Biogene Amine sind kleine organische Verbindungen, die sowohl bei Wirbeltieren als auch bei Wirbellosen als Neurotransmitter, Neuromodulatoren und/oder Neurohormone wirken können. Sie bilden eine bedeutende Gruppe von Botenstoffen und entfalten ihre Wirkungen über die Bindung an eine bestimmte Klasse von Rezeptorproteinen, die als G-Protein-gekoppelte Rezeptoren bezeichnet werden. Bei Insekten gehören zur Substanzklasse der biogenen Amine die Botenstoffe Dopamin, Tyramin, Octopamin, Serotonin und Histamin. Neben vielen anderen Wirkung ist z.B. gezeigt worden, daß einige dieser biogenen Amine bei der Honigbiene (Apis mellifera) die Geschmacksempfindlichkeit für Zuckerwasser-Reize modulieren können. Ich habe verschiedene Aspekte der aminergen Signaltransduktion an den „Modellorganismen“ Honigbiene und Amerikanische Großschabe (Periplaneta americana) untersucht. Aus der Honigbiene, einem „Modellorganismus“ für das Studium von Lern- und Gedächtnisvorgängen, wurden zwei Dopamin-Rezeptoren, ein Tyramin-Rezeptor, ein Octopamin-Rezeptor und ein Serotonin-Rezeptor charakterisiert. Die Rezeptoren wurden in kultivierten Säugerzellen exprimiert, um ihre pharmakologischen und funktionellen Eigenschaften (Kopplung an intrazelluläre Botenstoffwege) zu analysieren. Weiterhin wurde mit Hilfe verschiedener Techniken (RT-PCR, Northern-Blotting, in situ-Hybridisierung) untersucht, wo und wann während der Entwicklung die entsprechenden Rezeptor-mRNAs im Gehirn der Honigbiene exprimiert werden. Als Modellobjekt zur Untersuchung der zellulären Wirkungen biogener Amine wurden die Speicheldrüsen der Amerikanischen Großschabe genutzt. An isolierten Speicheldrüsen läßt sich sowohl mit Dopamin als auch mit Serotonin Speichelproduktion auslösen, wobei Speichelarten unterschiedlicher Zusammensetzung gebildet werden. Dopamin induziert die Bildung eines völlig proteinfreien, wäßrigen Speichels. Serotonin bewirkt die Sekretion eines proteinhaltigen Speichels. Die Serotonin-induzierte Proteinsekretion wird durch eine Erhöhung der Konzentration des intrazellulären Botenstoffs cAMP vermittelt. Es wurden die pharmakologischen Eigenschaften der Dopamin-Rezeptoren der Schaben-Speicheldrüsen untersucht sowie mit der molekularen Charakterisierung putativer aminerger Rezeptoren der Schabe begonnen. Weiterhin habe ich das ebony-Gen der Schabe charakterisiert. Dieses Gen kodiert für ein Enzym, das wahrscheinlich bei der Schabe (wie bei anderen Insekten) an der Inaktivierung biogener Amine beteiligt ist und im Gehirn und in den Speicheldrüsen der Schabe exprimiert wird. / Biogenic amines are small organic compounds that act as neurotransmitters, neuromodulators and/or neurohormones in vertebrates and in invertebrates. They form an important group of messenger substances and mediate their diverse effects by binding to membrane receptors that primarily belong to the large gene-family of G protein-coupled receptors. In insects, the group of biogenic amine messengers consists of five members: dopamine, tyramine, octopamine, serotonin, and histamine. Besides many other effects, some of these biogenic amines were shown, for example, to modulate gustatory sensitivity to sucrose stimuli in the honeybee (Apis mellifera). I have investigated various aspects of the aminergic signal transduction in the “model organisms” honeybee and American cockroach (Periplaneta americana). So far, I have characterized two dopamine receptors, a tyramine receptor, an octopamine receptor and a serotonin receptor of the honeybee, which is well-known for its learning and memory capacities. The receptors where expressed in cultivated mammalian cells in order to analyze their pharmacological and functional (i.e., second messenger coupling) properties. The spatiotemporal expression patterns of the respective receptor mRNA were investigated in the honeybee brain by using different techniques (RT PCR, Northern blotting, in situ-hybridization). The salivary glands of the American cockroach were used as a model object in order to investigate the cellular effects of biogenic amines. Both dopamine and serotonin trigger salivary secretion in isolated salivary glands. The quality of the secreted saliva is, however, different. Stimulation of the glands by serotonin results in the production of a protein-rich saliva, whereas stimulation by dopamine results in saliva that is protein-free. Serotonin-induced protein secretion is mediated by an increase in the intracellular concentration of cAMP. The pharmacological properties of dopamine receptors associated with cockroach salivary glands were investigated and the molecular characterization of putative aminergic receptors of the cockroach was initiated. Furthermore, I have characterized the ebony gene of the cockroach. This gene encodes an enzyme that is probably involved in the inactivation of biogenic amines in the cockroach (as in other insects). The ebony gene is expressed in the brain and in the salivary glands of the cockroach.

