A quantitative and qualitative study of the behavioral effects of an insecticide on various life stages of the German cockroach, Blattella germanica (L.), was conducted. Comparisons between a susceptible laboratory strain (VPI) and a resistant field strain (BP) were made to examine the relationship between physiological resistance and behavioral resistance. The effects of vapors from a formulation of 1% propoxur in oil were compared with vapors from a mineral oil control. Vapors from the control had little effect on cockroach behavior. Adult males, adult females, and middle instars were repelled from aquaria containing the insecticide vapors. Early instars sought shelter in the pores of water sponges. No differences in behavior due to reproductive state were observed as females with oothecae and females without oothecae were equally repelled. Responses by adult males included increased movement, antennal grooming, and tarsal grooming. Electroantennogram studies showed that the vapors of the propoxur formulation were detected by the antennae. Adults of the susceptible VPI strain responded more strongly to the insecticide vapors, relative to the controls, than did adults of the resistant BP strain. The negative correlation between physiological resistance and behavioral responses was not due to the development of behavioral resistance. Rather the resistance mechanism of the BP strain was probably responsible for inhibiting the behavioral response of that strain. Responses to the propoxur insecticide may have been compounded by solvents in the formulation, but differences between the two strains were probably due to the propoxur. The oil base had no effect on cockroach behavior. That wet sponges provided safe harborage for the early instars indicates the importance of even short range dispersal. Also, the use of repellent barriers to exclude cockroaches from sensitive areas may become less effective if physiological resistance in cockroach populations is accompanied by decreased avoidance responses. Knowledge of the levels of physiological resistance must be supplemented by an understanding of the behavioral characteristics of insect populations for the successful development of pest control programs. / Ph. D.

Separate field studies were started in fall 2005, which continued through fall 2007, to investigate the effect of different levels of European corn borer tunneling on yield in corn grown for grain and to predict spring infestation levels of early season soil insects, specifically white grubs (Coleoptera: Scarabaeidae) and wireworms (Coleoptera: Elateridae) in cornfields. In the first study, model variables included corn growth stage and larvae per plant. In both years of this study, larvae per plant had a significant effect on grain yield. Grain yield was reduced by 13.1 and 3.65% in plants infested with four larvae per plant in 2006 and 2007, respectively. For 2006, linear regression models provided average percent yield loss per larva per plant at 4.1, 6.8, and 1.8% during late vegetative (V12), early silking (R1), and blister (R2) growth stages, respectively. Economic injury levels (EILs) were calculated based on average percent yield reductions across each growth stage and year. In the second study, no significant differences were detected in both fall and spring between two sampling methods after correcting for differences in sampling volume. Strong correlations were observed between fall and spring grub densities in both years. In 2006, fields with grub densities above the spring nominal threshold had significantly greater stand and yield in the Poncho 1250 (1.25 mg clothianidin / kernel) treatment when compared to the Poncho 250 (0.25 mg clothianidin / kernel) and untreated plots. This information was used to develop fall EILs and economic thresholds for white grubs. / Master of Science in Life Sciences

Diseases such as malaria and dengue impose great economic burdens and are a serious threat to public health, with young children being among the worst affected. These diseases are transmitted by mosquitoes, also called disease vectors, which are able to transmit both parasitic and viral infections. One of the most important strategies in the battle against mosquito-borne diseases is vector control by insecticides and the goal is to prevent people from being bitten by mosquitoes. Today’s vector control methods are seriously threatened by the development and spread of insecticide-resistant mosquitos warranting the search for new insecticides. This thesis has investigated the possibilities of vector control using non-covalent inhibitors targeting acetylcholinesterase (AChE); an essential enzyme present in mosquitoes as well as in humans and other mammals. A key requirement for such compounds to be considered safe and suitable for development into new public health insecticides is selectivity towards the mosquito enzyme AChE1. The work presented here is focused on AChE1 from the disease transmitting mosquitoes Anopheles gambiae (AgAChE1) and Aedes aegypti (AaAChE1), and their human (hAChE) and mouse (mAChE) counterparts. By taking a medicinal chemistry approach and utilizing high throughput screening (HTS), new chemical starting points have been identified. Analysis of the combined results of three different HTS campaigns targeting AgAChE1, AaAChE1, and hAChE allowed the identification of several mosquito-selective inhibitors and a number of compound classes were selected for further development. These compounds are non-covalent inhibitors of AChE1 and thereby work via a different mechanism compared to current anti-cholinergic insecticides, whose activity is the result of a covalent modification of the enzyme. The potency and selectivity of two compound classes have been explored in depth using a combination of different tools including design, organic synthesis, biochemical assays, protein X-ray crystallography and homology modeling. Several potent inhibitors with promising selectivity for the mosquito enzymes have been identified and the insecticidal activity of one new compound has been confirmed by in vivo experiments on mosquitoes. The results presented here contribute to the field of public health insecticide discovery by demonstrating the potential of selectively targeting mosquito AChE1 using non-covalent inhibitors. Further, the presented compounds can be used as tools to study mechanisms important in insecticide development, such as exoskeleton penetration and other ADME processes in mosquitoes.

