Symbiosis brought important evolutionary novelties to life on Earth. Lichens, the symbiotic entities formed by fungi, photosynthetic organisms and bacteria, represent an example of a successful adaptation in surviving hostile environments. Yet many aspects of the lichen symbiosis remain unexplored. This thesis aims at bringing insights into lichen biology and the importance of symbiosis in adaptation. I am using as model system a successful colonizer of tundra and alpine environments, the worm lichens Thamnolia, which seem to only reproduce vegetatively through symbiotic propagules. When the genetic architecture of the mating locus of the symbiotic fungal partner was analyzed with genomic and transcriptomic data, a sexual self-incompatible life style was revealed. However, a screen of the mating types ratios across natural populations detected only one of the mating types, suggesting that Thamnolia has no potential for sexual reproduction because of lack of mating partners. Genetic data based on molecular markers revealed the existence of three morphologically cryptic Thamnolia lineages. One lineage had a clear recombination structure and was found in the tundra region of Siberia, shorelines of Scandinavia, and Aleutian Islands. The other lineage was allopatric with the previous, and was highly clonal; only two haplotypes were found across the alpine region of central and southeastern Europe. However, the third lineage was sympatric with the other two, had a worldwide distribution, and although highly clonal, showed a recombinant population structure. Our data could not reveal whether the signs of recombination resulted from rare recombination events due to the extreme low frequency of the other mating type or ancestral variation before the loss of sexual reproduction. However, investigation of Thamnolia’s green algal population showed that in different localities, different algal genotypes were associated with the same fungal genotype. Furthermore, data suggest that Thamnolia carried several algal genotypes within its thalli and shared them with other distantly related but ecologically similar fungal species.

This thesis presents genetic studies aiming at enlarging our knowledge regarding the genetic factors underlying two immune-mediated diseases, hypothyroidism and autoimmune Addison’s disease (AAD), in dogs and humans, respectively. Genetic and environmental factors are indicated to contribute to canine hypothyroidism, which can be considered a model for human Hashimoto’s thyroiditis (HT). In Paper I we performed the first genome-wide association (GWA) study of this disease in three high-risk dog breeds (Gordon Setter, Hovawart and Rhodesian Ridgeback). Using an integrated GWA and meta-analysis strategy, we identified a novel hypothyroidism risk haplotype located on chromosome 12 being shared by the three breeds. The identified haplotype, harboring three genes previously not associated with hypothyroidism, is independent of the dog leukocyte antigen region and significantly enriched across the affected dogs. In Paper II we performed a GWA study in another high-risk breed (Giant Schnauzer) and detected an associated locus located on chromosome 11 and conferring protection to hypothyroidism. After whole genome resequencing of a subset of samples with key haplotypes, we fine mapped the region of association that was subsequently screened for the presence of structural variants. We detected a putative copy number variant overlapping with the upstream region of the IFNA7 gene, which is located in a region of high genomic complexity. Remarkably, perturbed activities of type I Interferons have been extensively associated with HT and general autoimmunity. In Paper III we performed the first large-scale genetic study of human AAD, a rare autoimmune disorder characterized by dysfunction and ultimately destruction of the adrenal cortex. We resequenced 1853 immune-related genes comprising of their coding sequences, untranslated regions, as well as conserved intronic and intergenic regions in extensively characterized AAD patients and control samples, all collected in Sweden. We identified BACH2 gene as a novel risk locus associated with AAD, and we showed its independent association with isolated AAD. In addition, we confirmed the previously established AAD association with the human leukocyte antigen complex. The results of these studies will hopefully help increasing the understanding of such diseases in dogs and humans, eventually promoting their well-being.

