The bacteria in host-associated microbial communities influence host health through various mechanisms, such as immune stimulation or the release of metabolites. However, viruses that target bacteria, called bacteriophages (phages), may also shape the animal microbiome. Most phage lifecycles can be classified as either lytic or temperate. Lytic phages infect and directly kill bacterial hosts and can directly regulate bacterial population size. Temperate phages, in contrast, have the potential to undergo either a lytic cycle or integrate into the bacterial genome as a prophage. As a prophage, the phage may alter bacterial host phenotypes by carrying novel genes associated with auxiliary metabolic functions, virulence-enhancing toxins, or resistance to other phage infections. Lytic phages may also carry certain auxiliary metabolic genes, which are instead used to takeover bacterial host functions to better accommodate the lytic lifecycle. In either case, the ability to alter bacterial phenotypes may have important ramifications on host-associated communities. This dissertation focused on the genetic contributions that phages, and particularly prophages, provide to the bacterial members of two separate host-associated communities: the honey bee (Apis mellifera) gut microbiome and the amphibian skin microbiome. My second chapter surveyed publicly available whole genome sequences of common honey bee gut bacterial species for prophages. It revealed that prophage distribution varied by bacterial host, and that the most common auxiliary metabolic genes were associated with carbohydrate metabolism. In chapter three, this bioinformatic pipeline was applied to the amphibian skin microbiome. Prophages were identified in whole genome bacterial sequences of bacteria isolated from the skin of American bullfrogs (Lithobates catesbeianus), eastern newts (Notophthalmus viridescens), Spring peepers (Pseudacris crucifer) and American toads (Anaxyrus americanus). Prophages were additionally identified in publicly available genomes of non-amphibian isolates of Janthinobacterium lividum, a bacteria found both on amphibian skin and broadly in the environment. In addition to a diverse set of predicted prophages across amphibian bacterial isolates, several Janthinobacterium lividum prophages from both amphibian and environmental isolates appear to encode a chitinase-like gene undergoing strong purifying selection within the bacterial host. While identifying the specific function of this gene would require in vitro isolation and testing, its high homology to chitinase and endolysins suggest it may be involved in the breakdown of either fungal or bacterial cellular wall components. Finally, my fourth chapter revisits the honey bee gut system by investigating the role of geographic distance in bacteriophage community similarity. A total of 12 apiaries across a transect of the United States, from Virginia to Washington, were sampled and honey bee viromes were sequenced, focusing on the lytic and actively lysing temperate community of phages. Although each apiary possessed many unique bacteriophages, apiaries that were closer together did have more similar communities. Each bacteriophage community also carried auxiliary carbohydrate genes, especially those associated with sucrose degradation, and antimicrobial resistance genes. Combined, the results of these three studies suggest that bacteriophages, and particularly prophages, may be contributing to the genetic diversity of the bacterial community through nuanced relationships with their bacterial hosts. / Doctor of Philosophy / The microbial communities of animals, called "microbiomes", play important roles in the health of animals. The bacteria in these microbiomes can help strengthen the immune system, provide resistance to dangerous pathogens, and break down nutrients. However, bacteria are not alone in the microbiome; viruses are also present. Surprisingly, the vast majority of the world's viruses, even those living inside animals, infect bacteria. These viruses, called "bacteriophages" or "phages", can impact the bacterial communities in a microbiome. Phages can be grouped in to two broad categories based on lifecycle. Lytic phages kill the bacterial host directly after infection. Temperate phages, on the other hand, can either immediately kill the host like lytic phages or alternatively, become a part of the bacterial genome and live as prophages. Phages with both lifecycles can sometimes carry genes that, although not essential to the phage, may change the traits of the bacteria during infection. For example, some phages carry toxin genes, which bacteria use to cause disease in animals. Other phages might carry genes that provide antibiotic resistance or alter the metabolism of the infected bacteria. If a phage gene benefits the infected bacteria, the bacteria may begin interacting with its environment in a new way or may even become more abundant. Alternatively, phages that directly kill infected bacteria may have a negative effect on bacterial population sizes. To begin unraveling how phages influence bacterial species in microbiomes, I investigated two different animal systems: the Western honey bee (Apis mellifera) gut microbiome and the amphibian skin microbiome. I first identified prophages of several common