensity factors is compared with the convergence of strain energy with the aim of investigate the superconvergence of the methods. The contour integral (CIM), the cutoff function (CFM) and the J-integral methods are considered to compute stress intensity factors and energy release rate. The proposed implementation of the extraction techniques is completely independent of the discretization used. Several numerical examples demonstrating the convergence of the computed stress intensity factors and the energy release rate, with the increasing of p order of the GFEM solution, are presented

A oleuropeína é o mais abundante biofenol presente nas folhas de oliveira (Olea Europaea), com importantes funções antimicrobiana e antioxidante. Estudos visando à obtenção deste composto têm sido conduzidos, porém, muitos deles utilizam solventes tóxicos e métodos caros. A presente dissertação teve por objetivo estudar diferentes metodologias para a obtenção de extratos de folhas de oliva contendo quantidades significativas de oleuropeína. Os extratos foram obtidos a partir de folhas de oliva micronizadas, com ou sem pré-tratamento para redução do teor de clorofila, submetidas a contatos simples ou múltiplos com diferentes solventes, como dietil éter, clorofórmio, acetona, etanol, 1-propanol, 2-propanol, água e soluções hidroalcoólicas com diferentes concentrações. O contato das folhas micronizadas com os solventes foi promovido pelos seguintes métodos: agitação manual em temperatura ambiente, agitação mecânica a 50 ºC, ultrassom ou uma combinação desses dois últimos, totalizando 38 experimentos, sendo que em 17 destes os extratos foram produzidos na forma líquida e 21 na forma sólida. Os resultados mostraram que, de maneira geral, a etapa prévia de redução do teor da clorofila (realizada através de sucessivos contatos com hexano, diclorometano ou ainda, com CO2 supercrítico) não foi vantajosa, devido à elevada quantidade de solventes utilizados em relação às quantidades de extratos obtidos. Nestes experimentos, a maior concentração de oleuropeína, 1,88%, foi detectada no procedimento em que as folhas micronizadas foram previamente umedecidas com etanol e limpas com CO2 supercrítico, e posteriormente colocadas em contato com a mistura etanol e água, na proporção 1:1, utilizando o ultrassom combinado com a agitação mecânica como método de extração. Nos experimentos finais do trabalho, foi estudada ainda a adição de ácidos orgânicos (cítrico ou acético), juntamente com os solventes hidroalcoólicos (diferentes teores de água) no momento da extração da oleuropeína em banho de ultrassom, utilizando amostras que não foram previamente tratadas para remoção da clorofila. A combinação do ácido acético com a solução etanólica contendo 30 % de água resultou em um extrato com 2,17 % de oleuropeína, em apenas 1 contato com o solvente. Quando três contatos foram utilizados, nestas mesmas condições, o teor de oleuropeína aumentou para 4,8 %, maior do que alguns valores encontrados na literatura, utilizando o mesmo método de extração, indicando que processo ainda pode ser otimizado, utilizando técnicas simples e solventes que não agridam o meio ambiente. / Oleuropein is the most abundant biofenol present in olive leaves (Olea europaea), presenting important antioxidant and antimicrobial functions. Studies focusing on obtaining this compound have been conducted; however, many of them use toxic solvents and expensive methods. The present work aimed to study different methodologies for obtaining extracts from olive leaves containing significant amounts of oleuropein. The extracts were obtained from micronized olive leaves, with or without pretreatment for reducing the chlorophyll content, submitted to single and multiple contacts with different solvents such as diethyl ether, chloroform, acetone, ethanol, 1- propanol, 2 -propanol, water and hydroalcoholic solutions at different concentrations. The contact between the micronized leaves and the solvents was promoted by the following methods: manual shaking at room temperature, mechanical agitation at 50 °C, ultrasound or a combination of these last two methods, totaling 38 experiments, from which in 17 of them the extracts were produced in the liquid form, and in 21 in the solid form (as a powder). The results show that, in general, previous step of reducing the content of chlorophyll (performed by successive contacts with hexane, dichloromethane or with supercritical CO2) was not advantageous due to the high amount of solvent used in relation to amounts of extracts. In these experiments, the highest concentration of oleuropein, 1.88 % was detected in the procedure in which the micronized leaves were previously soaked with ethanol and cleaned with supercritical CO2 and then placed in contact with the mixture of ethanol and water in the proportion 1:1, using ultrasound combined with mechanical agitation as extraction method. In the last experiments of the work, the addition of organic acids (citric or acetic acid) together with hydroalcoholic solvents (different water contents) in the extraction of oleuropein in ultrasound was studied, using samples that have not previously been treated for removal of chlorophyll. The combination of acetic acid to the ethanolic solution containing 30 % of water resulted in an extract with 2.17 % of oleuropein, with only one contact to the solvent. When three contacts were used, under the same conditions, the oleuropein content increased to 4.8 %, larger than some values found in the literature, using the same extraction method, indicating that the process can be further optimized using simple techniques and solvents that do not harm the environment.