O manejo agrícola aplicado a um agrossistema pode determinar a qualidade e produtividade da área, além das interações biológicas que podem ser estabelecidas entre o cultivar e ecossistema local. A agricultura convencional é bastante reconhecida como um manejo eficiente e lucrativo. Por outro lado, a agroecologia tem ganhado visibilidade na agroindústria em reflexo do aumento na demanda por alternativas mais sustentáveis de produção. As diferenças entre ambos manejos podem refletir sobre a dinâmica microbiana, alterando a composição e estruturação das comunidades ali presentes. Os micro-organismos que habitam a superfície foliar da planta compõem o micro-ambiente denominado filosfera, descrito como um dos hábitats colonizáveis mais extensos. Devido a sua alta exposição a variáveis ambientais, diversos fatores podem interferir na comunidade bacteriana e definir a filosfera. Desta forma, o principal objetivo nesse estudo foi avaliar como o manejo agrícola interfere na composição bacteriana na filosfera, analisando ainda em escala temporal sua estrutura e abundância. A área experimental amostrada foi cedida pelo Centro de Pesquisa \"Mokiti Okada\", em Mogi Guaçu, São Paulo. As amostras foram coletadas de maneira representativa em diferentes linhas de tratamento, uma sob manejo convencional e outra sob manejo ecológico. Análises microbiológicas dependentes e independentes de cultivo permitiram identificar a comunidade bacteriana residente da filosfera de citros, a qual era compartilhada por ambos os manejos. Entretanto, análises de sequenciamento NGS (New Generation Sequencing) mostraram uma diferença significativa entre as comunidades bacterianas dos dois manejos, com o ecológico apresentando uma maior diversidade. Apesar do manejo ter se mostrado um importante fator na composição bacteriana, quando avaliado em função temporal, viu-se que as épocas de coleta interferem mais intensamente na estrutura das bactérias (p=0,0001), mostrando uma sobreposição dos diversos fatores ambientais que atuam sobre a filosfera. Os resultados ainda indicaram uma redução na abundância de bactérias, a qual pode estar relacionada com a aplicação extra de produtos cúpricos em ambas as áreas, em função do acometimento da \"pinta preta\" no pomar, o que instigou monitorar o cobre no tecido foliar. Quimicamente, micro-análises de XRF (X-Ray Fluorescence) mostraram que há uma maior concentração de cobre nas folhas provenientes da área convencional, o que é resultado das maiores quantidades do produto que são aplicadas nesse tratamento. Além disso, foi possível o isolamento de bactérias do gênero Enterococcus na filosfera, as quais apresentam mecanismos de tolerância ao cobre, demonstrando que os produtos cúpricos podem ter selecionado esses organismos. Logo, esse estudo apresentou uma importante perspectiva do efeito do manejo agrícola sobre a filosfera, contribuindo para a compreensão da dinâmica microbiana na agricultura. / The agricultural management applied to a agrosystem is an important determinant for the quality and productivity of the crop yield, also for the biological interactions that can be stablished between the plants and the local ecosystem. Conventional agriculture has being well known as an efficient and lucrative crop management. On the other hand, agroecology has gaining visibility in the agroindustry due to increasing demand for a more sustainable production alternative. The differences between both approaches can reflect on the microbial dynamic, affecting the composition and structure of these communities. Microrganisms inhabitating the foliar surface correspond to a microenvironment called phyllosphere, which is described as one of the most extensive habitats. Due to its constant exposition to environmental variables, several factors can influence on the bacterial community and modulate the phyllosphere. Thus, the main purpose of this study was to evaluate how the agricultural management can impact on the phyllospheric bacteria, also considering a temporal effect on structure and abundance of these organisms. The experimental area was provided by \"Mokiti Okada\" Research Center, located at em Mogi-Guaçu, São Paulo. The samples were representativelly collected from two treatment lines, one under convencional management, and the other under ecological management. Afterwards, culture-dependent and independent microbiological analysis allowed to identify the resident bacterial community in citros phyllosphere, which was greatly shared bewteen both treatments. However, NSG (New Generation Sequencing) analysis demonstrated a significative difference between the bacterial community under conventional and ecological management, where the second one demonstrated a higher diversity, which can be related to the different approaches applied. Although the agricultural method have demonstrated an important factor on bacterial composition, when temporally evaluated, it was observed a more intense interferance on the bacterial structure by the time of sampling (p=0,0001), representing a possible overlap of environmental factors on the phyllosphere. The data also indicate a decrease in the abundance of bacteria that might be resulted from the extra use of cupric products, related to the impairment of \"black spot\" on the crop, what lead to a copper monitoring in the foliar tissue. Chemically, XRF micro-analysis (X-Ray Fluorescence) demonstrated that there is a higher concentration of copper on the leaves from the conventional area, which is resulted of the higher application of its products by this method. Moreover, a search for copper-tolerant microrganisms was conducted, and it was possible to isolate Enterococcus bacteria, which have copper tolerance mechanisms. This result implicate that the use of cupric products may have selected these microrganisms on citros phyllosphere. Therefore, this study presented an important perspective of how the agricultural management can influence the phyllosfere, which can contribute to undertand about the microbial dynamic and its roles on the agriculture.