bacterial species that reside in the honey bee gut (Chapter 2). Prophages were more common in certain bacterial species than others, and some possessed genes associated with the breakdown of sugars or pollen, suggesting they help honey bees process their food. Using similar techniques, I then identified prophages in bacteria isolated from the skin microbiomes of several amphibian species common in the eastern United States (American bullfrogs, Eastern newts, Spring peepers, and American toads) (Chapter 3). Most notably, the bacteria Janthinobacterium lividum may benefit from prophages that carry genes for potentially antifungal chitinase enzymes that destroy the fungal cell wall. Finally, I returned to the honey bee gut microbiome system by investigating how honey bee bacteriophage communities change over large geographic distances (Chapter 4). This study, which examined honey bees from 12 apiaries sampled from the east to west coast of the United States, looks primarily at lytic phage and temperate phage that are not integrated as prophage, but are instead seeking a bacterial host to infect. I found that nearby apiaries tended to have more similar communities of bacteriophages, compared to apiaries far away. Additionally, most bacteriophage communities carry genes associated with the breakdown of sugars like sucrose. Overall, these three studies show that phages, and especially prophages, contribute to the genetic landscape of the microbiome by broadly providing bacterial hosts with access to a diverse set of genes.

La colonie d’abeille (Apis mellifera L.) est une société complexe où les individus interagissent entre eux, notamment par le biais de phéromones. L’étude de cette communication chimique est indispensable à la compréhension des régulations sociales mises en place dans la colonie. Chez l’abeille, plus de 50 substances chimiques avec des effets incitateurs ou modificateurs sur la colonie ont été identifiées. Malgré ces découvertes, de nombreux travaux sont à accomplir pour mieux comprendre ce système de communication particulier.La problématique de cette thèse vise à caractériser l’histoire de vie d’une phéromone majeure l’Oléate d’Ethyle (EO), qui permet d’optimiser l’équilibre nourrices / butineuses dans la colonie. Parallèlement, d’autres recherches ont été entreprises, notamment l’étude de la communication chimique de la reine et du couvain, chez qui seulement deux phéromones ont été identifiées avec des effets pléiotropiques dans la colonie.Nos résultats ont mis en évidence une production variable d’EO par les ouvrières, en fonction de l’environnement de la colonie. La production de cette molécule chimique dans la colonie peut également être modifiée par un stress : des abeilles parasitées par du Nosema spp. ont une production anormalement élevée d’EO. En outre, cette molécule phéromonale est transmise des butineuses vers les nourrices par contact cuticulaire et par le pollen.Pour la compréhension de la communication entre la reine et les ouvrières, nos résultats montrent que la reine utilise d’autres composés phéromonaux puissant en redondance de la QMP pour orienter la construction de cire, le phénomène de cour et l’inhibition des ovaires des ouvrières.Chez le couvain, nous avons identifié un composé phéromonal volatil, le E-ß-ocimène, produit majoritairement par les jeunes larves, inhibant le développement des ovaires des ouvrières et accélérant leur maturation comportementale.Ces études nous ont permis d’avoir une connaissance plus précise de la communication chimique au sein de la colonie. Ainsi nous expliquons par deux théories le rôle de la complexité et de la redondance phéromonale de la colonie d’abeilles / In the honeybee colony (Apis mellifera L.) studies of the chemical communication are essential to understand social regulations. In the honey bee colony more than 50 chemical substances with releaser and primer effects have been identified. Despite years of research on this type of communication, significant work remains to be done.In this thesis, the aim is to characterize the dynamics of a major pheromone: ethyl oleate (EO), which optimizes the balance between nurses and foragers in the colony. In addition, we initiated research on the queen and brood chemical communication in which only two pheromones have been identified in the colony.We have demonstrated that EO production by workers varies under different colony environment. EO production can also be modified by stress; honey bees parasitized by the Nosema spp. have abnormally high EO production. In addition, we identified that EO is transmitted from foragers to nurses by contact (cuticle and pollen).For the queen, our results indicate that the queen uses multiple redundant pheromones (QMP and other unknown compounds), that affect wax construction, retinue behaviour and worker ovary inhibition.For the brood we have identified a volatile pheromone E-ß-ocimene produced mostly by the young larvae to inhibit the development of workers ovaries and accelerate workers’behavioural maturation.With these studies we clarify some aspects of what is known about chemical communication in the honey bee colony. Then we try to explain the role of complexity and redundancy of pheromones in the honey bee colony by two theories

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.