O trabalho apresenta uma análise do desempenho de vários métodos de extração de fatores de intensidade de tensão a partir de soluções numéricas obtidas com o método dos elementos finitos generalizados (MEFG). A convergência dos fatores de intensidade de tensão é comparada com a da energia de deformação a fim de investigar a superconvergência dos métodos. Para extração dos fatores de intensidade de tensão e o cálculo da taxa de energia disponibilizada para propagação da fissura, implementam-se os métodos da integral de contorno (MIC), da função cutoff (MFC) e da integral-J no contexto do MEFG. Desenvolve-se a formulação dos métodos de extração de forma a obter uma implementação independente da malha utilizada na modelagem do problema. Aplica-se a extração dos fatores de intensidade de tensão, para modos puros e mistos, em problemas clássicos da mecânica da fratura. Verifica-se a convergência dos fatores de intensidade de tensão e da taxa de energia disponibilizada para a propagação da fissura, obtidos com cada método de extração, com o enriquecimento da ordem polinomial da solução do MEFG. Investiga-se a robustez dos métodos com relação ao tamanho dos domínios de extração / The performance of several techniques to extract stress intensity factors (SIF) from numerical solutions computed with the generalized finite element method (GFEM) is investigated. The convergence of the stress intensity factors is compared with the convergence of strain energy with the aim of investigate the superconvergence of the methods. The contour integral (CIM), the cutoff function (CFM) and the J-integral methods are considered to compute stress intensity factors and energy release rate. The proposed implementation of the extraction techniques is completely independent of the discretization used. Several numerical examples demonstrating the convergence of the computed stress intensity factors and the energy release rate, with the increasing of p order of the GFEM solution, are presented

Context The huge amount of existing digital video files needs to provide indexing to make it available for customers (easier searching). The indexing can be provided by text information extraction. In this thesis we have analysed and compared methods of text information extraction in digital videos. Furthermore, we have evaluated them in the new context proposed by us, namely usefulness in sports news indexing and information retrieval. Objectives The objectives of this thesis are as follows: providing a better understanding of the nature of text extraction; performing a systematic literature review on various methods of text information extraction in digital videos of TV sports news; designing and executing an experiment in the testing environment; evaluating available and promising methods of text information extraction from digital video files in the proposed context associated with video sports news indexing and retrieval; providing an adequate solution in the proposed context described above. Methods This thesis consists of three research methods: Systematic Literature Review, Video Content Analysis with the checklist, and Experiment. The Systematic Literature Review has been used to study the nature of text information extraction, to establish the methods and challenges, and to specify the effective way of conducting the experiment. The video content analysis has been used to establish the context for the experiment. Finally, the experiment has been conducted to answer the main research question: How useful are the methods of text information extraction for indexation of video sports news and information retrieval? Results Through the Systematic Literature Review we identified 29 challenges of the text information extraction methods, and 10 chains between them. We extracted 21 tools and 105 different methods, and analyzed the relations between them. Through Video Content Analysis we specified three groups of probability of text extraction from video, and 14 categories for providing video sports news indexation with the taxonomy hierarchy. We have conducted the Experiment on three videos files, with 127 frames, 8970 characters, and 1814 words, using the only available MoCA tool. As a result, we reported 10 errors and proposed recommendations for each of them. We evaluated the tool according to the categories mentioned above and offered four advantages, and nine disadvantages of the Tool mentioned above. Conclusions It is hard to compare the methods described in the literature, because the tools are not available for testing, and they are not compared with each other. Furthermore, the values of recall and precision measures highly depend on the quality of the text contained in the video. Therefore, performing the experiments on the same indexed database is necessary. However, the text information extraction is time consuming (because of huge amount of frames in video), and even high character recognition rate gives low word recognition rate. Therefore, the usefulness of text information extraction for video indexation is still low. Because most of the text information contained in the videos news is inserted in post-processing, the text extraction could be provided in the root: during the processing of the original video, by the broadcasting company (e.g. by automatically saving inserted text in separate file). Then the text information extraction will not be necessary for managing the new video files / The huge amount of existing digital video files needs to provide indexing to make it available for customers (easier searching). The indexing can be provided by text information extraction. In this thesis we have analysed and compared methods of text information extraction in digital videos. Furthermore, we have evaluated them in the new context proposed by us, namely usefulness in sports news indexing and information retrieval.