Les maladies infectieuses sont souvent étudiées à l'aide d'approches réductionnistes alors qu'elles sont fortement influencées par de nombreux facteurs hôtes et environnementaux en interaction. Ainsi, de nombreuses maladies d’étiologie complexe restent difficiles à caractériser. En développant une approche holistique pour aborder la complexité de l'interaction, (i) nous avons déchiffré les interactions complexes sous-jacentes au syndrome de mortalité des huîtres du Pacifique chez les juvéniles d’huîtres Crassostrea gigas, la principale espèce d'huître exploitée dans le monde et (ii) nous avons validé le mécanisme de pathogénèse quel que soit l’environnement infectieux et le génotype de l’huître. En utilisant une expérience d’infection écologiquement réaliste combinée à des analyses moléculaires (métabarcoding, transcriptomique, et suivi des agents pathogènes) et histologiques sur des familles d'huîtres aux susceptibilités contrastées à la maladie, nous avons démontré que la maladie est causée par une infection multiple avec comme première étape nécessaire l’infection des cellules immunocompétentes de l’huître (les hémocytes) par OsHV-1µvar. La réplication du virus induit un état immunodéprimé de l’huître qui conduit à une septicémie par des bactéries pathogènes opportunistes entraînant la mort des huîtres. En identifiant les interactions intra-hôtes entre les microorganismes et l'immunité de l'hôte, cette étude déchiffre le code du syndrome de mortalité des huîtres du Pacifique et fournit d'importantes données pour la conception de mesures prophylactiques et de programmes de sélection d'huîtres résistantes au syndrome de mortalité. Nous pensons qu'une telle approche de la biologie des systèmes pourrait être appliquée pour déchiffrer d'autres maladies multifactorielles qui affectent des espèces d'invertébrés non modèles dans le monde entier. / Infectious diseases are very often explored using reductionist approaches, despite repeated evidence showing them to be strongly influenced by numerous interacting host and environmental factors. Many diseases with complex etiology therefore remain misunderstood. In this thesis, by developing a holistic approach to tackle the complexity of the interaction, (i) we deciphered the complex intra-host interactions underlying the Pacific oyster mortality syndrome affecting juveniles of Crassostrea gigas, the main oyster species exploited worldwide and (ii) we validated this mechanism in different infectious environments and oyster genotypes. Using ecologically realistic experimental infections combined with thorough molecular (metabarcoding, transcriptomics, pathogen monitoring) and histological analyses on oyster families with contrasting susceptibilities, we demonstrated that the disease is caused by a multiple infection whose initial and necessary step is the infection of oyster haemocytes by a herpesvirus. Viral replication leads to an immune-compromised state of the host, evolving toward subsequent bacteremia by opportunistic bacteria. By identifying critical intra-host interactions between microorganisms and host immunity, this study cracks the code of the Pacific oyster mortality syndrome and provides important molecular data for the design of prophylactic measures and breeding programs dedicated to the production of oysters resistant to the mortality syndrome. We believe that such a systems biology approach could be applied to decipher other multi-factorial diseases that affect non-model invertebrate species worldwide.