O mel é um alimento de uso milenar, açucarado de fácil digestão, que constitui uma importante fonte de energia, contribuindo para o equilíbrio do processo biológico do corpo humano, sendo elaborado a partir da desidratação e transformação do néctar das flores nativas pelas abelhas produtoras. Para que o mel seja comercializado para o consumo humano, ele precisa atender aos requisitos mínimos de identidade e qualidade exigidos pela Legislação Brasileira. No Brasil a criação de abelhas é dividida em duas práticas distintas, a Apicultura tradicional, que utiliza as abelhas Apis mellifera e a Meliponicultura que utiliza as abelhas sem ferrão como a Jataí (Tetragonisca angustula). Os méis de abelhas sem ferrão tem maior valor comercial comparado ao mel tradicional, entretanto são comercializados sem uma legislação própria. Na literatura existem poucos trabalhos que tratam da composição destes tipos de méis que são popularmente conhecidos por suas propriedades benéficas à saúde. Em vista do exposto acima, o objetivo deste presente trabalho foi o de comparar a composição e a qualidade de méis de Apis mellifera com os de abelhas sem ferrão da espécie Tetragonisca angustula, popularmente conhecida como Jataí. Para tanto as amostras de méis foram obtidas de colméias de Apis mellifera e de Tetragonisca angustula de uma mesma região botânica, o que foi possível constatar que os méis de abelha Jataí apresentaram maior diversidade botânica em relação aos méis de Apis. Neste trabalho foram utilizados os métodos de avaliação estabelecidos pela Legislação Brasileira para qualidade de mel de Apis mellifera e os valores sugeridos para méis de mellponíneos do Brasil pe10s pesquisadores VILLAS - BOAS e MALASPINA (2005). A maioria das amostras de Apis apresentaram-se dentro da legislação vigente, enquanto méis Jataí apresentaram os parâmetros: umidade (23,40 -25,60%), acidez (21,65 - 63,85 mE/Kg) e açúcares redutores (44,78 - 67,54%) e sacarose aparente (0,43 - 1,60%) fora dos padrões estabelecido pela legislação vigente para os méis de Apis mellifera. No entanto, encontram-se dentro dos valores sugeridos para méís de meliponíneos brasileiros, pelos pesquisadores acima mencionados. Além das análises físico-químicas tradicionais e a análise polínica também foi determinada a composição nutricional, sendo que o mel de Apis apresentou maior de valor energético (43,58- 66,32 Kcal) em relação aos méis de Jataí (36,83 - 60,52 Kcal) (p<0,05). Também foram determinados os açúcares por CLAE, condutividade elétrica (uS/cm-1) e a análise de cor (mmPfund). As amostras de Apis mellifera apresentaram maior o teor glicose (%), frutose (%) e condutividade elétrica (uS/cm-1) em relação aos méis de Jataí Em relação as análises de cor notou-se maior predominância da coloração âmbar-claro, mas amostras analisadas. / Honey is considered as a food that provides energy, being elaborated from the dehydration and transformation of the nectar of the flowers by the bees. For the human consumption, honey needs to attend the minimum requirements of identity and quality demanded by the regulation. In Brazil beekeepers can be divided in two practical distinct ones: the traditional ones, which use Apis mellifera bees and the Meliponiculture which uses stingless bees such as Jataí bee (Tetragonisca angustula). There are no identity and quality parameters or regulation for this type of honey. Honey from of stingless bees are more expensive compared with the traditional honey, however their are commercialized without a proper regulation. In literature few works were found regarding the composition of these types of honey which are popularly known by its beneficiaI properties to human health. The objective of the present work is to compare the composition and quality of honey from Apis mellifera and from stingless bees (Tetragonisca angustula), popularly known as Jataí bee. Samples of honey were obtained from Apis mellifera and Tetragonisca angustula bees in the same botanical region. In this work the methods used were based on the