Une nouvelle méthode, simple et rapide, a été développée pour isoler dans le miel différents antimicrobiens usuellement recherchés en contrôle sanitaire et appartenant à quatre classes différentes: les sulfamides, les tétracyclines, les macrolides et lincosamides associés et les aminoglycosides. Ces molécules antimicrobiennes sont analysées par chromatographie liquide couplée à la spectrométrie de masse en tandem (CL-SM/SM) après avoir été extraites de l’échantillon de miel par une méthode d’extraction unique. Afin de définir les conditions optimales de séparation et de détection, l’influence de la nature et de la concentration d’un agent d’appariement d’ions tel que le HFBA ou le PFPA, introduit dans la phase mobile, a pu être évaluée sur une colonne analytique en chromatographie de phase inversée de type C18. Plusieurs paramètres ont été pris en compte et étudiés lors de l’élaboration de la méthode d’extraction tels que la nature du solvant d’extraction, le pH, l’étape d’hydrolyse acide, l’efficacité de l’extraction par ultrasons et enfin la purification de l’extrait avant injection. La méthode développée a ensuite été validée suivant les recommandations de la Décision de la Commission Européenne (CE) No 2002/657 puis a subi une étape de validation supplémentaire en participant à une comparaison inter-laboratoire organisée sur des matériaux de miel contaminés et gérée par un organisme extérieur accrédité suivant la norme ISO17043. Par la suite, une démarche de transfert de la méthode analytique validée en CL-SM/SM a été mise en place pour son utilisation en chromatographie liquide couplée à la spectrométrie de masse à haute résolution (CL-SMHR). Une validation de cette démarche a été menée par l’application d’une étude statistique descriptive basée sur la notion de profil d’exactitude. Finalement, un programme expérimental de surveillance a été entrepris sur une série d’échantillons de divers miels collectés sur des marchés locaux pour tester la qualité des produits commercialisés au Liban. Contrôlés au regard de leur contamination en résidus d’antimicrobiens en CL-SM/SM parmi la trentaine de molécules prédéfinies dans l’étude, la positivité et/ou la non-conformité de certains échantillons ont pu être confirmées par l’utilisation de la CL-SMHR. / A new, simple and rapid method has been developed for the determination of multiclass antimicrobial residues in honey (sulfonamides, tetracyclines, macrolides, lincosamides and aminoglycosides). All the compounds were extracted from honey within single extraction method and analyzed by liquid chromatography - tandem mass spectrometry (LC-MS/MS) operating in positive electrospray ionization mode. In our study, we examined the behavior of volatile perfluorinated carboxylic acids (HFBA and PFPA) used as ion-pairing reagents for the separation of multiclass of antibiotic residues by reversed phase Zorbax SB C18 column. Furthermore, the extraction and clean-up steps were investigated and optimized by using ultrasonic-assisted extraction and dispersive solid phase extraction (d-SPE). Different parameters affecting the extraction efficiency including type of solvent, pH, breaking efficiencies of N-glycosidic linkage by hydrochloric acid, ultrasonic extraction and its duration compared to shaking technique, along with dispersive SPE clean-up were examined prior sample injection. The method was then validated according to European Commission Decision (EC) No 2002/657. Furthermore, the method was tested for its validity through participation in proficiency testing scheme organized by FAPAS for the analysis of tetracycline in honey. Afterwards, a transfer of the validated LC-MS/MS analytical method has been applied for the determination of antimicrobial residues in honey from low resolution to High Resolution Mass Spectrometry (HRMS). For that purpose, descriptive statistical approach was performed to assess the performance of the method based on simultaneous evaluation of the trueness and the intermediate precision. Finally, the method was applied for the determination of antimicrobial residues in honey collected from local markets at different regions in Lebanon. Positive samples were then analyzed by the LC-HRMS to confirm the presence of analytes detected by LC-MSMS.

