Submitted by EDISON ANTONIO DE SOUZA null (esmidia@gmail.com) on 2016-01-06T16:51:45Z No. of bitstreams: 1 TESE_DOUTORADO_Edison_Souza.pdf: 947001 bytes, checksum: a8a8d690c84bc72a0b5e27d8ebeee92f (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-01-06T19:50:21Z (GMT) No. of bitstreams: 1 souza_ea_dr_bot_par.pdf: 573913 bytes, checksum: 30e1665c27c8c583a0b5ab8dc8bd89e0 (MD5) / Made available in DSpace on 2016-01-06T19:50:21Z (GMT). No. of bitstreams: 1 souza_ea_dr_bot_par.pdf: 573913 bytes, checksum: 30e1665c27c8c583a0b5ab8dc8bd89e0 (MD5) Previous issue date: 2015-12-16 / INFLUÊNCIA DO CONSUMO DA PRÓPOLIS NA EXPRESSÃO DE GENES RELACIONADOS AO SISTEMA IMUNOLOGICO DE ABELHAS Apis mellifera L. SUMETIDAS AO DESAFIO BACTERIANO. As abelhas Apis mellifera podem estar sujeitas a uma série de ameaças como parasitas e patógenos que acometem seu sistema imunológico. Tal fato torna necessária a busca por produtos naturais que possam contribuir com a melhora do sistema imune destes insetos, como a própolis. Diante do exposto, o objetivo foi analisar a influência do fornecimento da própolis em expressões de genes relacionados à imunidade de abelhas Apis mellifera L. submetidas ao desafio bacteriano. Ao longo de 30 dias, quatro colmeias receberam semanalmente os tratamentos com diferentes porcentagens de extrato alcoólico de própolis 30% (0%, 5%, 10%, 15%). O experimento foi casualizado em esquema fatorial 4 x 2 x 2x 3 (tratamentos x com ou sem bactéria x tempos x períodos), totalizando 48 amostras. Foram observadas as expressões dos genes abaecin, hymenoptaecin, apidaecin e defensin1. Como controle interno foi utilizado o gene actina. Os resultados foram comparados por ANOVA seguidos do teste de Tukey (P<0,05). Foram observadas alterações na expressão gênica das abelhas estudadas para todos os períodos e tratamentos, antes e após desafio bacteriano, para todos os genes propostos, sendo ainda verificada induções da expressão relativa nos três períodos. Conclui-se que nas condições do presente trabalho, a própolis pode induzir a expressão relativa dos genes abaecin, hymenoptaecin, apidaecin e defensin1, quando submetidas a desafio bacteriano. Entretanto, não se pôde caracterizar um tratamento com própolis que apresente uma maior expressão relativa para todos os genes simultaneamente. EFEITO DO CONSUMO DE PRÓPOLIS NAS ALTERAÇÕES COMPORTAMENTAIS E MORTALIDADE DE ABELHAS Apis mellifera L. SUBMETIDAS AO INSETICIDA FIPRONIL. Os inseticidas representam o maior risco direto para os polinizadores, como as abelhas Apis mellifera, sendo responsável pela redução das populações de abelhas e da produção apícola, tornando necessária a busca por substâncias que possam contribuir na melhora da saúde dessas abelhas, como a própolis. Este estudo verificou o efeito do consumo do extrato alcoólico de própolis no comportamento, quando submetidas a DL50 (0,2 µg/abelha) e subletal (1/500 DL50), e mortalidade das abelhas Apis mellifera L. quando submetidas a diferentes doses do inseticida fipronil (0,05; 0,1; 0,2; 0,3 e 0,4 µg/abelha). Ao todo foram utilizadas quatro colmeias, uma para cada tratamento (0, 5, 10 e 15% de inclusão de própolis), e em três períodos (0,15 e 30 dias) para verificar o efeito dos tratamentos ao longo do experimento. Para a realização dos testes de mortalidade e de alterações comportamentais foram selecionadas as abelhas campeiras das colmeias tratadas. Para o teste de mortalidade, os dados foram analisados em modelo linear misto generalizado, adicionado do Teste de Tukey (P<0,05) e a análise da avaliação das atividades comportamentais foi feita por (ANOVA) seguida do Teste de Tukey (P<0,05). As colmeias que consumiram própolis apresentaram menor taxa de mortalidade quando comparados com a colmeia controle (0%), com exceção no dia 0 em que a colmeia controle não diferiu do tratamento 10%. Verificou-se ainda que as abelhas que receberam o tratamento 10% apresentaram as menores taxas de mortalidade nos dias 15 e 30 de coleta. Além disso, foi observada mortalidade de 23% nas abelhas quando submetidas a DL50 do fipronil. Nos testes de alterações comportamentais e locomotoras não foram observadas