MgAl layered double hydroxides nanoparticles (LDHs) are known as the useful materials in agrochemsitry. LDHs can be used as a bio-insecticide carrier to enhance insecticide’s activity efficiency. In our study, to improve the insecticide activity of anacardic acid, an extract from cashew nut shell liquid, we intercalated it MgAl layered double hydroxides nanoparticles. Different hybridization between anacardic acid and LDHs (37, 74, 148, and 296μg/mL) (L-As) were made and tested on the survivals of cutworms (Spodoptera litura). L-As or free anacardic acid was sprayed directly on the leaves mustard to feed cutworms or directly on the skin of cutworms. Our results showed that in all L-As treatments, the worm killing efficiency was higher than the free anacardic acid treatment. / Hạt nano lớp đôi hydroxides MgAl (LDHs) được biết đến như là những vật liệu hữu ích trong nông ngành hóa học nông nghiệp. LDHs có thể được dùng như là một loại chất mang cho thuốc trừ sâu sinh học để tăng cường hiệu lực diệt sâu. Trong nghiên cứu này, để tăng cường hiệu lực diệt sâu của anacardic acid, một loại hoạt chất được chiết từ dầu vỏ hạt điều, chúng tôi đã gắn chèn nó lên hạt nano lớp đôi hydroxides MgAl. Các nồng độ khác nhau của dạng lai của anacardic và LDHs (37, 74, 148 và 296μg/mL) (L-As) đã được kiểm tra tỷ lệ sống của ấu trùng sâu khoang (Spodoptera litura). Các nghiệm thức L-As và dạng anacardic acid tự do đã được phun lên lá rau cải ngọt cho ấu trùng sâu ăn hoặc phun trực tiếp lên da ấu trùng sâu. Kết quả cho thấy, tất cả các công thức có xử lý bằng L-As, hiệu lực diệt ấu trùng sâu đều cao hơn so với dạng anacardic acid ở trạng thái tự do.

Orientador: Claudia Marcia Aparecida Carareto / Banca: Ricardo De Marco / Banca: André Luís Laforga Vanzela / Resumo: A resistência aos inseticidas é um modelo de processo evolutivo onde o inseticida atua como agente seletivo e, como resposta à seleção, ocorre a evolução da resistência nas populações de insetos. As enzimas citocromo P450 monooxigenases (CYP) formam uma família responsável pela resistência aos inseticidas. Tem sido proposto que a inserção de elementos transponíveis (TEs) em regiões reguladoras ou codificadoras dos genes da família Cyp pode alterar a expressão gênica e induzir a resistência aos inseticidas. No presente estudo foram realizadas análises in silico que permitiram identificar a ocorrência de inserções de fragmentos de TEs em 35 genes Cyps com diferentes funções, e em seus genes flanqueadores, em Drosophila melanogaster e D. simulans, além de 13 genes Cyps de seis espécies do grupo melanogaster de Drosophila. As inserções de TEs ocorreram principalmente nas regiões flanqueadoras 5' dos Cyps associados à resistência aos inseticidas e à função monooxigenase geral. Os resultados não indicaram qualquer relação entre a distância em relação ao gene e o número de inserções. As análises mostraram que a maioria das inserções pertence à classe de transposons de DNA, sendo o transposon DNAREP1_DM o que apresentou o maior número de cópias. O fato de essas seqüências apresentarem putativos sítios de ligação de fatores de transcrição sugere que possam desempenhar algum papel na regulação dos genes Cyps. Também foi analisada a ocorrência de polimorfismos de inserção de TEs em regiões flanqueadoras de genes da família Cyp, em diferentes linhagens geográficas resistentes e suscetíveis, de D. melanogaster e D. simulans. Análises evidenciaram a presença de polimorfismo interpopulacional de tamanho das regiões flanqueadoras dos genes Cyp6w1, Cyp6a2 e Cyp12d1, porém, não indicaram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Insecticide resistance is a model of evolutionary process where the insecticide acts as the selective agent and resistance in the insect populations evolves as an answer to selection. Cytochrome monooxygenases (CYP) is family of enzymes responsible for the insecticide resistance. It has been proposed that insertion of transposable elements (TEs) in regulatory or coding regions of the Cyp genes can alter gene expression and induce insecticide resistance. In the present study in silico analyses allowed identifying the insertion of TE fragments in 35 Cyp genes with different functions, in Drosophila melanogaster and D. simulans, as well as in 13 Cyps of six species of the melanogaster group of Drosophila. The TE insertions occurred mainly in the 5' flanking regions of Cyp genes associated to resistance and to those with a general monooxygenase function. The results did not indicate any relationship between the number of insertions and the distance in relation to the gene. The analyses showed that most of the insertions belong to the DNA transposon class, being DNAREP1_DM the most numerous. Since this element carry putative biding sites of