In 2004, a population of dwarf longnose sucker was discovered co-existing with the normal form within the Elk River Watershed of south-eastern British Columbia. This thesis evaluated morphological, genetic and life history characteristics of this dwarf longnose sucker form to determine whether the dwarf morphotype warranted designation as an evolutionary significant unit and to determine any special habitat requirements. In addition to size, distinct morphological differences were indicated between Elk River Watershed dwarf and normal adult longnose sucker, with dwarf adults appearing to retain morphological features of juveniles and sharing morphological features with Salish sucker, which is a separate dwarf longnose sucker form that is considered endangered. Slight, but significant, genetic differences were indicated between Elk River Watershed dwarf and normal longnose sucker forms, and compared to Salish sucker, suggesting some basis for separate designation of the dwarf form. Dwarf longnose sucker are widespread in the Elk River Watershed, and most abundant in small, cool lentic water bodies that contained dense vegetative cover, potential oxycline fluctuation and/or limited fish species diversity. Dwarf adult longnose sucker showed some habitat preference differences compared to normal longnose sucker, with the findings suggesting that dwarf longnose sucker have adopted a more opportunistic life-history strategy than normal longnose sucker. It is postulated that an ontogenetic niche shift has allowed dwarf longnose sucker to more successfully exploit habitats experiencing periodic disturbances (e.g., hypoxia) that, in turn, has led to the occurrence of two longnose sucker morphotypes in the Elk River Watershed.

Fusarium head blight (FHB), a fungal disease caused principally by Fusarium graminearum, and barley yellow dwarf (BYD) caused by BYD luteoviruses are two serious fungal and viral diseases of wheat resulting in high economic losses annually. Wuhan, a Chinese wheat cultivar resistant to FHB, and Maringa, a Brazilian cultivar tolerant to BYDV were inter-crossed and crossed with Roblin, a Canada western red spring wheat susceptible to both FHB and BYDV, to determine the genetic basis of resistance/tolerance and to combine the two traits. Four hundred ninety nine F1-derived doubled haploid (DH) lines were generated from reciprocal crosses using corn pollen-mediated DH technology. The DH lines and the parents were evaluated for disease symptoms, reduction in height and spike mass for BYD and for disease incidence, disease severity and Fusarium-damaged kernels for FHB in field and controlled environments. A subset (20/150) of the best performing DH lines from Wuhan/Maringa populations for both BYD and FHB were further evaluated. Plants were point inoculated with F. graminearum in greenhouse experiments, and macroconidial spray inoculations and spread of corn inoculum were used in field environments to evaluate FHB. BYDV inoculations were performed by placing ten to fifteen viruliferous aphids (Rhopalosiphum padi infected with BYDV-PAV isolate 9301PAV), at the one to two leaf stage for both greenhouse and field trials. The studies showed that both FHB and BYDV are quantitatively inherited. Transgressive segregants were observed and the broad sense heritability was high (0.90 to 0.97) for all traits evaluated. Results from independent testing of diseases on Wuhan/ Maringa populations showed fourteen DH lines were as, or more resistant than Wuhan for FHB and Maringa for BYDV tolerance and have combined both BYDV tolerance and FHB resistance. Identifying such lines facilitates the pyramiding of independent genes to obtain adequate levels of enduring resistance. A further experiment was conducted on the 14 lines by inoculating them with BYDV and F. graminearum successively on the same plant. Six out of 14 selected DH lines demonstrated high resistance to FHB and tolerance to BYDV. These six lines can be used in FHB/BYDV resistance/tolerance breeding programs.

