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

No description available.

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.

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.

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.

A produção de mel no Brasil tem aumentado consideravelmente e vem ganhando destaque no mercado internacional, consequentemente muitas exigências passaram a fazer parte da cadeia produtiva apícola. Além das análises previstas na legislação brasileira, o mercado externo procura produtos que atendam aos mais rigorosos padrões de qualidade. O mel possui características interessantes devido a sua constituição, porém, existem problemas que podem alterar seu padrão de identidade e qualidade. A aplicação da radiação gama é uma proposta viável que promove a diminuição da carga microbiana sem alterar a constituição e as características físico-químicas, com perdas nutricionais menores quando comparada a outros tratamentos usados em alimentos. Neste estudo, além dos parâmetros físico-químicos exigidos pela legislação brasileira (MAPA), também foram incluídas outras análises pertinentes, inclusive a técnica de espectroscopia no infravermelho com transformada de Fourier (FTIR-ATR). As análises microbiológicas foram realizadas nas amostras de méis puros, nas quais, posteriormente, foram inoculadas cargas microbianas conhecidas para avaliar a ação da radiação gama nas doses de 5 e 10 kGy. O teste triangular foi utilizado na análise sensorial para diferenciar as amostras não-irradiadas das irradiadas. As maiores modificações nas amostras de méis foram, principalmente, nas concentrações de HMF (hidroximetilfurfural) e na atividade diastásica, ocorrendo diminuição significativa em todas as amostras analisadas. Houve redução na carga microbiana a partir da aplicação da dose de 5 kGy, atingindo a ausência com a aplicação de 10 kGy de dose, com exceção do Paenibacillus larvae que mostrou ser mais resistente. A análise sensorial realizada nas amostras de mel e mel irradiado, com doses de 5, 10 e 15 kGy, apontou não haver diferença significativa (5%) entre as amostras controle e irradiadas. A aplicação da radiação gama em méis mostrou ser um método muito útil na descontaminação microbiológica, apresentando poucas alterações no produto com doses de até 10 kGy. / Honey production in Brazil has increased considerably and gaining proeminence on the international market, as a result many demands have become part of the productive honey chain. In addition to the analyses provided for the Brazilian legislation, international market demand products that meet the most stringent quality standards. Honey has interesting characteristics due to its constitution, but even so there are problems that can change some how your identity and quality. The application of gamma radiation is a viable proposal which promotes the reduction of bioburden without changing the constitution and most importantly the physicochemical properties, with smaller nutritional losses when compared to other treatments used in foods. This study beyond the physicochemical parameters required by Brazilian regulation also included other relevant analyses, including FT-IR ATR. Microbiological analyses were carried out on samples of pure honeys and subsequently inoculated microbial known loads to assess the action of gamma radiation in doses of 5 and 10 kGy. The triangular test was used to differentiate the irradiated and not irradiated. samples The major changes that have occurred in samples of honeys were mainly in concentrations of HMF (hydroxymethilfurfural) and diastase activity occurring a significant decrease in all analyzed samples. There was a reduction microbiological from the application of the 5 kGy, reaching absence with 10 kGy dose except Paenibacillus larvae. The sensory analysis conducted on samples of honey and honey irradiated with 5, 10 and 15 kGy doses, pointed out that there is no significant difference (5%) between irradiated and control samples. The application of gamma radiation in honeys proved a very useful method in microbiological decontamination, featuring few changes in the product at doses up to 10 kGy.

