Made available in DSpace on 2017-07-21T19:59:55Z (GMT). No. of bitstreams: 1 Jonathan Pena Castro.pdf: 3378425 bytes, checksum: 726a329ba31e1d12d715441e9de01e5a (MD5) Previous issue date: 2012-02-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The species complex Astyanax scabripinnis are composed by fish with wide geographic distribution. Their populations are often isolated in small streams, separated by thousands of years, making it a very interesting system to study evolution. The use of different markers is crucial, since they allow inferences on the identification of individuals and therefore the study of populations. Thus, three populations isolated from each other of Campos do Jordão - SP, and two of Maringá - PR were analyzed by geometric morphometrics, correlating with data from presence / absence of B chromosomes, karyotypic and cytogenetic data, and reproductive data. Canonical variate analysis (CVA) showed that there are significant differences between populations (p <0.0001). The discriminant function analysis (DFA) between males and females showed intra and interpopulational sexual dimorphism. The separate CVA of males and females from each population also showed significant difference between them. All populations have 2n = 50 chromosomes arranged in 6m+22sm+10st+12a, the only exception was a population from Maringá, where there was 2n = 48 chromosomes, differentiated in 8m+26sm+6st+8a. B chromosome were only found in the population of Campos do Jordão (1850m altitude), confirming previous studies. In a morphometric analysis separated for individuals with and without B chromosomes, the CVA showed difference in body shape. Individuals with B chromosomes have the ventral anterior region less dilated than those without of this chromosomal element. The CVA also showed differences in body shape between 2n = 50 and 2n = 48 populations in Maringa. The fluorescent in situ hybridization showed differences between markers 5S rDNA and 18S rDNA for all populations. The data suggest pre zygotic reproductive isolation between Campos do Jordao populations. The analysis of all data indicates that the populations are differentiated from each other, indicating adaptation to different environments. This reinforces, among other factors, that these populations should represent different species within the species complex A. scabripinnis, followed by independent evolutionary pathways. / O complexo de espécies Astyanax scabripinnis corresponde a peixes com ampla distribuição geográfica. Suas populações encontram-se geralmente isoladas em pequenos riachos, separadas por milhares de anos, tornando esses animais interessantes ao estudo evolutivo. O uso de diferentes marcadores é fundamental, uma vez que permitem inferências na identificação dos indivíduos e consequentemente o estudo das populações. Dessa maneira, três populações isoladas entre si de Campos do Jordão - SP, e duas de Maringá - PR foram analisadas por morfometria geométrica, correlacionando com dados de presença/ausência de cromossomos B, dados cariotípicos e citogenéticos, além de dados reprodutivos. A análise de variáveis canônicas (CVA) mostrou que há diferenças significativas entre as populações (p<0,0001). A análise de função discriminante (DFA) entre macho e fêmea mostrou que há dimorfismo sexual intra e interpopulacional. A CVA separada para machos e fêmeas de cada população também mostrou diferença significativa entre elas. Todas as populações possuem 2n=50 cromossomos organizados em 6m+22sm+10st+12a, a exceção de apenas uma população de Maringá onde foi observado um cariomorfo com 2n=48 cromossomos, diferenciados em 8m+26sm+6st+8a. Não foi encontrada a presença de cromossomos B apenas na população de Campos do Jordão (662m de altitude), corroborando estudos anteriores. Na análise morfométrica associada a indivíduos com e sem cromossomos B, a CVA mostrou que há diferença na forma do corpo, sendo que os indivíduos com cromossomos B possuem a região ventral anterior menos dilatada dos que não possuem este elemento cromossômico. A CVA também apontou diferença na forma do corpo entre o cariomorfo 2n=50 e o 2n=48, em Maringá. A hibridação in situ fluorescente mostrou diferença entre os marcadores de rDNA 18S e rDNA 5S para todas as populações. Os dados reprodutivos sugerem isolamento pré zigótico entre as populações de Campos do Jordão. A análise de todos os dados aponta que as populações encontram-se diferenciadas entre si, indicando indivíduos adaptados a ambientes distintos. Isso reforça, entre outros fatores, que essas populações devem representar espécies diferentes, dentro do complexo de espécies A. scabripinnis, seguindo por caminhos evolutivos independentes.

