Made available in DSpace on 2014-06-11T19:30:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-29Bitstream added on 2014-06-13T19:19:43Z : No. of bitstreams: 1 britto_fb_dr_rcla.pdf: 9470706 bytes, checksum: a24839a07881484e26f17ad6fe4f590b (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / As glândulas hipofaríngeas são estruturas presentes apenas na ordem Hymenoptera e apresentam-se como células epidérmicas modificadas em alguns grupos, enquanto em outros, apresentam morfologia mais elaborada, caracterizando-se como órgãos com anatomia bem definida. Muito bem estudadas em abelhas, as glândulas hipofaríngeas são glândulas exócrinas que fazem parte do sistema salivar e são responsáveis, em Apis, pela produção de substâncias que constituem a geléia real. Este produto é oferecido a todas as larvas como alimento, de maneira desigual, sendo este um fator responsável pela diferenciação das castas nas abelhas. Os objetivos do presente estudo foram analisar as glândulas hipofaríngeas de vespas, que carecem de estudos, procurando reconhecer seu papel na regulação social da colônia, bem como identificar a presença de aspectos glandulares que indicassem a ocorrência de morte celular. Utilizando Polistes versicolor, uma das espécies do gênero considerado como chave para o entendimento da evolução do comportamento social em insetos, foram realizadas análises histoquímicas, enzimológicas e ultra-estruturais das glândulas hipofaríngeas. Juntamente com estas análises foi realizado o estudo comportamental dos indivíduos analisados. Estes estudos auxiliaram no entendimento do ciclo de desenvolvimento das glândulas hipofaríngeas, bem como determinaram a constituição química da secreção glandular, representando uma contribuição inédita à identificação da morfologia e função dessas estruturas e de seu possível papel no contexto social das colônias de P. versicolor. Os resultados encontrados levam a crer que as glândulas destas vespas não estão envolvidas na produção de alimento para as larvas, como ocorre em abelhas. As características morfológicas... / The hypopharyngeal glands are structures present only in the order Hymenoptera. They are constituted by modified epidermal cells in some groups, while in others, they present a more elaborated morphology and are characterized as distinct organs with a defined anatomy. These exocrine glands are part of the salivary system and have been well studied in bees. In Apis, they are the main responsible for the production of components of royal jelly. Brood is not fed equally with royal jelly resulting in cast differentiation in this genus. The aims of the present work were to analyze the hypopharyngeal glands of the wasp Polistes versicolor in order to recognize their role in the social regulation of the colony, as well as to identify the presence of glandular features that could indicate cell death. Polistes is considered a key genus for the understanding of the evolution of social behavior in insects. In the present work we used histochemical, enzymological and ultrastructural analyses. Behavioral studies were also carried out to determine the relationship between glandular development and individual activities. These studies provided important information about the development of the hypopharyngeal glands in P. versicolor and determined the histochemical characteristics of the glandular secretion. It represents an important contribution to the identification of the morphology and the roles played by these structures in the social context of P. versicolor colonies. The results suggest that these glands are not involved in food production for the brood, unlike the observed in bees. The morphological General Abstract characteristics of the secretory cells and their secretion, as well as the behavioral characteristics of these wasps suggest that the hypopharyngeal glands of P. versicolor might be involved in the production of digestive enzymes.

