Nós avaliamos a atividade antimicrobiana do mel coletado de ninhos de 12 espécies de abelhas nativas sem ferrão comumente encontrados no Brasil e 25 amostras de mel de Apis mellifera não pasteurizados que foram identificadas como sendo unifloral e uma amostra de mel multifloral. A atividade antimicrobiana de cada amostra de mel foi testada contra cinco espécies de bactérias patogênicas, uma espécie de fungo patogênico e uma espécie de levedura patogênica, comparando esta atividade com o mel terapêutico de manuka produzido por abelhas Apis mellifera na Nova Zelândia a partir do néctar de Leptospermum scoparium (Myrtaceae). Cinco das treze amostras de mel das abelhas sem ferrão foram bactericidas e oito foram fungicidas contra o fungo patogênico Trichophyton rubrum. Somente a levedura Candida albicans foi resistente a todas as amostras de mel. As amostras de mel de Apis mellifera que apresentaram atividade bactericida foram caju, romã e cana, nenhuma das amostras de mel de A. mellifera afetou o fungo T. rubrum. Os meis de Nannotrigona testaceicornis, Plebeia remota, Tetragona clavipes e Scaptotrigona depilis todos com alto nível de atividade antimicrobiana, foram significativamente mais eficiente em termos de atividade antimicrobiana que o mel de manuka, especialmente contra Escherichia coli, Klebsiella pneumoniae e T. rubrum. O mel artificial feito com os principais açúcares encontrados no mel e com teor de água semelhante, não teve atividade antimicrobiana em nossas análises. Nós também fizemos análises de espectrometria de massas das amostras de mel e encontramos alguns \"fingerprint\" característicos que foram associadas com a atividade antimicrobiana. / We evaluated the antimicrobial activity of honey collected from nests of 12 species of native stingless bees commonly found in Brazil and 25 unpasteurized honey samples of Apis mellifera that were identified as being unifloral and one multifloral honey sample. The antimicrobial activity of each honey sample was tested against five species of pathogenic bacteria, a pathogenic fungal species and a pathogenic yeast species, comparing this activity with therapeutic manuka honey produced by Apis mellifera bees in New Zealand from the nectar of Leptospermum scoparium (Myrtaceae). Five of thirteen samples of honey from the stingless bees were bactericidal and eight were fungicidal against the pathogenic fungus Trichophyton rubrum. Only the yeast Candida albicans was resistant to all honey samples. The Apis mellifera honey samples that showed bactericidal activity were caju, romã e cana, none of the A. mellifera honey samples affected the fungus T. rubrum. The honeys from Nannotrigona testaceicornis, Plebeia remote, Tetragona clavipes and Scaptotrigona depilis all had a high degree of antimicrobial activity, they were significantly more efficient in terms of antimicrobial activity than manuka honey, especially against Escherichia coli, Klebsiella pneumoniae and T. rubrum. Artificial honey, made with the main sugars found in honey and with similar water content, had no antimicrobial activity in our analyses. We also made mass spectrometry analyses of the honey samples and found some fingerprint characteristics that were associated with antimicrobial activity.

Na primeira etapa deste trabalho avaliou-se o desempenho fermentativo de 6 cepas de leveduras comerciais utilizadas na produção de vinhos e cerveja, em mosto de mel 30°Brix suplementado com 0,5 g/L de peptona e 1 g/L de extrato de levedura. Como resposta determinou-se o crescimento das leveduras em número de células e as concentrações de glicose, frutose e etanol por HPLC, que permitiram determinar os parâmetros rendimento, eficiência de fermentação e produtifidade em etanol, bem como a produção de sulfeto de hidrogênio. Assim, selecionou-se a cepa Saccharomyces bayanus - Pasteur Champagne - Red Star, que teve seu comportamento avaliado em mosto de mel suplementado com diferentes nutrientes. Os resultados demonstraram que a suplementação do mosto de mel com sais ou com suplemento comercial Enovit contribuiu para o bom desempenho da referida cepa. Na etapa seguinte, estudou-se a produção de hidromel em escala piloto a 18°C em reatores de polipropileno contendo 130 L de mosto sem suplementação (controle), suplementado com Enovit e suplementado com pedaços de maçãs (10% m/v). Terminada a fermentação, após 60 dias, o hidromel foi devidamente caracterizado físico-quimicamente quanto aos padrões de qualidade e identidade para hidromel estabelecidos pela Legislação Brasileira. Os resultados demonstraram que, com exceção da acidez total, todos os parâmetros avaliados se encontravam em consonância com os referidos padrões. Assim, as formulações estudadas foram adequadas para a obtenção de hidromel. Terminada a fermentação o hidromel foi submetido ao processo de envelhecimento a 20°C, por 216 dias, em tonel de carvalho (50L); galão de plástico (20 L); e frasco de vidro tipo bolha (20L), sendo neste período, caracterizado físico-químicamente, bem como quanto aos teores de compostos fenólicos totais, capacidade antioxidante, presença de aminas bioativas e compostos aromáticos derivados do tonel de carvalho. O hidromel envelhecido em galão de plástico apresentou teores de extrato seco reduzido abaixo do mínimo estabelecido nos padrões da legislação. A utilização de maçãs como suplemento do mosto de mel resultou em hidromel com maior teor de compostos fenólicos e maior capacidade antioxidante. As concentrações de aminas bioativas encontradas não comprometem a qualidade do hidromel, nem oferecem risco à saúde do consumidor. A análise de compostos fenólicos dos hidroméis envelhecidos em tonel de carvalho revelou que o processo de envelhecimento promoveu a extração de furfural, vanilina e ácido gálico, compostos que contribuem para o perfil sensorial da bebida, ficaram abaixo do limiar de percepção sensorial, fato que pode ser alterado pelo aumento do tempo de envelhecimento. Quanto a intenção de compra, os hidroméis avaliados apresentaram mais de 50% de notas variando entre 3 (tenho dúvidas se compraria) e 5 (certamente compraria), sendo os de melhor aceitação envelhecidos em recipientes de plástico e vidro. A análise sensorial revelou pouca influência do recipiente de envelhecimento sobre os atributos avaliados e a intensão de compra mostrou que os provadores preferiram os hidroméis envelhecidos em recipientes inertes (vidro e plástico). Diante do exposto, conclui-se que a produção de hidromel e seu posterior envelhecimento, por 216 dias, em recipientes inertes ou em tonéis de carvalho recondicionados, resultam em bebidas que apresentam boa aceitação, sendo uma alternativa viável para apicultores diversificarem a sua produção. / In the first stage of this work it was evaluated the fermentation performance of six commercial strains of yeast used in the of wine and beer production in honey wort 30 °Brix supplemented with 0.5 g / L peptone and 1 g / L yeast extract. In response, it was determined the yeast growth by number of cells and concentrations of glucose, fructose and ethanol by HPLC, as well as fermentation parameters as yield, efficiency and productivity in ethanol, and the production of hydrogen sulfide. Saccharomyces bayanus -Pasteur Champagne - Red Star showed better fermentation performance and it was evaluated in honey wort supplemented with different nutrients. The results showed that must supplementation with salts or commercial supplement Enovit contributes to the yeast performance. In the next step, it was studied the production of mead on a pilot scale at 18 ° C in polypropylene reactors containing 130 L of wort without supplementation (control), supplemented with Enovit and supplemented with apple pieces (10% m/v). By the end of fermentation, after 60 days, mead was properly characterized physico-chemically regarding the standards of quality and identity for mead, established by Brazilian legislation. The results showed that, with the exception of total acidity, all parameters were in accordance with the standards. After the fermentation, mead was subjected to aging process at 20 ° C by 216 days in oak barrel (50L) plastic gallon (20 L) and glass bottle (20L), and was characterized physico-chemically, as well as regarding the content of total phenolics, antioxidant capacity, presence of bioactive amines and aromatic compounds derived from the oak barrel. The results showed that the mead aged in plastic gallon presented content of \"reduced dried extract\" below the minimum patterns stated. It was also observed that the use of apples as the must supplement resulted in honey mead with a higher content of phenolic compounds and antioxidant capacity. The concentrations of bioactive amines found do not compromise the quality of mead, neither consumer health. The analysis of phenolic compounds in meads aged in oak barrel revealed that the aging process promoted the extraction of furfural, vanillin, gallic acid, compounds which contribute to the sensory profile of the beverage. Concentrations of these compounds were below the threshold of sensory perception, which can be changed by increasing aging time. Regarding purchase intent, all meads evaluated obtained more than 50% of positive scores, ranging between 3 (I doubt if buy) and 5 (definitely would buy), and meads aged in plastic and glass containers showed better acceptance. Sensory analysis showed little influence of the aging container on the attributes evaluated and the tasters preferred the meads aged in inert containers. So,it is concluded that the production of mead and its subsequent aging in inert containers or reconditioned oak casks, resulted in beverages that have good acceptance and are a viable alternative for beekeepers to diversify their products.

