Il sinergizzante piperonil-butossido (PBO) usato in combinazione con insetticidi è in grado di contrastare i parassiti resistenti inibendo temporaneamente i sistemi di detossificazione. Nuovi sinergizzanti sono stati sintetizzati a partire dalla struttura molecolare del PBO, ma anche gli insetti utili come le api possono risentire di una maggiore efficacia degli insetticidi. In questo studio abbiamo osservato l’attività enzimatica in vitro (esterasi, acetilcolinesterasi, glutatione S-transferasi e monossigenasi P450) di api operaie Italiane e Carniche in presenza del PBO e dei suoi nuovi derivati del benzodiossolo e 2,3-diidrobenzofurano mediante saggi spettrofotometrici, per determinare se i sistemi di detossificazione e altri sistemi enzimatici sono influenzati dai sinergizzanti. I nuovi derivati del PBO possono inibire parzialmente le esterasi in alcune popolazioni di api più sensibili. Altri sistemi enzimatici non sembrano essere inibiti dai sinergizzanti studiati. Studi di tossicità acuta orale e di contatto sono stati condotti in laboratorio presso il Bee Research Institute (Dol, Repubblica Ceca) su api operaie Carniche per verificare il possibile incremento di tossicità degli insetticidi imidacloprid e alfa-cipermetrina in combinazione con PBO e i suoi derivati del benzodiossolo. Gli studi hanno mostrato una differenza significativa tra la tossicità dei due principi attivi da soli e in combinazione con i sinergizzanti. / The synergist piperonyl-butoxide (PBO) has been proved to successfully control resistant pests when combined with insecticides by temporarily inhibiting the detoxification systems which lead to resistance. New synergists, starting from the molecular structure of PBO, have been developed but pollinators may also suffer from an increased efficacy of insecticides. In this study we observed in vitro enzyme (esterases, acetylcholinesterases, glutathione S-transferases and P450 mixed function oxidases) activity of Italian and Carniolan honey bee workers in presence of the synergist PBO and its novel benzodioxole and 2,3-dihydrobenzofuran derivatives through spectrophotometric assays, to determine if detoxification systems and other enzymatic systems are affected by synergists. This data show that novel PBO derivatives may partially inhibit esterases in some honey bee populations which are more sensitive. Other enzymatic systems seem not to be targeted by the studied synergists. Acute oral and contact toxicity studies were conducted in laboratory conditions at the Bee Research Institute (Dol, Czech Republic) on adult Carniolan workers to investigate the possible increase of imidacloprid and alpha-cypermethrin toxicity when combined with PBO and its benzodioxole derivatives. Tests showed some significant difference between the toxicity of the two active principles alone and combined with the synergists.

No description available.

Einige Zuchtmerkmale der Honigbiene lassen sich sinnvoll an einzelnen Arbeiterinnen messen. Dabei kann der Zuchtfortschritt potentiell beschleunigt werden, wenn Söhne der selektierten Arbeiterinnen zur Besamung herangezogen werden. Ziel dieser Arbeit war deshalb die Schaffung von Methoden zur Erzeugung von Söhnen selektierter Arbeitsbienen. Um die Fruchtbarkeit der selektierten Tiere zu fördern wurde zunächst vergeblich versucht, eine Hormonbehandlung zur gezielten Induktion der Eibildung zu entwickeln. Dabei konnten die Mechanismen der Fortpflanzungssteuerung bei Arbeitsbienen teilweise aufgeklärt werden. Um die Fruchtbarkeit in allen Bienen außer den selektierten zu unterdrücken wurde zunächst die Sterilisation durch harte Röntgenstrahlung erprobt. Die erhaltenen Tiere legten keine Eier, zeigten aber starke somatische Schäden. Die Aufzucht von Arbeiterinnen-Larven in überalterten Völkern ergab keine sterilen, sondern besonders fruchtbare Tiere. Schließlich gelang es unter Verwendung von Flugbienen als Begleittieren, in Jungbienen in über 60% der Fälle volle Ovarentwicklung zu erzielen. Auch konnten alle erzeugten Eier Jungbienen zugeordnet werden. Um die Eignung von Arbeiterinnen-gelegten Eiern für Zuchtzwecke zu prüfen, wurden Qualitätsparameter gemessen. Arbeiterinnen-gelegte Eier zeigten sich empfindlicher gegenüber Trockenheit und mechanischer Beschädigung als Königinnen-gelegte Eier. Alle in Völker verbrachten Arbeiterinnen-gelegten Eier wurden ausgefressen. Also wurde eine Methode zur Anfütterung von Larven im Brutschrank entwickelt. Die so erzeugten Larven überlebten aber ebenfalls nicht in Völkern. Deshalb wurde ein Verfahren entwickelt, mit dem aus über 50% der eingesetzten Larven durch Brutschrankaufzucht adulte Tiere erhalten werden können. Ein vorläufiges Protokoll zur Erzeugung von Söhnen individuell selektierter Arbeitsbienen wurde erarbeitet. / Breeding of honey bees usually involves selection at the colony level. However, some important breeding traits exist that can also be measured at the scale of the individual worker bee. In these cases, genetic progress can potentially be accelerated by using drones which are sons of the selected individuals. The aim of this study was therefore to create methods for the production of offspring from individually selected workers. To enhance fertility of selected workers, an attempt was made to develop a hormonal treatment to induce worker fertility. It led to the partial elucidation of the endocrine regulation of reproduction in worker honey bees. Methods were tested to suppress fertility in all bees except the selected workers, in order to prevent competition among workers and enable identification of the eggs of selected bees. Method tested included the application of ultra-hard X-rays and rearing of larvae inside over-aged colonies. Finally, it could be shown that forager bees can be used as companion bees to selected workers. This method led to fully developed ovaries in more than 60% of bees representing the selected workers in these tests, who also were the only animals to produce eggs. The next subtask was the measurement of characteristics linked to viability in eggs from laying workers. Worker-laid eggs showed to be more sensitive to dryness and mechanical damage. When worker-laid eggs were inserted into queenless or queenright colonies for rearing, they were systematically cannibalised. Therefore, a method was developed to rear eggs into living larvae in an incubator. However, the larvae produced in this way were not accepted by colonies either. To circumvent this problem, a protocol was designed and validated that allows to rear worker-derived drone larvae into adults in vitro. The study led to a preliminary protocol for the production of drones that are sons of individually selected worker bees.