Les abeilles en libre vol montrent des capacités cognitives visuelles remarquables mais les bases neurales sous-jacentes ne peuvent pas être étudiées chez des insectes en vol. À l'inverse, le cerveau des abeilles immobilisées est accessible mais ne permet pas d'explorer l'apprentissage visuel. Pour dépasser cette limite, notre objectif a été d'établir un dispositif de réalité virtuelle pour pouvoir tester l'apprentissage visuel sur des abeilles attachées. Dans un premier temps, les abeilles ont été testées sur leur capacité à discriminer des couleurs en s'appuyant sur les renforcements positifs ou négatifs qui leur étaient associés. Ces expériences ont permis de mettre en évidence le rôle de la vision active dans la réalité virtuelle et l'importance de la phototaxie dans ce type de système. Grâce à ce dispositif, un apprentissage non élémentaire en réalité virtuelle, le patterning négatif, a été mis en place. Ainsi il a été montré que les abeilles étaient capables de résoudre cette tache en réalité virtuelle malgré sa complexité. Enfin, en s'appuyant sur le protocole de discrimination de couleurs, l'activation du cerveau a été étudiée au cours d'un test de mémoire des couleurs en analysant l'expression de gènes considérés comme des marqueurs de l'activité neurale. Les résultats de cette thèse, fournissant deux protocoles de réalité virtuelle solides pour étudier les apprentissages visuels élémentaires et non élémentaires, constituent une avancée considérable vers la possibilité de comprendre les bases neurales sous-jacentes à ces apprentissages chez l'abeille qu'ils soient simples ou complexes. / Free flying bees are known for their impressive visual cognition abilities, but the neural bases underlying those are poorly studied because of the difficulty to explore the brain in a flying insect. Conversely, it is possible to have access to the brain with tethered bees but, until now, no studies explored visual learning. To bypass this limitation, our aim was to establish a virtual reality device to test visual learning in tethered bees. First, bees were tested for their abilities to learn to discriminate colors according to the reinforcement associated to each of them. These experiments allowed to highlight the role of the active vision in virtual reality and the importance of phototaxis in this kind of device. Driven by these good results, we explored the non- elementary visual learning in bees and, more specifically, the negative patterning. The results suggest that bees were able to resolve this task in virtual reality despite its complexity. Finally, using the protocol of color discrimination, the brain activation has been explored during a color memory recall thanks to a quantification of immediate early genes considered as neural markers. This thesis provides two solid virtual reality protocols to study elementary and non-elementary visual learning in honey bees. This constitutes a huge advance and will allow to go further towards the understanding of the neural bases of simple and complex visual learning.

The remarkable olfactory power of insect species is thought to be generated by a combinatorial action of G-protein-coupled olfactory receptors (ORs) and olfactory carriers. Two such carrier gene families are found in insects: the odorant binding proteins (OBPs) and the chemosensory proteins (CSPs). In olfactory sensilla, OBPs and CSPs are believed to deliver hydrophobic air-borne molecules to ORs, but their expression in non-olfactory tissues suggests that they also may function as general carriers in other developmental and physiological processes. ¶ Bioinformatics and experimental approaches were used to characterise the OBP and CSP gene families in a highly social insect, the western honey bee (Apis mellifera). Comparison with other insects reveals that the honey bee has the smallest set of these genes, consisting of only 21 OBPs and 6 CSPs. These numbers stand in stark contrast to the 66 OBPs and 7 CSPs in the mosquito Anopheles gambiae and the 46 OBPs and 20 CSPs in the beetle Tribolium castaneum. The genes belonging to both families are often organised in clusters, and evolve by lineage specic expansions. Positive selection has been found to play a role in generating a greater sequence diversication in the OBP family in contrast to the CSP gene family that is more conserved, especially in the binding pocket. Expression proling under a wide range of conditions shows that, in the honey, bee only a minority of these genes are antenna-specic. The remaining genes are expressed either ubiquitously, or are tightly regulated in specialized tissues or during development. These findings support the view that OBPs and CSPs are not restricted to olfaction, and are likely to be involved in broader physiological functions. ¶ Finally, the detailed expression study and the functional characterization of a member of the CSP family, uth (unable-to-hatch), is reported. This gene is expressed in a maternal-zygotic fashion, and is restricted to the egg and embryo. Blocking the zygotic expression of uth with double-stranded RNA causes abnormalities in all body parts where this gene is highly expressed. The treated embryos are `unable-to-hatch' and cannot progress to the larval stages. Our ndings reveal a novel, essential role for this gene family and suggest that uth is an ectodermal gene involved in embryonic cuticle formation.

