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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Comparative Phenotypic and Genomics Approaches Provide Insight into the Tripartite Symbiosis of Xenorhabdus bovienii with Steinernema Nematode and Lepidopteran Insect Hosts

McMullen, John George II January 2015 (has links)
Nematodes are highly diverse animals capable of interacting with almost every other form of life on Earth from general trophic interactions to intimate and persistent symbiotic associations. Much of their recognition originates from their various parasitic lifestyles. From an agricultural standpoint, plant parasitic nematodes are widely known for the destruction they can cause to crop plants, such as the case of the root-knot nematode Meloidogyne incognita, or livestock animals, like the Trichinella spiralis, which infects pigs and other animals. From a human health perspective, nematodes can cause many debilitating diseases, for example Wuchereria bancrofti, which is a causative agent of lymphatic filariasis or elephantiasis. However, not all parasitic nematodes have bad implications for human health. For instance, the diverse interactions of insect parasitic nematodes can be used to our benefit. Many of these species have been considered as biological control alternatives to different insect pests that wreak havoc on human, animal, and plant health. There still remain many questions surrounding their evolution, ecology, and physiological capabilities. Many of these taxa are hard to cultivate in the lab due to their complex and intimate lifestyles. Entomopathogenic nematodes (EPNs) are of great interest in agriculture because they vector insect pathogenic bacteria, which are capable of causing death to an insect host within 48 hours post-infection. Much of the molecular underpinnings in this system still remain to be discovered, from understanding the basic ability of these two organisms to associate with one another to genetically engineering more robust and host specific pathogens for application in the field. The focus of the research presented herein is on Steinernematidae nematodes and their bacterial symbionts. Specifically, it focused on the relationship between Xenorhabdus bovienii and its Steinernema hosts. Bioassays were designed to investigate insect virulence of X. bovienii alone in two Lepidoptera insect species with known differential susceptibility to Steinernema-Xenorhabdus pairs. A comparative genomic analysis was performed to compare different Xenorhabdus bovienii strains with observed variation in insect virulence. Results from this analysis demonstrated that virulent strains possess a type VI secretion system (T6SS) locus that is completely absent in strains with attenuated virulence. Bacterial competition assays between T6SS+ and T6SS- strains suggest this locus is involved in bacterial competition. Additionally, symbiont preference assays were carried out to investigate whether Steinernema hosts are able to discern between virulent and attenuated X. bovienii strains. Results from these assays revealed that Steinernema nematodes are able to distinguish between cognate and non-cognate X. bovienii symbionts, giving preference to virulent strains over those with attenuated virulence. Altogether these results provide further evidence that supports the notion that symbiont-switching events have occurred over the Steinernema-Xenorhabdus co-evolutionary history. Specifically, the competitive virulence of certain X. bovienii strains may have conferred them the ability to be selected by different Steinernema hosts, therefore contributing to the success of the nematode-bacterium partnership in being pathogenic to diverse insect hosts.
2

Processos fisiológicos envolvidos na aquisição, perda e recolocação da zooxantela ligados ao branqueamento de cnidários / Physiological processes associated to acquisition, loss and recovery of zooxanthellae related to cnidarian bleaching

