O MHC (Major Histocompatibility Complex) é um sistema genético importante para a manutenção de espécies ameaçadas, uma vez que baixa variabilidade para locos MHC tem sido associada a uma menor capacidade de resposta a doenças e diminuição do sucesso reprodutivo. Deste modo, pesquisas sobre a variabilidade genética do MHC têm demonstrado ser bastante informativas em estudos populacionais voltados para aspectos referentes à conservação. No presente trabalho foi investigada a variabilidade genética do MHC para três espécies de mamíferos marinhos (toninha, baleia franca austral e lobo marinho sul-americano) do sul do Brasil, com intensa mortalidade provocada por atividades humanas atuais ou passadas. As amostras foram coletadas de animais mortos encalhados na costa, de animais capturados acidentalmente por barcos pesqueiros, e também através de um sistema de biópsia. A região variável do exon 2 do gene DQB do MHC foi amplificada por PCR (Polymerase Chain Reaction) em 109 amostras de toninhas (Rio de Janeiro n=32, Rio Grande do Sul n=52, Argentina n=25), 35 amostras de lobo marinho sul-americano e 30 amostras de baleia franca austral, utilizando-se um par de primers heterólogos. O fragmento resultante de 172 pares de bases foi analisado quanto ao polimorfismo de seqüência através da técnica de SSCP (Polimorfismo de Conformação de Fita Simples) em todas as amostras de toninha e de lobo marinho sul-americano e 14 amostras de baleia franca austral. Dificuldades associadas à amplificação resultaram em padrões de SSCP pouco informativos para as amostras de lobo marinho sul-americano e baleia franca austral Todas as amostras de toninha apresentaram um padrão de pelo menos 4 bandas por indivíduo. As 4 bandas de um único indivíduo do Rio Grande do Sul foram seqüenciadas, tendo sido possível verificar que 2 seqüências relacionadas ao genes DQB estão sendo amplificadas com estes primers. Pelas análises de SSCP foi possível detectar ausência de variabilidade para as amostras de toninha provenientes do Rio de Janeiro e diferenciá-las da população da Argentina, que é polimórfica. A população do Rio Grande do Sul parece apresentar níveis intermediários de variação em relação aos extremos da distribuição da espécie. Analisando as três populações amostradas, conclui-se que a espécie apresenta baixos níveis de variabilidade para o loco DQB, a exemplo do que é reportado para os genes de MHC de outros mamíferos marinhos.