Mycobacterium tuberculosis, la bactérie causant la tuberculose, est un pathogène d'importance majeure à l'échelle mondiale. Depuis sa découverte en 1882 par Robert Koch, de nombreuses études se sont penchées sur les caractéristiques de cette bactérie et des souches proches, connues sous le nom de complexe Mycobacterium tuberculosis (MTBC). Dans le cadre de ce travail nous avons commencé par nous intéresser à l'espèce proche "Mycobacterium canettii", qui avait été identifiée au milieu du XXème siècle comme étant également capable de causer des cas de tuberculose chez l'Homme, tout en possédant des caractéristiques phénotypiques propres. Par le biais de l'étude de certains marqueurs phylogénétiques, nous avons pu établir que cette bactérie n'appartenait pas au MTBC au sens strict et pouvait donc être utilisée comme point d'ancrage dans le cadre de l'étude de la phylogénie et de l'émergence de ce dernier. C'est pourquoi nous avons choisi d'étudier la diversité de la collection de souches de "Mycobacterium canettii", qui proviennent toutes d'une même région du globe, la Corne de l'Afrique. L'étude de cette collection, construite au fil des ans par le Service de Santé des Armées (SSA), a permis de mettre en évidence l'émergence d'un groupe particulier de souches au sein de cette espèce, ainsi que d'obtenir des éléments permettant de préciser le positionnement du dernier ancêtre commun (MRCA) du MTBC. Du fait de l'origine géographique exclusive de ce taxon, nous avons ensuite décidé d'évaluer la diversité génétique des souches de Mycobacterium tuberculosis provenant de cette même région du globe. Cette seconde partie de l'étude, menée sur une collection à nouveau constituée par le SSA, a conduit à l'identification d'une nouvelle lignée au sein du MTBC, jusqu'alors inconnue. Cette découverte a un impact important sur la compréhension de l'émergence de Mycobacterium tuberculosis, car elle permet d'envisager un nouveau modèle d'apparition en interprétant cette lignée comme le descendant contemporain de l'écotype fondateur du MTBC. L'évolution de Mycobacterium tuberculosis peut ainsi être comprise suivant une progression liant "Mycobacterium canettii", pathogène occasionnel supposé environnemental, et cette nouvelle lignée. Une fois ce nouveau modèle proposé, nous avons tenté de le dater en extrapolant le taux de mutations observé lors d'évènements épidémiques contemporains, ce qui nous a permis de dater le MRCA du MTBC à environ 10 000 ans. Enfin nous avons mis en parallèle ces éléments concrets avec les connaissances paléo-ethnographiques actuelles concernant la Corne de l'Afrique pour proposer un modèle historiquement argumenté permettant d'expliquer la structuration phylogénétique actuelle du MTBC. / Mycobacterium tuberculosis, the causative agent of tuberculosis, is a pathogen of world-wide impact. Since its discovery in 1882 by Robert Koch many studies have been focusing on the characteristics of this bacterium and of the most closely related strains known as the Mycobacterium tuberculosis complex (MTBC). In this work we started by studying the closest neighbor to the MTBC, the "Mycobacterium canettii" taxon, which is only found in one particular region of the world, the Horn of Africa. It t has been first identified in the middle of the XXth century as being able to cause tuberculosis in humans, but having at the same time peculiar phenotypic characteristics. Through the study of some phylogenetic markers we have been able to establish that this bacterium does not belong to the MTBC sensu stricto and can therefore be used as an outgroup in order to root the phylogeny to study the emergence of the MTBC. The next step was to study the genetic diversity of a collection of strains of "M. canettii",using the “next generation sequencing” (NGS) approach.. The analysis of this collection, built