En Afrique sud- saharienne, plusieurs méthodes de protection des stocks alimentaires sont utilisées ; parmi elles, les insecticides synthétiques qui représentent un risque réel pour la santé humaine. Pour apporter une alternative à ces insecticides chimiques potentiellement dangereux, le présent travail a pour objectif de développer des bio-insecticides en formulant des poudres par adsorption d’huiles essentielles sur des argiles. Deux argiles naturelles du Cameroun ont été utilisées comme adsorbants des composés terpéniques des huiles essentielles de Xylopia aethiopica et Ocimum gratissimum pour la préparation d’insecticides. Dans le but d’améliorer leur capacité d’adsorption, ces argiles ont été traitées par la soude et l’acide sulfurique. Elles ont également été modifiées par des solutions de polycations d’aluminium ou de fer de rapport molaire varié ainsi que par des cations d’alkylammoniums. Une bentonite commerciale a été utilisée comme référence dans le suivi des modifications. Ces différentes matrices ont été caractérisées avant et après modifications par diffraction des rayons X, adsorption-désorption d’azote, spectroscopie infra rouge à transformée de Fourier, analyses thermogravimétriques et différentielles, et photométrie de flamme. Il ressort de la caractérisation que l’échantillon de Wak est majoritairement constitué de kaolinite, et celui de Maroua de montmorillonite. Le traitement par l’acide sulfurique et par les polycations métalliques entraînent une augmentation de la surface spécifique des argiles, tandis que la soude et les cations d’alkylammoniums la réduisent fortement. La surface spécifique de la montmorillonite passe de 82 m².g-1 à 4,5 m².g-1 après traitement au céthyl triméthyl ammonium (CTMA). Les diffractogrammes montrent une augmentation de la distance interfoliaire des argiles étudiées après modification aux cations alkylammoniums. La distance interfoliaire augmente de 5,5 Å et de 10,6 Å respectivement pour la montmorillonite et la bentonite traitée par le CTMA. Les argiles de type smectite après modifications aux polycations métalliques, présentent en revanche un étalement du pic caractéristique de l’espace interfoliaire. Les cations alkylammoniums entraînent donc une intercalation effective des molécules de CTMA et phényl triméthyl ammonium (PTMA) entre les feuillets de montmorillonite et de bentonite tandis que les polycations métalliques entraînent une exfoliation du matériau argileux. Les argiles-alkylammoniums présentent les plus grandes capacités d’adsorption des composés terpéniques malgré les plus faibles surfaces spécifiques. Ainsi, face aux composés terpéniques, la capacité d’adsorption des argiles ne dépend pas uniquement de la surface spécifique mais également de l’espacement interfoliaire de l’adsorbant ainsi que de l’affinité des molécules d’adsorbât vis-à-vis de l’adsorbant. Des tests insecticides ont montré que les formulations à base de la Mont-CTMA présentent une toxicité plus stable que celle préparée à partir de l’argile brute (Mont-Na). La formulation Mont-Na-HE perd la totalité de son activité insecticide au bout de 30 jours de conservation dans les boîtes ouvertes. La formulation Mont-CTMA-HE par contre n’en perd qu’environ 60% dans les mêmes conditions. La rémanence de la formulation varie avec l’adsorbant utilisé ; car l’effet insecticide de l’huile essentielle d’O. gratissimum persiste pendant 107 jours lorsqu’elle est fixée sur la Mont-CTMA, tandis que fixée sur l’argile brute, elle perd son activité au bout de 45 jours. Ces résultats nous permettent d’affirmer que les argiles modifiées augmentent la durée de l’effet insecticide des huiles essentielles et peuvent être utilisées pour une application industrielle dans la production des bio-insecticides. / In sub-Saharan Africa, several methods to