transcription factors it can be suggested they play same role in gene regulation. The polymorphism of TE insertions in the flanking regions of Cyp6w1, Cyp6a2 and Cyp12d1, genes associated to resistance, found in resistant and as well as in susceptible geographical strains of D. melanogaster and D. simulans, does not indicate any relationship between the presence of TEs in those regions and the insecticide resistance. The results also showed that the insertions of TEs in the proximities of the Cyps associated to resistance is differential among six species of the melanogaster group, not following the genomic proportion of TEs in each species. These results also suggest that TEs inserted in the Cyp flanking regions can carry out an adaptive... (Complete abstract click electronic access below) / Mestre

Os objetivos do presente trabalho foram avaliar a ocorrência / persistência / degradação de resíduos do inseticida organofosforado pirimifós-metil em grãos de trigo, milho e milho pipoca e em alguns de seus produtos processados, bem como a ação residual desse inseticida sobre Sitophilus spp. (Coleoptera, Curculionidae). O tratamento dos grãos foi realizado de modo a se obter a concentração teórica de 12 mg.kg-1 (ppm) de pirimifós-metil, tendo sido aplicados o equivalente a 5 litros de solução por tonelada de grãos. As amostras para estudos de resíduos foram tomadas aos zero, 15, 30, 60, 120 e 240 dias após o tratamento. O método analítico consta da extração dos resíduos com acetona, limpeza com partição acetonitrila/hexano seguida de coluna cromatográfica de sílica eluída com mistura de hexano+acetona (9/1, v/v). A determinação quantitativa foi feita por técnica de cromatografia em fase gasosa, usando-se cromatógrafo equipado com detector fotométrico de chama (GLC/FPD). Foram analisadas 264 amostras no total, sendo 72 de grãos cereais e 192 de seus produtos processados. Para os estudos de ação residual do inseticida sobre adultos de Sitophilus spp. os insetos foram submetidos a testes em exposição aos grãos tratados após 15 dias decorridos do tratamento, 30 dias após, e, subseqüentemente, em intervalos mensais até 12 meses. Os limites de quantificação dos resíduos pelo método de análise, para os diferentes substratos variaram de 0,05 a 0,1 mg.kg-1. Os resíduos de pirimifós-metil não foram persistentes nos grãos nem em seus produtos processados, decrescendo de 4-8 vezes no período de amostragem. Eles foram maiores nos produtos processados ricos em óleos (farelo de trigo e de milho), menores nos grãos (estes semelhantes às farinhas integral e branca, e pipoca) e, ainda menores, no pão, farinha de milho e canjica. O inseticida pirimifós-metil mostrou-se bastante eficiente no controle de Sitophilus spp., no tratamento dos três tipos de grãos cereais durante todo o período de observação. / The purposes of this study were to evaluate the occurence, persistence and degradation of pyrimiphos-methyl organophosphorus insecticide residues on wheat, corn and popcorn grains in and some of their processed products, as well the residual action of this insecticide for the control the Sitophilus spp. (Coleoptera, Curculionidae). Grain treatment was performed to obtain the theoretic concentration of 12 mg.Kg-1 (ppm) of pyrimiphos-methyl, which was applied at the dosage of 5 L of solution/ton of the grain. Sampling was performed at 0, 15, 30, 60, 120 and 240 days after the treatment. The samples were analyzed for residues of pyrimiphos-methyl using a residue method based on extraction with acetone, clean up by partition with acetonitrile/hexane followed by silica gel column chromatography eluted with a mixture of hexane + acetone (9:1) (v:v) solution. Residue determinations were done by the gas chromatography technique using a flame photometry detector (GLC/FPD). Total samples analyzed were 264, being 72 of cereal grains and 192 of processed products. The studies of residual action of the insecticide for the control of Sitophilus spp adults were conducted with adults and exposing the insects to treated grains 15 days after the application, 30 days, and subsequently at monthly intervals up to 12 months. The limits of quantitation of the analytical method for different substracts ranged between 0.05 to 0.1 mg.kg-1 (ppm). Pyrimiphos-methyl residues were not persistent on the grains and processed products. A decay of 4 to 8 times during the sampling period was observed. The residues found were higher in the processed products with high oil content (wheat and corn brans), lower on grains (similar to whole and white flours and popcorn) and even lower in bread, corn flour and "canjica". Pyrimiphos-methyl was very efficient for the control of Sitophilus spp on the three grain types during the entire period of observation.

