Fourteen spore-forming bacterial strains were isolated and screened for entomopathogenic activity. Five displayed toxicity towards the common mealworm, Tenebrio molitor L., (Coleoptera: Tenebrionidae). The majority of the isolates were obtained from insect larvae and insect rich environments. The three bacterial species identified were Bacillus thuringiensis Berliner, Brevibacillus laterosporus Laubach and Bacillus cereus Frankland and Frankland. Bioassays were conducted using T. molitor larvae. The one isolate of B. cereus required the highest concentration of bacterial cells to achieve its LC50, whereas one of the isolates of B. laterosporus required the lowest cell concentration to achieve its LC50. Dose response curves were generated for the five best isolates, which showed that the isolate of B. laterosporus (NDR2) was substantially more toxic than the other isolates. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009.

Made available in DSpace on 2016-06-21T13:43:14Z (GMT). No. of bitstreams: 2 830.pdf: 984013 bytes, checksum: 404aa18d3e40dad94e177518d24b5c02 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2009 / Made available in DSpace on 2016-07-05T22:16:49Z (GMT). No. of bitstreams: 3 830.pdf.txt: 186951 bytes, checksum: c158594d1dae991c3436229e07b6cb30 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 830.pdf: 984013 bytes, checksum: 404aa18d3e40dad94e177518d24b5c02 (MD5) Previous issue date: 2009 / Fundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil / O principal fator larvicida do Bacillus sphaericus (Bsp) para culicídeos é a protoxina Bin, produzida sob a forma de um cristal, durante a esporulação. Quando ingerido pelas larvas o cristal é processado e a toxina Bin reconhece e liga-se a receptores específicos do epitélio intestinal. O receptor em Culex quinquefasciatus é uma alfa-glicosidase de 60 kDa, ligada à membrana intestinal por uma âncora GPI, denominado Cqm1. Larvas de Aedes aegypti são consideradas refratárias ao Bsp, pois a toxina Bin não reconhece receptores no microvilli intestinal. No entanto, a análise do genoma do Ae. aegypti, revelou a presença do gene aam1, que codificaria uma proteína ortóloga e com 83 por cento de similaridade ao receptor Cqm1. O principal objetivo deste estudo foi elucidar a base molecular da refratariedade do Ae. aegypti ao Bsp, determinada pela ausência de ligação da toxina Bin ao epitélio intestinal das larvas. Para tal, foi feita uma investigação da expressão da proteína Aam1 e do perfil de alfa-glicosidases de Ae. aegypti, tendo como referência o receptor Cqm1. Os resultados mostraram que larvas e adultos de Ae. aegypti expressam uma ?-glicosidase de membrana de 70 kDa, reconhecida pelo anticorpo anti-Cqm1, e que provavelmente trata-se da proteína Aam1. Tal proteína é expressa no microvilli intestinal das larvas em níveis superiores à Cqm1, no entanto, não apresenta capacidade de ligação à toxina Bin. Em uma segunda etapa, a avaliação de proteínas Aam1 e Cqm1 recombinantes, produzidas em lisado de reticulócitos de coelho, mostrou que ambas não foram capazes de se ligar específicamente à toxina Bin. A falha na ligação da proteína Cqm1 à toxina Bin pode ser decorrente da ausência do processamento pós-traducional adequado neste sistema de expressão, indicando que certas modificações podem ser críticas para a sua funcionalidade. O tratamento da proteína Cqm1 nativa à temperatura de 100 °C aboliu a sua capacidade de ligação à toxina Bin, indicando que a conformação da proteína pode ser essencial para a sua funcionalidade. Os resultados obtidos demonstram que, apesar dos altos níveis de expressão da Aam1 nas larvas de Ae. aegypti, a proteína não é capaz de ligar-se à toxina Bin. Tal fato estar relacionado a outros fatores críticos para sua funcionalidade, tais como diferenças conformacionais e/ou modificações pós-traducionais que determinem o status de refratariedade do Ae. aegypti