A oleuropeína é o mais abundante biofenol presente nas folhas de oliveira (Olea Europaea), com importantes funções antimicrobiana e antioxidante. Estudos visando à obtenção deste composto têm sido conduzidos, porém, muitos deles utilizam solventes tóxicos e métodos caros. A presente dissertação teve por objetivo estudar diferentes metodologias para a obtenção de extratos de folhas de oliva contendo quantidades significativas de oleuropeína. Os extratos foram obtidos a partir de folhas de oliva micronizadas, com ou sem pré-tratamento para redução do teor de clorofila, submetidas a contatos simples ou múltiplos com diferentes solventes, como dietil éter, clorofórmio, acetona, etanol, 1-propanol, 2-propanol, água e soluções hidroalcoólicas com diferentes concentrações. O contato das folhas micronizadas com os solventes foi promovido pelos seguintes métodos: agitação manual em temperatura ambiente, agitação mecânica a 50 ºC, ultrassom ou uma combinação desses dois últimos, totalizando 38 experimentos, sendo que em 17 destes os extratos foram produzidos na forma líquida e 21 na forma sólida. Os resultados mostraram que, de maneira geral, a etapa prévia de redução do teor da clorofila (realizada através de sucessivos contatos com hexano, diclorometano ou ainda, com CO2 supercrítico) não foi vantajosa, devido à elevada quantidade de solventes utilizados em relação às quantidades de extratos obtidos. Nestes experimentos, a maior concentração de oleuropeína, 1,88%, foi detectada no procedimento em que as folhas micronizadas foram previamente umedecidas com etanol e limpas com CO2 supercrítico, e posteriormente colocadas em contato com a mistura etanol e água, na proporção 1:1, utilizando o ultrassom combinado com a agitação mecânica como método de extração. Nos experimentos finais do trabalho, foi estudada ainda a adição de ácidos orgânicos (cítrico ou acético), juntamente com os solventes hidroalcoólicos (diferentes teores de água) no momento da extração da oleuropeína em banho de ultrassom, utilizando amostras que não foram previamente tratadas para remoção da clorofila. A combinação do ácido acético com a solução etanólica contendo 30 % de água resultou em um extrato com 2,17 % de oleuropeína, em apenas 1 contato com o solvente. Quando três contatos foram utilizados, nestas mesmas condições, o teor de oleuropeína aumentou para 4,8 %, maior do que alguns valores encontrados na literatura, utilizando o mesmo método de extração, indicando que processo ainda pode ser otimizado, utilizando técnicas simples e solventes que não agridam o meio ambiente. / Oleuropein is the most abundant biofenol present in olive leaves (Olea europaea), presenting important antioxidant and antimicrobial functions. Studies focusing on obtaining this compound have been conducted; however, many of them use toxic solvents and expensive methods. The present work aimed to study different methodologies for obtaining extracts from olive leaves containing significant amounts of oleuropein. The extracts were obtained from micronized olive leaves, with or without pretreatment for reducing the chlorophyll content, submitted to single and multiple contacts with different solvents such as diethyl ether, chloroform, acetone, ethanol, 1- propanol, 2 -propanol, water and hydroalcoholic solutions at different concentrations. The contact between the micronized leaves and the solvents was promoted by the following methods: manual shaking at room temperature, mechanical agitation at 50 °C, ultrasound or a combination of these last two methods, totaling 38 experiments, from which in 17 of them the extracts were produced in the liquid form, and in 21 in the solid form (as a powder). The results show that, in general, previous step of reducing the content of chlorophyll (performed by successive contacts with hexane, dichloromethane or with supercritical CO2) was not advantageous due to the high amount of solvent used in relation to amounts of extracts. In these experiments, the highest concentration of oleuropein, 1.88 % was detected in the procedure in which the micronized leaves were previously soaked with ethanol and cleaned with supercritical CO2 and then placed in contact with the mixture of ethanol and water in the proportion 1:1, using ultrasound combined with mechanical agitation as extraction method. In the last experiments of the work, the addition of organic