El presente trabajo tuvo como objetivo analizar los recursos nectaríferos y poliníferos utilizados por Apis mellifera en diferentes ecosistemas del Departamento Diamante. Para ello, sobre la base de la distribución florística regional, se seleccionaron 3 apiarios ubicados en zonas de influencia de las comunidades vegetales más representativas, asociadas a diferentes tipos y usos del suelo. A estos apiarios se les realizó un seguimiento anual, registrando sus actividades de recolección de recursos alimenticios; identificando el polen recolectado durante un período apícola y el contenido polínico de la miel producida durante el período apícola, a través de 4 temporadas apícolas (1999-2003). Paralelamente se realizó un estudio de las comunidades vegetales en el área de influencia de cada uno de los apiarios en cuanto a la oferta de recursos melíferos, en diversidad, cobertura-abundancia y período de disponibilidad. Se recolectó material vegetal a fin de conformar una palinoteca de referencia que facilitara la identificación de los tipos polínicos encontrados en los productos elaborados por la abeja. Se analizaron además, muestras provenientes de otros apiarios del departamento, obtenidas en el mismo período, a fin de lograr una caracterización botánica y geográfica más representativa de las mieles producidas en esta región. Se describieron y documentaron 22 tipos polínicos mediante microfotografías tomadas al microscopio óptico y electrónico de barrido. El relevamiento de la vegetación permitió determinar un total de 358 especies, pertenecientes a 83 familias botánicas. Las mejores representadas en diversidad de especies fueron Asteraceae, Poaceae y Leguminosae. Predominaron las especies nativas y herbáceas. El seguimiento de la fenología de 300 especies, permitió establecer la duración de la temporada apícola en la región, la cual se extiende desde agosto a mayo dependiendo de las condiciones meteorológicas. Fueron identificados 105 tipos polínicos en las cargas corbiculares, correspondientes a 41 familias botánicas. Las mejores representadas fueron Asteraceae, Leguminosae y Apiaceae. Los tipos polínicos dominantes en las cosechas corresponden a Salix humboldtiana, Baccharis spp., Rapistrum rugosum, Lotus corniculatus, Glycine max, Prosopis sp., Melilotus albus, Typha latifolia y Ammi visnaga. El promedio de cosecha de polen para la zona fue de 23,6 g/colmena/día. En las mieles, se identificaron 152 tipos morfológicos pertenecientes a 62 familias botánicas. Las mejores representadas en número de especies fueron Asteraceae y Leguminosae. Predominaron las mieles monoflorales (59%). El principal tipo de miel producida fue de Glycine max (21%). Otras mieles obtenidas fueron Lotus corniculatus (15%), “tréboles” (11%), Ammi spp. (5%), Melilotus albus, Salix humboldtiana (3%) y Medicago sativa (1%). Los tipos polínicos más importantes fueron Ammi spp., Lotus corniculatus, Melilotus albus, Glycine max, Brassicaceae, Baccharis spp., Eryngium sp. y Salix humboldtiana. El análisis cuantitativo permitió clasificar al 85% de las mieles en la Clase I y II. Las mieles del Departamento Diamante se caracterizaron por la asociación de polen de Ammi spp., Lotus corniculatus, Melilotus albus, Glycine max, Asteraceae Asteroideae (principalmente especies nativas como Baccharis spp., Solidago chilensis, Eupatorium spp., Bidens spp. y/o Grindelia pulchella) y Salix humboldtiana. / The objective of this research was to analyze the nectariferous and polliniferous resources used by Apis mellifera in different ecosystems of Diamante Department. For this purpose, on the basis of regional floristic distribution, 3 apiaries were selected, located in areas of influence of the most representative plant communities, associated with different types and land uses. These apiaries underwent annual monitoring, recording their activities of collecting food resources, identifying the pollen collected during a beekeeping period and pollen content of honey produced during the beekeeping period, by 4 beekeeping season (1999-2003). In parallel, a study of plant communities in the area of influence of each apiary were realized, for to analyse the supply of resources in diversity, cover-abundance and availability period. Plant material was collected to form a reference palynoteca that facilitate the identification of pollen types found in products manufactured by the bee. Were also analyzed, samples from other apiaries in the Department, obtained in the same period in order to achieve a more representative geographical and botanical characterization of the honey produced in this region. Twenty two pollen types were described and documented by photomicrographs taken of optical microscopy and scanning electron microscopy. The survey of the vegetation allowed to determine a total of 358 species belonging to 83 botanical families. The best represented in variety of species were Asteraceae, Poaceae and Leguminosae. Native and herbaceous species were predominant. The monitoring of the phenology of 300 species, allowed to establish the duration of the beekeeping season in the region, which extends from August to May depending on weather conditions. One hundred and five pollen types were identified in corbicular pollen loads, corresponding to 41 botanical families. The best represented were Asteraceae, Leguminosae and Apiaceae. The dominant pollen types were Salix humboldtiana, Baccharis spp., Rapistrum rugosum, Lotus corniculatus, Glycine max, Prosopis sp., Melilotus albus, Typha latifolia and Ammi visnaga. The average harvest of pollen to the area was 23.6 g/hive/day. In honey, 152 morphological types were identified belonging to 62 botanical families. The best represented in number of species were Asteraceae and Leguminosae. Unifloral honeys were predominant (59%). The main type of honey produced was of Glycine max (21%). Other honeys obtained were Lotus corniculatus (15%), "clover" (11%), Ammi spp. (5%), Melilotus albus, Salix humboldtiana (3%) and Medicago sativa (1%). The most important pollen types were Ammi spp., Lotus corniculatus, Melilotus albus, Glycine max, Brassicaceae, Baccharis spp., Eryngium sp. and Salix humboldtiana. The quantitative analysis allowed to classify the 85% of the honeys in the Class I and II. The honey of Diamante Department were characterized by the association of pollen of Ammi spp., Lotus corniculatus, Melilotus albus, Glycine max, Asteraceae Asteroideae (mainly native species such as Baccharis spp., Solidago chilensis, Eupatorium spp., Bidens spp. and / or Grindelia pulchella) and Salix humboldtiana.