Um dos inseticidas mais utilizados para o controle do psilídeo Diaphorina citri (Hemiptera: Liviidae) em pomares de citros no Brasil tem sido o neonicotinoide imidacloprid. Para subsidiar um programa de Manejo da Resistência de D. citri a imidacloprid foram realizados estudos de monitoramento da suscetibilidade a inseticidas em populações de D. citri provenientes das principais regiões citrícolas do Estado de São Paulo, interações de imidacloprid com inseticidas e fungicidas, e avaliação da viabilidade de associação de imidacloprid com o parasitoide Tamarixia radiata (Hymenoptera: Eulophidae). Para a caracterização de linhas básicas de suscetibilidade de D. citri aos inseticidas imidacloprid, deltamethrin e dimethoate foram utilizados bioensaios de contato residual. Foram definidas as concentrações diagnósticas de 56 mg de imidacloprid/L água (CL95), 32 mg de deltamethrin/L água (CL90) e 56 mg de dimethoate/L água (CL95) para o monitoramento da suscetibilidade de 25 populações de D. citri entre 2010 e 1012. Não ocorreram diferenças na sobrevivência aos inseticidas testados em populações de psilídeos coletados em pomares de citros com diferentes intensidades de pulverização com inseticidas. A maior sobrevivência de insetos nas concentrações diagnósticas foi observada em 2010, com valores de sobrevivência variando de 4,7% a 24,0% para imidacloprid, de 0,9% a 11,8% para deltamethrin e de 5,2% a 13,0% para dimethoate. Não foram observados aumentos significativos na sobrevivência de D. citri nos monitoramentos realizados em 2011 e 2012. As interações de imidacloprid com deltamethrin ou dimethoate foram testadas com as CL25 dos respectivos inseticidas em bioensaio de contato residual. As interações foram aditivas sobre a mortalidade de adultos em condições de campo e de casa de vegetação. A mistura de imidacloprid com buprofezin e pyriproxyfen sobre ninfas de 3º instar também foi aditiva. A atividade biológica das concentrações campo de imidacloprid (40?L/mL) e buprofezin (375?g/mL) e da mistura dos mesmos mostrou uma degradação da atividade similar para o controle de D. citri em condições de campo e casa de vegetação. A avaliação da persistência da mistura de imidacloprid e pyriproxyfen (avaliado nas concentrações de 6,25?g/mL e 100?g/mL) indicou que pyriproxyfen teve degradação mais rápida que imidacloprid. A mistura de imidacloprid na CL25 para D. citri com a concentração de campo dos fungicidas Benzimidazol (500?g/mL de tiofanato-metilico), estrubilurina (37,5?g /mL de piraclostrobina) e triazol (50?g/mL de difenenoconazol) não afetaram o desempenho de imidacloprid. Houve significativa redução na taxa instantânea de crescimento (ri) de D. citri quando expostas a diferentes idades de resíduos de imidacloprid pulverizado na concentração de 40?g/mL, mesmo sobre resíduos com 56 dias de idade, levando à extinção da população ou em processo de extinção. A exposição a CL5 (0,501?g/mL), CL10 (0,804?g/mL), CL25 (1,995?g/mL) e CL50 (5,213?g/mL) de imidacloprid para D. citri reduziu o crescimento populacional de D. citri, mas não causou sua extinção. Contudo, essas concentrações de imidacloprid afetaram a ri do parasitoide com a CL50 causando sua extinção. / One of the most widely used insecticides for control of Diaphorina citri (Hemiptera: Liviidae) in citrus groves in Brazil has been the neonicotinoid imidacloprid. To implement an Insect Resistance Management program of D. citri to imidacloprid, studies were conducted to monitor the susceptibility to insecticides in D. citri populations collected from main citrus production regions of São Paulo State, to evaluate the interactions of imidacloprid with insecticides and fungicides, and to assess the feasibility of imidacloprid association with the parasitoid Tamarixia radiata (Hymenoptera: Eulophidae). Residual contact bioassays were used to characterize the baseline susceptibility of D. citri to the insecticides imidacloprid, deltamethrin and dimethoate. Diagnostic concentrations of 56 mg of imidacloprid/L water (LC95), 32 mg of deltamethrin/L water (LC90) and 56 mg of dimethoate/L water (LC95) were defined for monitoring the susceptibility in 25 populations of D. citri from 2010 to 2012. There were no differences