Thesis (M.S.)--Mississippi State University. Department of Entomology and Plant Pathology. / Title from title screen. Includes bibliographical references.

A fundamental problem in developmental and evolutionary biology is understanding the developmental genetic basis of morphological diversity. The current paradigm holds that a genetic and developmental program, or developmental genetic "toolkit", conserved across hundreds of millions of years patterns development in all metazoans. However, outside of a few well-characterized signal transduction pathways and developmental processes, overly broad strokes have been used to paint this "toolkit" metaphor as a hypothesis. Arthropoda, one of the largest groups of metazoans, represent the most morphologically diverse groups of metazoans, making them of particular interest for studies of morphological diversity and its evolution. Arthropoda is also home to one of the most well-understood model systems for developmental and genetic studies, the fruit fly Drosophila melanogaster. However, Drosophila is highly derived among arthropods with respect to the molecular genetic mechanisms that function during its development. As it is expected that all arthropods have access to the same development "toolkit", some changes are expected based on the observable differences in morphology, making arthropods extremely powerful tools for comparative genomic and molecular genetic studies. In this dissertation I characterize how modifications to the developmental "toolkit" contribute to the evolution of morphological diversity using emerging model arthropod systems. First, as part of a collaboration, I show that several genes expected to be conserved in all arthropods, belonging to the Hox family of transcription factors, have been lost from the genome of a phylogenetically basal arthropod, the two-spotted spider mite Tetranychus urticae. Second, I perform a genomic survey and find an overall reduction in the conservation of Drosophila orthologs from several major signal transduction pathways in the Tetranychus genome in comparison with findings from previous insect surveys. Third, I show that arthropod Hox genes, expected to be found in a tightly linked genomic cluster in most arthropod genomes, are not as tightly clustered as previously thought. Fourth, I show that changes in the genomic arrangement of Tetranychus Hox genes correspond with shifts in their expression and morphological change. Finally, I show the terminal Hox gene Abdominal-B is required for proper axial elongation and segment formation (both segment identity and number) during embryogenesis and metamorphosis in the red-flour beetle Tribolium castaneum. Taken together, these findings advance our knowledge of the evolution of morphological change, with a primary focus on Hox genes and their contribution to axial patterning during development.

No presente trabalho procura-se caracterizar biológica e ecologicamente o Aedes albopictus (Skuse) (Diptera: Culicidae) no seu mais recente ambiente geográfico, isto é, nas Américas. São descritos aspectos relacionados com a chegada e dispersão da espécie no Continente Americano. Paralelamente, são discutidos aspectos relativos ao comportamento, biologia, bionomia, ecologia, hematofagia, interação com outras espécies do gênero Aedes, bem como a importância epidemiológica do Ae. albopictus. Esses aspectos foram descritos, por serem considerados importantes pela influência que exercem na competência vetora da espécie, bem como na sua adaptação ao ambiente antrópico. Ressaltando a importância epidemiológica deste vetor para a espécie humana, citam-se dados referentes à potencialidade do Ae. albopictus participar na transmissão de arboviroses. Assim, procura-se caracterizar a importância da possível participação do Ae. albopictus na veiculação do vírus dengue, febre amarela, bem como outros arbovírus que afetam o ser humano nas Américas. Paralelamente, discute-se a possibilidade do Ae. albopictus atuar como vetor da Dirofilaria immitis (Leidy) ao homem. / In the present work, biological and ecological features of Aedes albopictus (Skuse) (Diptera: Culicidae) are discussed based on literature records. The main focus of this study is the epidemiological importance of the presence of this species in Americas. Aspects relative to colonization and dispersion of Ae. albopictus in Americas are discussed. Dispersion, ethology, biology, ecology, haematophagy, bionomics as well as epidemiological importance, and relationships among Ae. albopictus and other members of the genus Aedes are discussed. These aspects are included because of their influence in the vector competence of Ae. albopictus, and also in its adaptation to the human environment. Because of the epidemiological importance of Ae. albopictus to humans, data relative to the potentiality of this species to participate in the transmission of arboviruses are discussed. In this sense, the importance of Ae. albopictus as vector of dengue, yellow fever, and other arboviruses that cause human diseases is described. Besides, it is discussed the potentiality of Ae. albopictus to act as a vector of Dirofilaria immitis (Leidy) to humans.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21826号 / 農博第2339号 / 新制||農||1067(附属図書館) / 学位論文||H31||N5198(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)准教授 髙橋 知之, 教授 北山 兼弘, 准教授 刑部 正博 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