The effects of miticides on the reproductive physiology of queens and drones were examined. The first study examined the effects of Apistan (fluvalinate), Check Mite+ (coumaphos), and Apilife VAR (74% thymol) on sperm production and viability in drones. Drones from colonies treated with each miticide were collected at sexual maturity. Sperm production was determined by counting the number of sperm in the seminal vesicles. Sperm for viability assays was analyzed by dual fluorescent staining. Apilife VAR and coumaphos significantly lowered (P<0.0001) sperm production and coumaphos treatments caused a significant decrease (P<0.0001) in the sperm viability. The effects of miticides on queens was examined by treating queen-rearing colonies and examining the number and viability of sperm in the spermathecae of newly mated queens. Queens from each treatment group were collected after mating and the spermathecae were removed and analyzed. Colonies treated with coumaphos failed to provide viable queens and were excluded. Apilife VAR was found to significantly decrease (P<0.0016) sperm viability. No significant differences in sperm numbers were found between treatments. The effect of miticides on sperm viability over time was also examined. Drones were reared as described, but the spermatozoa were collected as pooled samples from groups of drones. The pooled samples from each treatment were subdivided and analyzed periods of up to 6 weeks. Random samples were taken from each treatment (n = 6 pools) over a period of 6 weeks. The exposure of drones to coumaphos during development significantly reduced sperm viability for all 6 weeks, and caused a large decline in week 6. The potential impacts of these results on queen performance and failure are discussed. / Master of Science in Life Sciences

Host-associated microbial communities, also known as microbiomes, are essential to the health of their hosts, and disturbance of these communities can negatively impact host fitness. The honey bee gut microbiome is a relatively simple host-associated community that makes an excellent model system for studying microbiome stability. In addition, honey bees are essential agricultural pollinators, so factors that impact their health are important for food security. The presented research focused on the stability of the honey bee gut microbiome in response to disturbance from the antibiotic tetracycline. Tetracycline was chosen because it is the most commonly used antibiotic in beekeeping, and may have negative effects on bees through the disruption of their gut microbiomes. The first study presents a new fecal sampling method for studying the honey bee gut microbiome of individual bees over time. This method accurately represented bacterial community structure in the gut microbiome as determined with 16S rRNA gene amplicon sequencing, as fecal and whole gut samples did not differ significantly for individual bees. The fecal sampling technique was then used to examine changes to individual honey bee gut bacterial communities before and after tetracycline exposure. Minimal differences in gut community structure were detected prior to and five days after tetracycline treatment. However, there was variability in how individual gut microbiomes were affected by tetracycline treatment, highlighting the importance of intraspecific variation in response to disturbance. The second study investigated whether the timing of disturbance during a host's life impacts microbiome community stability. Newly emerged bees were treated with tetracycline, returned to their hive, and recollected 7 or 14 days later. The gut communities of the bees were then characterized using 16S rRNA gene amplicon sequencing. Gut microbiome structure of bees treated with tetracycline at emergence differed from controls both 7 and 14 days after emergence, with the antibiotic-treated bees having lower community richness overall. This study showed that early life disturbance of host-associated microbial communities can influence microbiome structure later in life. The final study describes the occurrence of antibiotic resistance genes (ARGs) in honey bee gut bacterial symbionts from hives across the US. Honey bee gut metagenomes were sampled from hives at 13 apiaries located in a transect from Virginia to Washington, and ARG presence was assessed across the sites. We also specifically quantified the abundances of two common tetracycline resistance genes (tet(B) and tet(M)) across apiaries. ARGs, both for antibiotics used in beekeeping and unrelated antibiotics, were detected in honey bee gut bacteria from all apiaries. Tetracycline resistance genes were the most common across all apiaries, and the abundance of two tetracycline resistance genes varied by apiary. Members of the honey bee gut microbiome contained different proportions of ARGs, but taxa within a single family contained similar proportions, possibly indicating phylogeny plays a role in ARG accumulation. In particular, Gilliamella and Frischella, both in the family Orbaceae, contained the highest percentages of ARGs. The results from this study suggest honey bee bacteria act as reservoirs of ARGs. Overall, the presented research contributes to the field of biology by highlighting the importance of intraspecific variation in host-associated microbial communities and presenting a new method for studying honey bee gut microbiome variation at the individual-level, showing that early life events in honey bees influence microbiome development, and suggesting that honey bee bacterial symbionts have adapted to deal with antibiotic disturbance through the accumulation of ARGs. / Doctor of Philosophy / Nearly all animals, including honey bees, have communities of bacteria that live on and in them. These communities, called microbiomes, are often essential to the health of their hosts. For instance, communities of gut bacteria can be important for breaking down food for digestion. Honey bees have approximately 10 bacterial species that consistently live in their guts and provide these types of services to their host. As with many bacterial communities, these beneficial bacteria can be impacted by exposure to antibiotics, even though antibiotics can also be important for treating or preventing dangerous bacterial infections. In honey bee hives, the antibiotic tetracycline is used to prevent bacterial disease. However, tetracycline may simultaneously be negatively