along the years by the French Army Health Service (SSA), has permitted to show the rapid emergence of a particular clone, as well as to get information enabling to precise the position of the most recent common ancestor (MRCA) of the MTBC. Because of the restricted geographic location of this species, it was also decided to assess the genetic diversity of strains of M. tuberculosis coming from the same part of the globe. This second part of the study, performed on a collection of strains also gathered by the SSA, has lead to the identification of a new, previously unknown, lineage of the MTBC. This discovery has a profound impact on the comprehension of the emergence of M. tuberculosis, as it permits to develop a new model of appearance by interpreting this lineage as the founder ecotype of the MTBC. The evolution of M. tuberculosis can therefore by understood along a path linking "M. canettii", opportunistic pathogen supposedly environmental, and this new lineage. After this proposal of a new model, we tried to date it by extrapolating

Squamous cell carcinoma of the head and neck (HNSCC) is an extremely common and devastating disease with a bleak prognosis. Despite intensive research, survival rates have not improved over the past 30 years principally due to untreatable recurrent/metastasising disease. Feline oral squamous cell carcinoma (FOSCC) is an equally common disease in cats with an even less favourable prognosis than humans. Human and feline squamous cell carcinomas share similar etiopathogenesis, molecular markers, tumour biology and treatment thus making FOSCC an excellent model for HNSCC. Epithelial to mesenchymal transition (EMT), under the direction microRNAs (miRNAs/mirs) could be a key driver in oncogenic transformation and chemoresistance. The aim of this study was to induce resistance to characterise the EMT/resistance phenotype and to investigate whether common miRNA-mediated pathways are present in HNSCC and FOSCC that drive this phenomenon. We used epidermal growth factor (EGFR)-inhibitor gefitinib to induce resistance in HNSCC and FOSCC and investigated the associated EMT-related molecular changes. In vitro and in vivo invasive and migratory properties of both species were explored to determine whether resistance and/or EMT status conferred a functional advantage. We determined the miRNA expression pattern during acquisition of resistance to gefitinib in both species by next generation sequencing and screened candidate miRNAs as potential therapeutics. We found that gefitinib-resistance produced a previously unrecognised biphasic response that consisted of two distinct phenotypes, a highly invasive mesenchymal phenotype during early resistance, and a more epithelial phenotype associated with established resistance. The biphasic nature of this transition may prove critical in establishing effective therapeutic targets and the timing of treatment to overcome resistance or in preventing local invasion or metastatic spread of squamous cell carcinoma. We found that the major anti-apoptotic PI3K/AKT pathway was activated in transitioning and resistant cells of both species as demonstrated upregulation of AKT, pAKT and c-FLIP together with inactivation of PTEN by phosphorylation. This indicates that avoidance of apoptosis may be a major pathway in resistance that could be targeted therapeutically. We showed that three miRNAs were differentially expressed in both gefitinib-resistant human and feline cell lines: miR-107 was downregulated, and miR-551b and miR-574 were upregulated. These microRNAs provide potential therapeutic targets in the fight against drug resistance in head and neck cancer although much further research needs to be conducted to elucidate the complex network of interactions that may be affected by targeting these powerful regulatory molecules.