acids (citric or acetic acid) together with hydroalcoholic solvents (different water contents) in the extraction of oleuropein in ultrasound was studied, using samples that have not previously been treated for removal of chlorophyll. The combination of acetic acid to the ethanolic solution containing 30 % of water resulted in an extract with 2.17 % of oleuropein, with only one contact to the solvent. When three contacts were used, under the same conditions, the oleuropein content increased to 4.8 %, larger than some values found in the literature, using the same extraction method, indicating that the process can be further optimized using simple techniques and solvents that do not harm the environment.

<p>U okviru ovog rada izvr&scaron;eno je ispitivanje različitih procesa ekstrakcije herbe industrijske konoplje. Za ekstrakciju određenih bioaktivnih jedinjenja primenjene su klasične tehnike ekstrakcije, maceracija i Soxhlet ekstrakcija, dok su od savremenih metoda primenjene ultrazvučna ekstrakcija, mikrotalasna ekstrakcija, ekstrakcija vodom u subkritičnom stanju i ekstrakcija ugljendioksidom u superkritičnom stanju. Praćenjem uticaja ulaznih promenjivih (procesnih parametara) na odabrane izlazne promenjive (sadržaj bioaktivnih jedinjenja ili antioksidativnu aktivnost) za svaku tehniku ekstrakcije pojedinačno utvrđeni su optimalni uslovi ekstrakcije. Nakon izvr&scaron;enog poređenja odabrana je najadekvatnija tehnika ekstrakcije za pripremu tečnog ekstrakta koji će biti transformisan u suvi ekstrakt primenom metode su&scaron;enja raspr&scaron;ivanjem. Prilikom dobijanja suvog ekstrakta ispitan je uticaj različite koncentracije nosača na kvalitet ekstrakata i efikasnost su&scaron;enja. Dobijeni ekstrakti su okarakterisani u pogledu fizičkih i hemijskih osobina. Sadržaj kanabinoida u tečnim ekstraktima suve herbe industrijske konoplje ispitanih u ovoj doktorskoj disertaciji je bio u opsegu od 0,4314 mg/mL CBD-a i 0,0364 mg/mL THC-a (u optimalnom ekstraktu dobijenom maceracijom) do 1,0420 mg/mL CBD-a i 0,0430 mg/mL THC-a (u optimalnom ekstraktu dobijenom mikrotalasnom ekstrakcijom). U ekstraktima koji su nepolarnog karaktera i koji su dobijeni primenom Soxhlet ekstrakcije sadržaj CBD-a i THC-a je bio 64,40 i 2,90 mg/g, dok je primenom superkritične ekstrakcije dobijen sadržaj CBD-a i THC-a u opsegu od 76,1193 do 163,1111 mg/g i od 4,1746 do 6,5803 mg/g. Tečni ekstrakti dobijeni primenom vode i etanola kao ekstragensa bili su bogati flavonoidima i fenolima, a utvrđeno je i da imaju značajnu antioksidativnu aktivnost, dok su ekstrakti dobijeni Soxhlet ekstrakcijom i ekstrakcijom ugljendioksidom u superkritičnom stanju bili bogati lipofilnim jedinjenjima i terpenima. Ovako dobijeni ekstrakti predstavljaju visokovredne proizvode koji imaju veliki potencijal za upotrebu u farmaceutskoj industriji.</p> / <p>Various extraction processes of the aerial parts of industrial hemp were examined. From classic extractions, maceration and Soxhlet extraction were applied, while from novel methods, ultrasound-assisted extraction, microwave-assisted extraction, subcritical water extraction and supercritical carbon dioxide extraction were applied. By monitoring the influence of the input variables on the selected output variables for each extraction, the optimal extraction conditions were determined separately. After a comparison of all the extraction processes tested, microwave-assisted extraction was selected to prepare the liquid extract for spray drying. The effect of different carrier concentrations was examined during the preparation of the dry extract. The extracts obtained were characterized in terms of physical and chemical properties. The content of cannabinoids in liquid extracts of dry Cannabis sativa herba examined ranged from 0.4314 mg/mL for CBD and 0.0364 mg/mL for THC in the optimum extract obtained by maceration to 1.0420 mg/mL for CBD and 0.0430 mg/mL for THC in the optimum extract obtained by microwave-assisted extraction. The CBD and THC content in extract obtained by Soxhlet extraction was 64.4000 mg/mL and 2.9000 mg/mL, respectively, while the highest content of CBD and THC in the supercritical carbon dioxide extract was 163.11 and 6.5803 mg/g. In addition to cannabinoids, the liquid extracts obtained were rich in phenols and flavonoids, and have considerable antioxidant activity, while the extracts obtained by Soxhlet extraction and supercritical carbon dioxide were rich in lipophilic compounds and terpenes. The extracts mentioned above are high value products which have great potential for implementation in the pharmaceutical industry.</p>