impacting colony health through disruption of the honey bee gut microbiome. The goal of the presented work was to understand how tetracycline impacts the honey bee gut microbiome. In my first chapter, I demonstrate a new fecal sampling method that will allow us to understand how gut microbiomes from individual bees change over time. I first compared the bacteria found in fecal samples to those in the whole guts of bees and found that the bacterial communities of the fecal samples and guts were very similar, indicating that fecal sampling is a good method for studying the honey bee gut microbiome. I then used my fecal sampling method to determine how individual honey bee gut microbiomes respond to antibiotic disturbance over time. I collected fecal samples from adult bees prior to treatment, treated the bees with tetracycline, and after five days of being maintained in the lab, recollected fecal samples. My results showed few changes to the bacterial communities before and after treatment, suggesting some honey bee gut microbiomes may be resistant to tetracycline. In my second chapter, I addressed whether exposure to antibiotics early in life had long-term impacts on the gut microbiome. I treated bees at the start of adulthood with tetracycline, returned the bees to their hive for 7 or 14 days, and assessed their microbiome. Tetracycline treatment at the beginning of adulthood changed the gut microbiome later in life, as the microbiomes of tetracycline-treated bees and controls differed from one another both 7 and 14 days after exposure. This chapter shows that disturbances to microbiomes during early life can also affect microbiomes later. My third chapter addressed how honey bee bacteria have adapted to antibiotic use by identifying antibiotic resistance genes (ARGs) in honey bee gut bacteria from 13 hives located in a transect across the US from the state of Washington to Virginia. I found a variety of antibiotic resistance genes in honey bee gut bacteria, both associated with beekeeping and likely environmental contamination. The prevalence of antibiotic resistance genes in honey bee bacteria may help us track antibiotic resistance in the environment. Ultimately, my dissertation contributes to our understanding of how antibiotic use affects honey bees by changing their gut microbiome.

Honey bees, Apis mellifera Linn., provide vital economic and ecological services via pollination while concurrently facing multiple interconnected stressors impacting their health. Many crops like apples, peaches, and cherries that add diversity and nutrition to our diet are wholly or partially dependent upon the pollination services of insects. Orchard crops are self-incompatible and commonly regarded as crops reliant on the pollination services of insects, and while previous studies have focused on the impact of bees to orchard crops during bloom, fewer studies have examined the reciprocal relationship of the orchards on honey bees, particularly across the entire foraging season. Here we investigated the foraging dynamics of honey bees in an orchard crop environment in Northern Virginia, United States. We decoded, mapped, and analyzed 3,710 waggle dances, which communicate the location of a valuable resource in the environment, for two full foraging seasons (April-October, 2018-2019), and, concurrent to the dance filming, collected pollen from returning foragers. We found that bees forage locally the majority of the time (< 2 km) throughout the season, with some long-range distances occurring in May after bloom (both 2018 and 2019) and in fall (2019). The shortest communicated median distances (0.50 km and 0.53 km), indicating abundant food availability, occurred during September in both years, paralleling the bloom of an important late season resource, goldenrod (Solidago). We determined, through plotting and analyzing the communicated forage locations and from the collected pollen from returning foragers, that honey bees forage more within apple orchards after the bloom (29.4% and 28.5% foraging) compared to during bloom (18.6% and 21.4% foraging) on the understory of clover and plantain. This post bloom foraging also exposes honey bees to the highest concentration of pesticides across the entire foraging season (2322.89 ppb pesticides versus 181.8 during bloom, 569.84 in late summer, and 246.24 in fall). Therefore, post bloom apple orchards supply an abundance of forage, but also the highest risk of pesticide exposure, which may have important implications for management decisions of bees in orchards. / Master of Science in Life Sciences / Honey bee hives have been declining significantly in the United States, driven by a multitude of issues and stressors including pesticide exposure, disease, pests such as varroa mites, and poor nutrition caused by natural land being converted into development or agriculture. Agricultural landscapes, in particular, are often monocultures are saturated with pesticides creating a potentially hazardous environment, yet reliant on bees to provide pollination for crops. Because of this interconnected relationship between bees and flowers and the effects of stressors agricultural systems cause have with pollinators, it is necessary to understand how honey bees forage in these environments and what potential health risks they face. We investigated honey bees foraging dynamics in an apple orchard in Northern Virginia, United States by observing honey bee waggle dance behavior, where bees literally waggle back and forth for a certain time and at a certain angle telling their nestmates where a resource is, and collecting pollen from returning forager bees to better understand when, where, and upon what honey bees forage throughout the foraging season, which is when flowers are available and the weather warm enough (April – October). We found that bees mostly forage locally near the hive throughout the season, indicating that sufficient amount of food was available even after short bloom time of the apple (April to mid-May). We determined, through plotting and analyzing the waggle dance locations, that honey bees forage more within apple orchards after the bloom on mostly clover and plantain. This abundance of post bloom foraging also exposes honey bees to the highest amounts of pesticides across the entire foraging season. Post bloom apple orchards supply an abundance of forage, but also the highest risk of pesticide exposure to honey bees.