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La première partie de ma thèse est une revue donnant un aperçu du répertoire génomique des bactéries intracellulaires et de leurs symbiotes. L'objectif de cette étude est d'explorer le processus permettant aux bactéries intracellulaires d'acquérir leur mode de vie spécifique. Nous avons commencé par examiner les données à propos de l'existence ancienne de bactéries intracellulaires, leur adaptation à leur hôte et les différences entre sympatrie et allopatrie. Une comparaison du contenu génomique de plusieurs bactéries avec différents modes de vie a révélé la capacité des bactéries à échanger des gènes à des degrés différents, en fonction de l'écosystème. La deuxième partie de ma thèse porte sur la séquence du génome de la souche Diplorickettsia massiliensis 20B qui est une bactérie intracellulaire obligatoire à Gram négatif isolée à partir des tiques de Slovaquie Ixodes ricinus. Dans ma troisième et dernière partie, nous exploré le répertoire du génome de Diplorickettsia massiliensis en le comparant aux génomes de bactéries phylogénétiquement très proches de Diplorickettsia massiliensis, issues de différentes niches. Ceci a permis de révélé son mode de vie allopatrique. Dans cette étude, nous avons comparé les caractéristiques du génome de Diplorickettsia massiliensis avec vingt-neuf espèces séquencées de Gammaproteobacteria (Legionella, Coxiella burnetii, Francisella tularensis et Rickettsiella grylli) en utilisant l'approche pangénomique multi-genre. Ce travail de thèse fournit des données originales et permet d’apporter plus de lumière sur la diversité des bactéries intracellulaires. / The initial purpose of my thesis is to understand with the help of comparative genomics, genomic variations based on coexistence, by examining data on the ancient existence of intracellular bacteria, their host adaptation and the differences between sympatry and allopatry. The first part of my thesis is a review giving insight into intracellular bacterial genome repertoire and symbionts. The goal of this review is to explore how intracellular microbes acquire their specific lifestyle. Due to their different evolutionary trajectories, these bacteria have different genomic compositions. We reviewed data on the ancient existence of intracellular bacteria, their host adaptation and the differences between sympatry and allopatry. A comparison of the genomic contents of bacteria with certain lifestyles revealed the bacterial capacity to exchange genes to different extents, depending on the ecosystem. The second part of my thesis present about the genome sequence of Diplorickettsia massiliensis strain 20B which is an obligate intracellular, gram negative bacterium isolated from Ixodes ricinus ticks collected from Slovak. In the third part, we investigated the genome repertoire of Diplorickettsia massiliensis compared to closely related bacteria according to its niche, revealing its allopatric lifestyle. In this study, we compared the genomic features of Diplorickettsia massiliensis with twenty-nine sequenced Gammaproteobacteria species (Legionella strains, Coxiella burnetii strains, Francisella tularensis strains and Rickettsiella grylli) using multi-genus pangenomic approach. This thesis work provides original data and sheds light on intracellular bacterial diversity.