In this study, some important aspects of the relationship between honey bees (Apis mellifera L.) and pesticides have been investigated. In the first part of the research, the effects of the exposure of honey bees to neonicotinoids and fipronil contaminated dusts were analyzed. In fact, considerable amounts of these pesticides, employed for maize seed dressing treatments, may be dispersed during the sowing operations, thus representing a way of intoxication for honey bees. In particular, a specific way of exposure to this pesticides formulation, the indirect contact, was taken into account. To this aim, we conducted different experimentations, in laboratory, in semi-field and in open field conditions in order to assess the effects on mortality, foraging behaviour, colony development and capacity of orientation. The real dispersal of contaminated dusts was previously assessed in specific filed trials. The results showed a significant effect on mortality of neonicotinoids and fipronil contaminated dusts, both in laboratory and in semi-field trials. However, no effects were evidenced in honey bees orientation capacity.In the second part, the impact of various pesticides (chemical and biological) on honey bee biochemical-physiological changes, was evaluated. Different ways and durations of exposure to the tested products were also employed. Three experimentations were performed, combining Bt spores and deltamethrin, Bt spores and fipronil, difenoconazole and deltamethrin. Several important enzymes (GST, ALP, SOD, CAT, G6PDH, GAPDH) were selected in order to test the pesticides induced variations in their activity. In particular, these enzymes are involved in different pathways of detoxification, oxidative stress defence and energetic metabolism. The analysis of different biochemical indicators highlighted some interesting physiological variations that can be linked to the pesticide exposure. We therefore stress the attention on the possibility of using such a methodology as a novel toxicity endpoint in environmental risk assessment.

O objetivo deste estudo foi estimar parâmetros genéticos relacionados ao peso à emergência, comprimento corporal, comprimento e largura do abdome, peso, diâmetro e volume de espermateca e peso dos ovários de rainhas Apis mellifera L. (Hymenoptera: Apidae) africanizadas. Os dados analisados se referem a pesagem corporal e medidas do comprimento e largura do abdome de 1056 rainhas, mensurações do comprimento corporal de 102 rainhas, peso da espermateca de 830 rainhas, diâmetro e volume da espermateca de 813 rainhas e peso dos ovários de 970 rainhas, totalizando 1133 animais na matriz de parentesco. Os efeitos fixos considerados foram família, época e mini-recria e os efeitos aleatórios, o genético aditivo e o resíduo. As análises foram realizadas por meio da inferência Bayesiana utilizando o modelo animal. Foram realizadas análises unicarater e bicarater para todas as características. Em análise unicarater, a proporção da variância genética aditiva foi responsável por mais de 50% da variância fenotípica total e as herdabilidades apresentaram estimativas de magnitudes médias a altas para todas as características, sendo 0,46 para peso à emergência, 0,59 para comprimento do abdome, 0,56 para largura do abdome, 0,80 para comprimento corporal total, 0,62 para peso da espermateca, 0,55 para diâmetro da espermateca, 0,64 para volume da espermateca e 0,62 para peso dos ovários. Correlações genéticas favoráveis de magnitudes entre 0,46 a 0,86 foram encontradas para as associações entre peso à emergência, comprimento corporal e comprimento e largura do abdome. Quando avaliadas as relações entre peso à emergência, comprimento corporal e comprimento e largura do abdome com peso, diâmetro e volume de espermateca e peso dos ovários, a maior correlação genética foi entre peso à emergência e peso dos ovários de 0,49. Correlação genética de 0,46 foi encontra para peso à emergência e comprimento do abdome, 0,86 para peso à emergência e largura do abdome e 0,70 para peso à emergência e comprimento corporal. Todas as características avaliadas em análise unicarater apresentam potencial de seleção e, as correlações genéticas entre o peso da rainha à emergência e características da espermateca e peso dos ovários são determinados, em parte, pelo mesmo conjunto de genes, indicando que essas associações podem ser utilizadas em programas de melhoramento genético de abelhas africanizadas, visando o maior potencial reprodutivo de rainhas A. mellifera africanizadas. / The objective of this study was to estimate the genetic parameters the weight at emergence, body length, length and width of abdomen, weight, diameter and volume the spermathecae and weight of ovaries of queen bees Apis mellifera L. (Hymenoptera: Apidae) africanized. The data analysed refer to weighing the body and length and width of abdomen measures of 1056 queen bees, measurements of body length the of 102 queen bees, weight of spermathecae of 830 queen bees, diameter and volume of spermathecae of 813 queen bees and weight of ovaries of 970 queen bees, amounting 1133 animals in the matrix parentage. The effects fixed considered were family, epoch and mini-recreates and the random effects, genetic aditive and the residual. The analyses realized were through the inference Bayesian using the animal model. Were conducted analyses unicharacter and bicharacter for all characteristics. In analyses unicharacter, the proportion the variance genetic aditive was responsible for more than 50% the total phenotypic variance and the heritabilities have submitted estimates of medium to high magnitudes the for all characteristcs, being 0.46 for weight at emergence, 0.59 for length abdomen, 0.56 for width of abdomen, 0.80 for body length, 0.62 for spermathecae weight, 0.55 for spermathecae diameter, 0.64 for spermathecae volume and 0.62 for weight of ovaries. Genetic correlations favorable of magnitude between 0.46 to 0.86 were found for the associations between weight at emergence, body length and length and width abdomen. When evaluated the relationship between the emergence weight, body length and width and length abdomen with weight, diameter and volume of spermatheca and ovarian weight, the greater genetic correlation was between the weight at emergence and weight of ovaries of 0.49. Genetic correlation of 0.46 was found for the emergence weight and length of the abdomen, 0.86 for weight at emergence and abdomen width and 0.70 for the emergence weight and body length. That all characteristics evaluated in analysis unicharacter have potential of selection and, genetic correlation the weight at emergence and characteristics of spermathecae and weight of ovaries are determined, in part, the same set of genes, indicating that these associations can be used in breeding programs of bees africanized, for the greater reproductive potential of queen bees A. mellifera africanized .

