Resumo: Recentemente as abelhas têm sido devidamente valorizadas como importantes polinizadoras de flores silvestres e cultivadas. A densidade populacional de muitos polinizadores tem diminuído devido, principalmente, à intensificação agrícola e ao uso de pesticidas, prejudicando os serviços de polinização. A metodologia clássica para estimar a toxicidade dos produtos químicos para insetos é determinar a dose letal média (DL50) ou a concentração letal média (CL50), podendo então estabelecer doses que sejam mais seguras aos organismos não-alvo ou benéficos. Além dos efeitos de toxicidade aguda, levando a morte das abelhas, doses subletais dos inseticidas podem provocar alterações comportamentais e fisiológicas nos indivíduos, que ao longo do tempo acarretarão em sérios prejuízos na manutenção da colônia. Um dos inseticidas amplamente utilizado é o fipronil, este atua ligando-se aos receptores do ácido gama-aminobutírico (GABA), interrompendo os canais de cloro, resultando na perda de sinalização inibitória neural. Na literatura pode-se encontrar diversos trabalhos que utilizam como modelo principal a abelha Apis mellifera, porém, é importante ressaltar a diversidade existente entre as abelhas nativas no Brasil, os meliponíneos, e sua participação na conservação da biodiversidade, assim como na polinização de áreas de cultivo, o que torna extremamente importante estudos com essa abelha. Com a finalidade de entender como o fipronil interfere morfo e fisiologicamente em abelhas sem ferrão, a região de interesse deste estudo foram os corpos pedunculados, já que estes são centros cerebrais complexos e tidos como local de convergência multisensorial. Para auxiliar no mapeamento metabólico, utilizou-se como marcador a enzima citocromo oxidase e a enzima caspase-3, técnicas utilizadas na observação de atividade neural... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: A few decades the bees are considered an important indicator of high environmental sensitivity, and appreciated as important pollinators of wildflowers and cultivated. The population density of many pollinators have declined to harmful levels to pollination services manly due to agricultural intensification and the use of pesticides. The classic methodology of estimating the effects of chemicals for insects is to determine the median lethal dose (LD50) or median lethal concentration (LC50) that can then establish doses that do not harm non-target organisms or beneficial. Besides the effects of acute toxicity, leading to death bees, sublethal doses of insecticides can cause physiological and behavior changes of individuals over time, resulting in serious harm to maintain the colony. One of the widely used insecticides is fipronil, its acts by binding to gamma-aminobutyric acid (GABA) disrupting chloride channels, resulting in loss of inhibitory neural signaling. In the literature one can find several works using as main bee model Apis mellifera, however, it is important to highlight the diversity of Brazilian native bees, the stingless bees, and their participation in biodiversity conversation, as well as in the pollination of cultivated land. In order to understand how fipronil affect morpho and physiologically the stingless bee S. postica, the region of interest in this study were the mushroom bodies, since these are complex brain centers and used as a place of multisensory convergence. This work established the contact LD50 and Ingestion LC50 to the fipronil insecticide for foragers workers stingless bee Scaptotrigona postica in 0.54ng/bee and 0.24ng/μL of the food after 24 hours, respectively, confirming the high toxicity of this phenylpyrazole, in the groups submitted to contact contamination, were identify morphological... (Complete abstract click electronic access below) / Orientador: Osmar Malaspina / Coorientador: Roberta Cornélio Ferreira Nocelli / Banca: Elaine Cristina Mathias da Silva Zacarin / Banca: Thaisa Cristina Roat / Mestre

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Over 80% of wild angiosperms are reliant upon animal pollination for fruit and seed set and bees and other insects provide a vital pollination service to around a third of the crops we produce. Habitat loss, climate change and disease spread all threaten pollinator populations, with local declines and range contractions in honeybees and bumblebees leading to concerns that crop production may suffer as a result of pollinator shortages. Whilst agriculture and wildlife are often presented as being at odds with one another, the relationship between farmers growing pollination dependent crops, and the bees and insects that service them could be mutualistic. Flowering crops planted by farmers can provide an important source of forage to wild bees, whilst in return wild bees can contribute to ensuring farmers achieve adequate yields of marketable crops. The potential of this mutualistic relationship can be maximised by farmers by adopting management practices that reduce harm to, and enhance the wellbeing of, the wild bees around their farm. A group of common pesticides (neonicotinoids) used by farmers have recently been linked to pollinator ill health. Sub-lethal effects resulting from exposure to the neonicotinoid imidacloprid have been reported in honeybees and bumblebees, with bumblebee reproductive success found to diminish as a result of exposure to field realistic doses of this agrochemical. Here, the mechanism behind the reduced queen production in bumblebee colonies is suggested, with bees exposed to imidacloprid showing reduced efficiency in foraging for pollen. Farmers dependent upon pollinating insects for crop production can opt to avoid the use of pesticides known to harm these insects, however future studies are needed to identify safer alternatives that can be use in their place. Farmers can choose to increase the number of bees at their farms by utilising domesticated honeybees and purchasing commercially reared bumblebees. The use of these pollinators can ensure a minimum number of bees in the vicinity of a crop, and facilitate the production of crops at times when wild bee numbers are low. Concerns have been raised, however, regarding the use of commercially reared bees, mostly in regard to pathogen and parasite transmission, but also in respect to the possibility of outcompeting native species. Here the frequency and severity of attacks on commercial Bombus terrestris colonies, by the wax moth, an understudied bumblebee pest, are examined. Wax moths were found to infest almost half of the bumblebee nests deployed at fruit farms, with around a third of infestations resulting in nest destruction. Farmers investing in commercial bees will want to reduce the impact of harmful pests that may result in a reduced pollination service being delivered. Wax moth infestation rates at the study farms using commercial bees were high and the potential of a ‘spill- back’ effect on wild bees was examined. No evidence was found to suggest that nests in close proximity to these farms were any more or less likely to suffer from an attack than nests situated further away. Nest size was found to be the most significant predictor of an infestation, with larger nests more prone to wax moth attacks. Whilst farmers can utilise domesticated and commercially reared bees, relying on one source of pollination is inherently risky, and the most robust service will likely be provided by a range of pollinators. As well as reducing the use of chemicals known to harm beneficial insects, farmers can improve the habitat around their farms to help encourage and sustain wild pollinator populations. Sowing flower strips has been found to increase the abundance and diversity of pollinating insects, however, studies linking the use of these strips to crop production are lacking. Here we demonstrate for the first time that sowing small flower strips, adjacent to strawberry crops serviced by both wild and managed bees, can increase the overall number of pollinators foraging on the crop. This thesis contributes to our understanding of the implications of farm management decisions on pollinator health. It provides experimentally based evidence to guide farmers in making informed decisions regarding the future of crop pollination services and highlights the need for an integrated approach to managing pollination services for sustainability.