Det här arbetet handlar om tambins inverkan på naturligt förekommande pollinatörer. Min frågeställning berör: tillgången på näring, hälsotillståndet och populationsnivån hos naturligt förekommande pollinatörer i förhållande till tambins närvaro. Det som framkommit av den här litteraturstudien är att introducerade tambin leder till att naturligt förekommande pollinatörer minskar i antal kring bikuporna, en del arter söker föda på andra blommor eller senare på dygnet än tidigare. Den minskade tillgången på föda som denna konkurrens innebär har en negativ inverkan på humlors kroppsstorlek och därmed deras överlevnad. Samt att tambin kan föra över patogener så som varroakvalster, nosemasjuka och deformed wing virus till humlor vilket leder till minskad livslängd och sämre fortplantning. / This study is about the impact of honeybees on native pollinators. My questions concerns: availability of food, the health and population level of native pollinators in relation to the presence of honeybees. What has emerged from this literature review is that the introduction of honeybees lead to a decline in numbers of bee and bumblebee pollinators in proximity of the hives and also alters their behaviour, some species choose to forage on other flowers, or later in the day than before honeybees where introduced. The reduced availability of food that this competition induces has a negative impact on the size of bumblebees body’s and thus their survival. What was also discovered was that honeybees can transfer pathogens such as varroa mites, the microsporidium Nosema ceranae and deformed wing virus to bumblebees, leading to reduced life expectancy and poor propagation.

The European honey bee (Apis mellifera) has been present in Australia for approximately 150 years. For the majority of that time it was assumed this species could only be of benefit to Australia‘s natural ecosystems. More recently however, researchers and conservationists have questioned this assumption. Honey bees are an introduced species and may be affecting native fauna and flora. In particular, native bees have been highlighted as an animal that may be experiencing competition from honey bees as they are of similar sizes and both species require nectar and pollen for their progeny. Most research to date has focused on indirect measures of competition between honey bees and native bees (resource overlap, visitation rates and resource harvesting). The first chapter of this thesis reviews previous research explaining that many experiments lack significant replication and indirect measures of competition cannot evaluate the impact of honey bees on native bee fecundity or survival. Chapters two and four present descriptions of nesting biology of the two native bee species studied (Hylaeus alcyoneus and an undescribed Megachile sp.). Data collected focused on native bee fecundity and included nesting season, progeny mass, number of progeny per nest, sex ratio and parasitoids. This information provided a picture of the nesting biology of these two species and assisted in determining the design of an appropriate experiment. Chapters three and five present the results of two experiments investigating the impact of honey bees on these two species of native bees in the Northern Beekeepers Nature Reserve in Western Australia. Both experiments focused on the fecundity of these native bee species in response to honey bees and also had more replication than any other previous experiment in Australia of similar design. The first experiment (Chapter three), over two seasons, investigated the impact of commercial honey bees on Hylaeus alcyoneus, a native solitary bee. The experiment was monitored every 3-4 weeks (measurement interval). However, beekeepers did not agist hives on sites simultaneously so measurement intervals were initially treated separately using ANOVA. Results showed no impact of honey bees at any measurement interval and in some cases, poor power. Data from both seasons was combined in a Wilcoxon‘s sign test and showed that honey bees had a negative impact on the number of nests completed by H. alcyoneus. The second experiment (Chapter 5) investigated the impact of feral honey bees on an undescribed Megachile species. Hive honey bees were used to simulate feral levels of honey bees in a BACI (Before/After, Control/Impact) design experiment. There was no impact detected on any fecundity variables. The sensitivity of the experiment was calculated and in three fecundity variables (male and female progeny mass and the number of progeny per nest) the experiment was sensitive enough to detect 15-30% difference between control and impact sites. The final chapter (Chapter six) makes a number of research and management recommendations in light of the research findings.