A document image, beside text, may contain pictures, graphs, signatures, logos, barcodes, hand-drawn sketches and/or seals. Further, the text blocks in an image may be in Manhattan or any complex layout. Document Layout Analysis is an important preprocessing step before subjecting any such image to OCR. Here, the image with complex layout and content is segmented into its constituent components. For many present day applications, separating the text from the non-text blocks is sufficient. This enables the conversion of the text elements present in the image to their corresponding editable form. In this work, an effort has been made to separate the text areas from the various kinds of possible non-text elements. The document images may have been obtained from a scanner or camera. If the source is a scanner, there is control on the scanning resolution, and lighting of the paper surface. Moreover, during the scanning process, the paper surface remains parallel to the sensor surface. However, when an image is obtained through a camera, these advantages are no longer available. Here, an algorithm is proposed to separate the text present in an image from the clutter, irrespective of the imaging technology used. This is achieved by using both the structural and textural information of the text present in the gray image. A bank of Gabor filters characterizes the statistical distribution of the text elements in the document. A connected component based technique removes certain types of non-text elements from the image. When a camera is used to acquire document images, generally, along with the structural and textural information of the text, color information is also obtained. It can be assumed that text present in an image has a certain amount of color homogeneity. So, a graph-theoretical color clustering scheme is employed to segment the iso-color components of the image. Each iso-color image is then analyzed separately for its structural and textural properties. The results of such analyses are merged with the information obtained from the gray component of the image. This helps to separate the colored text areas from the non-text elements. The proposed scheme is computationally intensive, because the separation of the text from non-text entities is performed at the pixel level Since any entity is represented by a connected set of pixels, it makes more sense to carry out the separation only at specific points, selected as representatives of their neighborhood. Harris' operator evaluates an edge-measure at each pixel and selects pixels, which are locally rich on this measure. These points are then employed for separating text from non-text elements. Many government documents and forms in India are bi-lingual or tri-lingual in nature. Further, in school text books, it is common to find English words interspersed within sentences in the main Indian language of the book. In such documents, successive words in a line of text may be of different scripts (languages). Hence, for OCR of these documents, the script must be recognized at the level of words, rather than lines or paragraphs. A database of about 20,000 words each from 11 Indian scripts1 is created. This is so far the largest database of Indian words collected and deployed for script recognition purpose. Here again, a bank of 36 Gabor filters is used to extract the feature vector which represents the script of the word. The effectiveness of Gabor features is compared with that of DCT and it is found that Gabor features marginally outperform the DOT. Simple, linear and non-linear classifiers are employed to classify the word in the feature space. It is assumed that a scheme developed to recognize the script of the words would work equally fine for sentences and paragraphs. This assumption has been verified with supporting results. A systematic study has been conducted to evaluate and compare the accuracy of various feature-classifier combinations for word script recognition. We have considered the cases of bi-script and tri-script documents, which are largely available. Average recognition accuracies for bi-script and tri-script cases