protect food stocks are used ; amongst them are synthetic insecticides. These synthetic insecticides pose high real risk to human health. This study aims at providing an alternative to the dangerous synthetic chemical insecticides, by developing bioinsecticides formulating powders through adsorption of essential oils on Cameroonian clays. Two natural clays collected in Cameroon were used as adsorbents of terpene compounds of Xylopia aethiopica and Ocimum gratissimum essential oils, for the preparation of insecticides. The clay fractions were obtained from soil aggregates by sedimentation. In order to improve their adsorption capacity, these clays were treated with sodium hydroxide and sulfuric acid. They were also modified with solutions of aluminum, iron polycations with molar ratio varied and alkylammoniums cations. A commercial bentonite was used as reference for these modifications. These different adsorbents materials were characterized before and after modifications by X-ray diffraction, nitrogen adsorption-desorption, infrared Fourier transform spectroscopy, thermal analysis, and flame photometry. According to the characterizations of the clay materials, Wak sample contained more kaolinite clay, while Maroua sample contained more montmorillonite clay. The sulfuric acid and metallic polycations caused an increase in the specific surface of clays, while sodium hydroxyl and alkylammoniums cations greatly reduced the SBET. For example, the Mont-Na+ SBET decreased from 82 m². g-1 to 4.5 m².g-1 after treatment with cetyl trimethyl ammonium (CTMA). Diffractogrammes showed spacing of smectites interlayer (montmorillonite and bentonite) after treatments with alkylammoniums cations at 5.5 Å and 10.6 Å respectively, for the interlayer space of Mont- CTMA and Bentonite- CTMA clays. After treatments with metallics polycations, smectite clays show a broadening of characteristic peak. Alkylammoniums cations led to an effective insertion of CTMA molecules and phenyl trimethyl ammonium (PTMA) between the interfoliar space of montmorillonite and bentonite while the metal polycations led to an exfoliation of the same clays. The interreticular spacings of kaolinite remain unchanged after any treatments. The kinetic model of pseudo second order is applicable to the adsorption of terpenic compounds by clays. Finding also showed that spread is not the only limiting factor in the process of adsorption of terpene compounds. Alkylammoniums-clays have the largest adsorption capacities of terpene compounds despite the lower specific surface they present. All this allows us to maintain that in the presence of terpenic compounds, the adsorption capacity of clays depends not only on the SBET but more on the interlayer spacing of the adsorbent and on the affinity of the adsorbate molecules toward the adsorbent. Concerning bioassays, it appears that the formulations made with Mont- CTMA presented a more stable toxicity than Mont-Na. Mont-Na-EO formulation loses all its insecticidal activity after 30 days of storage in the open boxes ; while Mont-CTMA- EO formulation only loses about 60% under the same conditions. Moreover, the Mont-Na-EO loses 50% of its insecticidal power (LD50) after 6 days ; while Mont-CTMA- EO loses the same quantity after 16 days when stored in open boxes. The remnance effect of the formulations based on essential oil varied with the adsorbent used. Because the insecticidal effect of O. gratissimum essential oil persisted during 107 days when it is adsorbed on Mont-CTMA, when adsorbed on Mont-Na, it loses all its activity for about 45 days. These results allow us to assert that modified clays increase the duration of the insecticidal effect of essential oils and can be used for industrial application in the production of bio-insecticides based on essential oils.