As operárias de Apis mellifera realizam diferentes tarefas de acordo com a sua idade (polietismo etário), sendo que operárias jovens cuidam da cria e quando mais velhas forrageiam. Entretanto, tal transição não segue uma cronologia fixa, mas se ajusta às necessidades da colônia, especialmente as condições nutricionais, tanto da colônia quanto dos indivíduos. Nesse sentido, abordamos a questão de como o metabolismo oxidativo no corpo gorduroso das operárias, i.e., o centro do metabolismo intermediário destes insetos, está relacionado com alterações da função das operárias na colônia e como o mesmo responde a um estresse nutritivo imposto individualmente em duas fases, na fase larval e na fase adulta. Como métodos para avaliar a atividade mitocondrial e o estado redox das células do corpo gorduroso utilizamos um sistema de respirometria de alta resolução para medir o consumo de O2, e realizamos ensaios bioquímicos para quantificar a geração de espécies reativas de oxigênio (EROs), os níveis de óxido nítrico, o número de unidades mitocondriais, e eventuais danos oxidativos resultantes. Ademais, por meio de PCR quantitativa medimos a expressão relativa de genes ligados à biogênese mitocondrial, ao sistema antioxidante e à via de sinalização por hipóxia. Na comparação destes parâmetros entre operárias nutridoras e forrageiras avaliamos separadamente os três principais compartimentos do corpo (cabeça, tórax e abdômen) uma vez que esses estão diferencialmente relacionados a tais funções. Para os tecidos da cabeça observamos que as amostras de nutridoras apresentaram maior consumo de O2, associado a uma maior produção de EROs e expressão elevada dos genes da via de hipóxia, enquanto forrageiras apresentaram maiores quantidades de transcritos dos genes do sistema antioxidantes e menores níveis de danos oxidativos. Para os tecidos do tórax observamos nas forrageiras uma maior capacidade de produção de ATP, acompanhada de uma elevada expressão de genes codificadores de enzimas do sistema antioxidante e menores níveis de danos oxidativos. Com relação aos tecidos abdominais, as abelhas nutridoras apresentaram maior atividade mitocondrial, enquanto as amostras de forrageiras apresentaram maior número de unidades mitocondriais, elevados níveis EROs e também uma expressão elevada dos genes do sistema antioxidante e da via de hipóxia. Assim, os dados revelaram uma clara mudança na atividade mitocondrial e nos padrões redox teciduais associados a esta transição de polietismo etário. Também notamos diferenças nestes parâmetros para cada compartimento do corpo analisado, refletindo uma demanda energética diferencial em cada tecido. A segunda parte do projeto teve como objetivo avaliar os mesmos parâmetros redox em uma situação de restrição alimentar em três situações distintas. O primeiro grupo experimental foi constituído de larvas que sofreram restrição calórica por umperíodo de 10 horas. O segundo grupo foi composto por abelhas recém emergidas que sofreram ou não tal restrição calórica no período larval. O terceiro grupo consistiu de abelhas que sofreram ou não restrição calórica no período larval, e após emergirem foram mantidas em caixas até atingirem a idade de 7 dias, quando foram submetidos novamente a uma restrição calórica. Para o primeiro grupo, as larvas, verificamos que após o período de restrição calórica os genes relacionados com a biogênese mitocondrial e o sistema antioxidante apresentaram níveis de expressão menores comparadas com larvas controle. Observamos também que larvas que sofreram restrição calórica apresentaram menor consumo de O2 e menores níveis de EROs. Para o segundo grupo, as abelhas recém emergidas, não observamos nenhuma diferença na expressão de genes mitocondriais e do sistema antioxidante entre as que sofreram ou não restrição calórica na fase larval, demostrando uma recuperação nestes parâmetros após o período de metamorfose. No terceiro grupo, as abelhas de 7 dias foram submetidas a uma nova restrição calórica para observar se indivíduos que passaram por a situação de estresse no período larval apresentariam uma resposta mais imediata a um estresse nutricional na fase adulta. Os resultados do terceiro grupo revelaram que abelhas que haviam sido expostas a uma situação prévia de restrição calórica, apresentaram uma redução dos transcritos de genes ligados a biogênese mitocondrial e do sistema antioxidante quando comparados ao grupo que sofreu restrição calórica apenas na fase adulta. Em conjunto nossos dados sugerem que a restrição calórica é capaz de diminuir o metabolismo oxidativo e que existe uma resposta aparentemente adaptativa em indivíduos adultos que passaram por essa situação previamente durante o desenvolvimento larval. Assim, esses resultados podem servir para direcionar estudos futuros