Chez de nombreuses Angiospermes, la pollinisation croisée est nécessaire pour le succès de la reproduction. La plupart de ces plantes ont évolué afin de favoriser la pollinisation entomophile, principalement assurée par les abeilles. C’est le cas des Cucurbitacées, dont le melon (Cucumis melo L.), importante culture du sud de la France et le concombre d’âne (Ecballium elaterium (L.) A. Rich.) plante spontanée endémique du bassin méditerranéen, les deux modèles de cette étude. Afin de mieux comprendre les relations plante-pollinisateur,nous avons choisi de caractériser le couple pollen de Cucurbitacées/abeille domestique (Apis mellifera L.) à travers la production de pollen, sa présentation sur la fleur et son transport sur l’abeille. Nous avons montré que, chez ces plantes, l’ontogenèse du pollen s’accompagne d’erreurs développementales conduisant à la formation de près de 3% de grains de pollen vides. De plus, nous avons mis en évidence que chez E. elaterium, des erreurs méiotiques entraînaient la formation de grains de pollen génétiquement anormaux qui pourraient être source d’autopolyploïdie. Chez C. melo, nos résultats ont révélé que la mise en contact du pollen avec le milieu environnant entraîne une baisse de 30% de sa viabilité, mais que celle-ci reste stable jusqu’à la fin de l’anthèse. L’aptitude à germer, quant à elle, finit d’être acquise juste avant anthèse et décroît de 12% en fin de période d’anthèse. Nous avons montré que le pollen de C. melo transporté sur le corps de l’abeille pouvait soit avoir une viabilité et une aptitude à germer préservée, probablement par protection contre la déshydratation, soit perdre quasi-totalement sa viabilité et son aptitude à germer. Enfin, nous avons détecté des composés biochimiques spécifiques des abeilles protégeant ou diminuant la qualité reproductrice du pollen / For many flowering plant species cross pollination is necessary to ensure reproductive success. Most of these plants have evolved to encourage insect pollination which is mainly carried out by bees. Such is the case of plants in the Cucurbitaceae family, including the cantaloupe (Cucumis melo L.), an important crop of the South of France, and squirting cucumber (Ecballium elaterium (L.) A. Rich.), a wild plant restricted to the Mediterranean Basin, the two vegetal models used in this study. To better understand plant-pollinator relations, we proposed to characterise the interaction between Cucurbitaceae pollen and the honey bee (Apis mellifera L.) through pollen production, its presentation and its transport on the honey bee body. Our results showed that developmental anomalies took place during pollen ontogenesis in these species leading to 3% of the pollen grains produced to be empty. Moreover, we demonstrated that meiotic abnormalities in E. elaterium leading to the production of genetically abnormal pollen grains could be a source of autopolyploidy. At anthesis in C. melo, our results showed that pollen viability decreased to 30% when pollen was exposed to environmental conditions, then remained stable during the period of anthesis.The ability for pollen to germinate was completed just before anthesis then decreased to 12% at the end of anthesis. We determined that viability and germinability of cantaloupe pollen carried on the honey bee body can be enhanced or decreased according to which specific honey bee biochemical compounds were detected

Nosema ceranae est un parasite émergeant d’Apis mellifera décrit dans certaines régions comme la cause majeure de la mortalité des abeilles. Dans d’autres cas, il est soupçonné d’affaiblir les colonies par l’interaction avec d’autres facteurs de pression de l’environnement. Dans le cadre du phénomène global de la mortalité des abeilles, nous avons orienté nos recherches vers l’étude des effets N. ceranae, en faisant l’hypothèse que ce parasite est capable d’induire des changements comportementaux chez A. mellifera dus à des altérations physiologiques, ce qui pourrait éventuellement perturber l’organisation sociale et aboutir à la mort de la colonie. Etant donné cette hypothèse, trois domaines d’étude ont été inclus dans notre recherche, (i) les effets de N. ceranae sur l’organisation sociale de la colonie, (ii) les mécanismes moléculaires à la base des effets chez les abeilles parasitées, et (iii) les différences en virulence d’isolats de N. ceranae ce qui pourrait expliquer la variation des effets du parasite chez l’abeille. Nous avons obtenu trois résultats majeurs. D’abord, nous avons constaté des modifications dans la structure sociale des abeilles après l’infection. Ces changements sembleraient contribuer à la survie de la colonie constituant probablement un mécanisme d’immunité sociale. Ce mécanisme géré par un signal phéromonal, permettrait de diminuer la transmission du parasite au sein de la colonie et prolonger la survie des abeilles saines. Ensuite, nous avons mis en évidence des effets sur la physiologie de l’intestin de l’abeille qui pourraient causer sa mort : l’induction du stress oxydatif et l’inhibition du renouvellement cellulaire de l’épithélium. Finalement, nos résultats suggèrent que certaines caractéristiques de l’hôte et conditions environnementales augmenteraient la probabilité de N. ceranae d’induire la mort. En conclusion, N. ceranae a le potentiel de causer la mort des abeilles, cependant, la colonie pourrait contrer l’infection, par exemple, par de mécanismes d’immunité sociale, or, la réponse générale à l’infection dépendrait des caractéristiques de l’hôte en combinaison avec les conditions de l’environnement. Le phénomène d’effondrement de colonies à l’échelle mondiale a mis en évidence la fragilité du système colonie d’abeilles – environnement. L’étude de chaque facteur participant au système, en autres, parasites, pesticides, changements dans l’environnement, pratiques apicoles, est essentielle pour une meilleure compréhension de toutes les interactions qui maintiennent l’équilibre écologique des colonies / Nosema