Les éléments transposables (ETs) sont des composants ubiquitaires des génomes eucaryotes, parfois prépondérants chez les plantes. Ce sont des séquences mobiles, potentiellement mutagènes, reconnues comme des acteurs de l’évolution des génomes. Cependant, la plupart des ETs sont aujourd’hui inactifs car réprimés par des mécanismes épigénétiques très efficaces. Néanmoins, ces derniers peuvent être relâchés par des stress, conduisant à la réactivation d’ETs. De tels stress sont-ils suffisants pour activer la transposition dans les populations naturelles? L’application répétée d’un stress peut-elle expliquer les pics d’activité transpositionnelle qui ont eu lieu en conditions naturelles? De récents travaux chez un mutant d’Arabidopsis thaliana, affecté dans une voie de répression d’ETs, le RdDM (RNA-directed DNA Methylation), ont démontré qu’un stress thermique conduisait à la réactivation transpositionnelle d’un ET. Mes travaux de thèse portent sur l’étude de riz sauvage et d’un mutant non décrit, affecté dans le RdDM, cultivés en conditions normales ou de stress thermique sur plusieurs générations. Les objectifs de mes travaux ont été de déterminer (1) l’impact de la mutation sur les différentes étapes d’activation rétrotranspositionnelle et (2) l’activation rétrotranspositionnelle en réponse à un stress thermique. Une part importante de ce travail a été consacrée au développement et à la comparaison de méthodes d’identification des mouvements d’ETs et différentes approches « omiques » ont été utilisées. La réactivation de 5 ETs dans les plantes mutantes, dont la mobilité n’avait pas encore été observée, suggère que la voie RdDM est impliquée dans le contrôle de leur répression. De plus, nos résultats confirment que les ETs ne sont pas tous réprimés par les mêmes voies de régulation. / Transposable elements (TEs) are ubiquitous among eukaryotic genomes sometimes overriding in plants. Due to their ability to replicate and transpose, they are potentially mutagenic and recognized as actors of genome evolution. However, the analysis of the transpositional activity of TEs in different plant species have shown that most of them are maintained in a transcriptionally inactive state through powerful and specific epigenetic mechanisms. These silencing processes can nevertheless be allievated under stress conditions, leading to TE reactivation. Are these stress sufficient to activate transposition in natural populations? Are repeated heat stress able to trigger transposition and therefore lead to bursts of transposition? In recent reports, reactivation of retrotransposons has been shown in Arabidopsis thaliana mutants impaired in the RdDM pathway (RNA-directed DNA Methylation) and submitted to heat stress. My PhD works reports the study of of a wild rice and a new rice mutant, affected in the RdDM, cultivated under optimal or heat stress conditions over generations. Here, we propose to determine (1) the impact of the mutation at the different levels leading to the retrotranspositional activation and (2) the retrotranspositional activity in response to heat stress. An important part of this work has been devoted to the development and the comparison of different methods to identify TE movements, and different -omics approaches have been used. The reactivation of 5 new TEs in mutants, suggests that the RdDM pathway is involved in the control of the repression of these TEs. Furthermore, our result confirm that all TEs are not regulated through the same pathways but are under the control of different lock.

Les entérovirus (EV) sont des Picornavirus (virus nus à génome ARN positif), caractérisés par une grande diversité génétique et antigénique (116 types classés en 4 espèces taxonomiques EV-A à D) et une évolution rapide. Les infections humaines sont très fréquentes, hautement contagieuses à partir des selles et épidémiques. La plupart des infections sont asymptomatiques ou bénignes ; elles peuvent être graves voire mortelles, en particulier chez les jeunes enfants. La poliomyélite, modèle d’infection à EV, est en voie d’éradication grâce aux programmes de vaccination et de surveillance sous l’égide de l’OMS. La détection de poliovirus sauvages dans des pays déclarés exempts de polio depuis plusieurs années et l’émergence récente de plusieurs EV non poliomyélitiques (EV-A71, EV-D68) associés à des manifestations cliniques sévères dans plusieurs régions du monde montrent l’importance de surveiller la circulation des EV dans la population humaine. Le but de la thèse était de rechercher et caractériser les EV dans les eaux usées de l’agglomération de Clermont-Ferrand et de comparer les données à celles de la surveillance clinique pour avoir une image plus complète de la circulation virale dans la population générale. Une méthode de concentration virale à partir des eaux usées prélevées en entrée (eaux usées brutes) et sortie (eaux usées