Orientador: Flávio Henrique Caetano / Banca: Elisa Aparecida Gregório / Banca: Vera Nisaka Solferini / Banca: Edilberto Giannotti / Banca: Maria Eliza Miyoko Tomotake / Resumo: As glândulas hipofaríngeas são estruturas presentes apenas na ordem Hymenoptera e apresentam-se como células epidérmicas modificadas em alguns grupos, enquanto em outros, apresentam morfologia mais elaborada, caracterizando-se como órgãos com anatomia bem definida. Muito bem estudadas em abelhas, as glândulas hipofaríngeas são glândulas exócrinas que fazem parte do sistema salivar e são responsáveis, em Apis, pela produção de substâncias que constituem a geléia real. Este produto é oferecido a todas as larvas como alimento, de maneira desigual, sendo este um fator responsável pela diferenciação das castas nas abelhas. Os objetivos do presente estudo foram analisar as glândulas hipofaríngeas de vespas, que carecem de estudos, procurando reconhecer seu papel na regulação social da colônia, bem como identificar a presença de aspectos glandulares que indicassem a ocorrência de morte celular. Utilizando Polistes versicolor, uma das espécies do gênero considerado como chave para o entendimento da evolução do comportamento social em insetos, foram realizadas análises histoquímicas, enzimológicas e ultra-estruturais das glândulas hipofaríngeas. Juntamente com estas análises foi realizado o estudo comportamental dos indivíduos analisados. Estes estudos auxiliaram no entendimento do ciclo de desenvolvimento das glândulas hipofaríngeas, bem como determinaram a constituição química da secreção glandular, representando uma contribuição inédita à identificação da morfologia e função dessas estruturas e de seu possível papel no contexto social das colônias de P. versicolor. Os resultados encontrados levam a crer que as glândulas destas vespas não estão envolvidas na produção de alimento para as larvas, como ocorre em abelhas. As características morfológicas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The hypopharyngeal glands are structures present only in the order Hymenoptera. They are constituted by modified epidermal cells in some groups, while in others, they present a more elaborated morphology and are characterized as distinct organs with a defined anatomy. These exocrine glands are part of the salivary system and have been well studied in bees. In Apis, they are the main responsible for the production of components of royal jelly. Brood is not fed equally with royal jelly resulting in cast differentiation in this genus. The aims of the present work were to analyze the hypopharyngeal glands of the wasp Polistes versicolor in order to recognize their role in the social regulation of the colony, as well as to identify the presence of glandular features that could indicate cell death. Polistes is considered a key genus for the understanding of the evolution of social behavior in insects. In the present work we used histochemical, enzymological and ultrastructural analyses. Behavioral studies were also carried out to determine the relationship between glandular development and individual activities. These studies provided important information about the development of the hypopharyngeal glands in P. versicolor and determined the histochemical characteristics of the glandular secretion. It represents an important contribution to the identification of the morphology and the roles played by these structures in the social context of P. versicolor colonies. The results suggest that these glands are not involved in food production for the brood, unlike the observed in bees. The morphological General Abstract characteristics of the secretory cells and their secretion, as well as the behavioral characteristics of these wasps suggest that the hypopharyngeal glands of P. versicolor might be involved in the production of digestive enzymes. / Doutor

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

Made available in DSpace on 2014-06-11T19:32:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-08Bitstream added on 2014-06-13T19:22:54Z : No. of bitstreams: 1 arauco_emr_dr_botfmvz.pdf: 810943 bytes, checksum: 6acc5913059689bd4615fef648271508 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O néctar é coletado pelas operárias coletoras e transportado em sua vesícula nectarífera para a colméia. Durante a sua transformação em mel, o néctar recebe enzimas provenientes das glândulas hipofaringeanas, as quais atuam em seu processamento. A invertase é uma das enzimas adicionadas ao néctar e promove a inversão da sacarose em frutose e glicose. Com o objetivo de se conhecer o pH ideal dessa enzima nas abelhas operárias foram extraídas as glândulas hipofaringeanas de abelhas com oito dias de idade. O método utilizado para a dosagem da invertase foi o do ácido 3-5þ dinitrosalicílico (DNS) para determinação de açúcares redutores, utilizando-se medida de leitura espectrofotométrica em absorbância da glicose em comprimento de onda (?) igual a 540nm. De acordo com os resultados, verificou-se que a atividade catalítica da enzima foi mais eficiente no pH 5,0 e em 40 minutos de reação em comparação com outros valores de pH e tempos de reação. Não foi possível a reutilização das glândulas. Desta forma, concluiu-se que a enzima invertase aumentou sua eficiência em função do tempo de reação de forma linear e significativa em pH 5,0. / The nectar is collected by the collecting workers that carry it in its nectar gallbladder to the beehive. During its transformation in honey, the nectar receives enzymes proceeding from the hypopharyngeal glands, which act in its processing. Invertase is one of enzymes added to the nectar and promotes the inversion of sacarose in fructose and glucose. With the aim of knowing the ideal pH, in the worker bees, the bee's hypopharyngeal glands with eight days of age had been extracted. The used method, for the dosage of invertase, had been the 3-5 ' dinitrosalisilic acid for determination of reducing sugars, using absorbance spectrophotometry data from the glucose in ? equal to 540nm. According to the results, it had been verified that the enzyme catalytic activity had been more efficient in pH 5,0 and in 40 minutes of reaction, comparing to other pH values and times of reaction. The glands reuse had not been possible. At this way, it was concluded that the invertase enzyme had increased its efficiency by the reaction time at linear and significant way in pH 5,0.