Luminous Blue compact galaxies (BCGs) are metal-poor actively star-forming systems, characterised by bright ultraviolet and blue luminosities. Hubble Space Telescope high-resolution data have revealed that the luminous star-forming knots in these galaxies are composed of hundreds of young massive star clusters. In this work we present a systematic study of the star cluster populations in BCGs with important implications for the formation history of their host systems. The studied galaxies show recently increased star formation rates and a high fraction of massive clusters, probably as a result of minor/major merger events. The age distributions have a peak of cluster formation at only 3 - 4 Myr, unveiling a unique sample of clusters still partially embedded. A considerable fraction of clusters (30 - 50 %), mainly younger than 10 Myr, shows an observed flux excess between 0.8 and 2.2 μm. This so-called near-infrared (NIR) excess is impossible to reproduce even with the most recent spectral synthesis models (that include a self-consistent treatment of the photoionized gas). The origin of the NIR excess, which still remains unexplained, challenges our understanding of the cluster formation process under extreme conditions. The results achieved in this work have produced important insights into the cluster formation process in BCGs. We suggest that the BCG environment has most likely favoured the compression and collapse of giant molecular clouds into compact massive star clusters. The cluster formation efficiency (i.e., the fraction of star formation happening in star clusters) in BCGs is higher than the reported 8 - 10 %, for quiescent spirals and local star-forming galaxies. Luminous BCGs have a cluster formation efficiency comparable to luminous infrared galaxies and spiral starburst nuclei (the averaged value is  about 30 %), suggesting an important role of the merger event in the cluster formation. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 4: Accepted. Paper 5: Manuscript. Paper 6: Manuscript.

Over the past decade, plants have been used as expression hosts for the production of pharmaceutically important and commercially valuable proteins. Plants offer many advantages over other expression systems such as lower production costs, rapid scale up of production, similar post-translational modification as animals and the low likelihood of contamination with animal pathogens, microbial toxins or oncogenic sequences. However, improving recombinant protein yield remains one of the greatest challenges to molecular farming. In-Plant Activation (InPAct) is a newly developed technology that offers activatable and high-level expression of heterologous proteins in plants. InPAct vectors contain the geminivirus cis elements essential for rolling circle replication (RCR) and are arranged such that the gene of interest is only expressed in the presence of the cognate viral replication-associated protein (Rep). The expression of Rep in planta may be controlled by a tissue-specific, developmentally regulated or chemically inducible promoter such that heterologous protein accumulation can be spatially and temporally controlled. One of the challenges for the successful exploitation of InPAct technology is the control of Rep expression as even very low levels of this protein can reduce transformation efficiency, cause abnormal phenotypes and premature activation of the InPAct vector in regenerated plants. Tight regulation over transgene expression is also essential if expressing cytotoxic products. Unfortunately, many tissue-specific and inducible promoters are unsuitable for controlling expression of Rep due to low basal activity in the absence of inducer or in tissues other than the target tissue. This PhD aimed to control Rep activity through the production of single chain variable fragments (scFvs) specific to the motif III of Tobacco yellow dwarf virus (TbYDV) Rep. Due to the important role played by the conserved motif III in the RCR, it was postulated that such scFvs can be used to neutralise the activity of the low amount of Rep expressed from a “leaky” inducible promoter, thus preventing activation of the TbYDV-based InPAct vector until intentional induction. Such scFvs could also offer the potential to confer partial or complete resistance to TbYDV, and possibly heterologous viruses as motif III is conserved between geminiviruses. Studies were first undertaken to determine the levels of TbYDV Rep and TbYDV replication-associated protein A (RepA) required for optimal transgene expression from a TbYDV-based InPAct vector. Transient assays in a non-regenerable Nicotiana tabacum (NT-1) cell line were undertaken using a TbYDV-based InPAct vector containing the uidA reporter gene (encoding GUS) in combination with TbYDV Rep and RepA under the control of promoters with high (CaMV 35S) or low (Banana bunchy top virus DNA-R, BT1) activity. The replication enhancer protein of Tomato leaf curl begomovirus (ToLCV), REn, was also used in some