Das Ziel der vorliegenden Arbeit war es, die Wirksamkeit der beiden biologischen Präparate NeemAzal MD5 und TRF 002 MD auf das Entwicklungspotential der Larven der großen Stubenfliege (Musca domestica) und der Güllefliege (Hydrotaea aenescens) zu untersuchen. Bei den Präparaten handelte es sich um den Extrakt aus den Samen des tropischen Niembaumes (Azadirachta indica) mit dem Hauptwirkbestandteil Azadirachtin A in einer Konzentration von 5 % und um den Extrakt aus dem Bitterholz (Quassia amara) mit dem Hauptwirkstoff Quassin in einer Konzentration von 0,61 %. Die Larvenstadien 1 und 3 jeder Fliegenart wurden dem jeweiligen Präparat, das in verschiedenen Konzentrationen entweder eingemischt im Brutsubstrat (orale Aufnahme) oder in einem Tauchbad (topische Applikation) getestet wurde, ausgesetzt. Der Einfluss der Wirkstoffe auf die Parameter Verpuppungsrate, Schlupfrate, Entwicklungsdauer, Fliegengröße und Vorkommen von Asymmetrien wurde dabei ermittelt. NeemAzal MD5 in 5 % Lösung verhinderte, eingemischt ins Brutsubstrat, vollständig den Schlupf der Fliegen unabhängig vom Alter der verwendeten Larven. Das jüngste Larvenstadium wies dabei noch eine deutlich höhere Empfindlichkeit gegenüber dem Wirkstoff auf, hier reichte bereits eine 1 % Lösung aus, um die Entwicklung von Fliegen zu unterdrücken. Unterschiede in der Wirksamkeit auf die beiden verwendeten Fliegenarten waren gering. Hydrotaea aenescens erwies sich hierbei als etwas empfindlicher gegenüber NeemAzal MD5 als die Larven von Musca domestica. Die topische Applikation des Präparates auf das Larvenstadium 3 führte in den verwendeten hohen Konzentrationen (5 %, 10 %) nur zu einer Reduktion der Fliegenanzahl. Für eine vollständige Verhinderung der Fliegenentwicklung ist eine möglichst frühzeitige orale Aufnahme des Wirkstoffes durch die Larven notwendig. Es kommt dabei neben der Fraßhemmung zu einer Beeinflussung der Wachstums- und Häutungshormone, was die Metamorphose der Insekten unterdrückt. Die unter Laborbedingungen bewiesene insektizide Wirksamkeit von NeemAzal MD5 und seine Umweltverträglichkeit prädisponieren das Präparat für Feldversuche gegen Fliegenplagen in der ökologischen Tierhaltung. TRF 002 MD bewirkte auch in der höchsten verwendeten Konzentration (10 %), eingemischt in das Brutsubstrat der Larvenstadien 1 und 3 der beiden Fliegenarten, nur eine Reduktion der Schlupfrate um bis zu 30 % gegenüber den Kontrollgruppen. Die topische Anwendung dieser Substanz erwies sich in den durchgeführten Versuchen als nahezu wirkungslos auf die untersuchten Parameter. Damit war das Präparat nicht geeignet, das Schlüpfen von Fliegen vollständig zu verhindern.

<p>Ispitivan je hemijski sastav etarskih ulja i vodenih ekstrakata tri vrste samoniklih biljaka familije Lamiaceae:&nbsp;<em> Satureja montana&nbsp;</em>L.,&nbsp; <em>Salvia sclarea&nbsp;</em>L.,<em>&nbsp; Clinopodium menthifolium</em>&nbsp; Host. Pored toga, ispitano je delovanje vodenih ekstrakata na korovske i ratarsko-povrtarske biljke, insekte i mikroorganizme, kao i etarskih ulja na insekte.<br />Potvrđeno je da ispitivane vrste imaju insekticidno dejstvoi da nemaju uticaj na rast korisnih mikroorganizama prisutnih u zemlji&scaron;tu.</p> / <p>The chemical composition of essential oils and aqueous extracts of three types of self-seeding plants of the Lamiaceae family are examined. Apart from the composition, the effects of aqueous extracts on weed and vegetable plants, insects and&nbsp; microorganisms are examined. The effects of essential oils on insects are also examined. Insecticidal activity of examined plants is confirmed. It is also confirmed that the examined plants have no effect on the growth of useful microorganisms present in the soil.</p>

