Os vírus possuem potencial de variabilidade genética muito alto, isso porque necessitam divergir seu material genético suficientemente para se adaptar às inúmeras mudanças às quais são submetidos. Portanto, a variabilidade genética é essencial para a sobrevivência desses organismos; é o primeiro passo para a adaptação em um novo hospedeiro, quebra de resistência, alterações nos sintomas e virulência, o que justifica o interesse em estudos nessa área. Os estudos de variabilidade consistem numa excelente ferramenta para a compreensão da evolução dos vírus e busca pelo manejo adequado de doenças virais. Por isso objetivou-se estudar a variabilidade genética da ORF p29 do CiLV-C, a fim de gerar informações relevantes acerca do patossistema e da preponderância de isolados, com possíveis implicações na epidemiologia da doença e seu manejo no campo, além de uma melhor compreensão sobre a evolução desse vírus, que até então nunca havia sido explorada. Neste trabalho foram avaliadas plantas de citros e outras hospedeiras potenciais do CiLV-C. Os resultados sugerem que as plantas de tangerina Cravo, Tardia da Sicília, Cleópatra, Vermelha, tangor Ortanique, laranja Azeda e trapoeraba são suscetíveis à doença e também podem servir como fontes de inóculo do vírus para citros. Já as plantas de limão Siciliano e Cravo, e limas ácidas Tahiti e Galego e Mimosa caesalpiniaefolia mostraram-se resistentes à doença, mas não à colonização do ácaro vetor. As plantas de Malvaviscus arboreus e Solanum violaefolium não apresentaram sintomas, mas mostraram-se possíveis fontes de inóculo do vírus para plantas de citros. Além disso, foram avaliadas as respostas de 62 genótipos de tangerinas e seus híbridos à doença, sendo que 15 mostraram-se resistentes e podem, posteriormente, ser utilizados em programas de melhoramento genético, que é uma das alternativas para reduzir o uso de pesticidas para o controle do vetor. Foi identificada baixa variabilidade genética entre os isolados do CiLV-C, independentemente do hospedeiro ou localidade, entretanto, o isolado de São José do Rio Preto pareceu ser o mais divergente e capaz de passar suas alterações durante sua transmissão a outros hospedeiros. Mais estudos devem ser feitos para que conclusões inquestionáveis sejam tiradas desse assunto, mas os resultados obtidos abriram um novo leque de possibilidades para futuros estudos nessa área até então pouco explorada. / Viruses have, potentially, broad genetic variability because of their need to adapt to several changes that they are exposed to. Therefore, genetic variability is essential for their survival; it is the first step to adapt to a new host, to break resistance down, to change symptoms and virulence, which justifies the interest in studies in this area. These studies consist in a great tool for a better understanding on the virus evolution and the search for a proper management of viral diseases. Hence, it was aimed to study the genetic variability of ORF p29 from CiLV-C in order to generate relevant information about the pathosystem and the predominance of isolates with possible implications on the epidemiology of the disease and its management in the field, besides a better understanding on the evolution of this virus, which has never been explored before. In this work, we evaluated citrus plants and potential hosts for CiLV-C. The results suggest that the plants of Cravo, Tardia da Sicília, Cleopatra, and Vermelha mandarin, Ortanique tangor, Sour orange and spiderwort are susceptible to the disease and can also serve as sources of inoculum of the virus to citrus. Siciliano lemon, Rangpur, Tahiti, and Mexican limes, and Mimosa caesalpiniaefolia were resistant to the disease, but not to the colonization of the mite vector. Malvaviscus arboreus and Solanum violaefolium plants did not present symptoms, but can be considered possible sources of CiLV-C inoculum to citrus plants. In addition, we evaluated the response of 62 mandarin genotypes and their hybrids to the disease. Fifteen of them were considered resistant and could be used in breeding programs with the objective to reduce the use of pesticides to control the vector. Low genetic variability was found amongst CiLV-C isolates, regardless of the host or geographic region; however, the São José do Rio Preto isolate was the most divergent and the changes in nucleotides were transmitted to the other hosts. Further studies should be conducted before unquestionable conclusions can be drawn from this issue, but the results obtained here have opened a new range of possibilities for future studies in this area so far almost unexplored.