As abelhas africanizadas (Apis mellifera L.) estão desaparecendo e acredita-se que seja devido ao intenso uso de inseticidas, poluição e alterações climáticas, no entanto muitas culturas dependem da polinização realizada por essas abelhas. Desta forma, o objetivo deste trabalho foi avaliar o efeito de três linhagens de Bacillus thuringiensis (Bt) (IPS 82, BR 81 e BR 147), entomopatógeno utilizado para o controle de lagartas desfolhadoras, e de três produtos fitossanitários sintéticos (PFS 1: Acetamiprido + Alfa-cipermetrina; PFS 2: Imidacloporido + Beta-Ciflutina e PFS 3: Fenitrotiona + Esfenvalerato), sobre a longevidade de operárias adultas de A. mellifera africanizada. Foram realizados três bioensaios com as linhagens de Bt e dos PFSs: A) Pulverização sobre A. mellifera; B) Contato de A. mellifera com superfície pulverizada e C) Pasta cândi incorporada com os produtos e análise histológica do intestino médio das abelhas. Nos três bioensaios a testemunha foi composta por água deionizada esterilizada. Cada tratamento foi composto por 60 abelhas, sendo cada abelha considerada uma repetição. Os bioensaios foram mantidos em câmara climatizada tipo BOD (34 ± 2ºC, U.R. de 60 ± 5%) e a longevidade das operárias avaliada a uma; duas; três; quatro; cinco; seis; nove; 12; 15; 18; 21; 24; 30; 36; 42; 48; 60; 72; 96; e 120 horas.Na pulverização de Bt sobre A. mellifera verificou-se que apenas a linhagem Bt 1 (IPS 82) causou redução da longevidade de operárias (88,7 horas), enquanto no bioensaio de contato a linhagem Bt 3 - BR147 provocou redução (101,2 horas). Todas as linhagens de Bt avaliadas, quando incorporadas à alimentação de A. mellifera reduziram a longevidade das operárias (Bt 1 – IPS 82: 64,50 horas; Bt 2 – BR 81: 64,56 horas e Bt 3 – BR 147: 60,00horas) e com a análise histológica pôde-se perceber a desintegração das células epiteliais do mesêntero das abelhas, causada pelas bactérias que elas ingeriram. Os três produtos fitossanitários sintéticos reduziram a longevidade das operárias de A. mellifera, no bioensaio de pulverização direta (PFS 1 e PFS 2 – 1,0 hora; PFS 3 – 2,21 horas). No contato de A. mellifera com superfície pulverizada com PFS, as abelhas ficaram duas horas em contato com os produtos na placa de Petri, e ao final dessas duas horas, 100% abelhas que entraram em contato com os três produtos fitossanitários sintéticos estavam mortas. Quando incorporados os PSFs na alimentação, ambos reduziram a longevidade das operárias (PFS 1 – 22 horas, PFS 2 – 21,72 horas e PFS 3 – 43,93 horas) e na análise histológica foi possível observar diferenças na estruturação do intestino médio das operárias de A. mellifera alimentadas com os três PFSs. Apesar das linhagens de Bt reduzirem a longevidade das operárias de A. mellifera não é possível afirmar que não sejam seletivas, pois a interferência é mínima quando comparada à interferência provocada pelos PFSs, os quais provocaram drástica redução na longevidade de A. mellifera. / Africanized bees (Apis mellifera L.) are disappearing and are believed to be due to the intense use of insecticides, pollution and climate change, however many crops depend on the pollination performed by these bees. The objective of this work was to evaluate the effect of three strains of Bacillus thuringiensis (Bt) (IPS 82, BR 81 and BR 147), entomopathogen used for the control of Acetamiprid + Alpha-cypermethrin, PFS 2: Imidacloporide + Beta-Cyflutin and PFS 3: Fenitrothione + Esfenvalerate), on the longevity of African female A. mellifera workers. Three bioassays were performed with the Bt and PFS lines: A) Spraying on A. mellifera; B) A. mellifera contact with pulverized surface and C) Candi paste incorporated with the products and histological analysis of the medium intestine of the bees. In the three bioassays the control was composed of sterilized deionized water. Each treatment consisted of 60 bees, each bee being considered a repetition. The bioassays were maintained in a BOD type chamber (34 ± 2ºC, 60 ± 5% U.R) and the workers longevity was evaluated at one; two; three; four; five; six; nine; 12; 13; 18; 21; 24; 30; 36; 42; 48; 60; 72; 96; And 120 hours. In Bt spraying on A. mellifera, it was verified that only the Bt 1 strain (IPS 82) caused a reduction in worker longevity (88.7 hours), while in the contact bioassay the Bt 3 - BR147 strain caused a reduction (101, 2 hours). All strains of Bt evaluated when incorporated into A. mellifera feed reduced workers longevity (Bt 1 - IPS 82: 64.50 hours; Bt 2 - BR 81: 64.56 hours and Bt 3 - BR 147: 60). And the histological analysis showed the disintegration of the epithelial cells of the bees mesentery, caused by the bacteria they ingested. The three synthetic phytosanitary products reduced the longevity of A. mellifera workers in the direct spray bioassay (PFS 1 and PFS 2 - 1.0 hour; PFS 3 - 2.21 hours). At the contact of A. mellifera with a PFS-sprayed surface, the bees stayed two hours in contact with the products on the petri dish, and at the end of these two hours, 100% bees that came in contact with the three synthetic phytosanitary products were dead. When the PSFs were incorporated in the diet, they reduced the longevity of the workers (PFS 1 - 22 hours, PFS 2 - 21.72 hours and PFS 3 - 43.93 hours) and in the histological analysis it was possible to observe differences in the workers from A. mellifera fed with the three PFSs. although the Bt lineages reduce the longevity of A. mellifera workers, it is not possible to assert that they are not selective, since the interference is minimal when compared to interference caused by PFSs, which caused a drastic reduction in the longevity of A. mellifera.