sobre a relação entre restrição calórica, metabolismo oxidativo e longevidade nesses insetos sociais. / Apis mellifera workers perform different tasks according to their age (age polyethism), where young workers care for the brood and older ones become foragers. However, this a transition does not follow a fixed chronology, but is fitted to the needs of the colony, especially the nutritional conditions of both, the colony and the individual bee. In this sense, we address the question of how oxidative metabolism in the fat body of the workers, e.g., the center of the intermediary metabolism of these insects, is related to changes in the function of the workers in the colony, and how these parameters responds to a nutritional stress imposed individually in two stages, in the larval and adult phase. For evaluating mitochondrial activity and the redox status of fat body cells we used a high resolution respirometry system to measure O2 consumption, and we performed biochemical assays to measure the generation of reactive oxygen species (ROS), levels of nitric oxide, the number of mitochondrial units, and eventual oxidative damages. Furthermore, using quantitative PCR, we measured the relative expression of genes linked to mitochondrial biogenesis, to the antioxidant system, and to the hypoxia signaling pathway. In the comparison of these parameters between nurse and forager workers, we evaluated separately the three main body compartments (head, thorax and abdomen), since these are differentially related to these functions. For the head tissues, we observed that nurses showed higher O2 consumption, associated with higher ROS production and elevated expression of hypoxia pathway genes, while foragers presented higher amounts of transcripts of the antioxidant system genes and lower levels of oxidative damages. For the thorax tissues we found in the foragers a greater capacity of ATP production, accompanied by a higher expression of genes encoding enzymes of the antioxidant system and lower levels of oxidative damage. With respect to the abdominal tissues, nurse bees presented higher mitochondrial activity, while foragers had a higher number of mitochondrial units, elevated ROS levels, and also a higher expression of genes related to the antioxidant system and hypoxia pathway. Thus, these data revealed a clear change in mitochondrial activity and redox tissue patterns associated with this transition of age polyethism. We also noticed differences in these parameters for each of the analyzed body compartment, reflecting a differential energy demand in each tissue. The second part of the project aimed to evaluate the same redox parameters in a food restriction condition in three distinct situations. The first experimental group consisted of larvae that experienced caloric restriction for a 10 hour period. The second group consisted of newly emerged bees that had or not experienced such a caloric restriction in the larval period. The third group consisted of bees that had experienced or not caloric restriction during the larval period, and after emergence were kept in boxes until reaching 7 days old, when they were resubmitted to another caloricrestriction. For the first group, the larvae, we found that after the period of caloric restriction, genes related to mitochondrial biogenesis and the antioxidant system presented lower levels of expression compared to control larvae. We also observed that larvae that had experienced caloric restriction presented lower O2 consumption and lower ROS levels. For the second group, newly emerged bees, we did not observe any differences in the expression of mitochondrial genes and the antioxidant system among those individuals that had experienced or not caloric restriction in the larval phase, indicating a recovery in these parameters after the period of metamorphosis. In the third group, the 7-day-old bees had experienced a new caloric restriction to see whether individuals that had been in a stressful situation during larval period would respond more readily to nutritional stress in adulthood. The results for this group showed that bees that had been exposed to a previous caloric restriction presented a reduction in the transcript levels of genes related to mitochondrial biogenesis and the antioxidant system when compared to the group that experienced caloric restriction only in adulthood. Taken together, our data suggest that caloric restriction is able to diminish the oxidative metabolism, and that there is a seemingly adaptive response in adult individuals that had previously experienced this situation during larval development. Thus, these results can serve to direct future studies on the relationship between caloric restriction, oxidative metabolism and longevity in these social insects.

