Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves 130-145). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves 130-145). Also available on the Internet.

Orientador: Juliana Campos Junqueira / Banca: Antonio Olavo Cardoso Jorge / Banca: Silvana Soléo Ferreira dos Santos / Banca: Renata de Oliveira Mattos Graner / Banca: Maria José Soares Mendes Gianinni / Resumo: O estudo da atividade inibitória de Lactobacillus pode contribuir na descoberta de novas estratégias terapêuticas nas infecções por Candida. Nesse contexto, o objetivo desse estudo foi isolar e identificar Lactobacillus da cavidade bucal de indivíduos livres de cárie e avaliar seu potencial de inibição de C. albicans por meio de estudos in vitro e in vivo. Primeiramente, foram avaliados os efeitos de 30 isolados clínicos de Lactobacillus sobre o número de células viáveis (UFC) em biofilme de C. albicans e sobre a formação de hifas. Os isolados que obtiveram os maiores efeitos inibitórios sobre C. albicans foram selecionados para os testes de determinação da biomassa total dos biofilmes pela absorbância do cristal violeta, análise da arquitetura dos biofilmes por microscopia eletrônica de varredura (MEV) e quantificação da expressão de genes de C. albicans (ALS3, HWP1, EFG1 e CPH1) por qPCR. Esses isolados também foram submetidos a estudos in vivo usando os modelos de Galleria mellonella e Caenorhabditis elegans. Para o estudo em G. mellonella, a infecção experimental foi avaliada pela curva de sobrevivência, quantificação da carga fúngica na hemolinfa, densidade hemocitária, quantificação da expressão gênica de peptídeos antifúngicos (Gallerymicina e Galiomicina) e monitoramento da infecção de C. albicans por análise de bioluminescência. No modelo de C. elegans, a infecção foi avaliada por meio dos ensaios de curva de sobrevivência e estudo da filamentação de C. albicans. Os ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The study of the antifungal activity of Lactobacillus may contribute to the discovery of new therapeutic strategies for Candida infections. In this context, the objective of this study was to isolate and identify Lactobacillus from the oral cavity of caries-free subjects and to evaluate its effects through in vitro and in vivo studies. First, the effects of 30 clinical isolates of Lactobacillus on the number of viable cells (CFU) in biofilms of C. albicans and on hyphae formation were evaluated. The isolates that obtained the highest inhibitory effects on C. albicans were selected for biofilm biomass determination by violet crystal absorbance, analysis of biofilm architecture by scanning electron microscopy (SEM) and quantification of the expression of C. albicans (ALS3, HWP1, EFG1 and CPH1) by real time PCR. These isolates were also submitted to in vivo studies using the Galleria mellonella and Caenorhabditis elegans models. For the study in the model of Galleria mellonella, the experimental infection was evaluated by the survival curve, quantification of the fungal load in the hemolymph, hemocitary density, the gene expression of antifungal peptides (Gallerymicin and Galiomicin) and monitoring of C. albicans infection by bioluminescence analysis. In the Caenorhabditis elegans model, the infection was evaluated by the survival curve assays and the study of C. albicans filamentation. The results of in vitro tests demonstrated that L. paracasei 28.4, L. rhamnosus 5.2 and L. fe... (Complete abstract click electronic access below) / Doutor

The success of an invasive alien species is often attributed to the ecological advantage gained from natural enemy release. Numerous factors have been suggested as contributing to the success of Harmonia axyridis as an invasive alien species, including enemy release. This thesis studied the interactions of several parasites with H. axyridis, investigating parasite transmission, growth and virulence as well as host immune responses, thereby shedding light on the potential role of enemy release in the invasion biology of this ladybird. Benefits gained by invasive alien species from enemy release diminish if parasites of native species shift hosts to exploit the novel invader. The fungal ectoparasite Hesperomyces virescens began infecting H. axyridis shortly after it invaded the UK, probably as a result of a host shift from Adalia bipunctata. This study found a rapid increase in H. virescens prevalence over three years in London H. axyridis populations. Laboratory study showed H. virescens transmission and growth to be more efficient on A. bipunctata than the novel host. In addition, reciprocal interspecific transfers of H. virescens strains isolated from A. bipunctata and H. axyridis revealed that the infection characteristics of the fungi from these two hosts differed, suggesting strains may have diverged after the initial shift from A. bipunctata to better exploit the host from which they were derived. Laboulbenialian fungi were previously thought to have negligible impacts on host fitness. A detailed examination of H. virescens infecting H. axyridis found distinct virulence, with infections resulting in a 50% reduction in host lifespan. In addition, chronic H. virescens infection in males caused acceleration in the age-associated decline in body condition while for females, infection triggered fecundity senescence and a faster age-related decline in fertility. While their role in accelerating ageing is debated, the results presented here provide evidence that infectious diseases can drive the ageing process in this insect species. In nature, multiple parasites affecting a single host are common. The effect of co-infection on the virulence caused by two fungal infections was characterised using H. axyridis and A. bipunctata hosts. The ability of two ladybird species to defend against an acute fungal parasite, while infected with the relatively avirulent H. virescens was found to be sex-specific. While for females, the presence of co-infection did not alter the virulence seen in singly infected females, a higher mortality rate existed for co-infected males compared with those infected singly. Previously, H. virescens has been considered to be avirulent, however, this study provides evidence that this chronic fungal parasite may be important when considering the mortality associated with co-infections in the field. The invasive success of H. axyridis has, in part, been attributed to a more vigorous immune ability compared with other competitor species. Previously, field studies have shown that the prevalence of the parasitoid wasp Dinocampus coccinellae in H. axyridis is considerably lower than in the UK primary host of this wasp, Coccinella septempunctata. The extent to which the prevalence asymmetry in the field is driven by differences in host encapsulation response was tested by first comparing the encapsulation ability of C. septempunctata and H. axyridis directed against an artificial implant. Following this, the encapsulation response of D. coccinellae parasitized individuals was assessed and compared between the two host species. While encapsulation ability did not differ between the host species, and D. coccinellae did not affect the immune response of H. axyridis, wasp parasitism did alter the encapsulation ability of C. septempunctata, although it was inconsistent across sexes and populations. Overall, this thesis furthers our understanding of the fungal parasite H. virescens and its association with the notorious invader H. axyridis. The research presented here also demonstrates the use of H. axyridis as a model system in areas other than invasion ecology and furthermore, contributes to understanding the role of infectious disease in the rate of ageing. Finally, sex-specific effects were found across the chapters of this thesis, demonstrating the use of H. axyridis in the study of sex-specific effects of infections.