As abelhas africanizadas (Apis mellifera L.) estão desaparecendo e acredita-se que seja devido ao intenso uso de inseticidas, poluição e alterações climáticas, no entanto muitas culturas dependem da polinização realizada por essas abelhas. Desta forma, o objetivo deste trabalho foi avaliar o efeito de três linhagens de Bacillus thuringiensis (Bt) (IPS 82, BR 81 e BR 147), entomopatógeno utilizado para o controle de lagartas desfolhadoras, e de três produtos fitossanitários sintéticos (PFS 1: Acetamiprido + Alfa-cipermetrina; PFS 2: Imidacloporido + Beta-Ciflutina e PFS 3: Fenitrotiona + Esfenvalerato), sobre a longevidade de operárias adultas de A. mellifera africanizada. Foram realizados três bioensaios com as linhagens de Bt e dos PFSs: A) Pulverização sobre A. mellifera; B) Contato de A. mellifera com superfície pulverizada e C) Pasta cândi incorporada com os produtos e análise histológica do intestino médio das abelhas. Nos três bioensaios a testemunha foi composta por água deionizada esterilizada. Cada tratamento foi composto por 60 abelhas, sendo cada abelha considerada uma repetição. Os bioensaios foram mantidos em câmara climatizada tipo BOD (34 ± 2ºC, U.R. de 60 ± 5%) e a longevidade das operárias avaliada a uma; duas; três; quatro; cinco; seis; nove; 12; 15; 18; 21; 24; 30; 36; 42; 48; 60; 72; 96; e 120 horas.Na pulverização de Bt sobre A. mellifera verificou-se que apenas a linhagem Bt 1 (IPS 82) causou redução da longevidade de operárias (88,7 horas), enquanto no bioensaio de contato a linhagem Bt 3 - BR147 provocou redução (101,2 horas). Todas as linhagens de Bt avaliadas, quando incorporadas à alimentação de A. mellifera reduziram a longevidade das operárias (Bt 1 – IPS 82: 64,50 horas; Bt 2 – BR 81: 64,56 horas e Bt 3 – BR 147: 60,00horas) e com a análise histológica pôde-se perceber a desintegração das células epiteliais do mesêntero das abelhas, causada pelas bactérias que elas ingeriram. Os três produtos fitossanitários sintéticos reduziram a longevidade das operárias de A. mellifera, no bioensaio de pulverização direta (PFS 1 e PFS 2 – 1,0 hora; PFS 3 – 2,21 horas). No contato de A. mellifera com superfície pulverizada com PFS, as abelhas ficaram duas horas em contato com os produtos na placa de Petri, e ao final dessas duas horas, 100% abelhas que entraram em contato com os três produtos fitossanitários sintéticos estavam mortas. Quando incorporados os PSFs na alimentação, ambos reduziram a longevidade das operárias (PFS 1 – 22 horas, PFS 2 – 21,72 horas e PFS 3 – 43,93 horas) e na análise histológica foi possível observar diferenças na estruturação do intestino médio das operárias de A. mellifera alimentadas com os três PFSs. Apesar das linhagens de Bt reduzirem a longevidade das operárias de A. mellifera não é possível afirmar que não sejam seletivas, pois a interferência é mínima quando comparada à interferência provocada pelos PFSs, os quais provocaram drástica redução na longevidade de A. mellifera. / Africanized bees (Apis mellifera L.) are disappearing and are believed to be due to the intense use of insecticides, pollution and climate change, however many crops depend on the pollination performed by these bees. The objective of this work was to evaluate the effect of three strains of Bacillus thuringiensis (Bt) (IPS 82, BR 81 and BR 147), entomopathogen used for the control of Acetamiprid + Alpha-cypermethrin, PFS 2: Imidacloporide + Beta-Cyflutin and PFS 3: Fenitrothione + Esfenvalerate), on the longevity of African female A. mellifera workers. Three bioassays were performed with the Bt and PFS lines: A) Spraying on A. mellifera; B) A. mellifera contact with pulverized surface and C) Candi paste incorporated with the products and histological analysis of the medium intestine of the bees. In the three bioassays the control was composed of sterilized deionized water. Each treatment consisted of 60 bees, each bee being considered a repetition. The bioassays were maintained in a BOD type chamber (34 ± 2ºC, 60 ± 5% U.R) and the workers longevity was evaluated at one; two; three; four; five; six; nine; 12; 13; 18; 21; 24; 30; 36; 42; 48; 60; 72; 96; And 120 hours. In Bt spraying on A. mellifera, it was verified that only the Bt 1 strain (IPS 82) caused a reduction in worker longevity (88.7 hours), while in the contact bioassay the Bt 3 - BR147 strain caused a reduction (101, 2 hours). All strains of Bt evaluated when incorporated into A. mellifera feed reduced workers longevity (Bt 1 - IPS 82: 64.50 hours; Bt 2 - BR 81: 64.56 hours and Bt 3 - BR 147: 60). And the histological analysis showed the disintegration of the epithelial cells of the bees mesentery, caused by the bacteria they ingested. The three synthetic phytosanitary products reduced the longevity of A. mellifera workers in the direct spray bioassay (PFS 1 and PFS 2 - 1.0 hour; PFS 3 - 2.21 hours). At the contact of A. mellifera with a PFS-sprayed surface, the bees stayed two hours in contact with the products on the petri dish, and at the end of these two hours, 100% bees that came in contact with the three synthetic phytosanitary products were dead. When the PSFs were incorporated in the diet, they reduced the longevity of the workers (PFS 1 - 22 hours, PFS 2 - 21.72 hours and PFS 3 - 43.93 hours) and in the histological analysis it was possible to observe differences in the workers from A. mellifera fed with the three PFSs. although the Bt lineages reduce the longevity of A. mellifera workers, it is not possible to assert that they are not selective, since the interference is minimal when compared to interference caused by PFSs, which caused a drastic reduction in the longevity of A. mellifera.