Estudos de ecologia de comunidade e conservação são importantes para entender quais áreas com diferentes fitofisionomias podem manter maior número de espécies coexistindo. Também é possível compreender o nicho trófico das abelhas, sendo este relacionado com a diversidade de recursos alimentares disponíveis e a sobreposição entre espécies de uma guilda. Uma ferramenta que atualmente vem sendo utilizada em estudos ecológicos de interações inseto-planta são as redes mutualísticas e suas métricas. Com essas ferramentas é possível inferir ou até prever as consequências da extinção de espécies e as perturbações ambientais, além de variações nos padrões para a comunidade como um todo, e a ecologia das abelhas e plantas. A partir disso o objetivo deste trabalho foi caracterizar e analisar a estrutura da comunidade de abelhas e vespas e o nicho trófico da comunidade de abelhas que nidificam em cavidades preexistentes em áreas de Cerrado e Mata semidecídua. A área de estudo foi a Estação Ecológica Jataí que apresenta diferentes fitofisionomias. Os dados foram obtidos com a utilização do método de ninhos-armadilha instalados em seis pontos, três no cerrado e três na mata semidecídua, em agosto de 2015. Foi utilizado o método de análise polínica e redes mutualísticas para o estudo do nicho trófico das abelhas. Foram amostradas 11 espécies de abelhas e oito de vespas, e essas comunidades apresentaram espécies com ampla distribuição geográfica o que confere a condição de generalistas por explorarem uma alta diversidade de ambientes. Elas tiveram mais nidificações no período chuvoso, refletindo na maior sobreposição temporal das abelhas nesse período. Entretanto na estação seca, a sobreposição temporal das vespas foi maior. Os dados obtidos de riqueza e abundância das abelhas amostradas na mata foram diferentes do cerrado, o que indica que, realmente, as espécies podem ter suas populações mudando significativamente de um ambiente para o outro como consequência de mudanças nos vários fatores abióticos e bióticos. A dieta das abelhas amostradas se mostrou bastante variada, um grande número de tipos polínicos de 14 famílias de plantas. A família mais importante para Anthidiini e Euglossini foi Fabaceae. Como já conhecida, a relação entre Centris e plantas que disponibilizam óleo, Malpighiaceae foi a família mais utilizada por C. analis. Já para as espécies de Tetrapediini, a família Euphorbiaceae foi mais frequente. As espécies se mostraram, de maneira geral, com comportamento generalista uma vez que suas amplitudes de nichos foram altas. Por outro lado, em alguns momentos, como foi possível perceber na sobreposição de nicho, essas fêmeas adotaram comportamentos mais especialistas. As Tetrapedia tiveram as maiores sobreposições de nicho entre elas mesmas, mas também utilizaram as espécies de plantas mais abundantes. Ludwigia foi o tipo polínico mais importante na dieta das abelhas para o cerrado enquanto que Gliricidia, Dalechampia, Serjania e Desmodium foram centrais na dieta da mata. As redes para ambas fitofisionomias e anos de coletas foram aninhadas e não-modulares, como esperado para redes mutualísticas. Além disso, provavelmente pelo bom estado de preservação, as redes foram robustas, indicando que a comunidade não está frágil à perda de espécies. A comunidade apresentou um comportamento mais especialista no cerrado e não alterou muito em relação aos dois anos. A especialização das populações foi maior, de maneira geral, para a área de cerrado, com exceção de Epanthidium erythrocephalum. A grande diferença nos parâmetros como composição, abundância e métricas de rede em comparação com outros estudos mostra a importância de se conhecer a dinâmica de cada área para pensar de maneira mais eficiente em estratégias específicas de recuperação e preservação de áreas naturais / Studies on community ecology and conservation are important to understand which areas with different phytophysiognomies can maintain a great number of species coexisting. It also understand the trophic niche of bees and this is associated with the diversity of food resources available and with the overlap between species of a guild. A tool that currently is being used in ecological insect-plant studies, mutualistic networks and their metrics. With these tools, it is possible to infer or even predict the consequences of the species extinction, the environmental disruption of the community as a whole, variations in community patterns and bee-plant ecology. Therefore, the aim of this study was to characterize and analyze the community structure of bees and wasps and the trophic niche of bees that nest in preexisting cavities in Brazilian savannah and semideciduous mesophytic forest. The study area was the Estação Ecológica de Jataí that presents different phytophysiognomies. The data were obtained from the method of trap-nests installed in six areas, three in the brazilian savannah and three in the semideciduous mesophytic forest, in August 2015. The methodology of pollen analysis and mutualistic networks was used to study the trophic niche of bees. Eleven species of bees and eight wasps species were sampled and this community showed species with high geographic distribution that confers the status of generalists by exploring a great variety of environments. They nested more in the rainy season reflecting in the larger temporal overlap of the bees in that period. Besides that, the wasps were higher time ovelap in the dry season. The richness and abundance of bees in the forest was different from Brazilian savannah, what can indicate that the species can have their populations changing significantly from one environment to another because of changes in the various abiotic and biotic factors. The diet of bees showed a variety of pollen types of 14 plants families. The most important family to Anthidiini and Euglossini was Fabaceae. As well documented, the relationship between Centris and plants that provide oil, Malpighiaceae was the most used family for C. analis. For the Tetrapediini species, the family Euphorbiaceae was the most frequent. All species showed a generalist behavior once their breath niche was high. On the other hand, in a few moments, as it was possible to observe in the niche overlap, the species adopted a more specialist behavior. Tetrapedia had the highest overlap niche among them, but they used the most abundant plants in the area. Ludwigia was the most important pollen type in the bees` diet in the Brazilian savannah, while Gliricidia, Dalechampia, Serjania e Desmodium were the most central in the diet of bees in the forest. The networks for both phytophysiognomies and year of sampling, was nested and not modular, as expected in mutualistic networks. Besides, probably, because of the good state of conservation of the study area, the networks were robust, indicating that the community is not fragile to the loss of species. The community showed a more specialist behavior in the Brazilian savannah and did not change much over the two years. The specialization of populations was greater, in the most cases, in the Brazilian savannah, except Epanthidium erythrocephalum. The important difference among the parameters such as composition, abundance and network metrics compared to other studies shows the importance of knowing the dynamics of each area to think more efficiently about specific strategies for recovery and preservation of natural areas