co-bombardment experiments to examine whether RepA could be substituted by a replication enhancer from another geminivirus genus. GUS expression was observed both quantitatively and qualitatively by fluorometric and histochemical assays, respectively. GUS expression from the TbYDV-based InPAct vector was found to be greater when Rep was expected to be expressed at low levels (BT1 promoter) rather than high levels (35S promoter). GUS expression was further enhanced when Rep and RepA were co-bombarded with a low ratio of Rep to RepA. Substituting TbYDV RepA with ToLCV REn also enhanced GUS expression but more importantly highest GUS expression was observed when cells were co-transformed with expression vectors directing low levels of Rep and high levels of RepA irrespective of the level of REn. In this case, GUS expression was approximately 74-fold higher than that from a non-replicating vector. The use of different terminators, namely CaMV 35S and Nos terminators, in InPAct vectors was found to influence GUS expression. In the presence of Rep, GUS expression was greater using pInPActGUS-Nos rather than pInPActGUS-35S. The only instance of GUS expression being greater from vectors containing the 35S terminator was when comparing expression from cells transformed with Rep, RepA and REnexpressing vectors and either non-replicating vectors, p35SGS-Nos or p35SGS-35S. This difference was most likely caused by an interaction of viral replication proteins with each other and the terminators. These results indicated that (i) the level of replication associated proteins is critical to high transgene expression, (ii) the choice of terminator within the InPAct vector may affect expression levels and (iii) very low levels of Rep can activate InPAct vectors hence controlling its activity is critical. Prior to generating recombinant scFvs, a recombinant TbYDV Rep was produced in E. coli to act as a control to enable the screening for Rep-specific antibodies. A bacterial expression vector was constructed to express recombinant TbYDV Rep with an Nterminal His-tag (N-His-Rep). Despite investigating several purification techniques including Ni-NTA, anion exchange, hydrophobic interaction and size exclusion chromatography, N-His-Rep could only be partially purified using a Ni-NTA column under native conditions. Although it was not certain that this recombinant N-His-Rep had the same conformation as the native TbYDV Rep and was functional, results from an electromobility shift assay (EMSA) showed that N-His-Rep was able to interact with the TbYDV LIR and was, therefore, possibly functional. Two hybridoma cell lines from mice, immunised with a synthetic peptide containing the TbYDV Rep motif III amino acid sequence, were generated by GenScript (USA). Monoclonal antibodies secreted by the two hybridoma cell lines were first screened against denatured N-His-Rep in Western analysis. After demonstrating their ability to bind N-His-Rep, two scFvs (scFv1 and scFv2) were generated using a PCR-based approach. Whereas the variable heavy chain (VH) from both cell lines could be amplified, only the variable light chain (VL) from cell line 2 was amplified. As a result, scFv1 contained VH and VL from cell line 1, whereas scFv2 contained VH from cell line 2 and VL from cell line 1. Both scFvs were first expressed in E. coli in order to evaluate their affinity to the recombinant TbYDV N-His-Rep. The preliminary results demonstrated that both scFvs were able to bind to the denatured N-His-Rep. However, EMSAs revealed that only scFv2 was able to bind to native N-His-Rep and prevent it from interacting with the TbYDV LIR. Each scFv was cloned into plant expression vectors and co-bombarded into NT-1 cells with the TbYDV-based InPAct GUS expression vector and pBT1-Rep to examine whether the scFvs could prevent Rep from mediating RCR. Although it was expected that the addition of the scFvs would result in decreased GUS expression, GUS expression was found to slightly increase. This increase was even more pronounced when the scFvs were targeted to the cell nucleus by the inclusion of the Simian virus 40 large T antigen (SV40) nuclear localisation signal (NLS). It was postulated that the scFvs were binding to a proportion of Rep, leaving a small amount available to mediate RCR. The outcomes of this project provide evidence that very high levels of recombinant protein can theoretically be expressed using InPAct vectors with judicious selection and control of viral replication proteins. However, the question of whether the scFvs generated in this project have sufficient affinity for TbYDV Rep to prevent its activity in a stably transformed plant remains unknown. It may be that other scFvs with different combinations of VH and VL may have greater affinity for TbYDV Rep. Such scFvs, when expressed at high levels in planta, might also confer resistance to TbYDV and possibly heterologous geminiviruses.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2006. / Härtill 4 uppsatser.