Les Cévennes sont depuis des siècles connues pour être des « terres de miel ». L'apiculture y est caractérisée par lo brusc, une ruche traditionnelle construite avec un tronc de châtaignier évidé, couvert d'une lauze de schiste. Les ruchers-troncs constituent une forme d'apiculture très ancienne et rustique, qui a contribué à l'histoire de l'occupation humaine et à la dynamique des paysages en Cévennes. Les ruches étaient à l'origine exclusivement sédentaires et peuplées d'abeilles noires (Apis mellifera mellifera), présentes en Cévennes bien avant l'arrivée des humains. Aujourd'hui, l'apiculture cévenole se fait principalement en ruches à cadres. Les pratiques apicoles actuelles — achat d'abeilles, transhumance, sélection, etc.— ont conduit à la diminution des populations de sous-espèces d'abeilles locales et à leur homogénéisation génétique. Afin de mieux connaître et de préserver ce patrimoine apicole naturel et culturel exceptionnel, le Parc national des Cévennes a initié cette étude pluridisciplinaire à travers le financement d'une thèse CIFRE. Ce travail est composé de trois parties.(I) L'objet de la première partie est l'habitat, la ruche. Les documents d'archives permettent d'affirmer que les ruches-troncs sont apparues en Cévennes à la fin du Moyen Âge, puis se sont développées et maintenues jusqu'à la première moitié du XXe siècle. Le passage de la ruche-tronc à la ruche à cadres moderne s'est réalisé progressivement au cours du siècle dernier. Les témoignages des anciens Cévenols ont permis d'appréhender les pratiques, les savoirs et les savoir-faire associés à ces ruchers traditionnels. Enfin, le micro-environnement des ruchers a été caractérisé grâce à des analyses spatiales qui viennent corroborer le discours local sur l'emplacement idéal d'un rucher.(II) L'abeille noire est au cœur de la deuxième partie. De l'abeille commune à l'abeille « noire agressive », les considérations du milieu apicole sur l'abeille locale ont évolué au cours du siècle écoulé. Un état des lieux de la population d'abeilles a été réalisé en utilisant la morphométrie géométrique, puis l'ADN mitochondrial. La morphométrie permet de dire que les 2/3 de la population d'abeilles des Causses et des Cévennes sont constitués d'abeilles noires. L'étude de l'ADN mitochondrial nous alerte néanmoins sur le taux élevé d'introgression dans ces populations. Cette introgression touche autant les populations d'abeilles élevées en ruches à cadres que celles maintenues en ruches-troncs. Ces populations ne se démarquent pas génétiquement l'une de l'autre.(III) La dernière partie de ce travail concerne la conservation du patrimoine apicole par l'établissement public du Parc national des Cévennes. Elle détaille les moyens et actions passés, présents, ainsi que les difficultés et les perspectives de conservation pour la ruche-tronc et l'abeille noire en contexte d'aire protégée. Cette partie met en exergue l'impérieuse nécessité d'une concertation multi-acteurs, axée sur une intégration de plusieurs types de savoirs — local, scientifique, d'expert — qui tienne compte des changements sociaux, économiques et écologiques auxquels la région des Cévennes est soumise. / For several centuries, the Cévennes region in Southern France has been renowned as a “land of honey”. Beekeeping in Cévennes is characterized by lo brusc, a traditional hive that is made of a hollowed chestnuts log which is covered with a schistous stone slab called “lauze”. Log hive apiaries are a very old and rustic form of beekeeping, which was a major driver of human occupation history and landscape dynamics throughout the Cévennes. From their origins, log hives were home most exclusively to black bees (Apis mellifera mellifera) that were settled in the region far before the rise of humankind. Nowadays, beekeeping in Cévennes is mainly carried out in frame hives. Current beekeeping practices — purchase of bees, transhumance, queen selection… — have led to a drastic decrease in populations of local bee subspecies, and to their genetic homogenization. In order to better understand and preserve this remarkable natural and cultural beekeeping heritage, the Cévennes National Park implemented a multidisciplinary study, through the funding of a CIFRE (Industrial contract for training through research) doctoral research.1- The first part of the study is dedicated to the hive. Archive documents strongly support the assertion that the very first log hives that were established in Cévennes date back to the end of the Middle Age. Afterwards they expanded and were maintained until the middle of the 20th century. The shift from log hive to frame hive occurred progressively throughout the past century. Testimonies by old Cévennes inhabitants helped assessing local practices, knowledge and know-how related to these traditional apiaries. Furthermore, spatial analyses were undertaken to characterize the micro-environment surrounding apiaries. These analyses corroborate local discourses about where an apiary should ideally be set up.2- The black bee is the epicenter of the second part of the study. From the common bee to the “black and aggressive” bee, views by the beekeeping community concerning the local bee in Cévennes have evolved over the past century. Genetic analyses using geometrical morphometry and mitochondrial DNA were successively implemented to establish a state of the art of local bee populations. Morphometric data tell us that nearly 2/3rd of the bee populations of Causses and Cévennes are composed of black bees. However, mitochondrial DNA data alert us on the high level of introgression within these populations. Such introgression equally affects bees kept in frame hives and those kept in log hives.3- The third part of the study addresses the sensitive issue of a conservation strategy of local beekeeping patrimony that is carried out by the public development agency of the Cévennes National Park. Past conservationist resources and actions are described and so are the perspectives and obstacles to a valuable conservation strategy of black bees and log hives in a context of protected area. In conclusion, the study advocates for an indispensable multi-stakeholder conciliation and a necessary integration of several types of knowledge — local ecological knowledge, knowledge from learned experts, scientific knowledge — that takes into consideration the social, economical and ecological changes affecting the overall Cévennes region.