traitées) de station d’épuration a été mise au point, permettant la détection moléculaire des EV et de 6 autres virus entériques humains. La présence de génomes viraux a été détectée dans tous les échantillons d’octobre 2014 à octobre 2015, avec une médiane de 6 virus différents en entrée de station et de 4 virus en sortie. L’analyse phylogénétique des séquences d’EV et des virus des hépatites A et E présents dans les eaux usées et les prélèvements cliniques des patients hospitalisés au CHU de Clermont-Ferrand pendant la même période, a validé l’approche mise en place pour surveiller la circulation communautaire d’un virus entérique. La diversité des EV présents dans les eaux usées brutes a été analysée par séquençage d’amplicons avec une technique haut débit Illumina (metabarcoding). Les résultats montrent la présence d’une grande diversité d’EV et la circulation silencieuse de 25 types (notamment 9 EV-C, dont des séquences de poliovirus 1 vaccinal) dans la population générale. L’analyse phylogénétique des variants intra-typiques a mis en évidence plusieurs profils épidémiques parmi les principaux types ayant circulé pendant la période d’étude. Les données obtenues montrent la faisabilité et la sensibilité de la stratégie développée pour détecter et caractériser les EV présents dans les eaux usées. Ils permettent de discuter la place de la surveillance environnementale dans la surveillance des infections à EV non polio (études épidémiologiques, prévention des épidémies, alertes sanitaires). Surveiller conjointement les virus entériques dans l’environnement et chez les patients permet une meilleure compréhension de leur prévalence. Cette approche globale de la circulation virale et de l’écologie de la santé représente un engagement important de la part des laboratoires et nécessitera une intégration dans des réseaux structurés de collaboration nationales et internationales dépassant la seule surveillance des EV. / Enterovirus (EV) are Picornaviruses (non-enveloped, positive-sense RNA viruses), characterized by a large genetic and antigenic diversity (116 types classified within 4 taxonomic species EV-A to D) and rapid evolution. Human infections are frequent, highly contagious from stools and occur as outbreaks. The infections are mainly asymptomatic or benign but severe or fatal cases can be reported in young children. Poliomyelitis is the model EV infection. Combined with clinical and virological surveillance, mass vaccination is closer than ever to achieve the WHO program of the Global Polio Eradication Initiative. However, the detection of wild type polioviruses in polio-free countries and the recent worldwide emergence of non-polio enteroviruses (EV-A71, EV-D68) associated with severe clinical manifestations underscore the importance of surveilling EV circulation in the general population. The aim of the PhD thesis was the detection and identification of EV strains in wastewater treated in the sewage treatment plant at Clermont-Ferrand (France). The viral data were compared with those reported through clinical surveillance to obtain a comprehensive picture of the viral circulation in the local population. A method was developed to concentrate viruses from raw and treated wastewater and molecular assays were used to detect EVs and 6 other human enteric viruses. The viral genomes were detected in all samples from October 2014 to October 2015, with a median of 6 and 4 different viruses in raw and treated wastewater respectively. Phylogenetic analysis of viral sequences (EV, hepatitis A and E viruses) determined in wastewater and reported in patients during the sampling period, showed the efficiency of the method for surveilling enteric viruses in the community. The EV diversity in raw wastewater was analyzed by sequencing of amplicons with the Illumina high throughput technology (metabarcoding). The analysis revealed a large viral diversity and the silent circulation of 25 types not detected from hospital data (in particular 9 EV-C, of which sequences of vaccine poliovirus 1). The phylogenetic analyses of intra-typic variants showed different epidemic patterns in the predominant EV types circulating over the study period. The data demonstrate the feasibility and sensitivity of the strategy developed for the detection and characterization of EV in wastewater and provide a future prospect for the implementation of environmental surveillance of non-polio EV infections in epidemiological studies, epidemic prevention, and for health alert. Combining the surveillance of enteric viruses in the environment and in the clinical setting allows a better understanding of their prevalence. This global approach of virus circulation and ecological health represents an important investment for laboratories, which will require integration in national and international collaboration networks beyond the scope of enterovirus surveillance.