influências dos tratamentos com própolis. Conclui-se que, para o presente estudo, a própolis influenciou na taxa de mortalidade e pode ter promovido um efeito protetor nas abelhas Apis mellifera L. quando submetidas a diferentes concentrações do inseticida fipronil, sendo ainda verificado que o tratamento 10% foi o mais eficaz, devido a ter apresentado as menores taxas de mortalidade nos dias 15 e 30. / INFLUENCE OF PROPOLIS CONSUMPTION IN GENES EXPRESSION RELATED TO IMMUNE SYSTEM OF Apis mellifera L. BEES SUBJECTED TO CHALLENGE BACTERIA. The Apis mellifera bees may be subject to a number of threats as parasites and pathogens that affect your immune system. This fact makes it necessary to look for natural products that can contribute to improving the immune system of these insects such as propolis. Given the above, the objective was to analyze the influence of the supply of propolis in gene expressions related to immunity of Apis mellifera L. bees subjected to bacterial challenge Over 30 days, four hives receive weekly treatments with different percentages of alcoholic extract of propolis 30% (0%, 5%, 10%, 15%). The experiment was randomized in a factorial 4 x 2 x 2x 3 (treatments x with or without bacteria x times x periods), totaling 48 samples. It was observed the expressions of abaecin, hymenoptaecin, apidaecin and defensing1 genes. As internal control was used actin gene. The results were compared by ANOVA followed by Tukey test (P <0.05). Alterations were observed in gene expression of bees studied for all periods and treatments before and after bacterial challenge for all proposed genes, were yet verified inductions of relative expression in the three periods. It is concluded that under the conditions of this study, propolis can induce the relative expression of genes abaecin, hymenoptaecin, apidaecin and defensin1, when subjected to bacterial challenge. However, it is not able to characterize a treatment with propolis presenting greater relative expression for all genes simultaneously. EFFECT OF PROPOLIS CONSUMPTION IN THE BEHAVIORAL CHANGES AND MORTALITY OF Apis mellifera L. BEES SUBMITTED TO PESTICIDE FIPRONIL. Pesticides pose the greatest direct risk for pollinators such as Apis mellifera bees, responsible for reducing bee populations and beekeeping, making it necessary to search for substances that can contribute to improving the health of these bees, such as propolis. Given this, the study found the effect of propolis alcoholic extract consumption behavior when subjected to DL50(0.2 µg/bee) and sublethal (1/500 DL50), and mortality of Apis mellifera L. bees submitted to different doses of the insecticide fipronil (0,05; 0.1, 0.2, 0.3 and 0.4 µg/bee). Four colonies were used, one for each treatment (0, 5, 10 and 15% inclusion of propolis), and in three periods (0,15 and 30 days) to determine the effect of the treatments throughout the experiment. To the achievement of mortality and behavioral changes tests the foraging bees from treated hives were selected. For the mortality test, the data were analyzed in generalized linear mixed model, added the Tukey test (P <0.05) and the analysis of the assessment of behavioral activities was made by (ANOVA) followed by Tukey test (P <0.05). Beehives who consumed propolis have a lower mortality rate compared to the hive control (0%), except on day 0 that the hive control did not differ from treatment 10%. It was also found that the hive that received treatment 10% was among the lowest mortality rates in the fifteenth and thirtieth collection day. Furthermore, the observed mortality was 23% in bees when subjected to LD50. In tests of locomotor and behavioral changes were observed influences of treatments with propolis. It concludes that, for this study, propolis influenced the mortality rate and may have promoted a protective effect on bees Apis mellifera L. when subjected to different concentrations of the insecticide fipronil, and also, that the treatment 10% was effective due to having presented the lowest mortality rates in the 15 and 30. However, treatments with propolis did not influence the behavioral and motor changes.