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work was divided into three parts focusing two main topics: the study of volatiles compouds of P. graveolens and the study of the essential oil of L. alba and its evaluation of insecticidal activity against Spodoptera frugiperda. The chapter 2 approuch the study of the volatile compounds of the leaves of P. graveolens extracted by dynamic headspace using Porapak Q® as adsorbent and in nature peat, a novel adsorbent in the extraction of plant volatiles, and the results were compared with those obtained by hidrodestilation. The results showed that hydrodistilation (HD) was more efficient for extracting linalool and citronellyl formate. While citroneol, geraniol and geranyl tiglate were obtained in greater quantity by dynamic headspace using in nature peat (HSD-T), isomenthone, 6,9- guaiadiene and - muurolene by headspace using Porapak Q® (HSD-P). The study of conversion proved that geraniol converts in linalool when geraniol is subjected to water vapor and high temperature, nevertheless a small ratio. This also prove, in parts, the high percentage of linalool obtained by HD compared to HSD-T and HSD-P. The chapter 3 approuch the study of the essential oils of 9 accessions of L. alba collected in 4 states of Brazil using statistical methods and checking the accessions with the best essential oil content. The statistical methods of hierarchical cluster analysis (HCA) and Principal component analysis (PCA) were used to prove the experimental data and confirmation of training groups. Three groups representatives of the three chemotypes were formed: Group I was characterized by linalool and 1,8-cineole; group II, by limonene and carvone; and group III by neral and geranial. In chapter 4 was approached the evaluation of insecticidal activity of the essential oils of P. graveolens (PEL-001) and L. alba (accessions LA-10, LA-22 and LA-57) and also of the main compounds (geraniol, linalool, 1,8-cineole, limonene, carvone) against S. frugiperda via topical aplication. The test of topical application showed that the essential oils of P. graveolens (PEL-001) and L. alba (accessions LA-10, LA-22, LA-57) showed acute toxicity against larvae of S. frugiperda, causing mortality of up to 100% at a dose 192 μg/larvae. / O presente trabalho foi dividido em 3 partes os quais abordam 2 temas centrais: estudo dos compostos voláteis de Pelargonium graveolens e do óleo essencial de Lippia alba e avaliação de atividade inseticida frente à Spodoptera frugiperda. O capítulo 2 abordou o estudo dos voláteis das folhas de P. graveolens extraídos por headspace dinâmico utlizando Porapak Q® como adsorvente e turfa in natura, um adsorvente inédito empregado na extração de voláteis de plantas, sendo os resultados comparados com àqueles obtidos por hidrodestilação. Os resultados mostraram que a hidrodestilação (HD) foi mais eficiente na extração de linalol e formiato de citronelila. Enquanto citronelol, geraniol e tiglato de citronelila foram obtidos em maior proporção por headspace dinâmico utilizando turfa in natura como adsorvente (HSD-T), isomentona, 6,9- guaiadieno e -muuroleno foram identificados em maior proporção por headspace dinâmico utilizando Porapak Q® (HSD-P). O estudo de conversão comprovou que geraniol se converte em linalol, quando o geraniol é submetido a vapor d água e altas temperaturas, porém a um pequeno percentual. Isso também comprova, em partes, o alto percentual de linalol obtido por HD em relação à HSD-T e HSD-P. A extração do hidrolato revelou variações na composição volátil em comparação com os outros métodos de extração. O Capítulo 3 abordou o estudo dos óleos essenciais de 9 acessos de L. alba coletados em 4 estados do Brasil empregando métodos estatísticos e verificando os acessos com melhores teor de óleo essencial. Os métodos estatísticos de análise hierárquica de agrupamentos (HCA) e análise de componentes principais (PCA) foram empregados para comprovar os dados experimentais e confirmar a formação dos grupos. Três grupos, representando os três quimiotipos, foram formados, dos quais o grupo I foi caracterizado por linalol e 1,8-cineol, grupo II por limoneno e carvona e grupo III por neral e geranial. No capítulo 4 foi abordado a avaliação de atividade inseticida dos óleos essenciais de P. graveolens (PEL-001) e L. alba (acessos LA-10, LA-22 e LA-57), bem como dos seus principais compostos majoritários (geraniol, linalol, 1,8-cineol, limoneno e carvona) frente à S. VII frugiperda. O ensaio de aplicação tópica mostrou que óleos essenciais de P. graveolens (PEL-001) e L. alba (acessos LA-10, LA-22, LA-57) apresentaram toxicidade aguda nas lagartas S. frugiperda, ocasionando mortalidade de até 100% na dose 192 μg/lagarta.