Casillas-Barragán, Isabel 06 February 2018 (has links)
O branqueamento por estresse térmico é a principal causa do declínio de cnidários simbióticos nos oceanos tropicais. Contudo, este processo pode ser revertido. Assim, o objetivo deste trabalho é o estudo integrado dos processos fisiológico da perda e recuperação das zooxantelas relacionado ao fenômeno de branqueamento por aumento de temperatura em E. pallida, mediante o estudo da resposta do holobionte ao estresse térmico, com ênfase na troca de população simbiótica. Um dos mecanismos de recuperação da pigmentação investigados nesse estudo é a proliferação dos endosimbiontes remanescentes no cnidário, onde foi encontrando um padrão de repigmentação funcional em diferentes partes do pólipo. Neste contexto é proposto um mecanismo de ruptura da simbiose na qual os endosimbiontes participam ativamente de sua expulsão e dispersão no meio, abrindo uma nova discussão sobre o agente diretor da simbiose. Uma vez que o branqueamento tem sido relacionado com estresse oxidativo e a fotossíntese é um processo oxidante, avaliamos a atividade redox induzida pela fotossíntese da alga hospedada, assim como a atividade antioxidante e reparadora do hospedeiro. Descobrimos no hospedeiro um mecanismo preventivo para lidar com a atividade fotossintética da zooxantela abrigada. Por outro lado, a perda da alga a partir do branqueamento pode fornecer uma oportunidade para o hospedeiro ser repopulado por outros simbiontes com diferentes capacidades fisiológicas. Assim, encontramos que existe uma troca de alga ideal para o frio (clado F), e para o calor (clado E). Entretanto, alguns clados podem estar indisponíveis no ambiente por serem termosensíveis em vida livre. Concluímos assim que, dentro dos cenários das mudanças climáticas globais, o mais vantajoso para o animal e para a alga é manter uma endopopulação mista e trocar a população simbiôntica predominante segundo as condições ambientais. Aliás, encontramos que o holobionte estudado está melhor preparado para condições de aquecimento que de esfriamento. Por outro lado, considerando que o peróxido de hidrogênio é um derivado de filtros solares e que ao mesmo tempo é um precursor do branqueamento, investigamos se o branqueamento da anêmona Anemonia sulcata produzido pela exposição a peróxido de hidrogênio é um bom modelo como bioindicador ambiental através da avaliação da cor e de biomarcadores fisiológicos. Porém, descartamos A. sulcata como espécie bioindicadora por apresentar uma alta tolerância ao estresse oxidativo, provavelmente fornecido pela atividade fotossintética da alga e por ser uma espécie intermareal que suporta grandes intervalos de oxido-redução. Desta forma, os resultados obtidos no presente estudo apontam a relevância do Symbiodinium dentro da fisiologia do holobionte, tanto na recuperação do branqueamento quanto no estabelecimento e ruptura da simbiose. Consequentemente, futuros estudos podem aprimorar as técnicas de identificação dos clados hospedados para entender a eventual sucessão endopopulacional de acordo com as condições ambientais / Thermal bleaching is the main cause of simbiotic cnidarian decay in tropical oceans. However, some cnidarians have shown to recover the pigmentation from such events. The main aim of this study is the holistic approach of the physiological processes associated to zooxanthela loss and recovery after thermal stress in the holobiont E. pallida. Firstm we considered pigmentation recovey from the remaining zooxanthelae within the cnidarian host. We found a functional pattern of pigmentation recovery. In this context, we propose a mechanism of symbiosis break in which the endosymbionts participate actively on its release and dispersion to the environment, bringing a new discussion about the role of each symbiotic partner and the driver of bleaching and recovery. Since bleaching has been related to oxidative stress and photosynthesis is an oxidizing process, we evaluated the redox activity of the host as well as the antioxidant and restorative activity, induced by photosynthesis of the hosted algae. We found in the host a preventive mechanism to deal with the photosynthetic activity of the sheltered zooxanthella. On the other hand, loss of endosymbionts from bleaching may provide an opportunity for the host to be repopulated by other symbionts with different physiological capabilities. Thus, we found that there is an ideal clade exchange for the cold (clade F), and for the heat (clade E). However, some clades may not be available in the environment because they are thermosensitive in free-living form. We conclude that, within global climate change scenarios, the most beneficial for the animal and the alga is to maintain a mixed endopopulation and to exchange the predominant symbiotic population according to environmental conditions. In fact, we found that the studied holobiont is better prepared for high than low temperatures. On the other hand, considering that hydrogen peroxide is a derivative of sunscreens and at the same time is a precursor to bleaching, we investigated whether the bleaching of anemone Anemonia sulcata produced by exposure to hydrogen peroxide is a good model as an environmental bioindicator through color evaluation and physiological biomarkers. However, we discard A. sulcata as a bioindicator species because it presents a high tolerance to oxidative stress, probably provided by algae photosynthetic activity and because it is an intertidal species that supports large oxido-reduction intervals. Thus, the results obtained in the present study point to the relevance of Symbiodinium within holobiont physiology, both in the recovery of bleaching and in the establishment and rupture of symbiosis. Consequently, future studies may improve the identification techniques of the host clades to understand the eventual endopopulation succession according to the environmental conditions

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