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Thesis (Ph.D)--Biology, Georgia Institute of Technology, 2009. / Committee Chair: Weissburg, Marc; Committee Member: Hay, Mark; Committee Member: Kubanek, Julia; Committee Member: Webster, Donald; Committee Member: Yen, Jeannette. Part of the SMARTech Electronic Thesis and Dissertation Collection.

Thesis (PhD)--Montana State University--Bozeman, 2008. / Typescript. Chairperson, Graduate Committee: David K. Weaver. Includes bibliographical references.

Thesis (M.S. (honours))--University of Western Sydney, 2003. / Title from PDF title page (viewed Sept. 19, 2007). Made available through Australian Digital Theses Program. Includes bibliographical references.

Thesis (M.S. (honours))--University of Western Sydney, 2003. / Title from PDF title page (viewed Feb. 2, 2005). Includes bibliographical references (p. 142-150). Issued also in print. Print ed. accompanied by 1 CD-ROM.

Master of Science / Department of Entomology / Thomas W. Phillips / The rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), is a very common worldwide pest of stored grains. The orientation of C. ferrugineus beetles to various semiochemicals was studied in both the laboratory and field. In laboratory experiments glass two-choice pitfall bioassay dishes were used. Mixed-sex populations of beetles responded positively and significantly to cucujolide I and II separately and in combination compared to controls. Bioassays using the two aggregation pheromones of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae). Dominicalure 1 and 2 together elicited unexpected significant attractive responses by C. ferrugineus as did assays with the synthetic pheromones from other species. Bioassays showed that C. ferrugineus were not attracted to either corn meal or rolled oats, but they were attracted to commercial grain-based lures and also to ethanol, a natural fermentation product from grains. Several bioassays were conducted to test the attractiveness of naturally produced beetle volatiles to C. ferrugineus, but these showed no orientation to volatiles from either the beetles or their food only. Field tests were conducted in Kansas to test attractiveness of the synthetic pheromones cucujolide I and II in lures together with wheat versus wheat only as a control using Lindgren funnel traps. Field tests in 2014 with cucujolide II pheromone lures showed a significant response by feral C. ferrugineus when tested against wheat. Other field tests to examine dispersal of C. ferrugineus relative to grain storages showed that more C. ferrugineus were caught on the western side of grain bins compared to other cardinal directions at two different locations. Results from field and laboratory studies suggest that semiochemical-based tools can be developed to study dispersal behavior in field populations of C. ferrugineus.

Cairomônios são semioquímicos envolvidos nas interações interespecíficas dos insetos, que beneficiam o receptor em detrimento do emissor. Uma destas interações envolve a busca hospedeira por parasitoides, que detectam estes compostos como estratégia para localizar seus hospedeiros. No caso do ectoparasitoide larval, Habrobracon hebetor Say (Hymenoptera: Braconidae), o \'frass\' de Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) libera um cairomônio que atrai as fêmeas do parasitoide. Considerando que A. kuehniella é muito utilizada para produção de ovos, visando à criação de inimigos naturais, este cairomônio é indesejável, pois acaba favorecendo altas infestações por H. hebetor, podendo causar prejuízos expressivos ao longo do processo. Uma das soluções para o controle de tais infestações poderia ser o emprego de armadilhas com o semioquímico sintético. Entretanto, como o cairomônio ainda não foi identificado, este trabalho teve como objetivo o seu isolamento - a etapa inicial para sua identificação. O cairomônio foi extraído por meio da lavagem com solvente e aeração do \'frass\' de A. kuehniella. Os extratos foram, então, testados em olfatometria de quatro vias com fêmeas do parasitoide, registrando-se o tempo de permanência em cada tratamento. O extrato de lavagem ativo foi fracionado em coluna de sílica em cinco frações, cuja atratividade também foi averiguada para proceder com o isolamento do cairomônio. Os extratos e frações foram então analisados por cromatografia gasosa por ionização em chama (GC-FID) e espectrometria de massas (GC-MS), a fim de localizar compostos exclusivos nas amostras ativas. Além disso, as frações provenientes do extrato de lavagem foram aeradas para a extração de seus compostos voláteis e também analisadas por GC-MS. O cairomônio foi isolado na fração 75% éter, indicando ser um composto polar. Por comparação dos cromatogramas, apenas um pico na coluna HP-5ms foi selecionado como pertencente ao cairomônio. Em coluna Rtx-1ms, o pico bifurcou-se, indicando que, pelo menos, um dos compostos faz parte do cairomônio. As combinações metodológicas deste trabalho conduziram ao isolamento do cairomônio presente no \'frass\' de A. kuehniella e à seleção de pelo menos um composto que compõe o semioquímico responsável pela atração do parasitoide H. hebetor, resultando nas primeiras informações de sua estrutura química para sua futura identificação. / Kairomones are semiochemicals involved in interspecific interactions of insects that benefit the receiver to the detriment of the emitter. One of these interactions is host searching by parasitoids which detect these compounds as a strategy to locate their hosts. In the case of the larval ectoparasitoid, Habrobracon hebetor Say (Hymenoptera: Braconidae), the \'frass\' of Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) releases a kairomone that attracts females parasitoids. Considering that A. kuehniella has been used for egg production in order to keep natural enemy rearing, this kairomone is undesirable, because it may lead to high H. hebetor infestation and causes important losses during the rearing process. One of the attempts to control these infestations could be the use semiochemical-baited traps. However, as the kairomone is still unidentified, the aim of this work was to isolate it - the first step for its identification. The kairomone was extracted by solvent washing and aeration of A. kuehniella \'frass\'. Then the extracts were tested in a four-way olfactometer with female parasitoids, registering the time taken in each treatment. The active washing extract was fractionated in five fractions in a silica column, and its attractivity was evaluated to proceed to isolation. The extracts and fractions were then analysed by gas chromatography with flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS), in order to locate exclusive compounds in active samples. Furthermore, the fractions from the washed extract were aerated to extract their volatile compounds that were also analysed by GC-MS. The kairomone was isolated in the 75% ether fraction, indicating that it is a polar compound. Comparing the chromatograms, only one peak on the HP-5ms column was selected as belonging to the kairomone. On the Rtx-1ms column, the peak bifurcated, indicating that at least one of the compounds is part of the kairomone. The methodological combinations of this work provided the isolation of the kairomone present in A. kuehniella \'frass\' and the selection of at least one compound that comprises the semiochemical that attracts H. hebetor parasitoid, resulting in the first source of information about its chemical structure for its future identification.