O Lobo-Marinho-do-Sul (Arctocephalus australis), carnívoro, caniforme, membro da superfamília dos Focóides, família Otaridae, é o mais comum mamífero marinho pertencente à fauna brasileira. É freqüentemente avistado, nos meses de outono e inverno na costa do Brasil, desde o Rio de Janeiro até o Rio Grande do Sul. Ainda é discutido se algum dia este animal já possuiu colônias reprodutivas neste país. É histórica a grande pressão antrópica que esta espécie recebeu, sendo explorado pelas populações litorâneas, para aproveitamento de sua pele, óleo e carne desde a antiguidade. No Uruguai, foram por muitos séculos abatidos em números que variavam em torno de 10 mil animais por ano, com o intuito de aproveitamento de sua pele para a confecção de casacos e testículos, para confecção de elixires no Oriente. Porém, deixou de ser explorado comercialmente naquele país na década de 80 e desde então seu número vem aumentando, e por isso especula-se sobre o surgimento de novos sítios reprodutivos em ilhas pedregosas, típicas de sua preferência. É possível até que estuários como o de Cananéia no litoral sul do Estado de São Paulo - BR, possam receber esta espécie nos meses de verão, o que, de acordo com a sazonalidade de sua reprodução, ocorre nesta época. Esse novo "pólo reprodutivo" pode representar um desbravamento de novas áreas (ou mesmo retorno a antigas áreas) pela espécie. É ainda muito escassa a literatura sobre esta espécie e poucos pesquisadores brasileiros a conhecem. Este trabalho buscou através da coleta de indivíduos (cinco jovens e um adulto) em óbito recente, por causas naturais, contribuir com conhecimentos anatômicos, histológicos, ultraestruturais e imuno-histoquímicos do sistema reprodutor masculino do Arctocephalus australis de animais jovens e adultos. Os resultados demonstraram muitas vezes a singularidade da espécie estudada: expressiva queratinização da camada córnea do epitélio escrotal e forte coloração negra nesta região; diferença estrutural existente entre as túnicas albugínea do testículo e do pênis: na primeira observaram-se fibras elásticas bem distribuídas e ausência destas fibras na segunda; ausência de ampolas nos ductos deferentes; uretra pélvica pequena; bulbo do pênis ventral aos pilares penianos e posicionamento singular do seu osso peniano; glande do pênis petaliforme. Encontramos um grande grau de adaptação do sistema reprodutor do macho ao da fêmea, nas duas idades estudadas, demonstrando que a evolução foi direcionada para o ciclo reprodutivo do tipo monoéstrico-estacional, de modo a otimizar a cópula e o volume do ejaculado, garantindo a deposição do sêmem o mais próximo possível do óstio uterino externo da fêmea, e conseqüentemente uma maior taxa de concepção. / The Southern-Fur-Seal (Arctocephalus australis) Carnivora, Caniform, member of the Superfamily Focoidae, Family Otaridae, is the most common marine mammal in brazilian sees, frequently seen in the months of autumn and winter in the coast from Rio de Janeiro until Rio Grande do Sul. There are controversies if some day A. australis already possessed reproductive colonies in this country. The great antropic pressure over the species is historical, being explored by cost populations because of its skin, oil, and meat. In Uruguay, it was slaughtered for many centuries in numbers that reached 10 000 animals per year: the skin was used for the confection of coats and testis, for elixir in the East. However, since the beginning of the decade of 80, commercial exploitation stopped and its number started increasing. Therefore one speculates on the sprouting of new reproductive places in stony islands, typical of its preference, and it is possible that estuaries like Cananéia, in the south coast of the State of São Paulo - BR, started to receiving this species in the summer months, what is in accordance with the reproductive season, which occurs at this time of the year. It can represent an opening of new areas (or even a return the old areas) for the species. Literature is still very scarce on this species and few brazilian researchers know about it. For developing this work, six recent dead for natural causes individuals (five young and one adult) were collected and the reproductive tract was submitted to anatomical, histological, ultrastructural and immunohistochemistry approaches aiming to improve the knowledge about the male reproductive system of the Arctocephalus australis of two ages. Results pointed towards peculiarities of the studied species: expressive keratinization of scrotal epithelium; structural differences between the tunica albuginea in the testis and penis: only in the testis elastic fibers could be observed; no ductus deferens ampulae, small pelvic urethra; glans penis in flower shape. We found a great degree of adaptation of the reproductive system of the male to the one of the female, in the two ages studied, demonstrating that the evolutive orientation was in direction of sazonal-monoestric reproductive cycle, in order to optimize copulation and the volume of the semen, guaranteeing the deposition of the semen as next as possible to the external female uterine ostium and consequently a bigger tax of conception.

Salsola australis is an introduced weed of crop and pasture systems in the Western Australian broad acre cropping and pasture region (wheat-belt). This thesis investigated the classification, biology and ecology of the genus Salsola in southwest Australia, as well as modelling the effectiveness of possible weed control practices. Prior to this research, S. tragus was the only recognised species of the Salsola genus within Australia. However, genetic analysis revealed that four genetically distinct putative taxa of the genus Salsola were found in southwest Australia, none of which were S. tragus. The taxa that is the most prevalent agricultural weed was classified as S. australis, but the other three putative taxa could not be matched to recognised species. All four taxa were diploid (2n = 18), as opposed to tetraploid (2n = 36) S. tragus. Within the agricultural system of southwest Australia, S. australis plants established throughout the year, although the majority of seed production occurred in late summer and autumn. Total seed production (138-7734 seeds per plant) and seed viability (7.6-62.8%) of S. australis were lower than that reported for other agricultural weed species of the Salsola genus. Seed dispersal occurred when the senesced plants broke free of their root system to become mobile. Wind driven plants travelled and shed seed over distances of 1.6 to 1247.2 m. Movement of approximately half the plants was restricted to less than 100 m by entanglement with other S. australis plants within the stand. Some seed was retained on the senesced plants, but the germinability of this seed fell to less than 2% in the two month period following plant senescence (i.e. a decline of 79%). Once seed shed into the soil seed bank, anywhere from 32.3 to 80.7% of the viable seeds germinated in the year following seed production, with the rest remaining dormant or degrading. A model of the life cycle of S. australis based on the population ecology data indicated that the dormant seed bank had very little effect on annual seedling recruitment, but seed dispersal from neighbouring populations had a large impact on population growth rate. Therefore, the most successful weed control measures were those that restricted seed dispersal from neighbouring populations, or those that were applied to all populations in the region rather than to a single population. Weed control techniques applied to a single population, without reducing seed dispersal, could not reduce population size.