in survival to the insecticides tested in D. citri populations collected in citrus groves with different regimes of insecticide use. The highest survival of insects at diagnostic concentrations was observed in 2010, with survival values ranging from 4.7% to 24.0% for imidacloprid, from 0.9% to 11.8% for deltamethrin and 5.2% to 13.0% dimethoate. There were no significant increases in survival of D. citri on monitoring conducted in 2011 and 2012. The interactions of imidacloprid with deltamethrin or dimethoate were tested with the LC25 of each insecticide with residual contact bioassays. The interactions of these insecticides were additive on D. citri adult mortality under field or greenhouse conditions. The interactions of imidacloprid with buprofezin and pyriproxyfen were additive on 3rd instar nymphs. The biological activity at field rates of imidacloprid (40?l/mL) and buprofezin (375?g/mL) as well as the mixture of these insecticides showed a similar degradation in the activity to control D. citri under field and greenhouse conditions. The evaluation of the persistence of the mixture of pyriproxyfen and imidacloprid (evaluated at concentrations of 6.25 ?g/mL and 100?g/mL) indicated that pyriproxyfen degradation was faster than imidacloprid. The mixture of LC25 of imidacloprid to D. citri with fungicides field rates of benzimidazole (500?g/mL of thiophanate-methil), strobilurin (37.5 ?g/mL of pyraclostrobin), and triazole (50?g/mL of difenoconazole) did not affect the performance of imidacloprid. A significant reduction in the instantaneous rate of increase (ri) of D. citri was observed when exposed to different residue ages of imidacloprid sprayed at concentration of 40?g/mL, even when exposed to residues of 56-day old, by leading to their extinction or in extinction process. The exposure to CL5 (0.501 ?g/mL), CL10 (0.804 ?g/mL), LC25 (1.995 ?g/mL) and LC50 (5.213 ?g/mL) of imidacloprid to D. citri did not cause their extinction. However, these concentrations of imidacloprid affected the ri of the parasitoid causing their extinction at LC50.

O presente trabalho foi subdividido em quatro capítulos com os seguintes objetivos: (i) apresentar revisão bibliográfica abordando as principais referências presentes na literatura sobre a cultura da batata, Solanum tuberosum L. (Solanaceae), os aspectos bioecológicos da traça-da- -batatinha, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), os hormônios no desenvolvimento dos lepidópteros e a atuação de alguns reguladores de crescimento de insetos (RCIs); (ii) avaliar a ingestão dos carboidratos por adultos de P. operculella e sua influência na reprodução da espécie; (iii) testar parâmetros reprodutivos devido à ingestão de alguns ingredientes ativos presentes em RCIs; (iv) avaliar a ingestão de carboidratos aplicados sobre plantas de batata de maneira a ser possível a aplicação de RCIs como método de redução populacional da praga. Inicialmente, para comprovação da ingestão dos carboidratos, foi utilizada a técnica de marcadores coloridos, em que se ofereceu, por capilaridade via rolete dental, solução de mel a 10% contendo corante líquido artificial de coloração azul anis. Os insetos foram confinados em um recipiente cilíndrico construído com tubo de PVC rígido. Após três dias de oferecimento da alimentação, analisou-se a presença do corante no sistema digestivo dos insetos. Para verificação da ingestão de carboidrato em condições mais próximas ao ambiente das culturas agrícolas, foi realizada nova avaliação em recipientes com diferentes dimensões: 0,0018; 0,054 e 9,6 m3. Posteriormente, simulando um borrifamento na cultura, foi realizada uma terceira avaliação no ambiente de 9,6 m3, na qual a solução de mel foi oferecida aos insetos em gotas grandes no limbo foliar de plantas de batata. Para análise da influência dos carboidratos na reprodução da espécie, foram verificados os seguintes parâmetros reprodutivos: ritmo de postura, fecundidade, viabilidade dos ovos e índice de ovigenia. Para análise dos efeitos dos RCIs, foram utilizados quatro compostos com diferentes ingredientes ativos: azadiractina, lufenurom, metoxifenozida e piriproxifem. Estes compostos foram oferecidos aos insetos em solução