Einleitung: Risiken für Honigbienen stellen heutzutage schwindende natürliche Flächen und der Einsatz von Pestiziden in der Landwirtschaft dar. Flupyradifuron ist der Wirkstoff eines neuen Pflanzenschutzmittels der Bayer AG, das unter dem Namen „Sivanto“ vermarktet wird. Flupyradifuron an den nikotinischen Acetylcholin-Rezeptor im Gehirn der Honigbiene. Zielstellung: Ziel dieser Arbeit ist es, subletale Effekte von Flupyradifuron auf Verhalten und Gehirnstrukturen der Honigbiene zu untersuchen. Material und Methoden: Der Effekt einer chronischen Flupyradifuron-Applikation in unterschiedlichen Konzentrationen über zehn Tage auf die Mortalität, wurde in frisch geschlüpften Sommerbienen und langlebigen Winterbienen untersucht (N = 30 pro Behandlung, jeweils vier Replikate). Die statistische Auswertung erfolgte mit der Kaplan-Meier-Methode mit log-Rank Test. Nachdem Flupyradifuron einmalig an Honigbienen (N = 46, 47, 48 bzw. 55 pro Behandlung bei Nektar-sammelnden; N = 54, 68, 56 bzw. 62 pro Behandlung bei Pollen-sammelnden Bienen) verfüttert wurde, wurde deren Geschmackswahrnehmung getestet Im Folgenden wurden die Bienen mittels klassischer olfaktorischer Konditionierung auf einen Duft konditioniert. Das Gedächtnis wurde am nächsten Tag getestet. Zur statistischen Auswertung wurde die logistische Regression mit Post-Hoc Least Significant Difference Test angewendet. Es wurde eine Videoanalyse durchgeführt, um den Einfluss von Flupyradifuron auf die motorischen Fähigkeiten von Sommer- und Winterbienen zu untersuchen. Dazu wurde Flupyradifuron einmalig (N = 19 pro Behandlung im Sommer; N = 17 bzw. 16 pro Behandlung im Winter) oder über 24 h (N = 15 pro Behandlung im Sommer; N = 18 pro Behandlung im Winter) verabreicht. Zum Vergleich wurde das Experiment mit dem Neonikotinoid Imidacloprid bei Winterbienen wiederholt (N = 17 bzw. 16 pro Behandlung bei einmaliger Gabe; N = 16 pro Behandlung bei 24 h Gabe). Die statistische Auswertung erfolgte mittels einer nicht-parametrischen Varianzanalyse (Kruskal-Wallis H Test) und dem Pearson Chi-Quadrat Test. Um die Effekte von Flupyradifuron auf das Sammelverhalten der Bienen zu untersuchen, wurde die RFID („radio frequency identification“) Technik angewandt. Frisch geschlüpfte Arbeiterinnen (N = 100) wurden in einem separaten Käfig im Bienenstock gehalten und mit Flupyradifuron behandelt. Nach sieben Tagen wurden die Bienen in die Kolonie entlassen und ihr Flugverhalten für 40 Tage verfolgt. Das Experiment wurde zweimal durchgeführt. Die Kaplan-Meier Methode mit log-Rank Test wurde angewandt um Sammelbeginn und -ende zwischen den Behandlungen zu vergleichen. Anzahl und Dauer der Sammelflüge zwischen den Behandlungsgruppen wurden mit dem Mann-Whitney-U-Test verglichen. Um den Einfluss von Flupyradifuron auf Gehirnstrukturen zu untersuchen, wurden die Bienen über zehn Tage oral mit Flupyradifuron behandelt. Gehirne (N =10 pro Behandlung) wurden präpariert, Schnitte von 5 µm wurden hergestellt und mit Hämatoxylin/Eosin (H/E) gefärbt. Bei allen Versuchen wurde ein Signifikanzniveau von P < 0,05 festgelegt. Ergebnisse: In einer mittleren Konzentration von 1,0 µg pro Biene pro Tag war die Mortalität von Sommer- und Winterbienen in drei bzw. zwei von vier Replikaten signifikant erhöht. Eine zehnfach höhere Konzentration führte zu 100 % Mortalität, eine zehnfach niedrigere Konzentration war subletal. Flupyradifuron reduzierte die Geschmackswahrnehmung und das appetitive Lernen. Dabei hatte nur die höchste verwendete Konzentration (8,3 *10 4 mol/l) einen signifikanten Einfluss, zwei zehnfach niedrigere Konzentrationen hatten keinen Effekt. Eine einmalige Flupyradifuron-Gabe störte das normale motorische Verhalten von Bienen und führte zu motorischen Ausfallerscheinungen. Dies war bei Winterbienen stärker ausgeprägt als bei Sommerbienen und wurde durch eine hohe Dosis (8.3 *10 4 mol/l) hervorgerufen. Nach einer chronischen Gabe über 24 h waren die Veränderungen weniger stark ausgeprägt. Imidacloprid führte nicht zu motorischen Ausfallerscheinungen. Insektizid-behandelte Bienen zeigten signifikant früheres Sammelverhalten. Dies galt für beide Replikate. Im zweiten Replikat zeigten die behandelten Bienen zudem mehr Sammelflüge und diese dauerten länger. Die Analyse von Gehirnstrukturen nach der Behandlung mit Flupyradifuron mit Hilfe von Lichtmikroskopier brachte keine Veränderungen zu Tage. Schlussfolgerungen: In hohen Konzentrationen beeinflusst das neue Insektizid Flupyradifuron Kognition und Motorik der Honigbiene in ähnlicher Weise wie die teilweise verbotenen Neonikotinoide. In niedrigeren Konzentrationen sind die Effekte weniger stark ausgeprägt. Zukünftige Studien sollten mögliche synergistische Effekte von Flupyradifuron in Kombination mit Pflanzenschutzmitteln, wie Fungiziden, aber auch in Kombination mit Parasiten und anderen