L'intensification de la concurrence et les exigences des clients pour des services toujours plus innovants et fiables conduisent l'opérateur à accélérer le renouvellement de son portefeuille de services et à vouloir devancer ses concurrents dans leur mise sur le marché. Mais dans le nouveau contexte NGN/NGS où l'utilisateur est nomade, change de terminal, change d'environnement et désire une continuité de service en tout lieu et selon ses préférences, la fourniture de services depuis la stratégie jusqu'à l'exploitation devient de plus en plus complexe et longue à maîtriser. Ainsi, avoir une gestion dynamique qui permet d'améliorer le TTM et le ROI devient un challenge pour les opérateurs. Dans cette thèse, nous nous intéressons à la résolution de problèmes de gestion du cycle de vie des services depuis le Déploiement jusqu'au Delivery dans un environnement ubiquitaire et mobile. Pour ce faire, dans un premier temps, nous commençons par analyser les travaux existants en termes de gestion du cycle de vie des services, puis nous identifions plus précisément les besoins du nouveau contexte NGN/NGS du point de vue de l'utilisateur et de l'opérateur. Les verrous à lever concernent les différentes phases du cycle de vie des services. Comment les repenser afin d'avoir une continuité de service et un maintien de la QoS dans ce contexte de mobilité et d'ubiquité. Nos contributions sont au nombre de quatre et concerne le Déploiement, le Provisioning, le Delivery et l'Assurance des services. Deux champs d'application sont directement impactés par nos propositions et permettent de valoriser nos travaux. Le premier se déduit de la gestion de la mobilité de la session de l'utilisateur par la QoS. Le deuxième est les standards télécom et plus particulièrement les processus business.

microRNAs (miRNAs) are small (20-23 nt), non-coding single stranded RNA molecules that play an important role in post-transcriptional regulation of protein-coding genes. miRNAs have been found in all animal lineages, and have been implicated as critical regulators during development in multiple species. The echinoderms, Strongylocentrotus purpuratus (sea urchin) and Patiria miniata (sea star) are excellent model organisms for studying development due to their well-characterized transcriptional gene networks, ease of working with their embryos in the laboratory and phylogenetic position as invertebrate deuterostomes. Literature on miRNAs in echinoderm embryogenesis is limited. It has been shown that RNAi genes are developmentally expressed and regulated in sea urchin embryos, but no study in the sea urchin has examined the expression of miRNAs. The goal of my work has been to study miRNA regulation in echinoderm developmental gene networks. I have identified developmentally regulated miRNAs in sea urchin and sea star embryos, using a combination of computational and wet lab experimental techniques. I developed a probabilistic model (named HHMMiR) based on hierarchical hidden Markov models (HHMMs) to classify genomic hairpins into miRNA precursors and random stem-loop structures. I then extended this model to make an efficient decoder by introduction of explicit state duration densities. We used the Illumina Genome Analyzer to sequence small RNA libraries in mixed stage population of embryos from one to three days after fertilization of S. purpuratus and P. miniata. We developed a computational pipeline for analysis of these miRNAseq data to reveal the miRNA populations in both species, and study their differential expression. We also used northern blots and whole mount in situ hybridization experimental techniques to study the temporal and spatial expression patterns of some of these miRNAs in sea urchin embryos. By knocking down the major components of the miRNA biogenesis pathway, we studied the global effects of miRNAs on embryo morphology and differentiation genes. The biogenesis genes selected for this purpose are the RNAse III enzyme, Dicer and Argonaute. Dicer is necessary for the processing of mature miRNAs from hairpin structures while Ago is a necessary part of the RISC (RNA interference silencing complex) assembly, which is required for the miRNA to hybridize to its target mRNA site. Knocking down these genes hinders normal development of the sea urchin embryo and leads to loss of the larval skeleton, a novel phenotype not seen in sea stars, as well as abnormal gastrulation. Comparison of differentiation gene marker expression between control and Ago knocked down sea urchin embryos shows interesting patterns of expansion and suppression of adjoining some embryonic territories, while ingression of larval skeletogenesis progenitors does not occur.