ceranae is an emergent parasite of the honey bee Apis mellifera. In some regions it has been found to be the main reason for bee mortality, while in others it is suspected of weakening honey bee colonies by interacting with other environmental stressors. In the context of worldwide colony losses, we focus our research on the study of N. ceranae, with the hypothesis that this parasite is able to induce behavioral changes in bees through physiological modifications, which could alter social organization and cause colony death. Given this hypothesis, the program of study falls into three areas; (i) N. ceranae effects on colony social organization, (ii) molecular mechanisms of N. ceranae infection underlying observed effects, and (iii) differences in virulence of N. ceranae strains which could explain the diversity of parasite effects. We obtained three main results. First, we observed modifications in honey bee social structure after infection. This mechanism under pheromone control, would reduce parasite transmission within the colony and increase the lifespan of healthy bees. These changes may contribute to colony survival as part of a mechanism of social immunity. Second, we found two mechanisms whereby the pathogen affects the physiology of bee midgut epithelium that could lead to host mortality: oxidative stress and the inhibition of cellular renewal. Finally, our results suggest that certain host and environmental conditions increase the probability that N. ceranae will cause bee mortality. In conclusion, N. ceranae has the potential to cause bee death, however at colony level bees might counteract infection through, for example, social immunity mechanisms; although, overall honey bee response to infection would depend on characteristics of the host in combination with environmental conditions. Worldwide colony losses phenomenon have highlighted the fragility of the “honey bee colony – environment” system. The study of each factor involve in this system, including parasites, pesticides, environmental changes and beekeeping practices, is essential to better understand all of the interactions that maintain the ecological balance of honey bee colonies

O Brasil apresenta grande potencial para a exploração da apicultura, dado ao seu vasto território e flora diversificada, o que permite diferentes variedades de méis com propriedades únicas. O estado do Paraná é um dos maiores produtores de méis do país e o investimento em produção que atenda aos mercados mais exigentes estimulou a produção do mel orgânico. O conhecimento desde a antiguidade sobre os efeitos benéficos à saúde pelo mel vem estimulando a pesquisa deste alimento nobre. Assim, este trabalho teve por objetivo estudar méis orgânicos brasileiros certificados (MO) para a caracterização do perfil fenólico, volátil, além da avaliação da capacidade de sequestro das espécies reativas de oxigênio e nitrogênio. Os méis foram coletados nos apiários de apicultores com certificação orgânica de dois municípios do sul do Paraná, General Carneiro e Turvo-PR. Nos ensaios foram utilizados extratos fenólicos dos méis, obtidos por meio da utilização da resina Amberlite® XAD®2, bem como méis brutos in natura. Os extratos apresentaram conteúdo de compostos fenólicos significativo, sendo o melato (MO5), de General Carneiro, o de maior teor (117,68± 4,40 mg EAG/g). Para as análises de sequestro das espécies reativas de oxigênio e nitrogênio, os extratos fenólicos foram sempre superiores aos méis brutos in natura. Os extratos fenólicos, de maneira geral, apresentaram alta capacidade de sequestro para o radical peroxila (ROOo), ácido hipocloroso (HOCl) e óxido nítrico (NOo). Em relação aos melatos, o extrato MO7 apresentou alta capacidade para o sequestro do HOCl (EC50= 4,83 ± 0,13 ?g/mL), enquanto que o MO5 foi melhor para o sequestro do NOo (EC50=2,16 ± 0,18 ?g/mL). Pelo método HPLC-ABTS on-line foi possível identificar e quantificar a contribuição para a atividade antioxidante do ácido ferúlico no extrato (MO1) e do flavonoide kanferol na amostra (MO4). O ácido ascórbico foi identificado e quantificado por HPLC somente nos melatos (MO3, MO5 e MO7). Pela técnica de LC-MS/MS foram identificados a presença dos seguintes compostos fenólicos: ácido caféico, rutina e hesperidina em todos os extratos. A análise de compostos voláteis por SPME-CG/EM mostrou a presença de dois compostos, encontrados apenas nos melatos, que foram o terpineno-4-ol, que possui ação antifúngica, antiparasitológica e anti-inflamatória; e o 3,4-dimetil-1-deceno, podendo assim serem utilizados como marcadores químicos destes méis. O conhecimento da composição química destes méis, bem como a composição fenólica bioativa, contribui para o fornecimento de antioxidantes naturais para a dieta, atenuando assim os efeitos negativos dos radicais livres / Brazil has a great potential to explore beekeeping due to its vast territory and diversified flora, what allows different varieties of honeys with unique characteristics. Parana state is one of the largest honey producers and the investment in production that meets the most demanding markets stimulated the organic honey production. The knowledge since early in history regarding the beneficial health effects promoted by honey is stimulating the scientific research of this noble food. Thus, this paper aimed to study certified Brazilian organic honeys (MO) in order to determine the phenolic and volatile profiles, and also the evaluation of radical scavenging capacity against Nitrogen and Oxygen Reactive Species (RNS and ROS, respectively). The honeys were collected from apiaries from beekeepers with the organic certification from two municipalities of southern Parana, General Carneiro and Turvo, PR. In the essays, phenolic extracts were obtained from honeys by using Amberlite® XAD®2 resin, as well as crude in natura honeys. The extracts showed a significant content in phenolic compounds, with honeydew (MO5), from General Carneiro, showing the highest