Brazilian Regulation for quality control of honey from Apis mellifera and the values suggested for honeys of meliponíneos of Brazil for researchers VILLAS-BOAS and MALASPINA (2005). The majority of the samples of Apis had presented in accordance with the regulation while the Jataí honeys had presented: humidity (23,40 - 25.60%), acidity (21,65 - 63,85 mE/Kg) and reducing sugars (44,78 - 67.54%) are out of the standards for honeys of Apis mellifera. However, they are in accordance with the values suggested for honeys of Brazilian meliponíneos. The physicochemical, polinic and nutritional analysis were determined, and the honey of Apis bee presented greater amount of energy value (43,58 - 66,32 Kcal) in relation to the honeys of Jataí bee (36,83 - 60,52 Kcal) (p<O,05). The sugars for HPLC, electric conductivity (uS/cm-1) and analysis of color (mmPfund) were determined. Samples of Apis mellifera had presented greater values for glucose (%), frutose (%) and electric conductivity (uS/cm-1) in relation to the honeys from Jataí (p<0,05). Regarding the color analyses, it was predominance of the coloration clear -amber.

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

O mel é um alimento de uso milenar, açucarado de fácil digestão, que constitui uma importante fonte de energia, contribuindo para o equilíbrio do processo biológico do corpo humano, sendo elaborado a partir da desidratação e transformação do néctar das flores nativas pelas abelhas produtoras. Para que o mel seja comercializado para o consumo humano, ele precisa atender aos requisitos mínimos de identidade e qualidade exigidos pela Legislação Brasileira. No Brasil a criação de abelhas é dividida em duas práticas distintas, a Apicultura tradicional, que utiliza as abelhas Apis mellifera e a Meliponicultura que utiliza as abelhas sem ferrão como a Jataí (Tetragonisca angustula). Os méis de abelhas sem ferrão tem maior valor comercial comparado ao mel tradicional, entretanto são comercializados sem uma legislação própria. Na literatura existem poucos trabalhos que tratam da composição destes tipos de méis que são popularmente conhecidos por suas propriedades benéficas à saúde. Em vista do exposto acima, o objetivo deste presente trabalho foi o de comparar a composição e a qualidade de méis de Apis mellifera com os de abelhas sem ferrão da espécie Tetragonisca angustula, popularmente conhecida como Jataí. Para tanto as amostras de méis foram obtidas de colméias de Apis mellifera e de Tetragonisca angustula de uma mesma região botânica, o que foi possível constatar que os méis de abelha Jataí apresentaram maior diversidade botânica em relação aos méis de Apis. Neste trabalho foram utilizados os métodos de avaliação estabelecidos pela Legislação Brasileira para qualidade de mel de Apis mellifera e os valores sugeridos para méis de mellponíneos do Brasil pe10s pesquisadores VILLAS - BOAS e MALASPINA (2005). A maioria das amostras de Apis apresentaram-se dentro da legislação vigente, enquanto méis Jataí apresentaram os parâmetros: umidade (23,40 -25,60%), acidez (21,65 - 63,85 mE/Kg) e açúcares redutores (44,78 - 67,54%) e sacarose aparente (0,43 - 1,60%) fora dos padrões estabelecido pela legislação vigente para os méis de Apis mellifera. No entanto, encontram-se dentro dos valores sugeridos para méís de meliponíneos brasileiros, pelos pesquisadores acima mencionados. Além das análises físico-químicas tradicionais e a análise polínica também foi determinada a composição nutricional, sendo que o mel de Apis apresentou maior de valor energético (43,58- 66,32 Kcal) em relação aos méis de Jataí (36,83 - 60,52 Kcal) (p<0,05). Também foram determinados os açúcares por CLAE, condutividade elétrica (uS/cm-1) e a análise de cor (mmPfund). As amostras de Apis mellifera apresentaram maior o teor glicose (%), frutose (%) e condutividade elétrica (uS/cm-1) em relação aos méis de Jataí Em relação as análises de cor notou-se maior predominância da coloração âmbar-claro, mas amostras analisadas. / Honey is considered as a food that