Severe population bottlenecks and the small size of many remnant habitats may render many bird populations prone to extinction from disease outbreaks. Bottlenecks may increase inbreeding which in turn may result in a low diversity of resistance and an immune system that is impaired or defective. Thus, bottlenecked populations may be less immunocompetent and more vulnerable to microbiological and parasitic perturbations. Few studies have assessed the effect of bottlenecks on the immunocompetence of birds. In this study, I used twelve saddleback (Philesturnus carunculatus) and two New Zealand robin (Petroica a. australis) populations, to determine if the severe bottlenecks reduce the immunocompetence of birds. When I experimentally challenged the immune system of two robin populations I found that despite the two populations having similar parasite loads, robins from the severely bottlenecked Motuara Island population exhibited a significantly lower T-cell mediated immune response than the source population (Nukuwaiata Island) suggesting that birds passing through severe population bottlenecks have a compromised immunocompetence. In the saddleback, severe bottlenecks, as well as high population densities and small island size, lead to individuals exhibiting higher stress levels and feather mite loads and lower immune function, as was evident by lower lymphocyte counts. I did not find levels of fluctuating asymmetry of saddlebacks to be directly influenced by bottleneck size. However, I did find that individuals with higher levels of fluctuating asymmetry had higher loads of hippoboscid flies and lower loads of coccidia suggesting a possible trade-off between growth and immune function. In contrast to previous studies looking at behavioural secondary sexual traits, I found no effect of founder number on the size of wattles in saddleback. I did however demonstrate that wattle size reflected the level of immune function in females as well as males, suggesting that females play a far greater role in offspring fitness than has been appreciated in traditional theories of sexual selection. Overall, my results indicate that severe bottlenecks can lead to reductions in immunocompetence in the resulting populations, especially in those populations that pass through the most severe bottlenecks. Based on the evidence from my thesis, I recommend conservation managers should aim to use at least 90 individuals to found new populations in order to reduce the deleterious effects of bottlenecks on immune function. If the costs of population bottlenecks and inbreeding are to be avoided, conservationists must adequately address the role of genetic factors in susceptibility to disease, and work towards minimising the risk of severe population bottlenecks in the management of endangered birds

The potential of kauri (Agathis australis) for paleoclimate research is well established. Multiple treering chronologies have been derived from living and sub-fossil material and growth-climate relationships have been identified. Work has progressed to the stage where raw ring-width data and chronologies covering the last half of the second millennium can confidently be placed in the public domain, to facilitate multiproxy paleoclimate studies. This paper outlines progress in deriving kauri tree-ring chronologies, summarises data availability and quality, and explores the scope for developing composite chronologies. Statistical quality control of the available data was undertaken, following application of an "optimum" standardisation technique. Variations in sample depth with time and between sites result in a complex evolving pattern of chronology quality across sites. Analysis of inter-site statistical relationships identified a pervasive regionalscale signal in kauri with some minor secondary patterns. In light of the strong common signal, a kauri master chronology was built by pooling tree-ring series. Analysis of the quality of this chronology indicates that high-quality master chronologies can be derived for A.D. 1597-1996 from as few as 25 trees from seven sites.