La plupart des toxines de Bacillus thuringiensis perméabilisent la membrane intestinale des insectes sensibles en formant des pores qui abolissent le potentiel électrique et les gradients ioniques. Plusieurs toxines ont été étudiées avec des vésicules purifiées de la bordure en brosse intestinale des insectes. Malheureusement, la membrane intestinale de beaucoup d’insectes ne forme pas des vésicules suffisamment étanches pour les expériences de perméabilisation. Une nouvelle technique utilisant des liposomes géants et une sonde de perméabilité membranaire a été développée pour caractériser deux nouvelles toxines particulièrement prometteuses pour le biocontrôle d’un des principaux ravageurs du maïs, la chrysomèle des racines du maïs (Diabrotica virgifera virgifera LeConte), Cry6Aa1 et la toxine binaire DS10/DS11. Les deux toxines perméabilisent efficacement les liposomes. La toxine binaire forme des pores qui sont légèrement sélectifs pour les cations, comme la plupart des toxines de B. thuringiensis. Bien que la Cry6Aa1 puisse former des pores sélectifs pour les anions, les résultats suggèrent aussi qu’elle pourrait, contrairement aux autres toxines de cette bactérie, ne former des pores qu’en présence d’une force ionique élevée. La formation des pores par ces deux toxines semble être sensible à la courbure de la membrane cible étant donné qu’elle est beaucoup plus efficace dans des liposomes géants que dans des liposomes de même composition, mais plus petits. Ce travail jette les bases de la mise au point d’une technique qui permettrait l’étude des toxines dans des liposomes géants enrichis avec des protéines et des lipides provenant de la membrane intestinale des insectes cibles. / Most Bacillus thuringiensis toxins permeabilize the intestinal membrane of susceptible insects by forming pores that abolish transmembrane electrical potentials and ionic gradients. Several toxins have been studied using brush border membrane vesicles purified from the insect midgut. Unfortunately, the intestinal membrane from many insects does not form vesicles that are tight enough to be used in permeabilisation experiments. A new technique using giant liposomes and a membrane permeability probe was developed to evaluate the pore-forming ability of two particularly promising toxins for the biocontrol of a major corn pest, the Western corn rootworm (Diabrotica virgifera virgifera LeConte), Cry6Aa1 and the binary toxin DS10/DS11. Both toxins permeabilized the liposomes efficiently. However, analysis of the permeabilisation rates under different experimental conditions indicates that these toxins differ in their biophysical properties. The binary toxin forms pores which are slightly selective for cations, like most B. thuringiensis toxins. On the other hand, although the results suggest that Cry6Aa1 could form anion-selective pores, they could also indicate that, in contrast with other toxins produced by this bacterium, it could form pores only under high ionic strength conditions. Pore formation by both toxins appears to be sensitive to membrane curvature since it is much more efficient in giant liposomes than in liposomes with identical composition, but smaller in size. This study sets the bases for the development of a technique that would allow the toxins to be studied in giant liposomes enriched with proteins and lipids from the intestinal membrane of target insects.

Maize is the staple food for most South Africans. This implies that any damage to the maize crop will affect food security of many South Africans. Although Eastern Cape Province is not a traditionally maize producing area, smallholder farmers in the province produce it mostly for subsistence purposes and some sell the surplus on the local market or use it to secure other good through barter trading. In South Africa, insect-resistant Bt maize/yieldgard has been used commercially for approximately 10 years now. Available impact studies on Bt maize reveal that, this technology is beneficial not only to farmers but consumers of maize products as well. Welfare gains as well as positive effects for human health are realised by both groups. Due to the costs and effectiveness associated with traditional and conventional maize stem borer control methods, Bt technology has the potential to be part of the solution. This thesis has attempted to investigate the economic viability of planting Bt maize seeds under smallholder farming conditions and identify factors as well as perceptions relating to attributes of Bt maize and to analyze the relationships between those perceptions and choices regarding use of Bt technology. Data was collected from 90 households who were selected using purposive sampling through the use of the snowball method. To collect data, a questionnaire was administered through face-to-face interviews. Gross margin analysis revealed that Bt maize is a more profitable option as compared to conventional maize seeds. Furthermore, econometric analyses, through use of the binomial regression model revealed that perceptions could be used to distinguish between users and non-users of Bt maize seed in the Eastern Cape Province. Results of inferential analysis indicate that the statistically significant variables at 5% level are gene erosion, quality and nutrition of products and food labels for Bt maize products perceptions. On the other hand, low expenses, seed market availability and farmers’ knowledge perceptions were significant at 10%. These findings suggest that an adjustment in each one of the significant variables can significantly influence the probability of Bt maize adoption. In view of the research findings, several policy proposals are suggested to support policy formulation. Key words: Bacillus thuringiensis (Bt) maize, yieldgard, smallholder farmers, perceptions, Flagstaff, gross margin analysis, binomial logistic regression model, Eastern Cape Province.