content (117,68 ± 4,40 mg AGE/g). For the analyzes to determine the radical scavenging capacity against RNS and ROS, the phenolic extracts always showed up superior results in comparison to crude in natura honeys. Phenolic extracts showed, in general, great capacity to scavenge peroxyl radical (ROOo), hypochlorous acid (HOCl) and nitric oxide (NOo). In relation to honeydews MO7 extract showed the highest capacity to scavenge HOCl (IC50= 4,83 ± 0,13 ?g/mL) while MO5 was the sample with better capacity to scavenge NOo (IC50=2,16 ± 0,18 ?g/mL). By using HPLC-ABTS on-line method it was possible to identify and to quantify the ferulic acid in MO1 extract, a compound with an important contribution to the antioxidant activity of this sample, as well as the flavonoid kaempferol in MO4 sample. Ascorbic acid was identified and quantified by HPLC only in the honeydew samples (MO3, MO5 and MO7). The analyzes developed by LC-MS/MS techniques indicated the presence of the phenolic compounds caffeic acid, rutin and hesperidin in all the extracts. The analysis of volatile substances developed by SPME-GC/MS promoted the identification of two compounds found only in the honeydew samples. The compounds were the terpinen-4-ol, which has antifungal, antiparasitological and anti-inflammatory activities; and 3,4-dimethyl-1-decene. Both compounds can be used as chemical markers of these honeys. The knowledge of the chemical composition of the studied honeys, as well as their bioactive phenolic composition, contributes to supply natural antioxidants to human diet, thus attenuating the negative effects of free radicals

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-18T19:21:27Z No. of bitstreams: 1 2010 - Luiza D'Oliveira Sant'Ana.pdf: 2146437 bytes, checksum: 5d8a20463f01fc73a88b5b0c2e56d28c (MD5) / Made available in DSpace on 2017-05-18T19:21:27Z (GMT). No. of bitstreams: 1 2010 - Luiza D'Oliveira Sant'Ana.pdf: 2146437 bytes, checksum: 5d8a20463f01fc73a88b5b0c2e56d28c (MD5) Previous issue date: 2010-07-05 / This work aimed at the evaluation of the total phenolic and flavonoid contents of sixty Apis mellifera honey samples, commercialized in different regions of the state of Rio de Janeiro (Metropolis, Southern, Central, Northwestern and Seaside). For each sample, the total phenolic and the total flavonoid contents were analyzed by the Folin-Denis and aluminum chloride (AlCl3) methods, respectively. Also, the antioxidant activity (%AA and EC50) for all the honey samples was evaluated, as well as for some phenolic acids and flavonoids, used as standards. The determination of the antioxidant capacity was carried out by the method of trapping the 2,2-diphenyl-1-picryl-hydrazyl organic radical (DPPH) by antioxidant compounds, resulting in an absorbance decrease at 520 nm. The analyzed honeys were classified as monofloral or heterofloral. For the monofloral samples, it was observed that Anadenanthera honeys showed the highest average value for total phenolics (112,60 mgEAG/100g) and also for the antioxidant capacity (59,93 (AA)mgEQC/500g or 18,92 mg/mL if expressed as EC50), while the Eucalyptus honeys showed the highest mean for total flavonoids (9,17 mgEQC/100g). With these results, a comparative study was carried out with diverse honey samples of different regions, in terms of total phenolic and flavonoid content, antioxidant capacity and some physico-chemistry parameters (HMF, acidity, pH, humidity, water activity and color intensity). Comparison of these results allowed to observe that darker honeys showed both higher antioxidant activity and the highest values for total phenolic and flavonoid contents, as well as for all the other physico-chemical parameters. / O objetivo deste trabalho foi avaliar os teores de fen?licos e flavon?ides totais de sessenta amostras de mel de Apis mellifera comercializadas em diferentes mesorregi?es do estado do Rio do Janeiro (Metropolitana, Sul, Centro, Noroeste Fluminense e Baixada Litor?nea). Para cada amostra foram analisados os totais de fen?licos e flavon?ides, atrav?s dos m?todos de Folin-Denis e cloreto de alum?nio (AlCl3), respectivamente. Foi determinado, tamb?m, o potencial da atividade antioxidante (%AA e CE50) para todas as amostras de mel, bem como para alguns padr?es de ?cidos fen?licos e flavon?ides. O m?todo utilizado para determinar a capacidade antioxidante foi o de captura do radical org?nico 2,2-difenil-1-picril-hidrazil (DPPH) por antioxidantes, produzindo um decr?scimo da absorb?ncia a 520 nm. Os m?is avaliados foram classificados de monoflorais e heteroflorais. Para os monoflorais foi observado que os m?is de Anadenanthera apresentaram a maior m?dia em totais de fen?licos (112,60 mgEAG/100g), e tamb?m a maior atividade antioxidante (59,93 (AA)mgEQC/500g ou 18,92 mg/mL em termos de CE50), enquanto os m?is de Eucalyptus apresentaram, em m?dia, o maior conte?do de flavon?ides totais (9,17 mgEEC/100g). A partir dos resultados obtidos foi feito um estudo comparativo entre as v?rias amostras de m?is de diferentes regi?es quanto ao teor de fen?licos e flavon?ides totais, a atividade antioxidante e algumas propriedades f?sico-qu?micas (HMF, acidez, pH, umidade, atividade de ?gua e intensidade de colora??o). Ao comparar esses resultados, foi poss?vel observar que os m?is mais escuros apresentaram uma maior atividade antioxidante e uma maior quantidade de fen?licos e flavon?ides totais, bem como valores mais altos para todos os outros par?metros f?sico-qu?micos comparados.