provides energy, being elaborated from the dehydration and transformation of the nectar of the flowers by the bees. For the human consumption, honey needs to attend the minimum requirements of identity and quality demanded by the regulation. In Brazil beekeepers can be divided in two practical distinct ones: the traditional ones, which use Apis mellifera bees and the Meliponiculture which uses stingless bees such as Jataí bee (Tetragonisca angustula). There are no identity and quality parameters or regulation for this type of honey. Honey from of stingless bees are more expensive compared with the traditional honey, however their are commercialized without a proper regulation. In literature few works were found regarding the composition of these types of honey which are popularly known by its beneficiaI properties to human health. The objective of the present work is to compare the composition and quality of honey from Apis mellifera and from stingless bees (Tetragonisca angustula), popularly known as Jataí bee. Samples of honey were obtained from Apis mellifera and Tetragonisca angustula bees in the same botanical region. In this work the methods used were based on the Brazilian Regulation for quality control of honey from Apis mellifera and the values suggested for honeys of meliponíneos of Brazil for researchers VILLAS-BOAS and MALASPINA (2005). The majority of the samples of Apis had presented in accordance with the regulation while the Jataí honeys had presented: humidity (23,40 - 25.60%), acidity (21,65 - 63,85 mE/Kg) and reducing sugars (44,78 - 67.54%) are out of the standards for honeys of Apis mellifera. However, they are in accordance with the values suggested for honeys of Brazilian meliponíneos. The physicochemical, polinic and nutritional analysis were determined, and the honey of Apis bee presented greater amount of energy value (43,58 - 66,32 Kcal) in relation to the honeys of Jataí bee (36,83 - 60,52 Kcal) (p<O,05). The sugars for HPLC, electric conductivity (uS/cm-1) and analysis of color (mmPfund) were determined. Samples of Apis mellifera had presented greater values for glucose (%), frutose (%) and electric conductivity (uS/cm-1) in relation to the honeys from Jataí (p<0,05). Regarding the color analyses, it was predominance of the coloration clear -amber.

Thesis (MScConEcol (Conservation Ecology and Entomology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Two populations of Acacia mellifera were noted in Pniel, which is a semi-arid savanna, near Kimberley in the Northern Cape province of South Africa. One population appeared on a rocky, andesitic laval ridges (soil pHKCL 6.5-7.0) along the Vaal river. The other appeared in a sandveld area (soil pHKCL 4). Bush encroachment by A. mellifera was found to be more extensive on the rocky areas than in the sandveld and the two habitats differed extensively on soil pH, clay and silt contents and also on water holding capacities. The rocky habitat was thus deduced to have a higher CEC. Seeds were sampled on a logarithmic scale for allozyme analysis and also randomly in each of the two habitats for local adaptation tests, in which case lime (CaCO3) and organic matter (cow-dung) were used in a completely-crossed design. Detected interaction effects (between population source and pH; population source and organic matter and between pH and organic matter) and significant differences could not separate the two populations as the differences occurred across populations. Random genetic differences leading to phenotypic plasticity in the two observed populations, might be responsible for the observed phenotypic differences. Allozymic data showed no significant differences between the two populations and the genetic distance between and within the populations also confirmed that the two populations had not genetically differentiated. The Mantel Test on the two populations, showed nonsignificant results. Nei‟s UPGMA dendrogram revealed that the game farm subpopulations were more primitive and genetically related to