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

Les écosystèmes forestiers naturels du Sud-Ouest de l’océan Indien (SOOI) sont fortement impactés par la déforestation et font face à de nombreuses invasions biologiques pouvant altérer leur processus, en particulier les interactions de type plantes-pollinisateur. C’est dans ce contexte que s’inscrit cette thèse, décrivant les relations entre un pollinisateur généraliste indigène : Apis mellifera unicolor et la flore de deux écosystèmes de Ranomafana : RA et de Mare Longue : ML. La description palynologique de 135 espèces issues de 52 familles de plantes mellifères de la formation de RA, a fait ressortir les caractères spécifiques des pollens liés à une pollinisation entomophile. Dans un second temps, des suivis phénologiques mensuels de 131 espèces (90% d’indigènes) à RA et 120 espèces (53% d’exotiques) à ML ont permis d’identifier et d’estimer les ressources florales disponibles. Pendant un an, l’analyse mensuelle des miels et pollens collectés a permis de dresser l’inventaire des ressources florales réellement exploitées. Les espèces indigènes ont été significativement plus butinées que les espèces exotiques, malgré une diversité de ressources exotiques supérieures à celles des indigènes (ML). Le comportement de butinage de l’abeille sur le genre Weinmannia a été analysé sur la base de 104 h de vidéo (W. bojeriana et W. rutenbergii à Madagascar, et W. tinctoria à La Réunion). Les fleurs ont été visitées par un cortège de pollinisateurs potentiels : coléoptères, diptères, lépidoptères et autres hyménoptères. A. m. unicolor était le visiteur le plus fréquent à Madagascar. L’ensemble de ces résultats a permis de confirmer le comportement généraliste d’A. m. unicolor, avec toutefois une préférence forte et significative pour les espèces florales indigènes des strates arborées et arbustives et permet d’émettre des hypothèses sur la complexité des interactions entre l’abeille et les écosystèmes du hotspot de biodiversité SOOI. / The natural forest ecosystems of the Southwest Indian Ocean (SOOI) have been strongly impacted by deforestation and face many biological invasions that are alter their balance, particularly plant-pollinator interactions. It is in this context that this thesis is set, describing the relations between an indigenous general pollinator: Apis mellifera unicolor and the flora of two ecosystems of Madagascar (Ranomafana: RA) and Reunion (Mare Longue: ML). The palynological description of 135 species from 52 families of melliferous plants from the RA formation, highlighted the specific criteria of pollens associated with entomophilous pollination. In a second time, monthly phenological monitoring of 131 species (90% native) to RA and 120 species (53% exotic) to ML allowed to identify and estimate the available floral resources. During one year, the monthly analysis of honeys and pollen collected enabled an inventory of the floral resources actually exploited. Native species were significantly more visited than exotic species, despite a diversity of exotic resources superior to that of the natives (ML). The foraging behaviour of the honeybee on the genus Weinmannia was analysed on the basis of 104 hours of video (W. bojeriana and W. rutenbergii in Madagascar, and W. tinctoria in Reunion Island). The flowers were visited by many potential pollinators: Coleoptera, Diptera, Lepidoptera and other Hymenoptera. A. m. unicolor was the most frequent visitor to the two species from Madagascar. The results confirmed the generalist behaviour of A. m. unicolor, with, however, a strong and significant preference for native floral species of tree and shrub strata and makes it possible to speculate on the important place of this bee in the ecosystems of the SOOI biodiversity hotspot.

Submitted by Deyse Queiroz (deysequeirozz@hotmail.com) on 2018-06-01T18:40:56Z No. of bitstreams: 1 PATRÍCIA BASTOS DE ANDRADE ALBUQUERQUE MARANHÃO - DISSERTAÇÃO PPGSA PROFISSIONAL 2016..pdf: 1396221 bytes, checksum: 593b4ca006185c4d3ec8656d6cf6eda0 (MD5) / Made available in DSpace on 2018-06-01T18:40:56Z (GMT). No. of bitstreams: 1 PATRÍCIA BASTOS DE ANDRADE ALBUQUERQUE MARANHÃO - DISSERTAÇÃO PPGSA PROFISSIONAL 2016..pdf: 1396221 bytes, checksum: 593b4ca006185c4d3ec8656d6cf6eda0 (MD5) Previous issue date: 2016 / O agronegócio vem se destacando na economia brasileira registrando grandes avanços quantitativos e qualitativos. Apesar de ser antiga, só nos últimos anos a criação de abelhas vem ganhando maior destaque, transformando a apicultura em uma das atividades da agropecuária mais lucrativas. Os estados da região nordeste, em 2014, ocuparam o segundo lugar em produção, no qual a Paraíba teve uma produção em torno de 650 toneladas de mel. Esse novo cenário requer dos apicultores uma nova postura perante a atividade apícola. Diante desse novo momento, nossa pesquisa teve como objetivo demonstrar de que forma a aplicação dos custeios por absorção e variável podem auxiliar os produtores de mel na Associação dos Apicultores do Vale do Rio do Peixe no município de São João do Rio do Peixe no estado da Paraíba – APIVALE. Esta pesquisa tem caráter exploratório e, devido ao elevado grau de detalhamento, é um estudo de caso, feito a partir de pesquisa documental e entrevista semiestruturada. Adotamos o método quantitativo que permite a demonstração de resultados em números para posteriormente analisá-los. Os resultados indicaram que os sistemas de custeio por absorção e variável podem vir auxiliar os apicultores da APIVALE revelando a margem de contribuição, o ponto de equilíbrio, a margem de lucro, direcionando pontos ineficientes, eliminando custos e otimizando a produção. Como também, servindo como ferramenta de controle para tomada de decisões. Foi possível identificar que das 1.145 colmeias existentes, 57,7% das colmeias não estão produzindo, o que corresponde uma perda de aproximadamente 8.714 kg de mel. E das 42,3% que estão produzindo, 88,88% estão produzindo abaixo da média geral/colmeia que é de 13,2kg/colmeia. Porém, mesmo diante dessa baixa produção, a APIVALE em 2015 apresentou lucro de R$10.840,79 (dez mil oitocentos e quarenta reais e setenta e nove centavos), o que evidencia o potencial da associação e que se os sistemas de custeios por absorção e variável forem utilizadas para tomada de decisão pelos associados da APIVALE, estes terão condição de produzirem de forma eficaz. / Agribusiness has stood out in the Brazilian economy recorded large quantitative and qualitative advances. Despite being old, only in recent years beekeeping is gaining greater prominence, turning to beekeeping in one of the most profitable agricultural activities. The states of the Northeast region in 2014, occupied the second place in production, where Paraíba had a production around 650 tons of honey. This new scenario requires beekeepers a new attitude towards beekeeping. In this new period, our research aimed to show how the application of costing by absorption and variable can help honey producers in the association of beekeepers of Vale do Rio do Peixe in the municipality of São João do Rio do Peixe in Paraíba - Apivale. This research is exploratory and due to the high degree of detail, is a case study, made from documentary research and semi-structured interview. We adopted the quantitative method that allows the income statement in numbers for later review them. The results indicated that the costing systems for absorption and variable can help Apivale beekeepers revealing the contribution margin, the balance, the profit margin, directing inefficient points, eliminating costs and optimizing production. As well as serving as a control tool for decision making. It was possible that the 1,145 existing hives, 57.7% of the hives are not producing, representing a loss of approximately 8714 kg of honey. And 42.3% of which are producing, 88.88% are producing below the overall average/hive is 13,2kg/hive. However, despite of this low production, Apivale in 2015 showed profit 10,840.79 (ten thousand eight hundred and forty reais and seventy-nine cents), which shows the potential of the association and that the costing systems for absorption and variable are used for decision making by the members of Apivale, these will condition to produce effectively.