Krankheitserregern untersuchen. / Introduction: Changing landscapes and pesticides resulting from intensified agriculture are two of the main threats for honeybees. Flupyradifurone is the active ingredient of a new pesticide released by Bayer AG under the name of “Sivanto”. It binds to the nicotinic acetylcholine receptor (nAchR) in the honeybee brain, similar to neonicotinoids. Aim: The aim of this study is to investigate effects of flupyradifurone on honeybee behavior und brain structure. Material and methods: The effect of a chronic application of flupyradifurone in different concentrations on mortality was studied in newly emerged summer bees and long lived winter bees (N = 30 per treatment, four replicates) over a period of ten days. Survival was analyzed using Kaplan-Meier –Method with log-rank Test. After feeding foraging honeybees (N = 46, 47, 48 respectively 55 per treatment in nectar foragers; N = 54, 68, 56 respectively 62 per treatment in pollen foragers) a single dose of flupyradifurone, gustatory responsiveness was quantified. Afterwards the bees were trained to an odor using classical olfactory conditioning. Memory was tested the next day. Statistical analysis was conducted using Logistic Regression. For post-hoc multiple comparisons we used the Least Significant Difference Test. Video analysis was conducted to test the effects of flupyradifurone on honeybee motor abilities in young summer bees and long lived winter bees. Flupyradifurone was administered once (N = 19 per treatment in summer; N = 17 respectively 16 per treatment in winter) or over the period of 24 h (N = 15 per treatment in summer; N = 18 per treatment in winter). For comparisons this experiment was repeated with the neonicotinoid imidacloprid (N = 17 respectively 16 per treatment for single administration; N = 16 per treatment for 24 h application). Non-parametric analysis of variance (Kruskal-Wallis H Test) and Pearson Chi-Square Test were applied to determine the effect of the insecticides on motor behavior between the different treatment groups. To test effects of flupyradifurone on honeybee foraging, RFID (radio frequency identification) technology was applied. Newly emerged worker bees (N = 100) were taken in a separated cage on top of the bee hive and treated with flupyradifurone. After one week the bees were released into the colony and their flight behavior was tracked for 40 days. The experiment was repeated twice. Kaplan-Meier-Method with log-rank Test was applied for comparing onset and end of foraging between the two treatment groups. Trips per active day and duration per trip between the different treatment groups were compared using Mann-Whitney U-Tests. Effects of flupyradifurone on honeybee brain structure were analyzed. Bees were treated with flupyradifurone for ten days. Brains (N = 10) were dissected and 5 µm sections were produced and stained with hematoxylin and eosin (H/E). For all experiments a significance level p < 0.05 was determined. Results: The mortality experiment revealed comparable results in newly emerged summer bees and long lived winter bees. The mortality of an intermediate concentration of approximately 1.0 µg flupyradifurone per bee per day was in three respectively two out of four replicates significantly increased. A tenfold higher concentration led to 100 % mortality, whereas a tenfold lower concentration was sublethal. Flupyradifurone reduced taste and appetitive learning performance in honeybees foraging for pollen and nectar. Only the highest concentration (8.3 *10 4 mol/l) had significant effects, whereas two tenfold lower concentrations had no effects. Flupyradifurone disturbed normal motor behavior and evoked motor disabilities after a single administration. The observed effects were stronger in long lived winter bees than in young summer bees. However, only a high dose (8.3 *10 4 mol/l) had these strong effects. After a continuous administration over 24 h the observed effects were less severe. Imidacloprid did not lead to motor disabilities. Pesticide-treated bees initiated foraging significantly earlier than control bees. This was true for both replicates. In the second replicate flupyradifurone treated bees furthermore showed more and longer foraging trips. Analyzing honeybee brains using light microscopy, no altered brain structures were observed after treating honeybees with flupyradifurone. Conclusion: High concentrations of flupyradifurone influence honeybee cognition and motor abilities in a similar way as the partly banned neonicotinoids. In lower concentrations, the observed effects are less severe. Future studies should examine possible synergistic effects of flupyradifurone between in combination with other pesticides, such as fungicides, but also in combination with parasites and other stressors, such as diseases.