Uma previsão fundamental do cenário L cold dark matter (LCDM) na formação de estruturas é que os halos galácticos de DM do tamanho da Via Láctea (MW) crescem pela acreção de subsistemas menores. Neste contexto, simulações deNcorpos prevêem um número de sub-halos de matéria escura, cujo tamanho e massa são comparáveis às galáxias anãs, que é muito maior do que as galáxias satélites conhecidas. Este é o que se convencionu chamar de problema das satélites faltantes (MSP). A interpretação mais popular do MSP é que os sub-halos de matéria escura menores são extremamente ineficientes na formação de estrelas, o que torna mais difícil detectá-los. Com a chegada dos grandes levantamentos fotométricos, nos últimos anos, a descoberta de uma nova população de galáxias satélites que orbitam a MW fornece evidências empíricas para acreditar que realmente existem várias galáxias pouco luminosas não detectadas ou que simplesmente habitam regiões no céu que ainda não foram observadas. Neste trabalho implementamos um algoritmo estatístico eficiente para detectar subestruturas ultra fracas da MW. O código, chamado SPARSEX, foi testado e otimizado usando um conjunto de objetos estelares previamente identificados nos dados do Sloan Digital Sky Survey (SDSS), conseguindo recuperar todos os objetos com sucesso. Além disso, a aplicação do algoritmo aos dados dos dois primeiro anos do Dark Energy Survey (DES) deu como resultado centenas de candidatos a sistemas estelares. Dezessete sistemas estelares detectados em comum com outras técnicas de busca por subestruturas, implementadas dentro do grupo de colaboração do DES, foram publicados em três artigos em 2015. Um dos candidatos publicados é o aglomerado estelar DES 1. DES1 foi detectado pelo código SPARSEX com uma alta significância estatística e aparece nas imagens do DES como uma concentração compacta de fontes pontuais azuis. O sistema estelar é consistente com uma população velha e pobre em metal. Assumindo dois diferentes perfis de densidade, os quais permitem estimar a probabilidade de que cada estrela realmente pertença ao sistema, determinamos uma distância heliocêntrica e uma magnitude absoluta total num intervalo de 77.6—87.1 kpc e 3.00 . MV . 2.21, respectivamente. O raio à meia-luz desse objeto, rh 9.88 pc, e a luminosidade são consistentes com um aglomerado estelar de baixa luminosidade do halo externo. DES1 também é x alongado (e 0.6), o que faz supor que este objeto está em estágio avançado de dissolução. Mais tarde, através de uma cuidadosa reanálise de nossos resultados, dois novos candidatos foram detectados, DES J01111341 e DES J0225+0304. Os candidatos estão localizados a uma distância heliocêntrica de 25 kpc e parecem também estar dominados por populações estelares velhas e pobres em metais. Suas distâncias ao plano orbital da galáxia anã de Sagitário, 1.73 kpc (DES J01111341) e 0.50 kpc (DES J0225+0304), indicam que eles estão possivelmente associados com a corrente da anã de Sagitário. O raio à meia-luz (rh ' 4.55 pc) e a luminosidade (MV ' +0.3) de DES J01111341 são consistentes com um aglomerado estelar ultra fraco, enquanto o raio à meia-luz (rh ' 18.55 pc) e a luminosidade (MV ' 1.1) de DES J0225+0304 colocam este objeto em uma região ambígua do plano tamanho-luminosidade entre aglomerados estelares e galáxias anãs. Determinações dos parâmetros característicos da corrente de Sagitário, tais como o espalhamento de metalicidade (2.18 . [Fe/H] . 0.95) e o gradiente de distância (23 kpc . D . 29 kpc), dentro da área amostrada do DES no hemisfério sul, também indicam uma possível associação com estes sistemas. Se esses objetos forem confirmados através de follow-up espectroscópico como sistemas ligados gravitacionalmente e compartilharem uma trajetoria Galáctica com a corrente de Sagitário, DES J01111341 e DES J0225+0304 seriam os primeiros sistemas estelares ultra fracos associados com tal corrente. Recentemente, nós reportamos a descoberta de um novo aglomerado estelar, DES 3. O novo sistema foi detectado como uma sobredensidade estelar nos dados do primeiro ano do DES e confirmado com follow-up fotométrico obtido com o Southerm Astrophysical Research (SOAR) Telecope. Nós determinamos que DES 3 está localizado a uma distância heliocêntrica de 76 kpc e é dominado por uma população velha (' 9.8Ganos) e pobre em metal ([Fe/H] ' 1.88). Embora os valores de idade e metalicidade de DES 3 são semelhantes aos aglomerados globulares, o seu raio à meia-luz (rh 6.5 pc) e a luminosidade (MV 1.9) são mais indicativos de um aglomerados estelar fraco. Com base no tamanho angular aparente, o DES 3, com um valor de rh 0.03, está entre os menores aglomerados estelares fracos conhecidos até à data. Estas novas detecções indicam que o censo de satélites da MW é ainda incompleto. A identificação e estudo de novos satélites em futuros surveys, como por