A produção de mel no Brasil tem aumentado consideravelmente e vem ganhando destaque no mercado internacional, consequentemente muitas exigências passaram a fazer parte da cadeia produtiva apícola. Além das análises previstas na legislação brasileira, o mercado externo procura produtos que atendam aos mais rigorosos padrões de qualidade. O mel possui características interessantes devido a sua constituição, porém, existem problemas que podem alterar seu padrão de identidade e qualidade. A aplicação da radiação gama é uma proposta viável que promove a diminuição da carga microbiana sem alterar a constituição e as características físico-químicas, com perdas nutricionais menores quando comparada a outros tratamentos usados em alimentos. Neste estudo, além dos parâmetros físico-químicos exigidos pela legislação brasileira (MAPA), também foram incluídas outras análises pertinentes, inclusive a técnica de espectroscopia no infravermelho com transformada de Fourier (FTIR-ATR). As análises microbiológicas foram realizadas nas amostras de méis puros, nas quais, posteriormente, foram inoculadas cargas microbianas conhecidas para avaliar a ação da radiação gama nas doses de 5 e 10 kGy. O teste triangular foi utilizado na análise sensorial para diferenciar as amostras não-irradiadas das irradiadas. As maiores modificações nas amostras de méis foram, principalmente, nas concentrações de HMF (hidroximetilfurfural) e na atividade diastásica, ocorrendo diminuição significativa em todas as amostras analisadas. Houve redução na carga microbiana a partir da aplicação da dose de 5 kGy, atingindo a ausência com a aplicação de 10 kGy de dose, com exceção do Paenibacillus larvae que mostrou ser mais resistente. A análise sensorial realizada nas amostras de mel e mel irradiado, com doses de 5, 10 e 15 kGy, apontou não haver diferença significativa (5%) entre as amostras controle e irradiadas. A aplicação da radiação gama em méis mostrou ser um método muito útil na descontaminação microbiológica, apresentando poucas alterações no produto com doses de até 10 kGy. / Honey production in Brazil has increased considerably and gaining proeminence on the international market, as a result many demands have become part of the productive honey chain. In addition to the analyses provided for the Brazilian legislation, international market demand products that meet the most stringent quality standards. Honey has interesting characteristics due to its constitution, but even so there are problems that can change some how your identity and quality. The application of gamma radiation is a viable proposal which promotes the reduction of bioburden without changing the constitution and most importantly the physicochemical properties, with smaller nutritional losses when compared to other treatments used in foods. This study beyond the physicochemical parameters required by Brazilian regulation also included other relevant analyses, including FT-IR ATR. Microbiological analyses were carried out on samples of pure honeys and subsequently inoculated microbial known loads to assess the action of gamma radiation in doses of 5 and 10 kGy. The triangular test was used to differentiate the irradiated and not irradiated. samples The major changes that have occurred in samples of honeys were mainly in concentrations of HMF (hydroxymethilfurfural) and diastase activity occurring a significant decrease in all analyzed samples. There was a reduction microbiological from the application of the 5 kGy, reaching absence with 10 kGy dose except Paenibacillus larvae. The sensory analysis conducted on samples of honey and honey irradiated with 5, 10 and 15 kGy doses, pointed out that there is no significant difference (5%) between irradiated and control samples. The application of gamma radiation in honeys proved a very useful method in microbiological decontamination, featuring few changes in the product at doses up to 10 kGy.