Dans un monde dynamique la survie des animaux dépend de leur capacité à intégrer des signaux environnementaux afin d'adapter leur comportement à la survenue de conséquences positives (nourriture) ou négatives (dangers) c'est-à-dire de leurs capacités d’apprentissages associatifs appétitif et aversif. Pendant ce travail de thèse, nous nous sommes intéressés aux bases comportementales, moléculaires et génétiques de l'apprentissage aversif et aux relations existant entre apprentissages aversif et appétitif au sein d'un groupe social. L'abeille est un insecte eusocial qui constitue un modèle de choix pour cette étude grâce à l'existence des protocoles de conditionnement appétitif de la réponse d'extension du proboscis (REP) et de conditionnement aversif de la réponse d'extension du dard (RED). Jusqu'à présent, le renforcement utilisé dans le conditionnement aversif de la RED était un choc électrique. Ce stimulus traversant la majeure partie du corps de l'abeille, il est peu aisé d'étudier les structures responsables de sa détection. Dans un premier chapitre, nous avons donc testé l’effet d’une forte température (65°C) sur la RED. Nous montrons qu’une stimulation thermique au niveau des pièces buccales, des pattes ou des antennes induit une RED. De plus, les abeilles parviennent à associer une odeur à la présentation concomitante d'une forte température. Dans un deuxième chapitre, nous avons cartographié la sensibilité thermique du corps des abeilles en mesurant la RED. Ce travail a montré que la stimulation de presque toutes les parties du corps induit une RED. De plus, ces stimulations peuvent jouer le rôle de renforcement aversif lors d’un conditionnement olfactif de la RED. Nous nous sommes ensuite intéressés aux récepteurs périphériques potentiellement impliqués dans la détection des fortes températures, et en particulier à HsTRPA (Hymenoptera specific Transient Receptor Potential A). Nous montrons que l’injection d'inhibiteurs exogènes de HsTRPA réduit les RED à la température. Ces résultats suggèrent l’implication possible d’HsTRPA dans la détection de la température chez l’abeille. Dans un troisième chapitre, nous nous sommes intéressés aux relations existant entre les capacités d’apprentissages aversif et appétitif des abeilles. En nous appuyant sur le protocole aversif thermique, combiné au protocole de conditionnement de la REP existant, nous avons étudié la distribution des capacités hédoniques appétitive et aversive au sein d'une ruche. La reine étant fécondée par 15-20 mâles, la ruche est segmentée génétiquement en autant de lignées paternelles différentes. Nos données montrent que la sensibilité des individus aux renforcements aversif (chaleur) et appétitif (sucre) détermine leurs performances d'apprentissage au sein de chaque modalité hédonique. Nous montrons de plus l’existence d’un trade-off, sous déterminisme génotypique, entre les capacités cognitives appétitive et aversive au sein de la colonie. Le quatrième chapitre a étudié la plasticité comportementale induite par les deux types de conditionnement. La REP et la RED étant des réponses de type "tout ou rien", nous nous sommes demandé si les mouvements antennaires des abeilles pouvaient procurer une mesure fine et intégrer des apprentissages appétitif et aversif. Nous avons développé un système de capture vidéo enregistrant les mouvements antennaires à haute vitesse. Nous montrons que les abeilles modifient leur réponse antennaire à une odeur après un apprentissage appétitif mais pas après un apprentissage aversif. Durant ce travail de thèse, nous avons ainsi développé deux nouveaux protocoles comportementaux en contention, et avons procuré de nouvelles données sur l’apprentissage aversif chez l’abeille. Nous avons observé un trade-off au sein de la ruche entre les capacités hédoniques appétitive et aversive, sous déterminisme génétique. De telles spécialisations cognitives pourraient jouer un rôle prépondérant dans l'évolution des groupes sociaux. / In a dynamic world, animals constantly face environmental changes that may affect them (Alock, 1997). Their survival depends on their ability to integrate this information in order to adapt their behaviors to subsequent positive or negative outcomes, in other words to their appetitive and aversive learning abilities. This thesis investigates the behavioral, molecular and genetic basis of aversive learning and its putative relationship with appetitive learning in a social group. The honeybee is a social insect which constitutes a profitable model for this study thanks to appetitive conditioning of the proboscis extension response (PER) and aversive conditioning of the sting extension response (SER) protocols that have been developed. To date, aversive conditioning of the SER has involved the use of electric shocks as a negative reinforcement. However, this stimulus is ecologically irrelevant for honeybees and electric currents pass through almost every part of the bee’s body making it difficult to study the dedicated structures and sensory pathways responsible for its detection. In the first chapter, we assessed the impact of high temperatures on SER, a stimulus that is also highly aversive. We showed that a SER may be triggered by heat stimulation of mouthparts, forelegs and antennae. In addition, we demonstrated that honeybees are able to associate an odor with high temperatures, resulting in the odor alone triggering the SER after conditioning. In the second chapter, we mapped the heat sensitivity of the bee’s body recording SER subsequent to heat stimulation. This work revealed that heat stimulation of almost every body structures (beside the wings and the tip of the abdomen) induces a SER. Furthermore, these stimulations may act as aversive reinforcement during an olfactory conditioning of the SER. We then focused on putative peripheral receptors involved in high temperature detection, specifically focusing on HsTRPA (Hymenoptra specific Transient Receptor Potential). Using a neuropharmalogical approach, we showed that HsTRPA exogenous inhibitor injections decrease SER triggered by high temperatures yet have no impact on sucrose PER. These results suggest a putative involvement of HsTRPA in the detection of high temperatures in honeybees. In the third chapter, we investigated the relationship between appetitive and aversive learning. By combining thermal aversive conditioning with the PER conditioning protocol, we studied the distribution of hedonic learning abilities in the hive. Since the queen mates with 15-20 males, the hive is, thus, genetically segmented in as many different patrilines. Our data shows that individual sensitivity to aversive (heat) and appetitive reinforcement varies among workers and determines their learning success in each hedonic modality. In addition, we observed that the better an individual (and therefore a patriline) is at appetitive learning, the less successful it will be in aversive learning, and vice versa. The forth chapter focused on the behavioral plasticity induced by the two types of conditioning. PER and SER are binary responses in an “all or nothing” fashion and subtle behavioral variations are difficult to observe. We therefore investigated if honeybee antennal movements may yield a finer measurement to integrate appetitive and aversive learning. We developed a motion capture system that records antennal movements at high speed. We showed that honeybees modify their antennal response to an odor previously associated with sucrose reward however thermal aversive conditioning did not induce any antennal movement modifications. During this PhD work, we developed two new behavioral assays in harnessed individuals, furthering our understanding on aversive learning in the honeybee. Within the hive, we observed a trade-off between appetitive and aversive hedonic learning, which was genetically influenced. Such cognitive specialization could play a key role in social evolution.