My dissertation research examined the effect of the cultivation of insect-resistant Bacillus thuringiensis (Bt) maize on the soil environment with a goal of understanding how to obtain a balance between technological advancement and maintenance of a healthy soil ecosystem. Although Bt plants may help to reduce pesticide use, conferring benefits to farm workers and the environment, there are still unresolved questions about how the cultivation of Bt plants affects soil organisms. For this dissertation project, I used 14 different genotypes of Bt maize and non-Bt maize (Zea mays) to investigate the effects of transgenic Bt plants on the colonization ability, abundance, and diversity of symbiotic arbuscular mycorrhizal fungi (AMF) in the soil ecosystem over time. My greenhouse studies demonstrated that Bt maize plants exhibited reduced AMF colonization across multiple Bt genotypes and that effects were most pronounced when fertilizer levels were limited and spore density was high. In addition, I found that although differences in AMF colonization between Bt and non-Bt maize were difficult to detect in the field, spore density was reduced in Bt field plots after just one growing season. When I tested the effect of plot history on AMF and plant growth, I found that Bt and non-Bt maize plants had higher leaf chlorophyll content when grown in plots previously cultivated with the same maize line as the previous year, indicative of a positive feedback effect. I also examined potential mechanisms contributing to the reduced AMF colonization observed in Bt maize in greenhouse studies and determined that follow-up experiments should continue to investigate differences in root apoplastic invertase activity and root permeability in Bt and non-Bt maize. Future investigations would also benefit from examining potential differences in root exudate profiles and volatile organic compounds between Bt and non-Bt cultivars. Taken together, my dissertation results suggest that, while difficult to detect in the field, reductions in AMF colonization in Bt maize roots may be ecologically significant as they could lead to a decrease in the abundance of AMF propagules in the soil over time, potentially impacting soil structure and function in areas where Bt crop cultivation is high.

No description available.

Depuis 1985, plus de 10 millions d’hectares de forêt ont été traités au Canada contre la tordeuse des bourgeons de l’épinette (TBE) avec l’insecticide biologique Btk. Toutefois, aucune étude ne s’est penchée sur les effets de ces traitements sur les sous-bois, bien qu’ils contiennent la majorité de la diversité végétale des forêts boréales et représentent une source importante de nourriture pour plusieurs animaux, dont le caribou forestier, une espèce menacée. Ainsi, il semble impératif de déterminer quelle fréquence de traitements de Btk serait idéale pour réduire les perturbations aux communautés végétales de sous-bois. À l’aide d’un dispositif expérimental où les populations de TBE sont contrôlées, nous avons évalué l’influence de différentes modalités d’épandage de Btk sur la défoliation des peuplements et sur les sous-bois. Nos résultats montrent qu’une diminution de la fréquence des traitements influence positivement la lumière disponible aux sous-bois, la richesse, la présence d’espèces intolérantes à l’ombre et la production de petits fruits, particulièrement dans les stations non traitées, où la défoliation était maximale. Or, un sous-bois diversifié comprenant des espèces intolérantes à l’ombre et de nombreux petits fruits est un habitat idéal pour l’orignal et ses prédateurs communs avec ceux du caribou, l’ours et le loup. Une diminution du contrôle des épidémies de TBE affecterait donc négativement le caribou forestier en augmentant potentiellement le risque de prédation, ce qui est un des principaux facteurs limitant ces populations. Des traitements bisannuels de Btk pourraient donc aider à la protection de l’habitat des populations résiduelles de caribou forestier. / Since 1985, more than 10 million hectares of Canadian forests were treated against spruce budworm (SBW) epidemics using the Btk biological insecticide, but no study has evaluated the effects of these interventions on understory vegetation. Since the forest floor host most of plant diversity in boreal forests and provides critical wildlife habitat for, amongst others, woodland caribou, it is crucial to determine the best Btk treatment frequency to reduce wood losses, application costs, and disturbances to understory communities. In an experimental design, we tested the influence of different spraying frequencies for SBW control on tree defoliation and understory vegetation. We found positive relationships between a decrease in treatment frequency and light availability in the understory, richness, shade-intolerant species cover, and fruit production, particularly in control plots, where defoliation was maximal. An increase in shade-intolerant species and fruit production could provide high-quality forage for moose and its predators, wolves and bears, which can increase predation pressure on caribou. A decrease in SBW control may thus negatively affect woodland caribou by increasing predation risks, the main factor limiting caribou populations in managed forests. Consequently, we suggest that SBW control could help maintain woodland caribou habitat during a SBW epidemic and recommend Btk applications every 2 years to prevent both wood loss and further degradation of woodland caribou habitat. Key