Orientador: Ricardo De Oliveira Orsi / Abstract: INFLUENCE OF PROPOLIS CONSUMPTION IN GENES EXPRESSION RELATED TO IMMUNE SYSTEM OF Apis mellifera L. BEES SUBJECTED TO CHALLENGE BACTERIA. The Apis mellifera bees may be subject to a number of threats as parasites and pathogens that affect your immune system. This fact makes it necessary to look for natural products that can contribute to improving the immune system of these insects such as propolis. Given the above, the objective was to analyze the influence of the supply of propolis in gene expressions related to immunity of Apis mellifera L. bees subjected to bacterial challenge Over 30 days, four hives receive weekly treatments with different percentages of alcoholic extract of propolis 30% (0%, 5%, 10%, 15%). The experiment was randomized in a factorial 4 x 2 x 2x 3 (treatments x with or without bacteria x times x periods), totaling 48 samples. It was observed the expressions of abaecin, hymenoptaecin, apidaecin and defensing1 genes. As internal control was used actin gene. The results were compared by ANOVA followed by Tukey test (P <0.05). Alterations were observed in gene expression of bees studied for all periods and treatments before and after bacterial challenge for all proposed genes, were yet verified inductions of relative expression in the three periods. It is concluded that under the conditions of this study, propolis can induce the relative expression of genes abaecin, hymenoptaecin, apidaecin and defensin1, when subjected to bacterial challenge. However, it i... (Complete abstract click electronic access below) / Resumo: INFLUÊNCIA DO CONSUMO DA PRÓPOLIS NA EXPRESSÃO DE GENES RELACIONADOS AO SISTEMA IMUNOLOGICO DE ABELHAS Apis mellifera L. SUMETIDAS AO DESAFIO BACTERIANO. As abelhas Apis mellifera podem estar sujeitas a uma série de ameaças como parasitas e patógenos que acometem seu sistema imunológico. Tal fato torna necessária a busca por produtos naturais que possam contribuir com a melhora do sistema imune destes insetos, como a própolis. Diante do exposto, o objetivo foi analisar a influência do fornecimento da própolis em expressões de genes relacionados à imunidade de abelhas Apis mellifera L. submetidas ao desafio bacteriano. Ao longo de 30 dias, quatro colmeias receberam semanalmente os tratamentos com diferentes porcentagens de extrato alcoólico de própolis 30% (0%, 5%, 10%, 15%). O experimento foi casualizado em esquema fatorial 4 x 2 x 2x 3 (tratamentos x com ou sem bactéria x tempos x períodos), totalizando 48 amostras. Foram observadas as expressões dos genes abaecin, hymenoptaecin, apidaecin e defensin1. Como controle interno foi utilizado o gene actina. Os resultados foram comparados por ANOVA seguidos do teste de Tukey (P<0,05). Foram observadas alterações na expressão gênica das abelhas estudadas para todos os períodos e tratamentos, antes e após desafio bacteriano, para todos os genes propostos, sendo ainda verificada induções da expressão relativa nos três períodos. Conclui-se que nas condições do presente trabalho, a própolis pode induzir a expressão relativa dos genes a... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Les insectes contribuent à la pollinisation de 70% des espèces cultivées aujourd’hui à travers le monde. Avec l’intensification de l’agriculture au début du XXème siècle, les agriculteurs se sont mis à introduire des colonies d’abeilles mellifères dans leurs cultures entomophiles pour assurer le service de pollinisation. Avec la reconnaissance croissante du rôle majeur joué par les insectes sauvages dans la pollinisation des cultures, le concept de pollinisation intégrée des cultures a récemment vu le jour, encourageant à combiner insectes pollinisateurs introduits et sauvages en adaptant les pratiques agricoles pour assurer une pollinisation durable des cultures. Mais l’introduction d’insectes pollinisateurs d’élevage est une pratique encore très empirique qui manque de références techniques pour pouvoir être mise en œuvre avec précision. L’objectif général de cette thèse était de fournir des éléments pour mettre au point une méthode objective pour définir la charge en unités