Diss. (sammanfattning)--Uppsala : Sveriges lantbruksUniversity, 2001. / Härtill 4 uppsatser.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-04-11T18:53:42Z No. of bitstreams: 1 IdalecioPS_TESE.pdf: 5907205 bytes, checksum: 814ed97201f439fa05e27b9c404b4908 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-04-13T15:20:53Z (GMT) No. of bitstreams: 1 IdalecioPS_TESE.pdf: 5907205 bytes, checksum: 814ed97201f439fa05e27b9c404b4908 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-04-26T12:13:04Z (GMT) No. of bitstreams: 1 IdalecioPS_TESE.pdf: 5907205 bytes, checksum: 814ed97201f439fa05e27b9c404b4908 (MD5) / Made available in DSpace on 2017-04-26T12:13:13Z (GMT). No. of bitstreams: 1 IdalecioPS_TESE.pdf: 5907205 bytes, checksum: 814ed97201f439fa05e27b9c404b4908 (MD5) Previous issue date: 2015-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Pollinators are among the essential components for the functioning of ecosystems in general, because it is essential for the reproduction and maintenance of the diversity of plant species and provide food for humans and animals, also influencing the qualitative aspect of the production. Currently, the population density of many pollinators are being reduced to levels that can impair pollination services. The decline of honeybee Africanized (Apis mellifera L.) populations impacts global agricultural production and has a clear impact on both food production and the economy. One of the probable causes for this decline is the indiscriminate use of pesticides. Thus, there was comparative levels of exposure to pesticides among bees (Apis mellifera L.) that are used to pollinate melon crops, bees (Apis mellifera L.) kept in the bush, and Jandaira bees (Melipona subnitida ) kept in the bush. To do this, honey samples were collected from 23 colonies of Apis mellifera used to pollinate melon palntações, 20 colonies of Apis mellifera foraging forest and 10 colonies of Melipona subnitida foraging in the forest. The level of exposure to pesticides was determined by measuring the residual levels of 152 compounds in honey using a multiresidue analysis technique in UFLC-MS system. Honey samples from the present study contained 19 different pesticides, 13 of which were present in honey from bees pollinating melon crops. The levels of some compounds were sufficiently high to promote adverse effects in the bees. Thus, crop pollination offers a great toxicological risk to bees that may contribute to colony collapse / Os polinizadores estão entre os componentes essenciais para o funcionamento dos ecossistemas em geral, pois é essencial para a reprodução e manutenção da diversidade de espécies de plantas e provê alimentos para humanos e animais, influenciando, também, o aspecto quantitativo e qualitativo da produção. Atualmente, a densidade populacional de muitos polinizadores está sendo reduzida a níveis que podem comprometer os serviços de polinização. O declínio das populações de abelhas (Apis mellifera L.) tem impactado na produção agrícola mundial e tem um impacto claro sobre a produção de alimentos e da economia. Uma das causas prováveis para este declínio é o uso indiscriminado de agrotóxicos. Desta forma, fez-se estudo comparativo dos níveis de exposição aos pesticidas entre as abelhas Apis mellifera L. (africanizadas) que são usadas para polinizar plantações de melão, as abelhas Apis mellifera L.(africanizadas) mantidas na caatinga, e as abelhas jandaíra (Melipona subnitida) mantidas na caatinga. Para isso, amostras de mel foram coletadas de 23 colônias de abelhas africanizadas Apis mellifera utilizadas para polinizar plantações de melão, 20 colônias de abelhas africanizadas Apis mellifera que forrageiam de na caatinga e 10 colônias de Melípona subnitida que forrageiam na caatinga. O nível de exposição aos pesticidas foi determinado medindo os níveis residuais de 152 compostos no mel, utilizando uma técnica de análise de multiresíduos no sistema UFLC-MS. As amostras de mel estudadas apresentaram 19 pesticidas diferentes, 13 dos quais estavam presentes no mel de abelhas polinizadoras das plantações de melão. Os níveis de alguns compostos foram suficientemente elevados para promover efeitos adversos em abelhas. Assim, a polinização das culturas oferece um grande risco de toxicidade para as abelhas que pode contribuir para o colapso da colônia / 2017-04-11

Les virus font partie des entités les plus abondantes sur Terre, mais la diversité des virus est très peu connue puisque biaisée en faveur d’hôtes animaux d’intérêts sociétal, agronomique et économique. L’apport des nouvelles techniques de séquençage permet actuellement d’obtenir des informations qui étaient tout simplement inaccessibles. Le but de mon travail de thèse a été l’étude de la diversité virale présente au sein d’un grand nombre d’animaux non-modèles. Pour répondre à cette problématique il m’a fallu mettre en place une méthodologie analytique innovante de découverte de nouveaux virus par une approche de méta-transcriptomique. Ce travail i) montre que la méthodologie bioinformatique mise en place est pertinente, ii) permet de découvrir de nouveaux virus ayant des caractéristiques génomiques particulières relevant de nouveaux genres ou familles de virus, iii) révèle de nouveaux hôtes pour des virus appartenant à des familles virales très étudiées et iv) montre que la gamme d’hôte de virus connus peut être plus étendue qu’attendu grâce à un focus sur la diversité des virus d’hyménoptères. D’une manière plus globale, mon travail permet de combler quelques lacunes existantes dans les connaissances liées à l’étude de la diversité virale et met en évidence l’importance de l’étude des animaux non-modèles. / Viruses are among the most abundant entities on Earth, but the viral diversity remains mostly unknown as currently biased in favour of animals of social, agronomic and economic interest. Next Generation Sequencing technologies provide access to so far inaccessible information. The aim of my PhD thesis was the study of the viral diversity within a large range of non-model animals. To address this question I set up an innovative analytical framework to discover new viruses based on a meta-transcriptomic approach. This work i) shows that this bioinformatics method is efficient and powerful, ii) allows the discovery of new viruses with particular genomic organisations suggesting they belong to new virus genera of families, iii) uncovered new viruses from new hosts in well-known viral families and iv) shows wider viral host range than previously expected based on a particular focus on hymenopteran viral diversity. Overall, my work allows to fill some gaps in the knowledge of viral diversity and shows the importance of studying non-model animal species in virology.