exemplo o Large Synoptic Survey Telescope (LSST) será crucial para a nossa compreensão das subestruturas existentes no halo Galáctico e a evolução da Galáxia como um todo. / A fundamental prediction of the L cold dark matter (LCDM) scenario of structure formation is that galactic DM haloes of the size of the Milky Way (MW) grow by the accretion of smaller sub-systems. In this context, Nbody simulations predict a number of dark matter subhalos, with size and mass comparable to dwarf galaxies, but which is much larger than currently known satellite galaxies. This is what is conventionally called the missing satellites problem (MSP). The most popular interpretation of the MSP is that the smaller dark matter subhalos are extremely inefficient in star formation, making it more difficult to detect them. With the arrival of large photometric surveys, in recent years, the discovery of a new population of satellite galaxies orbiting the MW provides empirical evidence to believe that there are actually several low luminosity galaxies that were not yet detected or that simply inhabit regions in the sky that have not yet been observed. In this work we present an efficient statistical algorithm to detect ultra-faint MW substructures. The code, called SPARSEX, was tested and optimized using a set of stellar objects previously identified in the Sloan Digital Sky Survey (SDSS) data. It has detected successfully all known objects. In addition, the algorithm was applied on the first two years of Dark Energy Survey (DES) data, resulting in hundreds of stellar system candidates. Seventeen stellar systems detected in common with other substructure search techniques implemented within the DES collaboration were published in three papers in 2015. In particular, one published candidate is a star cluster, DES 1. DES 1 was detected by the SPARSEX code with high statistical significance and appears in DES images as a compact concentration of blue point sources. The stellar system is consistent with being dominated by an old and metal-poor population. Assuming two different density profiles, based on which we may evaluate a membership probability for each star, we determined a heliocentric distance and total absolute magnitude in the ranges of 77.6—87.1 kpc and 3.00 . MV . 2.21, respectively. The half-light radius of this object, rh 9.88 pc, and luminosity are consistent with a low-mass halo star cluster. DES1 is also elongated (e 0.6), which suggests that this object is in advanced stage of dissolution. Later, through a careful reanalysis of our results, two new candidates were detected, DES J01111341 e DES J0225+0304. The candidates are located at a heliocentric xii distance of 25 kpc and appear to have old and metal-poor populations as well. Their distances to the Sagittarius dwarf orbital plane, 1.73 kpc (DES J01111341) and 0.50 kpc (DES J0225+0304), indicate that they are possibly associated with the Sagittarius dwarf stream. The half-light radius (rh ' 4.55 pc) and luminosity (MV ' +0.3) of DES J01111341 are consistent with it being an ultra-faint star cluster, while the half-light radius (rh ' 18.55 pc) and luminosity (MV ' 1.1) of DES J0225+0304 place it in an ambiguous region of size-luminosity space between star clusters and dwarf galaxies. Determinations of the characteristic parameters of the Sagittarius stream, such as its metallicity spread (2.18 . [Fe/H] . 0.95) and distance gradient (23 kpc . D . 29 kpc), within the DES footprint in the Southern hemisphere, also indicate a possible association with these systems. If theses objects are confirmed through spectroscopic follow-up to be gravitationally bound systems and to share a Galactic trajectory with the Sagittarius stream, DES J01111341 and DES J0225+0304 would be the first ultra-faint stellar systems associated with the Sagittarius stream. Recently, we reported the discovery of a new star cluster, DES 3. The new system was detected as a stellar overdensity in first-year DES data, and confirmed with deeper photometry from the Southern Astrophysical Research (SOAR) telescope. We determine that DES 3 is located at a heliocentric distance of 76 kpc and it is dominated by an old (' 9.8 Gyr) and metal-poor ([Fe/H] ' 1.88) population. While the age and metallicity values of DES 3 are similar to globular clusters, its half-light radius (rh 6.5 pc) and luminosity (MV 1.9) are more indicative of faint star clusters. Based on the apparent angular size, DES 3, with a value of rh 0.03, is among the smallest faint star clusters known to date. These new detections indicate that the MW satellites census is still incomplete. The identification and study of new satellites in future surveys, such as the Large Synoptic Survey Telescope (LSST), will be crucial to our understanding of substructures in the Galactic halo and the evolution of the Galaxy as a whole.