Cette recherche étudie la mise en exposition de la thématique du déclin des abeilles sauvages au sein du volet médiation du projet de recherche UrbanBees. Ce projet étudie l'hypothèse de l'espace urbain comme zone refuge pour les abeilles sauvages. Il a la particularité d'être adossé à une vaste opération de médiation qui associe des acteurs d'horizons variés (chercheurs, militants associatifs, médiateurs culturels) positionnés différemment sur la thématique concernée. Le déclin des abeilles sauvages constitue à l'heure actuelle ce qu'il est convenu d'appeler une question scientifique socialement vive (QSSV). Elle repose sur quatre dimensions étroitement liées : l'existence même du déclin, ses conséquences et causes potentielles, ainsi que les solutions pour y remédier. Notre étude vise à caractériser la nature de la prise en charge de cette QSSV dans l'exposition et les formes de médiation auxquelles ont recours les acteurs engagés dans sa conception. Les résultats obtenus montrent que la vivacité de la QSSV n'apparait que très ponctuellement et d'une façon relativement consensuelle, orientée par les finalités du projet. En revanche, l'étude de la conception de cette exposition révèle des oppositions marquées, en particulier entre les deux acteurs scientifiques. Les différends sont cependant résolus par les médiateurs qui recentrent le propos sur les points d'accord (les solutions portées par le projet) et le concept fédérateur de Nature en ville. Cela nous conduit à définir les spécificités du média exposition dans le traitement des QSSV par rapport à des dispositifs de médiation visant la participation des publics et à interroger les modes de production qui leurs sont associés / This research studies an exhibition and its conception about the decline of wild bees in the mediation component of the research project UrbanBees. This project explores the hypothesis of urban space as a refuge area for wild bees. It has the particularity of being support on a large operation of mediation that combines actors from diverse backgrounds (researchers, activists, cultural mediators) positioned differently on the subject concerned. The decline of wild bees is currently called a socioscientific controversy (SSC). It is based on 4 interrelated dimensions : the actual existence of decline, its potential causes and consequences, and solutions to overcome the decline. Our study aims to characterize how the SSC is supported in the exhibition and what forms of mediation are used by actors during its conception. The results show that the uncertain character of SSC appears only very occasionally and relatively consensual manner, guided by the goals of the project. Contrariwise, the study of its conception reveals disagreements, especially between two scientists. However, they are resolved by mediators : they refocus on the points of agreement (solutions brought by the project) and the unifying concepts of Nature in the City. This leads us to define the specificities of media exhibition in the treatment of SSC compared to communication actions aimed at the participation of public and examine in parallel conception processes

Objetivos Avaliar a composição e qualidade de méis de abelha Jandaira através de análises físico-químicas; avaliar se os parâmetros apresentados se encaixam nas legislações nacional e internacional para méis de Apis mellifera; analisar os efeitos de estocagens de méis de Jandaira em três diferentes temperaturas (ambiente, geladeira e freezer) e os efeitos de um ano de armazenamento de méis de Jandaira e Apis mellifera de mesma região botânica e a comparação da alteração de seus parâmetros de qualidade. Metodologia Revisão de literatura relativa aos padrões de identidade e qualidade dos méis de Apis mellifera e de abelhas sem ferrão com ênfase na Melipona subnitida. Realização das análises preconizadas pela legislação brasileira para méis de Apis mellifera nas amostras coletadas de Jandaira e Apis, além das análises qualitativas (Fiehe, Lund e Lugol), condutividade elétrica e análise dos açúcares glicose, frutose e sacarose por CLAE. Resultados As amostras de mel de Apis apresentaram-se monoflorais (pólen dominante de Althernanthera sp.) e com parâmetros dentro do preconizado pela legislação vigente, enquanto os de méis de Jandaira apresentaram-se heteroflorais (pólen de Mimosa verrucosa, Mimosa caeselpiniaefolia e Piptadenia moniliforme) a umidade (24,80%) e atividade diastásica (ausente) fora do estabelecido pela legislação vigente para os méis de Apis mellifera. A condutividade elétrica de Apis (284,00 µS.cm-1) foi superior a obtida no mel de Jandaira (102,77µS.cm-1) e a cor do mel de Apis apresentou-se mais escura (26,67 mmPfund), comparada ao mel de Jandaira (7,00 ± 0,00 mmPfund). Determinou-se maior concentração de glicose, frutose e sacarose nos méis de Apis (23,50%, 38,78% e 5,72% respectivamente) do que nos de Jandaira (21,76%, 29,21% e 4,86% respectivamente). Os parâmetros mais afetados em um ano de estocagem em temperatura ambiente foram: HMF, acidez livre, condutividade elétrica e cor. O método de estocagem que melhor conservou as características do produto foi o freezer. Conclusões Os méis de Apis mellifera e Melipona subnitida apresentam diversas diferenças em seus padrões físico-químicos, mais acentuadamente nos valores de umidade e atividade diastásica. As análises do Tempo 0 dos méis de Apis se encaixaram sem exceção nas legislações nacional e internacional vigentes, enquanto as amostras de Jandaira se encontram fora do preconizado para a umidade e atividade diastásica. Após um ano de estocagem em temperatura ambiente com incidência