[No Abstract]

<p>The use of transformed bacteria to stop or kill disease-causing agents in the gut of vector insects is called paratransgenics. Two of the major steps in creating a paratransgenic <i>Anopheles</i> mosquito, unable to spread the<i> Plasmodium</i> parasites that cause malaria, are to find a bacterium suitable for the purpose and a way to introduce the transformed bacterium into mosquitoes in the field. In this project, bacteria associated with malaria mosquitoes have been identified by phylogenetic analysis of their 16S rRNA genes. First, the midgut flora of field-caught <i>Anopheles</i> mosquitoes was examined using two pathways, one culture dependent and one culture independent. Second, six bacterial species from an<i> An. gambiae </i>laboratory colony, and third, ten isolates from <i>Anopheles</i> oviposition sites have been identified. Altogether, 32 bacterial species, representing 16 families, seven classes and four phyla were identified. Interestingly, several of them are related to bacteria known to be symbionts in other insects. Two possible ways of introducing bacteria into mosquitoes in the field in a paratransgenic approach were investigated in a laboratory setting. It was shown that sugar solutions with or without bacteria are equally attractive to <i>An. gambiae</i> mosquitoes and that the mosquitoes were able to take up bacteria from the water they emerged from. These results show that it may be possible to use sugar-baits and oviposition sites for distribution of genetically modified bacteria in the field. To facilitate the distribution of the modified bacteria mosquito attractants should be used. We investigated whether the bacterial isolates identified in this project produce attractants affecting mosquito sugar-feeding or oviposition site selection. While no responses were observed from the mosquitoes towards bacteria-containing sugar solutions, seven of the 19 isolates examined mediated positive oviposition responses. In total, 13 putative oviposition attractants were identified among the volatiles emitted by the attractive bacteria.</p>

The use of transformed bacteria to stop or kill disease-causing agents in the gut of vector insects is called paratransgenics. Two of the major steps in creating a paratransgenic Anopheles mosquito, unable to spread the Plasmodium parasites that cause malaria, are to find a bacterium suitable for the purpose and a way to introduce the transformed bacterium into mosquitoes in the field. In this project, bacteria associated with malaria mosquitoes have been identified by phylogenetic analysis of their 16S rRNA genes. First, the midgut flora of field-caught Anopheles mosquitoes was examined using two pathways, one culture dependent and one culture independent. Second, six bacterial species from an An. gambiae laboratory colony, and third, ten isolates from Anopheles oviposition sites have been identified. Altogether, 32 bacterial species, representing 16 families, seven classes and four phyla were identified. Interestingly, several of them are related to bacteria known to be symbionts in other insects. Two possible ways of introducing bacteria into mosquitoes in the field in a paratransgenic approach were investigated in a laboratory setting. It was shown that sugar solutions with or without bacteria are equally attractive to An. gambiae mosquitoes and that the mosquitoes were able to take up bacteria from the water they emerged from. These results show that it may be possible to use sugar-baits and oviposition sites for distribution of genetically modified bacteria in the field. To facilitate the distribution of the modified bacteria mosquito attractants should be used. We investigated whether the bacterial isolates identified in this project produce attractants affecting mosquito sugar-feeding or oviposition site selection. While no responses were observed from the mosquitoes towards bacteria-containing sugar solutions, seven of the 19 isolates examined mediated positive oviposition responses. In total, 13 putative oviposition attractants were identified among the volatiles emitted by the attractive bacteria.