Considerable research has occurred in recent years to build Agathis australis (D. Don) Lindley (kauri) tree-ring chronologies for paleoclimate applications and to identify statistical relationships between kauri tree rings and climate. This paper reports on a multi-year study of the seasonal growth of kauri, designed to assist in the interpretation of identified statistical relationships, and to determine if kauri’s seasonal growth characteristics are dependent on tree size. To achieve this, 43 kauri (0.09-2.00 m diameter) at Huapai Scientific Reserve were fitted with vernier bands to measure circumference change over 3-4 growing seasons. Absolute (mm) and relative (proportion of total ring) monthly growth rates were calculated for each tree and statistics characterizing the timing of growth were calculated (e.g. date corresponding to 50% of growth). Tree size-related differences were assessed by splitting the data into three subsets based on size, then comparing the monthly growth rates and growth timing statistics for the subsets. The growth timing statistics were also correlated with tree diameter. A key finding is the strong dominance of spring growth, with October and November alone accounting for 38-50% of the total ring width. This result is consistent across age cohorts, although the largest trees tended to peak in November, rather than October. This indicates that kauri tree rings are likely to have value in terms of reconstructing spring conditions; consistent with reported statistical relationships between kauri tree rings and the El Niño-Southern Oscillation phenomenon. High inter-tree variance in growth rates characterized the results, but little of this variance was accounted for by tree size. Although relationships between tree size and growth characteristics were generally weak and inconsistent, they are considered sufficient to warrant a precautionary approach in the development of tree-ring chronologies for climate reconstruction purposes.

The traditional approach to standardizing tree-ring time series is to divide raw ring widths by a fitted curve. Although the derived ratios are conceptually elegant and have a more homogenous variance through time than simple differences, residual heteroscedasticity associated with variance dependence on local mean ring width may remain. Incorrect inferences about climate forcing may result if this heteroscedasticity is not corrected for, or at least recognized (with appropriate caveats). A new variance stabilization method is proposed that specifically targets this source of heteroscedasticity. It is based on stabilizing the magnitude of differences from standardization curves to a common reference local mean ring width and uses data pooled from multiple radii. Application of the method to a multi-site kauri (Agathis australis (D. Don) Lindley) data set shows that (a) the heteroscedasticity issue addressed may be generic rather than radius-specific, at least for some species, (b) variance stabilization using pooled data works well for standardization curves of variable flexibility, (c) in the case of kauri, simple ratios do not appear to be significantly affected by this cause of heteroscedasticity, and (d) centennial-scale variance trends are highly sensitive to the analytical methods used to build tree-ring chronologies.