de luz, o mel de Apis conservou melhor suas características físico-químicas em relação ao mel de Jandaira em todos os parâmetros analisados ainda estando em conformidade com a legislação. Quando estocados em diferentes temperaturas, os méis de Jandaira conservaram melhor seus parâmetros nas condições de freezer e geladeira, enquanto sofreram alterações visíveis em temperatura ambiente. / Objectives Evaluate the quality and composition of Jandaira bee\'s honey through the use of physicochemical analyses; find out if the obtained parameters are in concordance with the Apis mellifera honey Legislations established both nationally and internationally; evaluate the effects of storage conditions for Jandaira Honey in three different temperatures (ambient, fridge and freezer) over one year, as well as the storage of Apis mellifera samples from the same botanical region over one year in ambient temperature and the comparison of both kinds of honeys and the alterations of their quality parameters. Metodology Review of the literature asserting the quality and identity parameters of Apis mellifera honey and stingless bees, with emphasis on Melipona subnitida honey. Procedure of the analyses contained in the Brazillian Legislation (2000) for Apis mellifera honey in the collected samples of Apis mellifera and Melipona subnitida honeys, as well as the qualitative analyses (Fiehe, Lund and Lugol), electric conductivity, and quantification of the glucose, fructose and sucrose sugars via HPLC. Results The Apis honey samples were classified as monofloral (Althernantera sp. as dominant pollen) and demonstrated parameters in accord with the current Legislation, while the Jandaira honey samples were classified as heterofloral (nearly even amounts of Mimosa verrucosa, Mimora caeselpinaefolia and Piptadenia moniliforme pollen) displaying moisture (24,80%) and diastase activity (null) in discordance with the established by the current legislation for Apis mellifera honeys. The Apis honey samples presented higher values of electric conductivity (284,00 µS.cm-1) than the obtained in the Jandaira honey samples (102,77µS.cm-1) as well as a darker color (26,67mmPfund) when compared to Jandaira honey (7,00mmPfund). The concentration of the glucose, fructose and sucrose sugars was higher in the Apis honeys (23,50%, 38,78% and 5,72% respectively) than in the Jandaira honey samples (21,76%, 29,21% and 4,86% respectively). The most affected parameters throughout one year of storage were HMF, free acidity, electric conductivity and color. The temperature that conserved better the original characteristics of the product was the freezer. Conclusions The Apis mellifera and Melipona subnitida honey samples displayed several differences in their physicochemical parameters, namely in moisture and diastase activity. At first (Time 0) the Apis honey samples were without exception in accord with the national and international legislations, whereas the Jandaira samples had moisture and diastase activity stray from the protocol, however, after one year of storage in ambient temperature, the Apis honey samples still was in concordance with the legislations in every analyzed parameter and had it\'s physicochemical characteristics better conserved then the Jandaira honey samples. Jandaira also had it\'s own characteristics better conserved in the fridge and freezer, when compared to the notable alterations in the ambient temperature storage.

Os objetivos deste estudo foram: avaliar a fauna de polinizadores potenciais do morangueiro em campos de produçaõ comercial; verificar a contribuição de diversos agentes de polinização na produtividade do morangueiro, conhecendo a biologia floral e reprodutiva dos cultivares mais utilizados para o plantio; testar, em cultivo fechado (estufa), diversas espécies de Meliponini (abelhas sem ferrão)como polinizador do morangueiro, avaliando a sua adaptação ao cultivo e sua efetividade nesta cultura. Os resultados obtidos revelaram que as flores do dos cultivares "Sweet Charlie" e "Oso Grande" necessitam de um polinizador para a produção de frutos sem deformação pois a morfologia de suas flores e o amadurecimento de suas estruturas reprodutivas não favorecem a ocorrência da autopolinização, nem a distribuição homogênea do pólen pelos estigmas da flor, resultando em frutos deformados que não alcançam o seu potencial máximo de desenvolvimento. A abelha jataí (Tetragonisca angustula) adaptou-se bem ás condições do cultivo fechado (estufa) e mostrou-se efetiva na polinização do morangueiro, resultando em frutos mais pesados e em uma porcentagem mais elevada de frutos bem formados. / The aims of this study were: to investigate potencial pollinators of strawberry crops in commercial fields; to verify the contribution of several pollination agents on strawberry productivity, studying the floral and reproductive biology of some commercial cultivars; to evaluate the possibility of the rational use of stingless bees in strawberry pollination inside greenhouses. The results showed that the floral morphology of "Sweet Charlie" and Oso Grande" cultivars and the maturation pattern of their stamens and pistils nor favour the occurrence of self pollination neither the regular pollen distribuction among stigmata, yielding deformed fruits that don´t complete their development. Therefore these cultivars need pollinators to commercial production. Tetragonisca angustula (Apinae, Meliponini)was remarkable for its relatively quick adaptation inside greenhouses and for its effectivity as strawberry pollinator, resulting in heavier berries and high percentage of marketable fruits.