are 98.4% and 98.2%, respectively. A hierarchical blind script recognizer, involving all eleven scripts has been developed and evaluated, which yields an average accuracy of 94.1%. The major contributions of the thesis are: • A graph theoretic color clustering scheme is used to segment colored text. • A scheme is proposed to separate text from the non-text content of documents with complex layout and content, captured by scanner or camera. • Computational complexity is reduced by performing the separation task on a selected set of locally edge-rich points. • Script identification at word level is carried out using different feature classifier combinations. Gabor features with SVM classifier outperforms any other feature-classifier combinations. A hierarchical blind script recognition algorithm, involving the recognition of 11 Indian scripts, is developed. This structure employs the most efficient feature-classifier combination at each individual nodal point of the tree to maximize the system performance. A sequential forward feature selection algorithm is employed to. select the most discriminating features, in a case by case basis, for script-recognition. The 11 scripts are Bengali, Devanagari, Gujarati, Kannada, Malayalam, Odiya, Puniabi, Roman. Tamil, Telugu and Urdu.

Les compartiments géologiques profonds suscitent un intérêt grandissant dans la communauté scientifique depuis les 50 dernières années. Néanmoins, ces écosystèmes demeurent largement méconnus du fait de leur difficulté d’accès. Le forage profond réalisé par l’ANDRA dans le Bassin parisien en 2008 a offert une opportunité unique de les étudier. Dans ce cadre, cette thèse avait deux objectifs majeurs ; i) caractériser, d’un point de vue microbiologique, quatre formations sédimentaires terrestres triasiques situées entre 1700 et 2000 m de profondeur et ii) étudier les effets combinés des paramètres de température, pression et salinité ainsi que de leur interaction sur l’activité métabolique de procaryotes anaérobies afin de mieux appréhender leur comportement au cours d’un enfouissement géologique.Malgré la recherche de microorganisme par la réalisation d’une gamme de milieux de culture diversifiée, ciblant préférentiellement les types trophiques fréquemment rencontrés en subsurface (méthanogènes, fermentaires, réducteurs de composés soufrés), aucun microorganisme viable et cultivable n’ait été isolé. En parallèle, une approche moléculaire complémentaire, composée (i) de l’étude comparative de l’efficacité de différentes méthodes d’extraction directe d’ADN et (ii) de l’analyse de la diversité bactérienne par la réalisation d’inventaires moléculaires, par DGGE (Denaturing Gel Gradient Electrophoresis) et clonage, a été réalisée sur le coeur des carottes de roches, conservées à pression atmosphérique ou sous pression, dans leurs états initiaux et post-incubation. L’exploration de ces formations sédimentaires profondes a indiqué la présence d’une très faible biomasse et d’une biodiversité microbienne pauvre principalement composée de membres aérobies et mésophiles appartenant au domaine Bacteria. Cette communauté bactérienne inattendue car a priori peu adaptée aux conditions régnant in-situ, également retrouvée dans divers écosystèmes de subsurface ainsi que dans des biotopes extrêmes, pourrait provenir en partie d’une paléo-recharge de l’aquifère du Trias par des eaux froides dérivées de la fonte des glaces formées lors de la dernière glaciation du Pléistocène.Le second objectif a été abordé à travers l’élaboration d’un plan factoriel complet dans le but d’identifier les effets des paramètres sur les activités microbiennes. Ainsi, les activités métaboliques de huit souches microbiennes halophiles et thermo-tolérantes ont été mesurées sous trente conditions distinctes de température (40, 55 et 70°C), pression (1, 90 et 180 bars) et salinité (13, 50, 110, 180 et 260 g.l-1). Toutes les souches originaires d’environnements profonds se sont révélées être au minimum piézo-tolérantes et capables de maintenir leur activité métabolique sous pressions hydrostatiques. Les métabolismes fermentaires (Thermovirga lienii et Halothermothrix orenii) et thiosulfato-réducteurs (Petrotoga mexicana et Thermosipho japonicus) se sont avérés particulièrement bien adaptées, d’un point de vue métabolique, aux hautes pressions, les plus hautes activités ayant été détectées sous