de mel a 10%, nas concentrações de 5, 10 e 20 ppm. Durante 10 dias após o acasalamento, foram analisados os parâmetros reprodutivos. O corante colorido mostrou-se adequado para os estudos visuais do sistema digestivo de P. operculella. A ingestão dos carboidratos pelos insetos ocorreu somente quando estes foram confinados em ambientes menores, 0,0018 e 0,054 m3. No ambiente maior, de 9,6 m3, a ingestão ocorreu somente quando a solução foi oferecida via gotas em plantas de batata. Quanto aos efeitos dos RCIs, verificou-se diminuição significativa na fecundidade das fêmeas de P. operculella que ingeriram solução de mel contendo azadiractina e lufenurom à concentração de 5 ppm e piriproxifem à concentração de 20 ppm. Quanto aos parâmetros viabilidade dos ovos e período embrionário não foram verificadas diferenças significativas entre os tratamentos. Este fato sugere que a fertilidade não foi alterada, demonstrando que os RCIs não afetam os machos. A comprovação da ingestão dos carboidratos oferecido via gotas no limbo foliar de plantas de batata e os resultados obtidos na redução da fecundidade de P. operculella indicaram que esta técnica pode ser utilizada em programas de manejo integrado da traça-da-batatinha. / The present work has been subdivided into four chapters, with the following objectives: (i) to present a bibliographic review of the main aspects related to the potato crop, Solanum tuberosum L. (Solanaceae), the bioecological aspects of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), the hormones related to lepidopterans development and the role of some insect growth regulators (IGRs); (ii) to evaluate the carbohydrate intake by P. operculella adults and its influence on the species reproduction; (iii) to test the effects of active ingredients of IGRs on reproductive parameters; (iv) to evaluate the ingestion of carbohydrates applied on potato plants, in order to allow IGRs application as a population reduction strategy for this pest. Initially, confirmation of carbohydrate consumption was by a means colored marker technique, which offers a 10% honey solution containing an artificial liquid food dye, blue anise, by capillarity via a dental roll wick. The insects were caged in a cylindrical container made of a rigid PVC tube. After three days of food offering, dye presence was analyzed within the insects digestive system. For confirmation of carbohydrate intake under simulated field conditions, an additional evaluation was conducted in containers with different sizes: 0.0018, 0.054 and 9.6 m3. Later, a third evaluation was carried out in the 9.6 m3 container, in which the honey solution was offered to insects as big drops applied on the leaf blade of potato plants, simulating a spray application. To analyze carbohydrate influence on species reproduction, the following reproductive parameters were assessed: oviposition rate, fertility, egg viability and ovigeny index. Four compounds with different active principles were utilized to analyze IGR effects: azadirachtin, lufenuron, methoxyfenozide and pyriproxyfen. These compounds were offered to the insects in a 10% honey solution at concentrations of 5, 10 and 20 ppm. During 10 days after mating the reproductive parameters were evaluated. The colored dye method proved to be efficient for visual studies of P. operculellas digestive system. Carbohydrate ingestion by the insects occurred only when they were caged in small environments, 0.0018 and 0.054 m3. In larger ones 9.6 m3, feeding occurred when the solution was offered by dripping over potato plants. Results showed a significant reduction of P. operculella fertility among females that ingested the honey solution containing azadirachtin and lufenuron, at 5 and 20 ppm, respectively. Regarding egg viability and embrionary period, no significant differences were found between the treatments. This fact suggests that fertility was not affected, thus demonstrating that IGRs did not affect males. Confirmation of carbohydrate ingestion offered by dripping over potato leaf blades and the results of fertility reduction in P. operculella, indicated that this technique may be utilized in integrated management programs against the potato tuber moth.