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This thesis investigates aspects of the community ecology of rabbit parasites with particular emphasis upon the gut helminths, utilising a 23 (later extended to 26) year time series of rabbits and their parasites. A clearer understanding of parasite communities can lead to more effective biological control strategies. Rabbits are regarded as a serious pest species throughout Europe and the Antipodes and the use of the myxomatosis virus, as a biological control agent, has already been tried and failed. However, a clearer picture of the parasite community may offer future possibilities for control. Additionally, the rabbit is a good model for other grazing species, as it carries a similar gut helminth community. Drug resistance is an increasing problem in a wide range of parasites. A clearer appreciation of parasite communities could also aid in the search for effective and environmentally sound pathogen control strategies (e.g. via cross immunity or competition with benign species). Theoretical models have revealed the importance of aggregation to the stability of the host parasite relationship, to parasite evolution and to interspecific parasite interactions. A number of models have considered the effect of varying aggregation upon these dynamics with differing outcomes to those where aggregation was a fixed parameter. Here the stability of the distribution for each of the rabbit helminths was examined using Taylor's power law. The analyses revealed that aggregation was not a stable parameter but varied with month, year, host sex, host age, and host myxomatosis status. Evidence for the existence of interspecific parasite interactions in natural systems has been equivocal. Factors influencing parasite intensity were evaluated for the gut helminth. A network of potential interactions between the parasites was revealed. Only month was shown to be of greater influence on the community. Following, from the above analyses, a community model was constructed which incorporated both seasonal forcing and interspecific parasite interactions, with interaction mediated via host immunity. One unexpected emergent property was an interaction between the seasonality and the immune decay rate with slower immune decay resulting in a shift of the immune response out of phase with the species against which it was produced. The model was also used to assess the potential effects of two control strategies, an anticestodal and a single species vaccine. The vaccine had greater effects on the whole community than the anticestodal because of the immune- mediated interactions. The host is also an integral part of the community as the parasite dynamics are linked with that of their host. Therefore an assessment of the parasites' impact upon host condition and fecundity was also undertaken. This revealed a variety of positive and negative associations between the parasites and their host, with potential implications for future host control strategies. This study has shown that ignoring parasite-parasite or parasite-host interactions and interactions of both the host and the parasite with the external environment, could result in a poor description of the community dynamics. Such complexities need to be considered and incorporated into theory if future control strategies for either host or parasites are to be effective.

This study involves a measure of the variability of descendants from a cross between Ustilago hordei race 7 and race 11, on two varieties of barley, Trebi and Odessa. Components of variability were defined, statistically described and compared. Biometrical analyses uncovered the action of significant additive and non-additive genetic effects. Differential interactions between treatments and varieties revealed the existence of at least one virulence gene. Specific polygenes and the virulence gene were found to produce significant interactions with different environmental conditions. Homogeneity of variance of the genetic components of the F2 from three randomly chosen F1 dikaryotic lines demonstrated the highly homozygous condition of the parental teliospores. Covariance - variance regression analysis was used to study the dominance and epistatic differences between treatment dikaryons. There is evidence for ambidirectional dominance. The number of effective factors operating against the varieties, Trebi and Odessa, were estimated to be between 4-6 and 1-2 respectively. / Science, Faculty of / Botany, Department of / Graduate

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