opérationnelles d’abeilles mellifères à introduire par unité de surface de culture cible pour complémenter la faune pollinisatrice sauvage pour que la pollinisation ne soit pas un facteur de production limitant.Nous avons travaillé sur une lignée mâle fertile et une lignée mâle stérile de colza (Brassica napus L.), les productions de semence hybride dépendant entièrement des insectes pour la pollinisation chez cette espèce. Nous avons pu déterminer (i) le nombre minimum de grains de pollen viables devant être déposés par stigmate pour que la grenaison puisse être complète en fonction de la température, (ii) la durée après anthèse pendant laquelle le pollen doit être déposé sur le stigmate pour que la grenaison puisse être complète en fonction de la température, (iii) la durée pendant laquelle une fleur sécrète du nectar en fonction de la température pour chacune des deux lignées, et (iv) la vitesse de cette sécrétion nectarifère en fonction de la température pour chacune des deux lignées. Nous avons également validé une méthode d’évaluation rapide de la taille des cheptels d’abeilles mellifères introduits dans les cultures entomophiles, couramment utilisée dans certains pays, en tenant compte de la température.A partir de ces éléments, nous avons proposé d’introduire les concepts d’offre et de demande en pollinisation d’une culture cible, afin de quantifier les déficits de pollinisation et le nombre d’insectes pollinisateurs à introduire pour compléter ces déficits. Ces premiers éléments fournissent une base pour construire un modèle mécaniste de gestion intégrée de la pollinisation, pour prédire le nombre d’insectes pollinisateurs à introduire dans une culture étant donné son contexte climatique, paysager et variétal. / Insects contribute to the pollination of 70% of the species cultivated today around the world. With the agriculture intensification in the early twentieth century, farmers began to introduce honey bees colonies into their entomophilous crops to provide pollination service. With the growing recognition of the major role played by wild insects in crop pollination, the concept of integrated crop pollination recently emerged, encouraging the combination of introduced and wild pollinating insects by adapting agricultural practices to ensure sustainable pollination of crops. But the introduction of managed pollinating insects is still an empirical practice that lacks technical references to be implemented with precision. The general objective of this thesis was to provide elements to develop an objective method to define the operational unit load of honey bees to be introduced per unit area of target crop to complement wild pollinating fauna so that pollination is not a limiting factor for production.We worked on a male fertile and a male sterile oilseed rape (Brassica napus L.) lines, hybrid seed productions being entirely dependent on insects for pollination in this species. We were able to determine (i) the minimum number of viable pollen grains to be deposited on stigma so that seed set was complete depending on the temperature, (ii) the duration after anthesis during which the pollen must be deposited on the stigma so that seed set was complete depending on the temperature, (iii) the length of time a flower secretes nectar as a function of temperature for each of the two lines, and (iv) the rate of this nectar secretion as a function of temperature for each of the two lines. We also validated a method for rapid assessment of the size of honey bee stocking rate introduced into entomophilous crops, commonly used in some countries, taking into account temperature.From these elements, we proposed to introduce the concepts of supply and demand in pollination of a target crop, in order to quantify the pollination deficits and the number of pollinating insects to introduce to complete these deficits. These first elements provide a basis for constructing a mechanistic model of integrated pollination management, to predict the number of pollinating insects to be introduced into a crop given its climatic, landscape and varietal context.

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