abstract: Over the past decade, several high-value proteins have been produced using plant-based transient expression systems. However, these studies exposed some limitations that must be overcome to allow plant expression systems to reach their full potential. These limitations are the low level of recombinant protein accumulation achieved in some cases, and lack of efficient co-expression vectors for the production of multi-protein complexes. This study report that tobacco Extensin (Ext) gene 3' untranslated region (UTR) can be broadly used to enhance recombinant protein expression in plants. Extensin is the hydroxyproline-rich glycoprotein that constitutes the major protein component of cell walls. Using transient expression, it was found that the Ext 3' UTR increases recombinant protein expression up to 13.5- and 6-fold in non-replicating and replicating vector systems, respectively, compared to previously established terminators. Enhanced protein accumulation was correlated with increased mRNA levels associated with reduction in read-through transcription. Regions of Ext 3' UTR essential for maximum gene expression included a poly-purine sequence used as a major poly-adenylation site. Furthermore, modified bean yellow dwarf virus (BeYDV)-based vectors designed to allow co-expression of multiple recombinant genes were constructed and tested for their performance in driving transient expression in plants. Robust co-expression and assembly of heavy and light chains of the anti-Ebola virus monoclonal antibody 6D8, as well as E. coli heat-labile toxin (LT) were achieved with the modified vectors. The simultaneous co-expression of three fluoroproteins using the single replicon, triple cassette is demonstrated by confocal microscopy. In conclusion, this study provides an excellent tool for rapid, cost-effective, large-scale manufacturing of recombinant proteins for use in medicine and industry. / Dissertation/Thesis / Ph.D. Plant Biology 2012

White dwarf stars are the most common final stage of stellar evolution, corresponding to 99% of all stars in the Galaxy. White dwarf models can be used to obtain the age of stellar populations, to build an initial to final mass relation to understand the connection between the properties of white dwarfs and their progenitors, determine the upper mass limit that separates white dwarfs progenitors from Type II supernovae, enhance the comprehension of the physical properties of high density matter and derive ages and masses for observed white dwarfs from the cooling tracks. The literature is populated with low mass and intermediate mass white dwarf models, however the massive white dwarfs are often forgotten and the evolutionary sequences are incomplete. In this dissertation, we compute full evolutionary sequences for massive white dwarfs, exploring the evolution of hydrogen-rich and hydrogen-deficient white dwarfs stars with masses between 1.012 and 1.307 M , and initial metallicity of Z = 0.02. These sequences are the result of main sequence stars with masses between 8.8 and 11.8 M . The simulations were performed with the Modules for Experiments in Stellar Astrophysics - MESA code, starting at the zero-age main sequence, through thermally pulsing and mass-loss phases, ending as the white dwarfs at the cooling sequence. Our simulations are full evolutionary, in which we consider the entire evolutionary history of the progenitors. We present reliable nuclear chemical profiles for the whole mass range considered, covering the different expected central compositions, i.e. C/O, O/Ne and Ne/O/Mg, and their dependence with the stellar mass. In addition, we present detailed chemical profiles of hybrid C/O-O/Ne core white dwarfs, found in the mass range between 1.024 and 1.150 M . We present the initial-to-final mass relation, mass-radius relation, and cooling times with improved crystallization limits, considering the effects of atmosphere and core composition.