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-06-06T18:24:16Z No. of bitstreams: 1 ZILMAR FERNANDES NÓBREGA MELO - DISSERTAÇÃO PPGEA 2002..pdf: 14604622 bytes, checksum: 92a403c722a2a592fbb333f263c9b64f (MD5) / Made available in DSpace on 2018-06-06T18:24:16Z (GMT). No. of bitstreams: 1 ZILMAR FERNANDES NÓBREGA MELO - DISSERTAÇÃO PPGEA 2002..pdf: 14604622 bytes, checksum: 92a403c722a2a592fbb333f263c9b64f (MD5) Previous issue date: 2002-08 / Estudou-se o armazenamento de méis de abelhas africanizadas (Apis mellifera L.) coletados no brejo e cariri do estado da Paraíba. Utilizou-se, quanto à florada, dois tipos de mel de abelha (mel de florada silvestre e mel de florada de baraúna), armazenados sob 3 diferentes condições de embalagens (El - recipiente de polietileno opaco e exposto à luz e temperatura ambiente; E2 - recipiente de polietileno exposto à temperatura ambiente e ao abrigo da luz; E3 - recipiente em vidro translúcido exposto à temperatura e luz ambiente), ao longo de 180 dias, tendo sido estes submetidos a análises físico-químicas mensalmente, com a finalidade de observar as possíveis alterações nas características físico-químicas (°Brix, Umidade, Cinzas, Hidroximetilfurfural-HMF, Açúcares redutores, Sacarose aparente, Atividade diastásica e pH), objetivando verificar a eficiência destas embalagens no índice de qualidade dos méis. As análises físico-químicas dos méis foram feitas seguindo o recomendado pela Instrução Normativa do Ministério da Agricultura e do Abastecimento (Brasil, 2000). Os valores médios encontrados, ao longo do Armazenamento, para o °Brix do mel silvestre ficaram no intervalo de variação de 78,03 a 78,42 °Brix e o da florada de baraúna de 81,31 a 81,63 °Brix e não houve diferença significativa entre as embalagens ao longo do armazenamento. Os níveis de Umidade (%) para os méis silvestres e de baraúna foram 19,48 a 20,48 % e 16,25 a 16,70 %, respectivamente. Os valores Cinzas e de Sólidos Insolúveis não apresentara diferenças significativas no tempo e/ou nas embalagens. Os níveis no índice de HMF ficaram no intervalo de variação de 4,57 a 10,17 mg/kg de mel e 1,08 a 7,12 mg/kg, no mel Silvestre e no mel de baraúna, respectivamente. Pelas análises estatísticas, verificou-se, para os dois méis, diferença significativa a 1% de probabilidade e que as médias diferem entre si. Houve para o HMF, diferença significativa entre as embalagens e a E2 foi a mais eficiente no controle do aumento deste índice nos dois méis estudados. Os valores médios de Açúcares Redutores ficaram entre 74,09 e 76,41 % no mel Sivestre e entre 62,51 e 64,80 % para o mel de baraúna. Os valores de Sacarose Aparente ficaram entre 2,18 a 3,01 % para o mel silvestre e entre 2,40 e 2,89 % no mel de baraúna. Os méis sofreram pequena variação no tempo e as embalagens não influenciaram sobre a sacarose, nos dois méis. A Atividade Diastásica (DN) no mel Sivestre ficou no intervalo de variação de 13,37 a 18,71 DN e 9,14 a 13,25 DN no mel de baraúna. As análises estatísticas para os dois méis foram significativas ao nível de 1% de probabilidade e no teste de Tukey, as médias diferiram entre si, e a embalagem E2, nos dois méis analisados, mostrou-se mais eficiente na desaceleração do índice de Diástase em relação a El e E3. Quanto ao pH, o mel Sivestre foi mais ácido (3,42 a 3,55) do que o mel de baraúna (3,85 a 4,15). Todas as amostras encontram-se dentro dos padrões exigidos pelo Ministério da Agricultura e do Abastecimento. Do resultado final, concluiu-se que houve maior eficiência na embalagem E2, que contribuiu para a desaceleração do ritmo no índice de diástase. Observou-se que o índice de acidez livre aumentou ao longo do armazenamento. / The storage of african bees honeys (Aipis mellifera L.) which were collected from slough and cariri in Paraíba state was studied. Considering the bloomed, two kinds of bee honey (wild bloomed honey and baraúna bloomed honey) were used and storaged under three different conditions of packages ( Ei - opaque polyethylene container, that was under environment light and temperature; E2 - polyethylene container that was under environment temperature but it wasn't under light environment; E3 - translucent glass container which was exposed to environment temperature and light). These honeys stayed 180 days in packages and their physiochemical characteristics were analysed monthly to observe the possible alterations of them ( 0 Brix, Humidity, Ashes, Hydroximethylfurfurol - HMF, Reducers Sugar, Apparent Saccharose, Diastasic Activity and pH) to verify the efficiency of these packages in the honeys qualities index. The physiochemical analyses of the honeys were done according to the Normative Instruction of the Agriculture and Provisioning Ministry (Brazil, 2000). During the storage time, the médium found values to the 0 Brix of the wild bloomed honey were between 78.03 and 78.42 0 Brix and the ones of the baraúna bloomed honey were between 81.31 and 81.63 0 Brix and there wasn't significant difference among the packages during the storage period. The Humidity rates (%) were between 19.48 and 20.48% to the wild honey and between 16.25 and 16.70 to the baraúna honey. There were no significant differences of the Ashes and Insoluble Solids values on their time and packages. The rates in the HMF index varied from 4.57 to 10.17 mg/Kg of honey and from 1.08 to 7.12 mg/Kg, to the wild and baraúna honeys, respectively. According to the statistics analyses, a significant difference of 1% of probability for both honeys was verified, besides the averages are different among them. To the HMF, there was a significant difference among the packages and the E2 was the most efficient to the control of the increase of this index in both studied honeys. The médium values of the Reducers Sugar were between 74.09 and 76.41% to the wild honey and they were between 62.51 and 64.80% to the baraúna honey. The Apparent Saccharose values were between 2.18 and 3.01% to the wild honey and they were between 2.40 and 2.89 to de baraúna honey. The honeys had a little time variation and the packages had no influence on the saccharose, in both honeys. The Diastasic Activity (DN) on the Wild honey varied from 13.37 to 18.71 DN and the variation of the baraúna honey was from 9.14 to 13.25 DN. The statistics analyses were significative at 1% of probability and at the Tukey test for both honeys, the averages were diferent among them and the E2 package was more efficient than the Ei and E3 to the unacceleration of the diástase index for both honeys. Analysing the pH, the wild honey was more acid (3.42 to 3.55) than the baraúna honey (3.85 to 4.15). Ali the samples are according to the Agriculture and Provision Ministry patterns. According to the final result, it's conclued that there was a bigger efficiency of the E2 package and it contributed to the rhythm unaceleration in the distasic index. It was observed that the free acidity index increased during the storage period.