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Brazilian lemon balm (Lippiaalba (Mill.) N. E. Brown) is a medicinal and aromatic species widely used to fight diseases. Its essential oil, rich in carvone, has insecticidal activity on pests of agricultural interest. Cutting ants cause severe economic damage to forest and horticultural species and hinder cultivation worldwide. Plant aging is one of the main factors that alter the biological activity of essential oils. The objective of this work was to evaluate the influence of plant age on the essential oil of L. alba accessionsand of a recurrent selection cycle onformicidal activity. The experiments were carried out at the Experimental Farm - Campus Rural UFS.The experiment consisted of a randomized blocks design with three replications. For the first experiment, 18 L. albaaccessions were used at two plant ages: one-year-old plants (February2006), and ten-year-old plants (February 2016). In the second experiment, a recurrent selection cycle was carried out with three accessions of the limonene/carvonechemotype. For the progeny competition experiment, three parent genotypes and 11 progenies were taken to the field. In the first experiment, plant age influenced the essential oil production and the chemical composition.The highest essential oil content increase was observed for accessions LA-41 (0.69-2.48%) and LA-53 (0.76-2.84%). The compounds with the greatest variation in the second harvest were p-cymene (0.00-5.12%), limonene (0.00-11.08%), and elemol (0.00-10.54%), because of plant aging. In the second experiment, the highest dry weight of aerial part was observed for parental LA-56 (57.54 g plant-1). The essential oil content was higher in the progeny LA-57-10 (2.844%) and the parental LA-57 (2.664%). Carvone concentration was higher for LA-57 (59.02%), with a significant difference when compared with the evaluated progenies. Among the progenies, the highest carvone concentration was observed for LA-56-04 (57.78%) and the lowest for LA-57-01 (17.71%) and LA-57-02 (17.27%).One recurrent selection cycle resulted in increased formicidal activity of some progenies. Higher mortility of Acromyrmex balsani was caused by the essential oils of the progenies LA-56-04 and LA-70-03. / A erva-cidreira-brasileira [Lippia alba (Mill.) N. E. Brown] é uma espécie medicinal e aromática que apresenta vários quimiotipos. O óleo essencial rico em carvona possui atividade inseticida sobre pragas de interesse agrícola. Diante disso, o objetivo do trabalho foi avaliar a influência da idade da planta nos óleos essenciais de acessos de L. alba e de um ciclo de seleção recorrente na atividade formicida. Os ensaios foram conduzidos na Fazenda Experimental “Campus Rural da UFS”. Para o primeiro ensaio foram testados 18 acessos de L. alba, que foram colhidos quando as plantas estavam com um ano de idade (fevereiro/2006) e dez anos (fevereiro/2016). No segundo experimento foi realizado um ciclo de seleção recorrente com três acessos com alto teor de carvona no seu óleo essencial. Para o ensaio de competição de progênies foi implantado um ensaio testando 11 progênies e os três parentais. A idade da planta influenciou na produção e composição química dos óleos essenciais dos acessos de L. alba testados. Observou-se maior aumento doteor de óleo essencial para os acessos LA-41 (0,69-2,48%) e LA-53 (0,76-2,84%). Com o envelhecimento das plantas notou-se maior variação nos óleos essenciais para os compostos p-cimeno (0,00-5,12%), limoneno (0,00-11,08%) e elemol (0,00-10,54%). No segundo experimento a maior produção de massa seca da parte aérea foi observada para o genótipo LA-56 (57,54 g planta-1). O teor de óleo essencial foi superior na progênie LA-57-10 (2,844%) e o parental LA-57 (2,644%). A carvona foi superior para o LA-57 (59,02%) com diferença significativa em comparação as progenies avaliadas. Entre as progênies, o maior teor foi observado para a progênie LA-56-04 (57,78%) e o menor para o LA-57-01 (17,71%) e LA-57-02 (17,27%). Um ciclo de seleção recorrente resultou no aumento da atividade formicida de algumas progênies.Maior mortalidade de Acromyrmex balsani foi causada pelos essenciais das progênies LA-56-04 e LA-70-03. / São Cristóvão, SE