each other. Despite differences in allozyme frequencies, between the sampled sites, genetic differentiation was found to be low (FST = 0.337). Nei‟s (1972) original measures of genetic distance ranged between 0.871 and 1.000 with a mean of 0.949 ± 0.053. The study concluded that the two observed populations had not genetically differentiated and no local adaptation could be established rather phenotypic plasticity was evident and resulted in the observed divergent growth forms. Nonetheless, the overall direction of spread of encroachment appeared to be the eastward. / AFRIKAANSE OPSOMMING: Twee bevolkings van Acacia mellifera is gevind in Pniel, wat „n semi-ariede savanna is naby Kimberley in die Noord-Kaap provinsie van Suid-Afrika. Een bevolking het voorgekom op klipperige andesitiese lava riwwe (grond pHKCL 6.5-7.0) al langs die Vaalrivier. Die ander het voorgekom in „n sandveld area (soil pHKCL 4). Bos-oorskryding deur A. mellifera was meer uitgebreid op die klipperige areas as in die sandveld en die twee habitats het noemenswaardig verskil ten opsigte van grond pH, klei en silt inhoud asook waterhoukapasiteit. Dit kan was dus afgelei word dat die klipperige habitat „n hoër CEC het. Die sade was versamel op „n logaritmiese skaal vir allosiem-analise en ook ewekansig in die twee habitats vir lokale aanpassings toetse. In dié gevalle was kalk (CaCO3) en organiese material (koeimis) gebruik in „n totaal-gekruisde ontwerp. Bespeurde interaksie effekte (tussen bevolkings bron en pH; bevolkings bron en organiese material en tussen pH en organiese material) en noemenswaardige verskille kon nie die twee bevolkings skei nie, aangesien die verskille voorgekom het regdeur die twee bevolkings. Ewekansige genetiese verskille wat lei tot fenotipiese plastisiteit tussen die twee waargeneemde bevolkings mag dalk verantwoordelik wees vir die waargeneemde fenotipiese verskille. Allosiem-data het geen beduidende verskille gelewer tussen die twee bevolkings nie en genetiese afstand binne en tussen die bevolkings het ook bevestig dat die twee bevolkings nie geneties gedifferensiëer is nie. Die Mantel toets op die twee bevolkings het geen beduidende resultate gelewer nie. Nei se UPGMA dendogram get gewys dat die wildsplaas bevolkings was meer primitief en geneties verwant aan mekaar. Ten spyte van die allosiem frekwensies tussen die gemonsterde gebiede, was die genetiese differensiasie laag (FST = 0.337). Nei (1972) se oorspronlike meeting van genetiese afstand het tussen 0.871 en 1.000 beloop met „n gemiddeld van 0.949 ± 0.053. Die studie het bepaal dat die twee waargeneemde bevolkings nie geneties gedifferensiëer het nie en dat geen lokale aanpassing teenwoordig was nie. Fenotipiese plastisiteit was duidelik waarneembaar en het gelei tot die divergerende groeivorme. Nieteenstaande, was die algehele rigting van oorskryding ooswaarts.

Thesis (MScConEcol (Conservation Ecology and Entomology))--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Insect pollinators play an important role in producing crops in global agriculture. Pollinatordependent crops contribute to maintaining a healthy variety in the human diet and often have a high market value, beneficial for local or regional economies. Insect crop pollinators can either be from natural areas adjacent to orchards, or they can be brought into orchards by beekeepers that manage them. Pollination by wild pollinators is an ecosystem service, while managed pollinators (mostly honeybees) is a humanly managed service, considered not to be related to the ecosystem. Ecosystem services and their economic value have often been used as an incentive for conservation, although it is sometimes difficult to characterise and quantify them. Wild and managed pollinators have been reported to be threatened in several regions around the world, and there is concern about the effect a pollination deficit may have on crop production. Different crops and cultivars have different levels of dependence on insect pollination due to a combination of biological, physical