Les mutations sont à l'origine des nombreux "variants" présents dans les populations naturelles. Les variants adaptatifs sont propagés par sélection naturelle. Cependant, une mutation bénéfique sur un trait peut affecter négativement d’autres traits (coût sélectif): un compromis émerge alors entre les avantages et les coûts qu’elle induit. Cette thèse vise à comprendre comment des modifications de l’environnement peuvent affecter les compromis évolutifs de différents types de mutations adaptatives (substitutions, duplications hétérogènes, amplifications). Chez les moustiques, l’utilisation d’insecticides organophosphorés (OPs) et carbamates (CXs) a sélectionné trois réponses adaptatives majeures : une amplification de gènes au locus Ester (codant pour des enzymes détoxicantes), une substitution au locus ace-1 (codant pour la cible des insecticides), et des duplications associant une copie sensible et une copie résistante du locus ace-1. Un premier axe de ma thèse a été de mieux comprendre le rôle de ces duplications hétérogènes (qui associent deux copies divergentes d’un même gène) dans l’adaptation. En caractérisant leurs compromis évolutifs nous avons montré qu'elles confèrent un phénotype proche de celui d’hétérozygotes standards. Toutefois, l’étude de leur distribution mondiale et des analyses en laboratoire ont révélé que ces duplications, avantageuses à l’état hétérozygote, sont majoritairement sublétales à l’état homozygote. Le second axe de cette thèse a été l’étude de l’influence des variations de pression de sélection sur la dynamique des allèles adaptatifs. Une étude d’évolution expérimentale a montré que des pressions de sélection intermédiaires pouvaient générer des situations de superdominance au locus ace-1, favorables à la sélection de duplications hétérogènes. Par ailleurs, l’analyse d’échantillons montpelliérains récoltés sur une trentaine d’années nous a permis de relier quantitativement les variations de la pression de sélection et les variations de la valeur sélective des différents allèles du locus Ester. Enfin, l’étude de trois zones géographiques (Mayotte, Martinique, et Montpellier) a permis de montrer que les différentes adaptations ne répondaient pas de la même façon à une modification environnementale majeure liée au retrait de la pression de sélection (interdiction des OPs et CXs en 2007) : alors que les allèles de résistance du locus ace-1 tendent à disparaitre, ceux du locus Ester se maintiennent en fréquence non négligeable dans les populations naturelles. / Mutations are the origin of the many "variants" present in natural populations. Adaptive variants are propagated by natural selection. However a mutation beneficial for a trait can negatively affect other traits (selective cost): a trade-off thus emerges between the benefits and the costs it induces. This PhD aimed at understanding how environmental changes could affect the evolutionary trade-offs of various types of adaptive mutations (substitutions, heterogeneous duplications, amplifications). In mosquitoes, organophosphate (OPs) and carbamates (CXs) insecticides usage has selected three major adaptive responses: gene amplifications at the Ester locus (encoding detoxifying enzymes), a substitution at the ace-1 locus (encoding the target of the insecticides), and gene duplications pairing susceptible and resistance ace-1 copies. The first axis of my PhD aimed at understanding the role of these heterogeneous duplications (combining two different copies of the same gene) in adaptation. Characterizing their evolutionary trade-offs, we showed that they confer a phenotype similar to standard heterozygotes. However, the study of their worldwide distribution and laboratory analyzes showed that these duplications, advantageous at the heterozygous state, are mostly sublethal when homozygous. The second axis of this PhD was the study of the impact of selection pressure variations on the dynamics of adaptive alleles. An experimental evolution study showed that intermediate selective pressures could generate overdominance situations at the ace-1 locus, promoting the selection of heterogeneous duplications. Furthermore, analyzing Montpellier samples collected over a 27 years period allowed us establishing the quantitative relationship between selective pressure variations and fitness variations for the different Ester resistance alleles. Finally, by studying three different geographical areas (Mayotte and Martinique islands and Montpellier) we showed that the various adaptations were not responding similarly to a major environmental change resulting from the selection pressure withdrawal (OPs and CXs were banned in 2007): while the ace-1 locus resistance alleles tended to disappear, those of the Ester locus remained at a significant frequency in natural populations.