pression. Certaines souches ont montré une résistance accrue aux hautes températures sous pression (Petrotoga mexicana). Toutefois une résistance variable à la salinité dans les différentes conditions de température et de pression a été observée pour chacune des souches, suggérant que certains mécanismes de résistance contre la pression osmotique seraient également efficaces pour lutter contre les températures et les pressions hydrostatiques élevées.Ce travail souligne que l’étude des écosystèmes terrestres profonds d’un point de vue microbiologique ne doit pas se restreindre à la recherche et à l’analyse de la diversité présente. L’étude des activités métaboliques de souches de subsurface en conditions profondes ouvre la voie à une meilleure compréhension des rôles joués par les communautés microbiennes en milieu extrême. / Over the past 50 years, the scientific community has shown a growing interest for deep geological compartments. However, these ecosystems remain largely unknown due to their inaccessibility. The aim of the present thesis was double; the first aim was to characterize, from a microbiological perspective, four terrestrial Triassic sedimentary formations located between 1700 and 2000 m depth in the Parisian Basin and collected by the ANDRA during a deep drilling campaign in 2008, and the second aim was to study the combined effects of temperature, pressure and salinity on the metabolic activity of anaerobic prokaryotes in order to predict their reaction to geological burial.Incubations in a large variety of media were carried out in order to stimulate the growth of the main trophic types found in such environments such as methanogens, fermenters and bacteria reducing sulphur compounds, however, no viable and cultivable microorganisms could be isolated. In parallel, a molecular approach was used to i) compare the efficacy of several DNA extractions methods and ii) analyse the bacterial diversity, using DGGE (Denaturing Gel Gradient Electrophoresis) and cloning, present in rock inner cores conserved either at atmospheric pressure or under pressure, in their initial states and following incubations in various media. The genetic exploration of these samples revealed a very low biomass and a poor diversity composed mainly of aerobic and mesophilic members of the Bacteria domain, a priori unadapted to such a deep, hot, saline and anoxic environment. This unexpected microbial community also found in many subsurface ecosystems as well as in extreme ecosystems could have partially originated from a paleorecharge of the Trias aquifer with cold waters coming from the melting of ice formed during the last Pleistocene glaciation.The second objective was to study the combined effects of temperature (40, 55 and 70°C), pressure (1, 90 and 180 bars) and salinity (13, 50, 110, 180 and 260 g.l-1) on the metabolic activity of anaerobic prokaryotes. In order to identify effects of parameters on microbial activities, a complete factorial plan was constructed from the metabolic activities measured for eight halophile and thermo-tolerant bacterial strains exposed to 30 distinct temperature, pressure and salinity conditions. All the strains issued from deep environments were at the least piezo-tolerant and capable of maintaining their activity under hydrostatic pressures. The fermenting (Thermovirga lienii and Halothermothrix orenii) and thiosulfate reducing strains (Petrotoga mexicana and Thermiosipho japonicus) were particularly well adapted, from a metabolic point of view, to high pressures; indeed the highest activities were measured under pressure. Also, several strains (such as Petrotoga mexicana) showed an increased resistance to high temperatures under pressure. However, resistance to an increase in salinity was variable for each strain under the different temperature and pressure conditions. This suggests that the resistance mechanisms for osmotic pressure also enable resistance to high temperatures and hydrostatic pressures.This work underlines that the microbiological characterization of deep terrestrial ecosystems must not be limited to the search and analyses of the existing diversity. Moreover, such upstream studies of the metabolic activities of subsurface bacterial strains in deep terrestrial conditions are a necessary beginning towards understanding the role of microbial communities in extreme environments.