Resumo: O néctar é coletado pelas operárias coletoras e transportado em sua vesícula nectarífera para a colméia. Durante a sua transformação em mel, o néctar recebe enzimas provenientes das glândulas hipofaringeanas, as quais atuam em seu processamento. A invertase é uma das enzimas adicionadas ao néctar e promove a inversão da sacarose em frutose e glicose. Com o objetivo de se conhecer o pH ideal dessa enzima nas abelhas operárias foram extraídas as glândulas hipofaringeanas de abelhas com oito dias de idade. O método utilizado para a dosagem da invertase foi o do ácido 3-5þ dinitrosalicílico (DNS) para determinação de açúcares redutores, utilizando-se medida de leitura espectrofotométrica em absorbância da glicose em comprimento de onda (?) igual a 540nm. De acordo com os resultados, verificou-se que a atividade catalítica da enzima foi mais eficiente no pH 5,0 e em 40 minutos de reação em comparação com outros valores de pH e tempos de reação. Não foi possível a reutilização das glândulas. Desta forma, concluiu-se que a enzima invertase aumentou sua eficiência em função do tempo de reação de forma linear e significativa em pH 5,0. / Abstract: The nectar is collected by the collecting workers that carry it in its nectar gallbladder to the beehive. During its transformation in honey, the nectar receives enzymes proceeding from the hypopharyngeal glands, which act in its processing. Invertase is one of enzymes added to the nectar and promotes the inversion of sacarose in fructose and glucose. With the aim of knowing the ideal pH, in the worker bees, the bee's hypopharyngeal glands with eight days of age had been extracted. The used method, for the dosage of invertase, had been the 3-5 ' dinitrosalisilic acid for determination of reducing sugars, using absorbance spectrophotometry data from the glucose in ? equal to 540nm. According to the results, it had been verified that the enzyme catalytic activity had been more efficient in pH 5,0 and in 40 minutes of reaction, comparing to other pH values and times of reaction. The glands reuse had not been possible. At this way, it was concluded that the invertase enzyme had increased its efficiency by the reaction time at linear and significant way in pH 5,0. / Orientador: Sílvia Regina Cunha Funari / Coorientador: Carlos Ducatti / Doutor

La nutrition est l'ensemble des processus par lesquels un être vivant transforme des aliments pour assurer son fonctionnement. Etudier la nutrition d’un organisme permet de comprendre les rapports entre la nourriture consommée et la santé de l’individu. L'ensemble des organes assurant l'extraction d'énergie est le système digestif, qui transforme les sucres en glucose, les corps gras en acides gras, et les protéines en acides aminés. L’objectif de cette étude est de déterminer l’influence que peut avoir la nutrition pollinique sur la santé de l’abeille domestique, Apis mellifera L.. Pour assurer cette nutrition, l’abeille est inféodée aux ressources florales. Du pollen récolté sur les plantes à fleurs, l’organisme puise les corps gras (ou lipides), les vitamines et les protéines. Or l’accès à ces ressources est variable en qualité, en quantité, et en diversité selon le temps et le milieu. Des problèmes de mortalités et affaiblissements des colonies sont observés depuis une vingtaine d’années, et l’une des causes suspectées est le manque de disponibilité et la faible valeur nutritionnelle des ressources polliniques en zones agricoles céréalières. Nous avons donc testé en conditions contrôlées les effets de la quantité, la qualité et la diversité pollinique sur la physiologie d’abeilles nourrices, sur leur immunité, ainsi que sur leur survie. La nutrition jouant un rôle essentiel dans la prévention de nombreuses maladies, les effets de l’alimentation pollinique ont été déterminés en présence ou non d’un stresseur biotique, Nosema ceranae, dont la prévalence dans les colonies est très élevée. Il s’agit d’une microsporidie invasive qui se développe dans l’intestin de son hôte, provoquant divers effets sublétaux et létaux. Etant donné que la valeur nutritionnelle d’un aliment dépend de sa composition mais aussi de sa digestibilité, une troisième partie porte sur l’influence que peut avoir le stresseur sur les capacités de digestion des protéines contenues dans le pollen par l’abeille. Nos résultats apportent des connaissances sur l’impact d’une déplétion en pollen que peuvent subir les colonies dans un paysage d’agriculture intensive. En effet, des abeilles carencées à plus ou moins 60 %, comme cela peut être observé entre deux cultures à floraison massives, subissent des perturbations au niveau individuel (perturbations du développement physiologique des abeilles nourrices, diminution