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-04-11T16:57:51Z No. of bitstreams: 1 RenataVRSG_TESE.pdf: 4187159 bytes, checksum: 1407c0df06b55e7b10b6605b6b21cbe7 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-04-13T15:06:15Z (GMT) No. of bitstreams: 1 RenataVRSG_TESE.pdf: 4187159 bytes, checksum: 1407c0df06b55e7b10b6605b6b21cbe7 (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-04-13T15:06:23Z (GMT) No. of bitstreams: 1 RenataVRSG_TESE.pdf: 4187159 bytes, checksum: 1407c0df06b55e7b10b6605b6b21cbe7 (MD5) / Made available in DSpace on 2017-04-13T15:06:34Z (GMT). No. of bitstreams: 1 RenataVRSG_TESE.pdf: 4187159 bytes, checksum: 1407c0df06b55e7b10b6605b6b21cbe7 (MD5) Previous issue date: 2016-05-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work aimed to evaluate africanized bee (Apis mellifera L.) phenotypic characteristics, important in selecting matrix queens for honey production. Thirty africanized bee (Apis mellifera L.) colonies were transferred from partner beekeeping apiaries in coastal zone and South Zona da Mata of Pernambuco to experimental apiary of Animal Science Department (AEDZ/UFRPE). Colonies were fortnightly evaluated in the period from January 2014 to February 2016 and data of 89 queens of colonies or families were recorded. Energy and protein food was provided in off-season period. Infestation rate in colonies with Varroa destructor and infection degree with Nosema were verified, as well as hygienic behavior, queen egg laying, defensiveness and honey production tests were carried out in the first experimental phase and selection index (IS) was obtained of analyzed colonies. For the 2nd experimental phase, 1st, 2nd and 3rd generation queens were produced; daughters of matrixes with IS ≥ 8.0 and honey production was evaluated. Sixty-two out of 89 queens observed died naturally, presenting average longevity of 6.4 ± 2.9 months. Natural reproduction peak period is between September and December. The queen perform egg laying all months of the year, and as temperature increases and humidity and rainfall decrease, queen oviposition intensity is higher; thus brood comb increases. The diet used showed economic viability, avoided swarm loss and stimulated queen egg laying. Low level of Varroatose (2.92% ± 2.02%) and low intensity of Nosema infection were found. Success in queen cell production was 98% in acceptance of transferred larvae. Genetic gain was observed of 0.7 Kg in honey production at colonies with 3rd generation queens; which represents increasing of 5.4% of honey production, related to matrix colonies and 49.2% compared to production in partner beekeeping apiaries. These results are very encouraging, leading us to believe in breeding program success, using the variables and calculation format for selection index used in this work and give us subsides for possible adjustments in management of production apiaries, in accordance with environmental conditions of this region, aiming at professional beekeeping growth and increased honey production / Este trabalho teve como objetivo avaliar as características fenotípicas de abelhas africanizadas (Apis mellifera L.) importantes na seleção de rainhas matrizes para a produção de mel. Foram transferidas 30 colônias de abelhas africanizadas (Apis mellifera L.) dos apiários de apicultores colaboradores da região do litoral e zona da mata sul de Pernambuco para o apiário experimental do Departamento de Zootecnia (AEDZ/UFRPE). As colônias foram avaliadas quinzenalmente no período de janeiro de 2014 a fevereiro de 2016 e foram registrados dados de 89 rainhas de colônias ou famílias. Foi administrado alimentação energética e proteica no período de entressafra. Foi verificado o nível de infestação das colônias com Varroa destructor e Nosema e realizado os testes de comportamento higiênico, postura da rainha, defensividade e produção de mel na 1ª fase experimental e obteve-se o índice de seleção (IS) das colônias analisadas. Para 2ª fase experimental foram produzidas rainhas de 1ª, 2ª e 3ª geração filhas das matrizes com IS ≥ 8,0 e avaliou-se a produção de mel. Das 89 rainhas observadas, 62 morreram naturalmente apresentando longevidade média de 6,4 ± 2,9 meses. A época de pico da reprodução natural é entre os meses de setembro a dezembro. A rainha realiza postura todos os meses do ano, sendo que à medida que a temperatura aumenta e a umidade e o índice pluviométrico diminuem, maior é a intensidade de ovoposição da rainha, consequentemente aumenta o número de quadros com crias. A dieta utilizada apresentou viabilidade econômica, evitou a perda dos enxames e estimulou a postura da rainha. Foram encontrados um nível baixo de varroatose (2,92% ± 2,02%) e baixa intensidade de infecção por Nosema. O sucesso na produção de realeiras foi de 98% na aceitação das larvas transferidas. Observou-se um ganho genético de 0,7 Kg na produção de mel nas colônias com rainhas de 3ª geração, o que representa aumento de 5,4% na produção de mel em relação as colônias matrizes e 49,2% comparada a produção nos apiários dos apicultores parceiros. Esses resultados são muito estimulantes levando-se a acreditar no sucesso do programa de melhoramento aplicado, utilizando-se as variáveis e a forma de cálculo para o índice de seleção utilizados no presente trabalho e nos dá subsídios para possíveis adequações no manejo dos apiários de produção, em conformidade com as condições ambientais dessa região, visando o crescimento profissional da atividade apícola e o aumento da produção de mel / 2017-04-11