Les compartiments géologiques profonds suscitent un intérêt grandissant dans la communauté scientifique depuis les 50 dernières années. Néanmoins, ces écosystèmes demeurent largement méconnus du fait de leur difficulté d'accès. Le forage profond réalisé par l'ANDRA dans le Bassin parisien en 2008 a offert une opportunité unique de les étudier. Dans ce cadre, cette thèse avait deux objectifs majeurs ; i) caractériser, d'un point de vue microbiologique, quatre formations sédimentaires terrestres triasiques situées entre 1700 et 2000 m de profondeur et ii) étudier les effets combinés des paramètres de température, pression et salinité ainsi que de leur interaction sur l'activité métabolique de procaryotes anaérobies afin de mieux appréhender leur comportement au cours d'un enfouissement géologique.Malgré la recherche de microorganisme par la réalisation d'une gamme de milieux de culture diversifiée, ciblant préférentiellement les types trophiques fréquemment rencontrés en subsurface (méthanogènes, fermentaires, réducteurs de composés soufrés), aucun microorganisme viable et cultivable n'ait été isolé. En parallèle, une approche moléculaire complémentaire, composée (i) de l'étude comparative de l'efficacité de différentes méthodes d'extraction directe d'ADN et (ii) de l'analyse de la diversité bactérienne par la réalisation d'inventaires moléculaires, par DGGE (Denaturing Gel Gradient Electrophoresis) et clonage, a été réalisée sur le coeur des carottes de roches, conservées à pression atmosphérique ou sous pression, dans leurs états initiaux et post-incubation. L'exploration de ces formations sédimentaires profondes a indiqué la présence d'une très faible biomasse et d'une biodiversité microbienne pauvre principalement composée de membres aérobies et mésophiles appartenant au domaine Bacteria. Cette communauté bactérienne inattendue car a priori peu adaptée aux conditions régnant in-situ, également retrouvée dans divers écosystèmes de subsurface ainsi que dans des biotopes extrêmes, pourrait provenir en partie d'une paléo-recharge de l'aquifère du Trias par des eaux froides dérivées de la fonte des glaces formées lors de la dernière glaciation du Pléistocène.Le second objectif a été abordé à travers l'élaboration d'un plan factoriel complet dans le but d'identifier les effets des paramètres sur les activités microbiennes. Ainsi, les activités métaboliques de huit souches microbiennes halophiles et thermo-tolérantes ont été mesurées sous trente conditions distinctes de température (40, 55 et 70°C), pression (1, 90 et 180 bars) et salinité (13, 50, 110, 180 et 260 g.l-1). Toutes les souches originaires d'environnements profonds se sont révélées être au minimum piézo-tolérantes et capables de maintenir leur activité métabolique sous pressions hydrostatiques. Les métabolismes fermentaires (Thermovirga lienii et Halothermothrix orenii) et thiosulfato-réducteurs (Petrotoga mexicana et Thermosipho japonicus) se sont avérés particulièrement bien adaptées, d'un point de vue métabolique, aux hautes pressions, les plus hautes activités ayant été détectées sous pression. Certaines souches ont montré une résistance accrue aux hautes températures sous pression (Petrotoga mexicana). Toutefois une résistance variable à la salinité dans les différentes conditions de température et de pression a été observée pour chacune des souches, suggérant que certains mécanismes de résistance contre la pression osmotique seraient également efficaces pour lutter contre les températures et les pressions hydrostatiques élevées.Ce travail souligne que l'étude des écosystèmes terrestres profonds d'un point de vue microbiologique ne doit pas se restreindre à la recherche et à l'analyse de la diversité présente. L'étude des activités métaboliques de souches de subsurface en conditions profondes ouvre la voie à une meilleure compréhension des rôles joués par les communautés microbiennes en milieu extrême.