The beekeeping, especially the honey production is pointed as one of the alternatives with respect to the reversion of unfavourable the social and ambient conditions in the agricultural way of the Northeast Region it Brazil and Been it of Alagoas. Having in the agricultural space not more only activities exclusively agriculturists, however the other activities like the small agro-industry, of familiar characteristic, can stimulate the generation, direct and indirect, of new ranks of work and income mainly for the familiar beekeepers, promoting its (re) economic inclusion. We search here to argue through the concepts of Sustainable Development, Productive Management of Chain and Management of the agribusiness, modern, the dynamics of the production of honey of bees in Alagoas. The objective is to describe the functioning of the chain of agro-industrial production of the honey of bees, being aimed at to identify what it hinders its development and sustainability so that in fact, if constitutes in an inclusion alternative economic partner of involved actors and, at the same time where the activity is viable inside of the current economic model. This quarrel if relates, also, with the actions and public politics necessary to stimulate the cited model of production, with sustainability. / A apicultura, especialmente a produção de mel é apontada como uma das alternativas para a reversão das condições sociais e ambientais desfavoráveis no meio rural da Região Nordeste do Brasil e do Estado de Alagoas. Tendo no espaço rural não mais apenas atividades exclusivamente agrícolas, porém a pluriatividade, a pequena agroindústria, de característica familiar, pode impulsionar a geração, direta e indireta, de novos postos de trabalho e de renda principalmente para os apicultores familiares, promovendo a sua (re) inclusão econômica. Buscamos aqui discutir através dos conceitos de Desenvolvimento Sustentável, Gestão de Cadeia Produtiva e Gestão do Agronegócio, moderno, a dinâmica da produção de mel de abelhas em Alagoas. O objetivo é descrever o funcionamento da cadeia de produção agroindustrial do mel de abelhas, visando identificar o que impede o seu desenvolvimento e sustentabilidade para que de fato, se constitua numa alternativa de inclusão sócia econômica dos atores envolvidos e, ao mesmo tempo em que a atividade seja viável dentro do modelo econômico atual. Essa discussão se relaciona, também, com as ações e políticas públicas necessárias para impulsionar o referido modelo de produção, com sustentabilidade.

The aim of this thesis was to monitor the Varroa destructor parasitic mite in correlation with the microclimatic conditions of the Carniolan honey bee (Apis mellifera carnica). The rate of infestation of selected bee colonies at different locations was assessed in three-day intervals. At the same time, the microclimate in the hives was observed. The monitoring took place from April to mid-October. Also, the effect of the microclimate on the mite fall count was evaluated. During the whole evaluation, the highest average daily fall count was 2.08 mites per day at honeybee colonies at location 1 and 2; 1.97 at location 3. There was no statistically significant difference (P > 0.05) between those locations. The comparison of fall count between the moths of observation revealed, that highest fall count was during September (3.03 mites per day) and the lowest fall count was in April (0.41 mites per day; P 0.001). The rates of dependence (assessed by correlation analysis) between the fall count and microclimatic conditions in individual colonies varied. The strongest correlation between hive temperature and fall count (r = -0.45, P 0.05) was found at location 2. A low correlation was found at location 1 (r = -0.17, P 0.05). On the other hand, location 3 showed an insignificant and inconclusive correlation between hive temperature and fall count (r = 0.003, P > 0.05). The aggregate data (without distinction of location or month) showed significant (P 0.05) correlation between fall count and hive temperature (r = -0.14). The correlation between relative air humidity in hive and the fall count was statistically insignificant and low (r = -0.02, P > 0.05). The results revealed that the degree of correlation between the hive microclimate and the development (fall count respectively) of the Varroa destructor population is different for each colony. The monitoring of the dead Varroa destructor females is an appropriate complementary tool to diagnose a colony's infestation. The statistical analysis confirmed that with the decreasing summer and end-of-summer temperatures the Varroa destructor population grows and it is necessary to take measures to suppress its growth due to the development of the honey bee long-term winter generation.