O Complexo Principal de Histocompatibilidade (MHC) é um sistema genético reconhecidamente polimórfico em populações humanas. Em porções específicas dos genes que compõem o MHC de populações humanas também foi evidenciada a ação da seleção natural em nível molecular. Em várias espécies de vertebrados onde estes genes foram estudados diferentes níveis de variação, composição gênica e organização foram detectados, mantendo, entretanto, as características que os tornam tão interessante de ser estudados. Para mamíferos marinhos, as questões sobre a variação no MHC servem de pano de fundo para o entendimento sobre o sistema imunológico destas espécies e suas particularidades em relação ao ambiente marinho e a evolução destes genes em organismos não-modelos. No presente trabalho, duas espécies de cetáceos, a toninha ou franciscana (Pontoporia blainvillei) e a baleia-franca-austral (Eubalaena australis), foram investigadas quanto aos níveis de variação no gene DQB1 do MHC. O segundo exon da cadeia beta do gene DQ foi amplificado em 25 amostras de toninha e 18 amostras de baleia franca. Os fragmentos de 212pb (pares de base) resultantes do processo de amplificação foram clonados e, tiveram analisados por seqüenciamento 171pb correspondentes ao sítio de ligação ao peptídeo antigênico (PBR). Uma amostra de toninha e um par mãe-filhote de baleia-franca-austral foram analisadas quanto à expressão do gene DQB a partir de amostras de pele. A evidência de seleção natural a nível molecular nesta porção do gene é confirmada por um excesso de substituições não sinônimas em relação às substituições sinônimas (em um contexto de neutralidade as substituições sinônimas acumulam-se na mesma taxa de mutações) através da razão dN (não sinônimas) e dS (sinônimas). Para confirmar os desvios da neutralidade (Ho= dN/dS=1), foi usada a estatística z. Os valores de dN/dS para toninha e baleia-franca-austral foram, respectivamente, dN/dS = 2,32 (p=0,014) e dN/dS =3,75 (p= 0,001). Estes valores, as características das seqüências e a evidência de expressão desses genes na pele de cetáceos estão de acordo com os pressupostos de seleção positiva para esta região do gene e sugerem funcionalidade para estes genes nestas duas espécies. Há evidências de mais de uma cópia do gene para baleia-franca-austral. Foram descritos 6 alelos para toninha e 17 alelos para baleia-franca-austral sendo que um dos alelos é compartilhado entre as duas espécies, sugerindo que essa linhagem alélica é anterior à divergência entre odontocetos e misticetos. Paralelamente a este trabalho, o cariótipo de toninha foi descrito através de um processo de cultivo celular a partir da córnea de dois exemplares recém-mortos. Seu número cromossômico 2n=44 corrobora a estabilidade cariotípica descrita para cetáceos. / The Major Histocompatibility Complex is a highly polimorphic genetic system described to human populations. It is clear that a particular region from these genes is under natural selection which results in different patterns of variation from those expected under neutrality. In other vertebrates these genes were studied, these special characteristics seems to be preserved despite specific genomic organization and variation levels. In marine mammals, questions adressed to MHC molecular variation are focused in questions concerned on environmental restrictions and molecular evolution in non-model organisms. In the present study, two marine mammal species were surveyed concerning genetic variability using the gene DQB exon 2. Samples from 25 franciscana dolphins (Pontoporia blainvillei) and 18 southern right whales (Eubalaena australis) were used to amplify a 212bp (base pair) fragment from DQB exon 2. The 171bp resulting fragment was sequenced and analized in relation to the molecular level evidence of selection. Rates of nonsynonymous (dN) and synonymous (dS) base pair substitution and departures from neutrality for dN/dS (z statistics) were calculated. For both species examined, molecular evidence of selection was confirmed. The dN/dS ratios to franciscana and southern right whale were 2.32 (p=0.014) and 3.75 (p=0.001), respectively. These values, combined to data on DQB gene expression in franciscana and southern right whale skin and less restrictive aminoacid usage in both species, suggest a functional signature to sequences cetacean derived. Additionally, 6 alleles were described to franciscana dolphins and 17 to southern right whales. One more evidence of selection acting in this particular genomic region is provided from the fact that both species share one allele, suggesting that this allelic lineage was present before the splitting between mysticeti and odontoceti. To provide addtional information on franciscana dolphins the karyotype of P. blainvillei is presented. The karyotype was obtained from corneal culture of two dead animals. The diploid number 44 confirms the clear prevalence of this pattern among cetaceans.

Phragmites australis is a plant that is causing problems in wetlands by outcompeting native plants that provide food and shelter for millions of migratory birds. Currently, managers try to control Phragmites australis by spraying herbicide, burning, and mowing, but these methods are costly, time consuming, and have low levels of success. Adding grazing as a tool to control Phragmites australis provides a cheap and low labor alternative. However, there are many concerns regarding if grazing will cause nutrient loading in our wetlands that will decrease water quality and alter beneficial functions of wetlands. To better understand the effects of grazing in wetlands, we proposed a two-year study and received funding from many organizations including the Utah Department of Fire, Forestry, and State Lands, South Davis Sewer District, and the Utah Department of Environmental Quality and Water Quality. Also, the Utah Department of Natural Resources helped tremendously in allowing access to the sites, in the actual implementation of the project, coordinating with local ranchers who allowed for their cattle to be in the study, managed their cattle during the study, and assisted with fence installation, and many volunteers from Utah dedicated hunters helped with the fence installation. We collected water, manure, soil, and leaf samples over time to analyze nutrient changes and measured changes in the plants, water levels, soil cover, and litter cover over time. We then compiled and analyzed this information to better understand how grazing impacts our wetlands. As a result, we were able to make some recommendations for future research and how best to graze in wetlands with minimal impacts according to the information we found.

Copies of author's previously published articles inserted. Bibliography: p. 199-254. In eutherian mammals, the size of the testes and number of spermatozoa produced and stored in the excurrent ducts vary widely between species, with the hydromyine rodents of Australia exhibiting a greater range of interspecific variation than any other closely related group of species. This study compared the efficiency of germ cell production and sperm storage capacity in the extra-testicular ducts of two arid zone species, the plains rat, Pseudomys australis, and the spinifex hopping mouse, Notomys alexis, that have vast differences in testes size and number of stored spermatozoa. Results are discussed.