Une fois ingérées par un insecte sensible, les toxines insecticides du bacille de Thuringe doivent être activées par les protéases intestinales de cet insecte. Leur premier domaine, un ensemble de sept hélices-α amphipathiques, est responsable de leur insertion dans la membrane luminale de certaines cellules de l’intestin médian, ce qui crée des pores peu sélectifs. La toxicité et la capacité à former des pores d’une telle toxine, la Cry9Ca, de ses mutants simples R164A et R164K et d’un fragment de 55 kDa résultant d’un clivage protéolytique au niveau de son résidu 164 ont été étudiées à l’aide d’une combinaison de modélisation par homologie, de bioessais, d’expériences de gonflement osmotique avec des vésicules de membrane en bordure en brosse de larves de sphinx du tabac et de mesures électrophysiologiques sur des intestins isolés. Ni les mutations simples ni le clivage protéolytique n’ont altéré la toxicité de la Cry9Ca. Dans une solution à faible force ionique, toutefois, la formation des pores dépend fortement du pH : une augmentation de celui-ci de 6,5 à 10,5 a entraîné une baisse irrégulière et par étapes successives de la perméabilité membranaire. Les quatre préparations de toxine ont néanmoins dépolarisé la membrane apicale d’intestins médians fraîchement isolés baignant dans une solution contenant 122 mM de KCl à pH 10,5. L’activité de la Cry9Ca, et des mutants R164A et R164K, a été grandement stimulée lorsque les expériences ont été effectuées en présence de suc intestinal, de lipides extraits d’un volume équivalent de suc intestinal ou d’un cocktail d’inhibiteurs de protéases solubles dans l’eau. De plus, le rôle des boucles inter-hélicales du Domaine I lors de l’insertion dans la membrane a été étudié avec des mutants doubles de la Cry9Ca dont les mutations introduisaient, neutralisaient ou renversaient une charge électrique. À l’exception de trois d’entres eux, tous ces mutants ont conservé une toxicité et une capacité à former des pores comparables à celles de la toxine parentale. L’ensemble de ces résultats suggère que le micro-environnement de l’intestin médian contribue à minimiser l’influence des charges de surface portées par les résidus des boucles inter-hélicales du Domaine I sur la capacité des toxines du bacille de Thuringe à former des pores. Il indique aussi que, d’une part, selon le site de clivage et les conditions expérimentales utilisées, des protéolyses supplémentaires de la toxine Cry9Ca activée peuvent soit stimuler, soit nuire à son activité et que, d’autre part, le suc intestinal du sphinx du tabac contient probablement un inhibiteur de protéases qui pourrait jouer un rôle important dans l’activité des toxines du bacille de Thuringe. / Once ingested by susceptible insects, Bacillus thuringiensis insecticidal toxins must be activated by the insect’s intestinal proteases. Their first domain, a bundle of seven amphipathic -helices, is responsible for their insertion into the luminal membrane of midgut cells, thereby creating poorly selective pores. The toxicity and pore-forming ability of one such toxin, Cry9Ca, its single-site mutants, R164A and R164K, and of the 55-kDa fragment resulting from its proteolytic cleavage at residue 164 were investigated using a combination of homology modeling, bioassays, osmotic swelling experiments with Manduca sexta larval midgut brush border membrane vesicles and electrophysiological measurements on isolated midguts. Neither the single mutations nor the proteolytic cleavage altered Cry9Ca toxicity. In low ionic strength solutions however, pore formation was highly dependent on pH: increasing pH from 6.5 to 10.5 resulted in an irregular step-wise decrease in membrane permeabilization. All four toxin preparations nevertheless depolarized the apical membrane of freshly isolated midguts bathing in a solution containing 122 mM KCl at pH 10.5. The activity of Cry9Ca, R164A and R164K was greatly enhanced when the experiments were conducted in the presence of midgut juice, the lipids extracted from an equivalent volume of midgut juice or a cocktail of water-soluble protease inhibitors. Additionally, the role of the interhelical loops of Domain I in membrane insertion was investigated with Cry9Ca double mutants with mutations that either introduced, neutralized or reversed an electrical charge. All but three mutants retained a toxicity and a pore-forming ability that were comparable to those of their parental toxin. Overall, the results suggest that the midgut microenvironment contributes to minimizing the influence of surface charges carried by Domain I interhelical loop residues on B. thuringiensis toxins pore-forming ability. They also indicate that, depending on the cleavage site and on the experimental conditions used, further proteolysis of the activated Cry9Ca toxin can either stimulate or be detrimental to its activity and that M. sexta midgut juice probably contains protease inhibitors that could play a major role in the activity of B. thuringiensis toxins in the insect midgut.