de leur survie), ce qui peut les rendre moins performantes et plus sensibles aux stress présents dans l’environnement. De plus, nous avons pu démontrer que la valeur nutritionnelle des pollens, influence la tolérance des abeilles à Nosema ceranae. La qualité d’un pollen se définit par sa composition chimique totale (protéines, acides aminés, lipides, vitamines, sucres, etc…), et non pas uniquement par sa teneur en protéines ou acides aminés essentiels. Notre étude met d’ailleurs en évidence l’effet négatif d’une alimentation à base de pollen de maïs sur le développement des glandes hypopharyngiennes, l’expression du gène de la vitellogénine, et la survie des abeilles. Au regard de la présence élevée de cette culture dans les zones d’agriculture intensive et sa haute exploitation par les butineuses, nos résultats sont discutés en fonction des répercussions envisagées sur les colonies. Dans ces milieux, la diversité des ressources polliniques, par l’apport de pollens de qualité, compense la pauvreté nutritionnelle d’autres pollens. Dans nos conditions, nous pouvons observer la plus-value d’une alimentation pollinique polyflorale chez des abeilles infestées par Nosema. Les mesures agro-écologiques œuvrant pour régulariser dans le temps les apports polliniques aux colonies, et pour leur offrir une diversité alimentaire participent donc à renforcer leur capacité à lutter contre les stress présents dans l’environnement. / Nutrition is the set of processes by which a living organism transforms food for its functions. Studying nutrition allows to understanding the relationship between food intake and health of the individual. All organs for extracting energy are the digestive system, which converts sugars into glucose, fats into fatty acids, and proteins into amino acids. The objective of this study is to determine the possible influence of pollen nutrition on the honeybee health, Apis mellifera L. To ensure that nutrition, the bee is subservient to floral resources. Of pollen collected from the flowering plants, the body draws fats (or lipids), vitamins and proteins. Access to these resources is variable in quality, quantity, and diversity depending on the time and the environment. Problems of mortality and weakening of colonies are observed for some twenty years, and one of the suspected causes is the lack of availability and low nutritional value of pollen resources in agricultural areas. We therefore tested under controlled conditions the effects of quantity, quality and diversity of pollen on nurse worker physiology and on their immunity, as well as their survival. Nutrition plays a vital role in the prevention of many diseases, the effects of pollen diet were determined in the presence or absence of a biotic stressor, Nosema ceranae, whose prevalence in the colonies is very high. It is an invasive microsporidia that develops in the intestine of its host, causing various sublethal and lethal effects. Since the nutritional value of a food depends on its composition but also its digestibility, a third part focuses on the possible influence of the stressor on bee ability digestion of the proteins provided by the pollen.Our results provide knowledge on the impact of pollen depletion that may undergo the colonies in an intensive agricultural landscape. Indeed, bees starved to roughly 60%, as can be observed between two cultures massive flowering undergo disturbances at individual level (disturbances of nurse worker physiological development, reduced survival), which may make them less efficient and more susceptible to stress from the environment. In addition, we demonstrated that the nutritional value of pollen influences the Nosema ceranae‘s bee tolerance. The pollen quality is determined by the total chemical composition (proteins, amino acids, lipids, vitamins, sugars, etc ...), not only by its content of essential amino acids or proteins. Our study also demonstrated the negative effect of a diet based on maize pollen on hypopharyngeal glands development, the vitellogenin gene expression, and the bee survival. In view of the high presence of this culture in intensive agriculture areas and high exploitation by foragers, our results are discussed in terms of their impact on the proposed settlements. In these environments, the diversity of pollen resources by providing pollen of quality compensates nutritional poverty of other pollens. In our conditions, we can observe the gain of power of a polyfloral pollen in Nosema infested bees. Agro-ecological measures working to regularize, in time, the pollen contributions to the colonies, and to provide them with food diversity thus participate in building their capacity to fight against stress in the environment