O objetivo desta tese é analisar aspectos do sistema de satélites da Via Láctea de relevância cosmológica. Dentre estes aspectos destacam-se dois: o censo de satélites da Galáxia – onde constata-se que a quantidade destes objetos é muito inferior ao predito por modelos cosmológicos do tipo Matéria Escura Fria – e a frequência anômala de satélites luminosos, como a Pequena e Grande Nuvem de Magalhães (SMC e LMC respectivamente). Além disso, a determinação dos parâmetros estruturais da LMC pode impor vínculos a sua formação, histórico orbital e sobre a massa de nossa Galáxia. Neste trabalho é desenvolvida uma técnica de busca por satélites da Via Láctea. Esta técnica foi otimizada para utilizar dados da nova geração de grandes surveys de maneira eficiente. Este código, o FindSat, foi validado em uma amostra de galáxias anãs conhecidas e se mostrou eficiente em detectar as galáxias anãs mais tênues de que se tem registro. A aplicação desse código a uma região do Sloan Digital Sky Survey (SDSS) ainda não explorada nesse sentido revelou uma série de candidatos a novos satélites. Foram selecionados os candidatos mais promissores para observação de follow-up. Estas observações revelaram que um destes candidatos é de fato um novo satélite da Via Láctea. Este novo objeto é muito provavelmente um aglomerado globular do halo em estágio avançado de dissolução, porém, seu tamanho e magnitude integrada colocamno em um domínio limítrofe entre aglomerado e galáxia anã. Através da cuidadosa análise dos demais candidatos, constatou-se que nenhum outro é de fato um novo satélite da Galáxia. Além disso, foi realizado o estudo do perfil de densidade e geometria da LMC. Este estudo utilizou dados de verificação científica do Dark Energy Survey (DES). Constatase que o perfil de densidades para estrelas jovens (< 3 Gyr) possui um raio de escala cerca de 50% menor que o da população velha (> 3 Gyr), favorecendo o cenário de formação tipo outside-in. O estudo da extensão da componente estelar da LMC revela um raio de maré de cerca de 18 kpc, permitindo o cálculo da massa dinâmica total da LMC. O valor de massa obtido favorece a hipótese onde as Nuvens de Magalhães estariam por sua primeira passagem pelo perigaláctico. Além disso, a distância heliocêntrica e espessura do disco da LMC foram determinadas utilizando estrelas do Red Clump (RC). Notou-se que regiões no extremo norte da LMC estão sistematicamente mais próximas de nós do que o esperado, este efeito evidencia o warp no disco dessa galáxia. Observou-se que a espessura do disco aumenta na periferia da LMC, caracterizando o fenômeno de flare. O aumento na espessura juntamente com a maior extensão da população velha da LMC é interpretado como a presença de dois componentes discoidais. Esta é a primeira evidência desse tipo baseada apenas em métodos de contagem de estrelas. / The goal of this thesis is to analyse comologically relevant aspects of the Milky Way (MW) satellite system. Among these we may highlight two: the census of MW satellites – where the observed number of these objects is much less than what is expected by Cold Dark Matter (CDM) models – and the anomalous frequency of luminous satellites, such as the Small and Large Magellanic Clouds (SMC and LMC respectively). Besides the cosmological importance of the Clouds, the determination of its structural parameters may help to constraint models for their formation, orbital history, and ultimately the mass assembly in our Galaxy. In this work a technique to search newMWsatellites is developed. This technique was optimized to run efficiently on large datasets, such as the ones being generated by the new generation of surveys. The code, FindSat was validated in a sample of well known MW satellites and has proven to be well succeeded even for the most faint of these objects. The application of this code to an unexplored region of the Sloan Digital Sky Survey (SDSS) revealed a large amount of new dwarf galaxy candidates, some of which where selected for follow-up observation. These observations led to the discovery of a new MW satellite. This new object is most likely a globular cluster in an extreme stage of dissolution. However, its integrated magnitude and size makes it difficult to discern it from a dwarf galaxy. By a careful analysis of the remaining candidates, it was shown that no other new satellite was in the sample. The density profile and geometry of the LMC was also analysed. This study used the recent science verification data from the Dark Energy Survey (DES). It was found that the density profile for young stars (< 3 Gyr) has a scale radius 50% smaller when compared to the one obtained for older stars (> 3 Gyr). This result favours the outsidein galaxy formation scenario. The total extension of the LMC stellar component was measured, allowing the estimate of a truncation radius of about 18 kpc. Assuming that this truncation has tidal origins the dynamical mass of the LMC is inferred. The mass value found favours the case for the first perigalactic passage of the Clouds. Besides that, the heliocentric distance and thickness of the LMC disk was determined using Red Clump (RC) stars. Evidence for warp was found in the North edge of the LMC, in the sense that the disk is systematically more distance than expected. While the thickness of the disk increases towards the outer parts of the LMC, which is a phenomena known as flare. This effect joined with the fact that the older LMC stellar population is more extended, favours the presence of two disk components in this galaxy. This is the first evidence of this kind based only on star counts.