and management factors. In this study, the pollination dependence of the Granny Smith apple cultivar and the respective contributions of wild and managed pollinators are investigated in the Western Cape province of South Africa. Granny Smith apples show a significant increase in production with insect pollination (wild and managed). Managed honeybees are more abundant in orchards than wild honeybees, and also provide a better pollination service. This difference between the pollination service of wild and managed honeybees are specifically noted in the quality, where managed honeybees pollination result in significantly more seeds per fruit and consequently produce a better shaped apple. The study goes further by quantifying the ecosystem services to the managed honeybee industry through a questionnaire completed by beekeepers. It was found that 49% of the managed hives in the Western Cape rely to some extent on natural vegetation as a forage source. Furthermore 18% of honey produced is also from natural vegetation and the wild honeybee population replenish managed honeybee stocks if they become depleted. Although managed honeybees are not usually considered an ecosystem service, it is clear that they are still linked to the ecosystem via these pathways. It is thus obvious that all pollination sources are linked to the environment, not just wild pollinators. A further economic valuation of the ecosystem service studied, and to the argument for conservation of pollinators and the resources they depend on. / AFRIKAANSE OPSOMMING: Insek bestuiwing speel ‘n belangrike rol in die produksie van gewasse in landbou wêreldwyd. Gewasse wat bestuiwing-afhanklik is, dra by tot ‘n gesonde verskeidenheid in die mens se dieët en hul hoë mark waarde is voordelig vir plaaslike en streeks ekonomieë. Insek bestuiwers kan of van natuurlike areas langs boorde afkomstig wees, of bestuurde bestuiwers kan deur byeboere in boorde ingebring word. Bestuiwing deur wilde bestuiwers is ‘n ekosisteem diens, maar die byeboere verskaf ‘n bestuurde diens, wat nie altyd gereken word om aan die ekosisteem verwant te wees nie. Ekosisteem dienste en hul ekonomiese waarde word gereeld gebruik as insentief vir bewaring, alhoewel dit soms moeilik is om dit te karaktariseer en te kwantifiseer. In sekere streke wêreldwyd is dit bewys dat wilde, asook bestuurde bestuiwers, bedreig is en daar heers bekommernis dat ‘n tekort aan bestuiwers gewas produksie negatief sal beïnvloed. Verskillende gewasse en kultivars het verskillende vlakke van bestuiwing-afhanklikheid as gevolg van verskillende biologiese en fisiese faktore en bestuurspraktyke. In hierdie studie is die bestuiwings-afhanklikheid van die Granny Smith appel kultivar ondersoek, asook die bydrae van wilde en bestuurde heuningbye in die Wes-Kaap provinsie van Suid Afrika. Granny Smith appels toon ‘n betekenisvolle produksie verbetering met insek bestuiwing (wilde en bestuurde bye). Daar is ‘n groter hoeveelheid bestuurde bye in ‘n boord as wilde bye, en hulle verskaf ook dus ‘n beter bestuiwingsdiens. Die voordeel van bestuurde bye bo wilde bye word veral in vrug kwaliteit opgemerk. As bestuurde bye gebruik word, is daar betekenisvol meer sade per vrug en gevolglik het die appels ook ‘n beter vorm. Verder fokus die studie ook op die kwantifisering van ekosisteem dienste wat aan die bestuurde heuningby industrie verskaf word, deur inligting van byeboere te gebruik. Daar is bevind dat 49% van die kolonies bestuurde bye in die Wes- Kaap is tot ‘n mate afhanklik van natuurlike plantegroei vir voedsel. Verder is 18% van die geproduseerde heuning ook afkomstig van natuurlike plantegroei se nektar en byeboere vang wilde kolonies om uitgestorwe bestuurde kolonies te vervang. Dit is dus duidelik dat alle bestuiwings bronne gekoppel is aan die omgewing, nie slegs wilde bestuiwers nie. ‘n Verdere ekonomiese waardasie van die onderskeie ekosisteem dienste wat bestudeer is, voeg motivering by tot die bewaring van bestuiwers en die hulpbronne waarvan hulle afhanklik is.