A perda de polinizadores no mundo todo, especialmente abelhas, tem sido atribuída, entre outros fatores, à intensificação do uso de inseticidas em agro ecossistemas. A nova classe química de inseticidas, os neonicotinóides, tem sido o principal alvo de investigações, sobretudo em decorrência de sua ação sistêmica que transloca a substância ativa pela planta e atinge o pólen e néctar como resíduos. Dessa forma, o presente estudo teve como objetivo verificar os efeitos do neonicotinóide tiametoxam em abelhas sociais e avaliar a possibilidade da espécie Scaptotrigona aff. depilis ser utilizada como bioindicadora de contaminação ambiental por inseticidas. Para tanto, (i) operárias forrageiras de Bombus terrestris audax, Apis mellifera e Scaptotrigona bipunctata foram submetidas à exposição contínua de alimento contaminado; (ii) operárias de Scaptotrigona aff. depilis foram expostas ao inseticida, durante o estágio larval, através do alimento e; (iii) operárias forrageiras de Scaptotrigona aff. depilis foram coletadas no meliponário da Universidade de São Paulo e arredores, e submetidas a análises químicas, para a checagem de contaminação pelos neonicotinóides tiametoxam e/ ou imidacloprido. (i) Dentre as três espécies de abelhas sociais adultas, B. terrestris audax mostrou ser a mais sensível à ação de tiametoxam, seguida de S. bipunctata e A. mellifera, verificados através do decréscimo no consumo de alimento e da longevidade. Nesse sentido, os efeitos subletais não foram influenciados pelo tamanho do corpo dos indivíduos. (ii) As operárias de Scaptotrigona aff. depilis, quando expostas a doses do inseticida em fase larval, apresentaram desenvolvimento tardio e originaram indivíduos menores e assimétricos. Tanto em abelhas adultas como em imaturas, os efeitos subletais foram evidenciados nos indivíduos expostos às maiores doses de tiametoxam nos experimentos (39.37 e 98.43µg de substância ativa/kg de xarope em adultos, ou 0.044 e 4.375ng de substância ativa/larva). (iii) A detecção de tiametoxam e imidacloprido em operárias forrageiras de Scaptotrigona aff. depilis permitiu considerar essa espécie como bioindicadora de contaminação. Nesse estudo, a mortalidade não foi parâmetro conclusivo quanto aos potenciais efeitos adversos de tiametoxam para as abelhas. O potencial de Scaptotrigona aff. depilis como espécie bioindicadora é uma importante etapa para avaliar a contaminação ambiental por neonicotinóides em abelhas sem ferrão, podendo ser utilizada em políticas públicas. / Pollinator losses all over the world, especially bees disappearing, have been attributed, among others, to the intensification of insecticide use on crops. The new chemical class of insecticides, the neonicotinoids, has been the main target of investigation, mainly due to its systemic action, which translocates the active substance for the plant and to reach the pollen and nectar as residues. Thus, this study evaluated the effects of thiamethoxam neonicotinoid on bees and the possibility of the use of Scaptotrigona aff. depilis species as bioindicator of environmental contamination by insecticides. For this, (i) Bombus terrestris audax, Apis mellifera and Scaptotrigona bipunctata forager workers were induced to continue exposure of contaminated food; (ii) Scaptotrigona aff. depilis workers were exposed to the insecticide, during its larval phase, through the larval food and; (iii) Scaptotrigona aff. depilis forager workers were collected from University of São Paulo meliponary and surroundings and analyzed, in order to check for contamination by thiamethoxam and/ or imidacloprid neonicotinoids. (i) Among the three social bee species, B. terrestris audax showed to be more sensible to the thiamethoxam action, followed by S. bipunctata and A. mellifera, verified through decreasing food consumption and longevity. Thus, the sub lethal effects were not influenced by individual body size. (ii) Scaptotrigona aff. depilis workers, when exposed to insecticide doses on larval food, caused individuals with developmental delay and were smaller and asymmetric. Both adult bees and in immature, the sub lethal effects of thiamethoxam were evidenced on individuals exposed to the highest doses in the experiments. (iii) The detection of thiamethoxam and imidacloprid in Scaptotrigona aff. depilis forager workers allowed to consider this species as bioindicator of environmental contamination. In this study, the mortality was not a conclusive parameter as to the potential adverse effects of thiamethoxam for the bees. The potential of Scaptotrigona aff. depilis species as bioindicator is an important step to assess environmental contamination by neonicotinoids on stingless bees, may be used for public policy.