As leishmanioses e particularmente a leishmaniose visceral (LV) são doenças transmitidas por vetores artrópodes candidatas a experimentar uma grande expansão territorial em virtude de problemas relacionados ao aquecimento global. Este evento climático deverá causar grande impacto sobre a distribuição geográfica do artrópode transmissor no Brasil e no mundo. Com efeito, nos últimos 20 anos a situação epidemiológica da LV no Brasil vem se modificando de um padrão esporádico prevalente eminentemente em áreas rurais para uma condição de epidemias peri-urbanas que pode afetar todos os estratos sociais da população, tornando-se uma séria ameaça à saúde pública. As leishmanioses são consideradas até o momento doenças não preveníveis e seu padrão epidemiológico vêm se alterando de forma flagrante, o que demanda urgência para o desenvolvimento de novas ferramentas de controle e tratamento. Dentre as diversas questões levantadas sobre as demandas em pesquisa relacionadas ao controle desta enfermidade, destaca-se a importância de avanços em estudos de epidemiologia quantitativa e modelagem matemática que permitam prever efeitos de vacinações de populações empregando-se imunógenos com eficácia e/ou cobertura vacinal menor que 100%, o que parece ser uma realidade com as vacinas contra leishmanioses desenvolvidas até então pelos laboratórios no mundo todo. O sucesso de estratégias eficazes para o controle da LV depende do conhecimento de diversos parâmetros da dinâmica de infecção nas diferentes populações e espécies que atuam na cadeia epidemiológica da doença. Esse estudo teve objetivo de avaliar a efetividade de vacinas contra leishmaniose em cães bem como da utilização de coleira impregnada com inseticida através de um estudo de coorte realizado em uma região de transmissão moderada de leishmaniose visceral canina. Foram construídas seis coortes compostas por animais não reagentes ao teste rápido TR-DPP® e ao teste EIE-ELISA®. Todos os animais apresentaram estado clínico normal, conforme avaliação semiológica. As coortes compreendem grupos de animais sem qualquer medida de controle (grupo N), grupos de animais com aplicação de coleira (grupo C), grupos de animais vacinados com vacina de subunidade (grupo V1) e grupos de animais vacinados com vacina recombinante (grupo V2) e grupos de animais vacinados e com coleira (grupos V1C e V2C). Foram colhidas amostras de todas as coortes em três tempos com intervalo de seis meses cada, para sorodiagnóstico. A efetividade encontrada ao final de 12 meses de observação para os grupos C, V1, V2, V1C e V2C foram 38,2%, 58,1%, 35%, 68,6% e -36,5% respectivamente com base nos cálculos estatísticos feitos por regressão de Cox para riscos proporcionais. Todas as coortes, mesmo tendo desempenhando alguma efetividade exceto V2C, os resultados dos intervalos de confiança do risco relativo não foram significativos quando comparados ao grupo controle (N). / Leishmaniasis and particularly the LV are diseases transmitted by arthropod vectors candidates to experience a wide territorial expansion because of problems related to global warming. This weather event should cause great impact on the geographical distribution of the arthropod transmitter in Brazil and worldwide. Indeed, the past 20 years the epidemiological situation of LV in Brazil has been changing a prevalent sporadic pattern predominantly in rural areas to a condition of peri-urban epidemics that can affect all social strata of the population, making it a serious threat public health. Leishmaniasis are considered so far not preventable disease and its epidemiological pattern have been changing blatantly, which requires urgency to the development of new tools for control and treatment. Among the many questions raised about the demands on research related to the control of this disease, it highlights the importance of advances in quantitative epidemiological studies and mathematical modeling to anticipate vaccinations effects of employing immunogens effectively and / or lower vaccination coverage to 100%, which appears to be a reality with vaccines against leishmaniasis developed so far by laboratories worldwide. The success of effective strategies to control the LV depends on the knowledge of many aspects of the dynamics of infection in different populations and species that act in the epidemiological chain of the disease. This study aims to evaluate the effectiveness of vaccines against leishmaniasis in dogs as well as the use of insecticide impregnated collar with through a cohort study in a high transmission of canine visceral leishmaniasis region. It was built six cohorts composed of non-reactive animals to the rapid test DPP® and EIE-ELISA® test. All the animals had normal clinical status, as symptomatic evaluation. The cohorts include groups of animals without any measure of control (group N), group of animals with collar application (group C), groups of vaccinated animals with subunit vaccine (group V1), group of animals vaccinated with recombinant vaccine (group V2) and groups of animals vaccinated and collar application (V1C and V2C). Samples were collected from all cohorts in three times at intervals of six months each for serodiagnosis. The effectiveness found after 12 months of observation for groups C, V1, V2, V1C and V2C were 38.2%, 58.1%, 35%, 68.6% and -36.5% respectively based on the statistical calculations done by Cox proportional hazards regression to. All cohorts, even though playing some effectiveness except V2C, the results of risk relative confidence intervals were not significant when compared to the control group (N).