Une fois ingérées par un insecte sensible, les toxines insecticides du bacille de Thuringe doivent être activées par les protéases intestinales de cet insecte. Leur premier domaine, un ensemble de sept hélices-α amphipathiques, est responsable de leur insertion dans la membrane luminale de certaines cellules de l’intestin médian, ce qui crée des pores peu sélectifs. La toxicité et la capacité à former des pores d’une telle toxine, la Cry9Ca, de ses mutants simples R164A et R164K et d’un fragment de 55 kDa résultant d’un clivage protéolytique au niveau de son résidu 164 ont été étudiées à l’aide d’une combinaison de modélisation par homologie, de bioessais, d’expériences de gonflement osmotique avec des vésicules de membrane en bordure en brosse de larves de sphinx du tabac et de mesures électrophysiologiques sur des intestins isolés. Ni les mutations simples ni le clivage protéolytique n’ont altéré la toxicité de la Cry9Ca. Dans une solution à faible force ionique, toutefois, la formation des pores dépend fortement du pH : une augmentation de celui-ci de 6,5 à 10,5 a entraîné une baisse irrégulière et par étapes successives de la perméabilité membranaire. Les quatre préparations de toxine ont néanmoins dépolarisé la membrane apicale d’intestins médians fraîchement isolés baignant dans une solution contenant 122 mM de KCl à pH 10,5. L’activité de la Cry9Ca, et des mutants R164A et R164K, a été grandement stimulée lorsque les expériences ont été effectuées en présence de suc intestinal, de lipides extraits d’un volume équivalent de suc intestinal ou d’un cocktail d’inhibiteurs de protéases solubles dans l’eau. De plus, le rôle des boucles inter-hélicales du Domaine I lors de l’insertion dans la membrane a été étudié avec des mutants doubles de la Cry9Ca dont les mutations introduisaient, neutralisaient ou renversaient une charge électrique. À l’exception de trois d’entres eux, tous ces mutants ont conservé une toxicité et une capacité à former des pores comparables à celles de la toxine parentale. L’ensemble de ces résultats suggère que le micro-environnement de l’intestin médian contribue à minimiser l’influence des charges de surface portées par les résidus des boucles inter-hélicales du Domaine I sur la capacité des toxines du bacille de Thuringe à former des pores. Il indique aussi que, d’une part, selon le site de clivage et les conditions expérimentales utilisées, des protéolyses supplémentaires de la toxine Cry9Ca activée peuvent soit stimuler, soit nuire à son activité et que, d’autre part, le suc intestinal du sphinx du tabac contient probablement un inhibiteur de protéases qui pourrait jouer un rôle important dans l’activité des toxines du bacille de Thuringe. / Once ingested by susceptible insects, Bacillus thuringiensis insecticidal toxins must be activated by the insect’s intestinal proteases. Their first domain, a bundle of seven amphipathic -helices, is responsible for their insertion into the luminal membrane of midgut cells, thereby creating poorly selective pores. The toxicity and pore-forming ability of one such toxin, Cry9Ca, its single-site mutants, R164A and R164K, and of the 55-kDa fragment resulting from its proteolytic cleavage at residue 164 were investigated using a combination of homology modeling, bioassays, osmotic swelling experiments with Manduca sexta larval midgut brush border membrane vesicles and electrophysiological measurements on isolated midguts. Neither the single mutations nor the proteolytic cleavage altered Cry9Ca toxicity. In low ionic strength solutions however, pore formation was highly dependent on pH: increasing pH from 6.5 to 10.5 resulted in an irregular step-wise decrease in membrane permeabilization. All four toxin preparations nevertheless depolarized the apical membrane of freshly