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Aloe mites are herbivores of the genus Aloe (ALOACEAE) and are associated with hyperplastic growth in various aloe species, but the biochemistry of this interaction is poorly understood. In an effort to characterize plant defense responses to herbivory in the genus Aloe, a salivary extract was isolated from aloe mites (Aceria aloinis Keifer) and its bioactivity was tested using a hypocotyl elongation assay. Subsequently, Aloe striata plants were treated with jasmonic acid (JA), salicylic acid (SA), and the mite salivary extract. Using water and methanol, compounds of different polarity were extracted from aloe tissues that had been frozen and crushed at 4, 12, and 24h after treatment. Extracts were analyzed by HPLC and three compounds were found. One of these compounds was SA (mean concentration of 4µg/mL), and this is the first time that this aloe species has been found to produce SA. Two additional peaks of unknown identity were observed in JA- and SA-treated plants. These results suggest that A. striata may in fact undergo a JA-mediated change in secondary metabolism as part of a plant defense response.

Viruses in the Flavivirus genus (e.g., Zika, yellow fever, dengue, West Nile, and Japanese encephalitis viruses) are arthropod-borne, globally distributed, and can cause a range of neurological or hemorrhagic diseases. The ongoing epidemics of flaviviral disease consistently demonstrate the need for new vaccines capable of outbreak control. However, safe, effective, and easy-to-produce vaccines remain relatively elusive due to limitations of conventional vaccine development that make it difficult to balance safety and efficacy. Insect-specific flaviviruses (ISFVs) are emerging as a novel method to overcome this challenge. Herein, we develop a new flavivirus vaccine platform based on the novel insect-specific flavivirus called Aripo virus, which we used to create a preclinical Zika virus (ZIKV) vaccine named Aripo/Zika virus (ARPV/ZIKV). ARPV/ZIKV is a live recombinant virus consisting of the ZIKV pre-membrane and envelope protein genes expressed on an Aripo virus backbone. In this work, we quantify the safety and efficacy of ARPV/ZIKV in multiple murine models, and begin to elucidate the mechanisms of humoral and cell-mediated immune induction for this new platform. Overall, the vaccine showed no evidence of pathogenicity in immunocompromised or suckling mice, and demonstrated a complete inability to replicate in various vertebrate cell lines. Despite this lack of replication, a single dose of live, unadjuvanted ARPV/ZIKV completely prevented ZIKV disease in mice and prevented in utero ZIKV transmission in gravid mice. Direct protection post-ZIKV challenge appears to be primarily mediated by neutralizing antibodies based on passive transfer, adoptive transfer, and T-cell depletion studies. However, vaccination studies in Rag1 KO, Tcra KO, and muMt- mice demonstrate the critical role of T-cell responses in developing immunity post-vaccination. In summary, ARPV/ZIKV is a promising vaccine candidate that induces robust adaptive immune responses, and this success is a positive indication of ARPV's potential as a new resource for flavivirus vaccine development. This body of work contributes to the rapidly expanding field of insect-specific virus-based vaccines and generates new insights into their optimization. Ultimately, this work may help protect the health of millions of people worldwide that are currently at risk of flavivirus infection. / MPH / Arthropod-borne viruses (especially flaviviruses such as Zika virus (ZIKV), yellow fever virus, West Nile virus) represent a major global health threat and a significant burden on human life. Vaccination is a critical tool for controlling the often unpredictable outbreaks of flavivirus diseases. However, licensed flavivirus vaccines remain relatively elusive. This is, in part, because the same characteristics of traditional live-attenuated vaccines that make them highly effective can also reduce their safety. Insect-specific flaviviruses (ISFVs) are emerging as a novel method to overcome this challenge. ISFVs only replicate in insects and thus are safe in humans. They do not cause disease in vertebrates, eliminating the need for the chemical or physical inactivation methods required by traditional whole inactivated vaccines and which can result in reduced efficacy. Herein, we develop a new flavivirus vaccine platform based on a novel ISFV called Aripo virus (ARPV). As proof of concept, we used ARPV to create a preclinical ZIKV vaccine named Aripo/Zika virus (ARPV/ZIKV). ARPV/ZIKV expresses immune system-stimulating ZIKV structural proteins on its virus particle. However, it remains highly safe because the genetic material from ARPV makes it incompatible for replication in human cells. Here, we demonstrate the safety and protective ability of ARPV/ZIKV, and begin to elucidate its mechanisms of protection. Overall, ARPV/ZIKV shows promise as a ZIKV vaccine candidate, which supports the potential of ARPV as a platform for new flavivirus vaccines and the potential to protect the health of the millions of people currently at risk of flavivirus infection.

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