This thesis investigates the design of atria for daylighting in large scale buildings. Athree dimensional test building with a central atrium was constructed and various parameters of the atrium altered. The impact of these changes was studied through computer simulations of annual daylight distribution by implementing state of theart software. Daylight autonomy is simulated for an annual climate file for Stockholm, Sweden. In the thesis, notion is made of basic daylighting concepts, the importance of bringing daylight into buildings is argued, and the daylighting criteria of three environmental certification tools introduced. Furthermore, a detailed comparison is made on several well known daylight simulation tools. A newly developed, state of the art, daylight simulation tool called Honeybee, is used in the simulation process. The tool utilizes the calculation engines of wellknown daylight simulation software Radiance and Daysim, which apply backward ray-tracing to reach accurate results. Honeybee is coupled to the graphical algorithmeditor Grasshopper for Rhinoceros 3D, which allows for an efficient way of parametric modelling. The comparison of five different daylight simulation tools showed that Honeybee outweighs the capabilities of many of them by offering a wast range of simulation capabilities and also giving the user exceptional control of result data within multiple zones of the test building. The results of the daylight study have been compiled into a document which purpose is to serve as early stage design guidelines of atria for architects. Many factors have been shown through simulation to have a dramatic impact on daylighton an annual basis, and several suggestions have been made on how to maximize the quantity of daylight within buildings containing atria.

The EU building sector is a main contributor to greenhouse gas emissions, which need to be cut as part of the global response to anthropogenic climate change. This cut can be realised through improvements in building energy performance, such as optimisation of facade design. The early stage of the architectural process has been identified as the ideal time to implement such sustainable design choices. There is need for simple guidelines and tools to provide quantitative data to support these architectural decisions. BIM and parametric design can provide this, by facilitating model-based analysis and simulation, as part of an unbroken flow of information through the design process. This study uses Dynamo (the visual programming add-in for Autodesk Revit) together with the Honeybee and Ladybug environmental plugins, to conduct daylight, solar heat gains and thermal losses analyses and simulations. The aim was to identify limitations and opportunities in using Dynamo-Revit, to establish an optimal range for glazing-to-wall ratio (GWR) and to provide some simple room-sizing guidelines for architects in the early stage of the design process. The Dynamo-Revit workflow was found to be effective for specific projects, but difficult to perfect for multiple different projects. An optimal range for GWR was found as 30-40% for east, south and west-facing rooms and around 50% for rooms facing only north. Results were tabulated, linking room orientation and depth with estimated daylight access, solar heat gains and thermal losses. The results were reasonable, but could be improved by the use of more sophisticated analysis and simulation techniques, which should be facilitated by forthcoming development of Honeybee and Ladybug in Dynamo.

L’abeille mellifère (Apis mellifera) est une espèce clé dans son aire d’origine tant pour les services écosystémiques rendus que pour les productions apicoles. Toutefois, ses populations sont soumises à différents facteurs de stress biotiques responsables de mortalités importantes. Dans les îles du Sud- Ouest de l’Océan Indien (SOOI) la sous-espèce d’abeille endémique est A. m. unicolor. Toutefois, aucune étude n’avait été réalisée sur sa pathosphère et ce n’est que depuis 2010 où l’ectoparasite Varroa destructor a envahi certaines de ces îles, menaçant cette sous-espèce, que de premières études ont été conduites. Dans ce travail de thèse, nous avons réalisé un état des lieux général de la santé des cheptels d’abeilles grâce à des enquêtes épidémiologiques descriptives dans la majorité des îles : La Réunion, Maurice, Rodrigues, Madagascar et les Seychelles. Nous avons pu montrer des faciès épidémiologiques assez similaires entre les îles, marqués par une dominance de Nosema ceranae particulièrement dans les petites îles (80-100%) et la présence de 3 virus : BQCV (4-89%), CBPV (2-51% excepté à Rodrigues) et DWV (4-40% excepté à La Réunion). D’autres agents pathogènes n’ont été retrouvés que sur certains territoires comme Aethina tumida, Braula pretoriensis, Acarapis sp. ou Melissococcus plutonius. L’analyse de la diversité génétique réalisée sur les 3 virus a montré une proximité des souches virales au sein du SOOI. Les enquêtes réalisées dans un contexte sans varroa ont montré une bonne santé des colonies avec une absence de signes cliniques. Les mortalités constatées n’ont concernées que les territoires envahis par varroa : Madagascar, Maurice, La Réunion. Varroa semble donc être le principal responsable des mortalités aiguës de colonies dans la zone bien avant les autres agents pathogènes ou les causes environnementales. Son impact sur les populations d’abeilles et in fine sur les écosystèmes indigènes sera à évaluer dans les années futures. / The honeybee (Apis mellifera) is a key species in its native range for both ecosystem services offerred and for bee products. However, its populations are subject to various biotic stressors responsible for significant mortalities. In the South-West Indian Ocean region the endemic bee subspecies is A. m.unicolor. However, no studies had been carried out on its pathosphere and it is only since 2010 where the ectoparasite Varroa destructor invaded some of these islands, threatening this subspecies, that first studies were conducted. In this thesis work, we carried out a general inventory of the health of honeybee herds through descriptive epidemiological surveys in the majority of islands: Réunion, Mauritius, Rodrigues, Madagascar and Seychelles. We were able to show quite similar epidemiological facies between islands, characterized by a dominance of Nosema ceranae especially in small islands (80-100%) and the presence of 3 viruses: BQCV (4-89%), CBPV (2- 51% except in Rodrigues) and DWV (4-40% except in Reunion). Other pathogens have only been found in certain territories such as Aethinatumida, Braula pretoriensis, Acarapis sp. or Melissococcus plutonius. The analysis of the genetic diversity carried out on the 3 viruses showed a proximity of viral strains within the SOOI. Surveys conducted in a context without varroa showed good colony health with no clinical signs. The observed mortalities concerned only the territories invaded by varroa: Madagascar, Mauritius, Reunion. Varroa therefore appears to be the main cause of acute colony mortality in the area long before other pathogens or environmental causes. Its impact on bee populations and ultimately on native ecosystems will be evaluated in future years.