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.

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 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.

Depuis le début du XXème siècle, l’évolution de l’agriculture et l'intensification des pratiques agricoles ont conduit à des modifications profondes du paysage induisant des pertes importantes d’habitats naturels et semi-naturels. Ces changements, entraînant notamment la diminution des ressources végétales et impliquant l'utilisation systématique de pesticides, ont des répercussions sur les colonies d’abeilles. C’est dans ce contexte que, durant deux saisons apicoles, des inventaires botaniques ont été réalisés au sein de l’aire de butinage de deux ruchers situés en paysage de grande culture du nord-ouest de la France et que des échantillons de pelotes de pollen et de miels prélevés dans ces ruchers ont été analysés. L’objectif était de comparer les ressources floristiques disponibles et celles réellement exploitées par les abeilles et d’entrevoir les stratégies de butinage mises en place dans ce cadre paysager. Les relevés floristiques ont permis de distinguer d’une part, de larges surfaces d’espèces cultivées, à floraison ponctuelle et d’autre part, des surfaces non cultivées, de plus petite taille, occupées par de nombreuses espèces sauvages dont la floraison s’étale sur la totalité de la saison apicole. Les analyses palynologiques montrent que ces espèces floristiques sauvages sont exploitées en continu y compris pendant la période de floraison des espèces cultivées. Bien que cette étude ne soit pas en mesure de le démontrer, il est possible que les espèces sauvages procurent aux colonies un apport nutritionnel que la floraison ponctuelle des plantes cultivées ne peut totalement compenser. Une meilleure connaissance de la valeur nutritionnelle des différents pollens d’essences végétales, ainsi que des besoins alimentaires basiques nécessaires au développement et à la pérennisation des colonies devraient aider à éprouver cette hypothèse. / Since the beginning of the XXth century, the evolution of agriculture and the intensification of farming practices have led to landscape changes with the loss of natural and semi-natural habitats. These changes, involving a decrease of plant resources and a use of pesticides, may have noticeable impact on honey bee colonies. In this framework of weakened biodiversity, the initiative was taken of recording botanical species growing in the foraging area of two apiaries in cultivated landscapes of Western France and sampling pollen pellets and honey for pollen species determination during two successive beekeeping seasons. The aim of this study was to compare the available flora resources with those really exploited by the honey bees, to get a glimpse of the foraging strategies developed by the colonies in this landscape context. The floristic readings enabled to distinguish wide areas of cultivated plant species characterized by temporary blossoms from areas of wild species characterized by much smaller superficies and by flowerings spread over the year. Palynological analyses indicate that wild floristic species are exploited throughout the beekeeping season including during blooms of cultivated plant species. Even though not conclusively stated by the present study, it is possible that wild species provide colonies with nutritional intakes that cannot be totally compensated by temporary blooms of cultivated plants. A more extended knowledge of the nutritional values of pollens produced by plant species together with a better understanding of basic food needs for development and perennisation of colonies should help to test this hypothesis.

No description available.

European honey bee (Apis mellifera) is major pollinator for agriculture and vital for food production. Large number of viruses infecting A. mellifera have been discovered over the years, but it isn't yet known if they are pathogenic for their host. However, presence of non-viral pathogens like Varroa destructor can greatly increase their virulence and have fatal consequences for the colony. The aim of this study was to test and verify robustness of the method for virome detection on healthy honey bees from the Czech Republic. Last but not least we aimed to detect non-viral parasites and correlate their presence with detected viruses. We have successfully identified large number of viral sequences from different viral families. Viral composition was found to be influenced mainly by colony from where the honey bees were collected. That was mainly given by a large amount of bacteriophages in the samples. However, analysis of individual viruses, known to infect honey bee, indicated that viral prevalence and viral loads of specific viruses is quite different among individual honey bees from the same colony. Interestingly we were able to find highly diverse Lake Sinai viruses. We were able to observe correlations either between individual viruses or viral other non-viral pathogens. Further analysis is...