isolated midguts bathing in a solution containing 122 mM KCl at pH 10.5. The activity of Cry9Ca, R164A and R164K was greatly enhanced when the experiments were conducted in the presence of midgut juice, the lipids extracted from an equivalent volume of midgut juice or a cocktail of water-soluble protease inhibitors. Additionally, the role of the interhelical loops of Domain I in membrane insertion was investigated with Cry9Ca double mutants with mutations that either introduced, neutralized or reversed an electrical charge. All but three mutants retained a toxicity and a pore-forming ability that were comparable to those of their parental toxin. Overall, the results suggest that the midgut microenvironment contributes to minimizing the influence of surface charges carried by Domain I interhelical loop residues on B. thuringiensis toxins pore-forming ability. They also indicate that, depending on the cleavage site and on the experimental conditions used, further proteolysis of the activated Cry9Ca toxin can either stimulate or be detrimental to its activity and that M. sexta midgut juice probably contains protease inhibitors that could play a major role in the activity of B. thuringiensis toxins in the insect midgut.

Text in English / Malaria is endemic throughout Uganda and the leading cause of morbidity and mortality. Malaria causes complications in 80.0% of all pregnancies in Uganda. This study attempted to identify factors that influence pregnant women’s utilisation of anti-malaria services in the Buikwe district of Uganda. These factors were contextualised within the Social Learning Theory’s major concepts. The target populations comprised pregnant women attending antenatal clinics (phase 1) and midwives providing antenatal services (phase 2) at 16 clinics in the Buikwe district of Uganda during the data collection phase of the study. Structured interviews were conducted with a sample of 400 randomly selected pregnant women and with the accessible population of 40 midwives. Pregnant women, who had progressed beyond primary school level education, were more likely to take intermittent preventive treatment (IPT) drugs and to use long lasting insecticide treated nets (LLINs) to prevent malaria. Pregnant women were more likely to implement malaria-preventive actions if they lived within five kilometres of clinics, were satisfied with available health services, were knowledgeable about the malaria preventive measures and had used IPT during previous pregnancies. Pregnant women who implemented one malaria-preventive action were likely to implement other actions as well (Pearson’s correlation coefficient was 0.65; p<0.05). Midwives’ provision of malaria-preventive services to pregnant women were influenced by the availability of IPT drugs, accessibility of safe drinking water, frequency of giving health education to pregnant women, cooperation with village health teams, malaria-related in-service training, midwives’ education level and experience. Although 97.9% of the pregnant women had taken IPT and 84.2% of those who had received LLINs, utilised these nets, malaria prevention during pregnancy could be improved. All pregnant women should attend antenatal clinics at least four times during each pregnancy, commencing during the first trimester of pregnancy to receive adequate health education and prenatal services, including IPT and LLINs. All midwives should receive malaria-related in-service training. Regular audits of midwives’ records should identify and address strengths and weaknesses related to the prevention and management of malaria during pregnancy. Such actions could enhance the prevention and management of malaria, estimated to affect 80% of pregnant women in Uganda. / Health Studies / D. Litt. et Phil. (Health Studies)