La malnutrition est identifiée comme l’un des facteurs potentiellement responsables des mortalités élevées de colonies d’abeilles des dernières années au Québec. Pour contrer cela, les apiculteurs donnent des suppléments de pollen à leurs colonies, mais les impacts d’une telle pratique à diverses périodes sont méconnus. Les effets de la disponibilité du pollen sur le développement de colonies d’abeilles ont été mesurés pendant 3 différentes périodes : au printemps, durant la pollinisation de la canneberge et à la fin de l’été. À chacune des périodes correspondait une expérience distincte utilisant 40 colonies. Pour chaque expérience, des conditions d’abondance de supplément de pollen et de restriction de pollen naturel étaient créées chez les colonies pendant un mois selon un plan d’expérience factorielle 2x2. L’élevage du couvain et la récolte de miel ont été mesurés jusqu’à la fin de l’été (début de l’été suivant pour l’expérience de fin d’été). Au printemps, les colonies restreintes en pollen naturel ont élevé 18% moins de couvain (p<0.05) pendant la période de restriction et 11% de moins à la fin de l’été alors que l’utilisation du supplément n’a eu aucun effet (p>0.05). Les colonies supplémentées durant la pollinisation des canneberges ont élevé moins de couvain (p<0.05) à la fin de l’été. Pour l’expérience de fin d’été, les colonies supplémentées ont eut une meilleure reprise printanière (p<0.05) de l’élevage du couvain (60% de plus) alors qu’une restriction en pollen naturel avait un effet négatif (p>0.05). Les récoltes de miel ont été augmentées (p<0.05) de 1,3 kg pendant la pollinisation de la canneberge alors qu’elles ont été diminuées (p<0.05)par une restriction en pollen naturel de 4,2 kg à la fin de l’été et de 15 kg au printemps. / The use of pollen supplement is a countermeasure to honey bee malnutrition which is identified as one of the factors causing high colonies losses over the past few years in Quebec. There is little documentation on the results of using pollen supplement during different periods. The effects of pollen availability and supplementation on the development of honey bee colonies were examined during 3 different periods: in spring, during cranberry pollination and in late summer. Each period was a distinct study using 40 different colonies. In each study, pollen supplemented and pollen restricted conditions were created for one month in 10 colonies per treatment group in a 2x2 factorial design experiment. Brood rearing and honey yield were monitored until the end of summer for the spring and the cranberry pollination studies and until the end of the following spring for the late summer study. In the spring study, pollen restricted colonies reared 18% less brood (p<0.05) during the restriction period and 11% less brood (p<0.05) by the end of summer while pollen supplement had no effect (p>0.05). Colonies supplemented during cranberry pollination study reared less brood (p<0.05) by the end of summer. In the late summer study, supplemented colonies had a greater (p<0.05) spring build-up (60% more brood) and pollen restriction negatively influence (p<0.05) brood rearing. Honey yield was decreased (p<0.05) by 15 kg in colonies exposed to a pollen restriction in spring. It was increased by 1.3 kg (p<0.05) in pollen supplemented colonies during the cranberry pollination study and was reduced by 4.2 kg (p<0.05) in pollen restricted colonies in the late summer study. In conclusion, pollen supplement improved colonies population when fed in late summer and not during spring or in cranberry pollination and was without impact on honey yield.

Immunodeficient honey bee (Apis mellifera) colonies suffer from broad range of parasites including eukaryotic protozoa. Despite this fact, the eukaryotic parasites are still poorly documented in the Czech Republic. The presence of eukaryotic parasites (Nosema ceranae, Nosema apis, Crithidia mellificae and Apicystis bombi) was observed in different apiaries in the Czech Republic. The samples were taken in 9 apiaries in 53 beehives during the 2014/2015 season. From each beehive, 10 adult of honey bees were taken from the peripheral comb in triplicate. DNA was isolated from every sample of honey bees. The parasites were detected by polymerase chain reaction (PCR) with specific primers. The treatment fall of parasitic mite Varroa destructor was obtained from beekeepers for season of 2014. Crithidia mellificae was detected by 5 types of specific primers (SEF, SER; SSU, SSU rRNA, Cyt b, Tryp cyt b) and positive amplicons were cloned and sequenced. The obtained sequences were compared with GeneBank and showed similarity from 98-100% to sequences of Lotmaria passim (Trypanosomatid). Crithidia mellificae was not detected. L. passim had prevalence of 79,2% and is reported in the Czech Republic for the first time. Primer Tryp-cyt b is recommended for the routine detection of L. passim. Nosema ceranae was...