This study focuses on a secondary contact zone between two sister species of woodrat, Neotoma fuscipes (dusky-footed woodrat) and N. macrotis (big-eared woodrat). Along the Nacimiento River, on the border of southern Monterey and northern San Luis Obispo counties, the ranges of these sister species of woodrats meet and overlap forming a secondary contact zone. The zone of secondary contact is estimated to include a 500-meter (~1,650 linear feet) portion of the Nacimiento River riparian corridor. This research examines quantifiable morphological change that is likely associated with heightened inter-specific competition within the contact zone. When in sympatry the sister species may compete for resources indirectly through exploitative competition, or directly through contest competition, or through a combination of these two processes. The prediction that heightened competition has resulted in distinctive morphological character shifts between allopatric and sympatric populations was tested my examining size and shape of adult woodrats along a 20-kilometer transect. It was confirmed that adults woodrats of the two sister taxa are morphologically distinct (N = 602) and that the phallus morphology was indeed a reliable means to identify adult male woodrats as to species (p < 0.0001, N = 331). A two model approach was used to examine convergence and divergence in size and shape of woodrats across the transect. Neotoma fuscipes exhibited a statistically significant divergerence from N. macrotis with regard to breadth of rostrum (p < 0.0001, N = 414) in a region of sympatry along the Nacimiento River. Based on the results on one statistical model, N. macrotis exhibited a statistically significant convergence with regard to body-size (p = 0.0240, N = 587) and length of hind foot (p < 0.0001, N = 563) towards those of N. fuscipes between zones of sympatry and allopatry. Alternatively, based on the results of a second statistical model that accounted for environmental variation within the system both species exhibited a statistically significant divergence with regard to body-size (p = 0.0054, N = 587) and towards that of N. fuscipes between zones of sympatry and allopatry. Also, N. macrotis exhibited a statistically significant convergence with regard to length of ear (p = 0.0022, N = 563) towards that of N. fuscipes. Based on the results of both models, detectable re-patterning of size-independent traits was observed to varying degrees. The morphological character shifts between sympatric populations and allopatric populations of woodrats suggest that ecological interactions between the species are occuring. Specifically, across the contact zone, patterns of variation in body-size and other morphological character traits are consistent with expectations of a combination of contest and exploitative competition.

Une identification précise des espèces exotiques est essentielle afin de déterminer la nature et l’ampleur des impacts que ces espèces auront sur leurs nouveaux habitats. Le complexe d’Arion subfuscus s. l., originaire d’Europe, fait partie des limaces les plus abondantes dans le nord-est de l’Amérique du Nord et plusieurs impacts dus à leur présence ont été rapportés. Cependant, l’identité des espèces introduites demeure inconnue dans la plupart des régions. L’objectif de ce projet est donc de déterminer la répartition récente, la diversité taxonomique et l’origine des membres du complexe d’A. subfuscus s. l. au Québec en se basant sur leur identité mitochondriale (16S rDNA). Un total de 526 spécimens provenant de 68 sites à travers le Québec et un site en Nouvelle-Écosse ont été analysés à l’aide de la technique des SSCP et leurs séquences ont été déterminées. Huit haplotypes de deux espèces allopatriques, A fuscus et A. subfuscus s. s. (lignées S1 et S2) ont été détectés. Les résultats confirment que des limaces provenant de régions distinctes d’Europe ont été introduites à de multiples reprises. Une comparaison avec des données historiques de répartition a révélé une expansion fulgurante de la répartition depuis les 50 dernières années. Arion fuscus est la principale espèce envahissante qui a été détectée dans toutes les régions échantillonnées, ce qui contraste avec les études antérieures réalisées ailleurs en Amérique du Nord. Le rôle potentiel des échanges commerciaux internationaux dans l’histoire d’introduction des espèces exotiques est discuté. / Accurate identification of exotic species is required to assess the magnitude and nature of consequences on their new habitats. The Arion subfuscus s. l. species complex comprised slugs of European origins that are amongst the most abundant slug species in northeastern North America and various impacts of their presence are reported. However, the identities of the species introduced remain unknown in most regions. This study aims at determining the current distribution, taxonomic identity and the origins of the members of the A. subfuscus s. l. complex in Quebec (Canada) based on mitochondrial 16S rDNA. A total of 526 specimens from 68 locations throughout Quebec and one site in Nova Scotia were SSCP analysed and their sequences were determined. Eight haplotypes of the allopatric A. fuscus and A. subfuscus s. s. (lineages S1 and S2) were detected. Results confirmed that slugs from distinct European regions were introduced multiple times. Comparison with previous survey revealed an impressive expansion of the distribution during the last 50 years. Arion fuscus is the major invasive species found throughout Quebec, contrasting with previous North American studies. The potential role of international trade in the introduction history of exotic species is discussed.