Submitted by William Justo Figueiro (williamjf) on 2015-07-06T23:37:20Z No. of bitstreams: 1 10a.pdf: 1719495 bytes, checksum: 2ed1add8423e889edf7e44b9eb1498f1 (MD5) / Made available in DSpace on 2015-07-06T23:37:20Z (GMT). No. of bitstreams: 1 10a.pdf: 1719495 bytes, checksum: 2ed1add8423e889edf7e44b9eb1498f1 (MD5) Previous issue date: 2014-07 / Nenhuma / A baleia-franca-austral (Eubalaena australis) foi alvo de grande exploração comercial entre os séculos XVII a XX no Hemisfério Sul. Estimativas recentes sugerem que a população mundial da espécie é de 12.000 indivíduos, o que representaria 17 a 21% do seu tamanho original. Apesar de aparente recuperação, o declínio drástico no tamanho da população, resultante dos 400 anos de atividade da caça, pode ter gerado altos níveis de endogamia, baixa capacidade reprodutiva e perda de variabilidade genética da espécie, podendo ter seu potencial adaptativo comprometido, aumentando assim a probabilidade de extinção. Frente ao exposto é de extrema importância a avaliação comparativa da variabilidade genética entre as populações atuais de E. australis e amostras representativas do período de caça, em especial no Brasil. Sendo assim, os objetivos deste estudo foram: I. Otimizar e comparar métodos de extração e amplificação de DNA antigo (aDNA) de ossos de baleia-franca-austral, em diferentes estados de degradação; II. Caracterizar geneticamente segmentos da região D-Loop do DNA mitocondrial (mtDNA) a partir de ossos de espécimes de E. australis coletados na Península Valdés na Argentina e em Santa Catarina no Brasil; e III. Comparar as sequências nucleotídicas da região recuperada nas amostras de aDNA com as descritas para as populações atuais. Este estudo comparou cinco métodos diferentes para obter material genético de amostras degradadas. O método de extração que mostrou ser mais eficiente foi o que continha o reagente Dextran Blue. Com a amplificação de 20 fragmentos curtos, obtidos pelo Nested-PCR, com clonagem e posterior sequenciamento, foi possível recuperar pequenos fragmentos, que variaram de 7 a 59 pb, da região D-Loop do mtDNA de 11 amostras de ossos de E. australis das 88 tentativas realizadas. Ao analisar o material recuperado com dados atuais da espécie, o programa Network 4.6 revelou uma rede contendo 19 haplótipos, sendo quatro haplótipos restritos as populações antigas e até então não descritos na literatura. Os resultados são preliminares e seus significados devem ser avaliados com muita cautela. Contudo, deve-se ressaltar que os quatro novos haplótipos encontrados podem sustentar a hipótese de uma maior diversidade haplotípica em E. australis quando a mesma era caçada comercialmente. / The Southern Right Whale (Eubalaena australis) was object of great commercial exploitation between the XVII and XX centuries in the Southern Hemisphere. Recent surveys suggest that the world’s population is of 12,000 specimens, which would represent 17% to 21% of its original size. Despite the supposed recovery, the drastic populational decline, resultant from 400 years of hunting activities, could have generated high levels of endogamy, low reproduction capacity and loss of genetic variability of the species, compromising their potential of adjustment, thus increasing their probability of extinction. Due to the exposed, it is of extreme importance the comparative evaluation of the genetic variability between the current E. australis population and representative samples from the hunting period, especially from Brazilian coast. Therefore, the purpose of this study was: I. To optimize and compare extraction and amplification methods of aDNA from Southern Right Whale’s bones in different stages of deterioration; II. To genetically characterize segments of the mtDNA’s D-Loop region from E. australis bones collected at Península Valdés in Argentina and Santa Catarina in Brazil, and III. To compare nucleotide sequences of the recovered region from aDNA samples with those described for the current populations. This study compared five different extraction methods in order to obtain genetic material from deteriorated samples. The extraction method containing Dextran Blue reagent showed the higher efficiency. With the amplification of 20 short fragments and overlapping of aDNA through Nested-PCR technique, and further cloning of the latter fragment, it was possible to sequence small fragments, varying from 7 to 59 pb, from the mtDNA’s D-loop region from 11 bones samples of E. australis, out of the 88 attempts performed. Analyzing the recovered material together with the updated data of the species, the Network 4.6 program revealed a network containing 19 haplotypes, 4 of them generated only with ancient population samples, which are new for the species. The results are still preliminary and their meanings should be evaluated with caution. However, it should be noted that the four new haplotypes found could support the hypothesis of a higher haplotype diversity in E. australis when it was hunted.

Un dispositif expérimental a été développé pour l'étude du couplage de la croissance d'un biofilm de Shewanella oneidensis MR-1 et du transport conservatif de l'Erioglaucine. La croissance du biofilm a été suivie par mesure de conductivité hydraulique et par acquisition d'images à l'aide d'une caméra digitale. La fraction volumique du biofilm a été caractérisée par des essais d'élution d'une macromolécule (i.e. : le Bleu Dextran) par analogie avec les méthodes de chromatographie d'exclusion ou de filtration sur gel. Ainsi au bout de 29 jours, un biofilm quasi-homogène sur l'ensemble de la cellule d'écoulement (0,1×0,1×0,05m3) et équivalent à 50% du volume poral a été formé. L'influence de la croissance du biofilm sur les propriétés de transport du milieu a été évaluée. Les essais de transport conservatif de l'Erioglaucine effectués pour deux vitesses d'injection et à deux stades de croissance du biofilm (17 et 29 jours) ont montré l'influence d'hétérogénéités locales sur les paramètres de transport (i.e. : la porosité, la perméabilité et la dispersion hydrodynamique). Ainsi après 17 jours de culture quand le biofilm occupe partiellement le milieu poreux (moitié inférieure) un modèle à deux équations ou double milieu permet de caractériser le transport conservatif. A contrario après 29 jours de culture où le biofilm occupe tout le milieu poreux, un comportement fickien classique caractérise le transport. Les valeurs théoriques du coefficient de dispersion longitudinale prédites par la méthode de prise de moyenne volumique ont permis de reproduire de manière satisfaisante le comportement observé expérimentalement / An experimental device was performed for the study of coupling the growth of a Shewanella oneidensis MR-1 bacterial biofilm and the non reactive transport of Brilliant Blue FCF. The biofilm growth was monitored by hydraulic conductivity measurements and by image acquisition with a digital camera. The biofilm volume fraction was estimated through tracer experiments with a macromolecular tracer (i.e., Dextran Blue) as in size-exclusion chromatography or gel filtration chromatography. Then after 29 days of bacterial culture a quasi-homogenous biofilm was grown in the whole flow cell (0,1×0,1×0,05m3) occupying about 50% of void space volume. The influence of biofilm growth on porous media transport properties was evaluated. Conservative tracer experiments with Brilliant Blue FCF run at two hydrodynamic conditions and at two growth steps of biofilm (17 and 29 days) showed the influence of local heterogeneities on transport parameters (i.e., porosity, permeability and hydrodynamic dispersion). Then at 17 days of growth when the biofilm partially covers the porous medium (bottom half of the flow cell) a two-equation model or double-layer model was suitable to characterize the conservative transport. A contrario after 29 days of growth, when the biofilm covers the whole porous medium, a classical fickian model was convenient. Numerical values of longitudinal dispersion coefficient from volume averaging well fitted experimental results