The White-bellied Sea-Eagle Haliaeetus leucogaster is widespread in Australia, but has been the subject of conservation concern due to suggested localised declines and extinctions. Regionalised monitoring programmes have addressed some aspects of local concern, however a broader approach is needed to gain an understanding of large-scale processes affecting long-term persistence at scales equivalent to the species Australian range. Ultimately, the ability to predict change in population size over time accurately depends on the scale of analysis. By necessity, ecological studies using direct sampling techniques are often made across spatial scales smaller than a species geographic range and across relatively short time frames. This seems counter-intuitive considering that long-term species persistence is often dependent on large-scale processes. The principal aim of this thesis was to identify historical and contemporary forces responsible for the current pattern of population structure in H. leucogaster. This required a multi-scale approach, and the resulting research uses genetic, distributional and morphometric data. Haliaeetus leucogaster is a large territorial raptor that historically has been associated with coastal regions, lakes and perennial river systems. It has an extensive worldwide distribution from the western coast of India throughout the Indomalaysian region, Papua New Guinea and Australia. By virtue of the species' large-scale distribution, in Australia it is fairly cosmopolitan in its use of habitat and prey types. Haliaeetus leucogaster is monomorphic for adult plumage colouration, but in body size displays reversed sexual dimorphism with female birds significantly larger. A discriminant function based on 10 morphometric characters was 100% effective in discriminating between 19 males and 18 females that had been sexed using molecular genetic methods. Re-classification using a jackknife procedure correctly identified 92% of individuals. The discriminant function should be a viable alternative to genetic sexing or laparoscopy for a large proportion of individuals within the Australo-Papuan range of this species; and can also be used to identify a small proportion of "ambiguous" individuals for which reliable sexing will require those other techniques. I used mitochondrial (mtDNA) control region sequence data to investigate the current distribution of genetic variation in this species at the continental level and within and between specified regional units. I was specifically interested in identifying breaks in genetic connectivity between the west and east of the continent and between Tasmania and the Australian mainland. Overall, genetic diversity was low and there was no significant level of genetic subdivision between regions. The observed genetic distribution suggests that the population expanded from a bottleneck approximately 160 000 years ago during the late Pleistocene, and spread throughout the continent through a contiguous range expansion. There is insufficient evidence to suggest division of the population into different units for conservation management purposes based on the theoretical definition of the 'evolutionary significant unit'. It is clear from the analysis that there are signatures of both historical and contemporary processes affecting the current distribution. Given the suggestion that population expansion has been relatively recent, additional sampling and confirmation of the perceived pattern of population structure using a nuclear marker is recommended to validate conservation monitoring and management at a continental scale. To determine the existence of perceived population declines across ecological time scales, I analysed the Australian Bird Atlas Data to identify the extent and pattern of change in range and density of the species between three Atlas Periods (1901-1976, 1977-1981 and 1998-2001) using a new standardised frequency measure, the Occupancy Index (OI) for 1° blocks (approx. 100km2) across the continent. At the continental scale, there was no significant difference in the spatial extent of occupancy between Atlas Periods. However, there were considerable changes in frequency and range extent between defined regions, and there were distinct differences in the pattern of change in OI between coastal and inland blocks over time. Coastal blocks showed much more change than inland blocks, with a clear increase in the use of coastal blocks, accompanied by a decrease in inland blocks, during the 1977-1981 Atlas Period, relative to both other Atlas Periods. The over-riding factor associated with distributional shifts and frequency changes was apparently climatic fluctuation (the 1977-1981 period showing the influence of El Nino associated drought). The impression of abundance was strongly dependent on both the temporal and spatial scale of analysis. To test for correspondence between geographic variation in morphology and geographic variation in mtDNA I analysed morphometric data from 95 individuals from Australia and Papua New Guinea. First, the degree of morphometric variation between specified regions was determined. This was then compared with the pattern of genetic differentiation. There was a strong latitudinal cline in body dimensions. However, there was no relationship between morphometric variation and patterns of genetic variation at least for mtDNA. Females showed a pattern of isolation by distance based on morphometric characters whereas males did not. Three hypotheses to explain the pattern of morphometric variation were considered: phenotypic plasticity, natural selection and secondary contact between previously isolated populations. I conclude that the pattern of morphometric variation is best explained by the suggestion that there is sufficient local recruitment for natural selection to maintain the observed pattern of morphometric variation. This implies that gene flow may not be as widespread as the mtDNA analysis suggested. In this instance either the relatively recent colonisation history of the species or the inability of the mtDNA marker to detect high mutation rates among traits responsible for maintaining morphometric variation may be overestimating the levels of mixing among regions. As might be expected given the physical scale over which this study was conducted, the pattern of genetic, morphometric and physical distribution varied dependent on the scale of analysis. Regional patterns of genetic variation, trends in occupancy and density and morphometric variation did not reflect continental patterns, reinforcing the contention that extrapolation of data from local or regional levels is often inappropriate. The combined indirect methodologies applied in this study circumvent the restrictions imposed by direct ecological sampling, because they allow survey across large geographic and temporal scales effectively covering the entire Australian range of H. leucogaster. They also allow exploration of the evolutionary factors underpinning the species' current distribution.

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