No presente trabalho, primeiramente avaliou-se o desempenho fermentativo de 10 cepas de leveduras isoladas de frutas, de mel centrifugado e de resíduo do processamento de mel após a etapa de sedimentação (borra do mel), em tubo de ensaio contendo mosto constituído de Mel 15 °Brix suplementado, individualmente, com diferentes nutrientes, a saber: extrato de levedura comercial (10,0 g/L), farelo de arroz (20,0 g/L), mistura (1,0 g/L) constituída de farelos de milho, arroz e soja na proporção 5:2:5 e nutriente comercial (D&R) (0,15 g/L). Os parâmetros avaliados coexistiram da determinação do número de células em câmara de Neubauer e das concentrações de açúcares e etanol por HPLC, o que permitiu a determinação dos parâmetros rendimento, eficiência de fermentação e produtividade. Estes resultados sustentaram a seleção do mosto suplementado com extrato de levedura comercial e da melhor cepa de levedura que foram posteriormente avaliados em escala piloto. A cepa selecionada foi a levedura isolada a partir da borra de mel, a qual foi posteriormente identificada como Saccharomyces cerevisiae e nomeada Saccharomyces cerevisiae EEL 2009. A avaliação da produção de aguardente de mel em escala piloto foi realizada nas instalações do Alambique da Associação Rural de Canas, Canas-SP e o destilado obtido armazenado em tonel de carvalho de 200 L por 6 meses. Amostras foram coletadas a cada 30 dias para caracterização físico-química em conformidade com os parâmetros estabelecidos na Legislação Brasileira para aguardente de frutas. Em paralelo, as respectivas amostras foram devidamente avaliadas sensorialmente por 120 provadores não treinados, por meio de testes de aceitação, em escala hedônica, considerando os quesitos aparência, aroma, sabor, corpo e impressão global, além da atitude de compra. Estes resultados foram analisados estatisticamente por ANOVA e teste de Tukey, sendo os resultados de aceitação em relação à impressão global, analisados por meio da análise multivariada o que permitiu traçar o Mapa de Preferência Interno - MDPREF. Os resultados referentes à caracterização físico-química das respectivas amostras demonstraram que todas apresentaram os parâmetros de avaliação em conformidade com os padrões estabelecidos pela legislação. Os resultados da análise sensorial revelaram que o tempo de armazenamento de 180 dias em tonel de carvalho não foi suficiente para a ocorrência de reações desejadas, o que influenciaria nas características sensoriais da bebida, tornado-a mais agradável e suave. No entanto, apesar do pouco tempo de armazenamento, a aguardente de mel apresentou uma boa aceitação por parte dos provadores, cuja maioria manifestou sua aceitação em termos de \"gostei ligeiramente\" e \"não gostei, nem desgostei\". No tocante ao MDPREF, este revelou que a amostra referente a 180 dias de armazenamento foi preferida por um maior número de provadores. Em relação à atitude de compra, as amostras armazenadas por 60 (79,17%) e 180 dias (75,83%) apresentaram os melhores resultados. Diante do exposto, conclui-se que o mel se apresenta como uma alternativa viável para a formulação de mosto para produção de aguardente no período de entre safra de cana-de-açúcar, contribuindo para melhor aproveitamento das instalações e como fonte alternativa de renda para o produtor rural. / In this work, first it was evaluated the fermentation performance of 10 yeast strains isolated from fruits, centrifuged honey and residue of honey processing after the sedimentation (sludge honey) step. This experiment was carried out in test tube containing wort made from honey 15 °Brix supplemented individually with different nutrients, namely: commercial yeast extract (10.0 g/L), rice bran (20.0 g/L), mixture (1.0 g/L) consisting of bran corn, rice and soy in the ratio 5:2:5 and commercial nutrient (D&R) (0.15 g/L). The parameters evaluated included the cell number determination in Neubauer chamber, and sugars and ethanol concentrations by HPLC, which allowed to determine the fermentation parameters as yield, fermentation efficiency and productivity. These results supported the selection of the wort supplemented with commercial yeast extract and the best yeast strain, which were subsequently evaluated in a pilot scale. The selected strain was the yeast isolated from the sludge honey, which was later identified as Saccharomyces cerevisiae and named Saccharomyces cerevisiae EEL 2009. The production evaluation of honey spirit on a pilot scale was conducted at the Canas Rural Association Pilot Plant for cachaça production - Canas-SP and distillate was stored in oak barrel of 200 liters per 6 months. Samples were collected every 30 days for physic-chemical characterization in accordance with the guidelines established in the Brazilian Legislation for fruit spirit. Beyond that, the respective samples were properly sensory evaluated by 120 untrained consumers regarding to acceptance testing employing a hedonic scale, considering characteristics as appearance, aroma, flavor, and overall impression, and the attitude of purchase, as well. These results were statistically analyzes by ANOVA and Tukey`s test and the results of acceptance regarding to the overall impression, were analyzed by multivariate analysis which allowed tracing the Internal Preference Map - MDPREF. The results concerning the physic-chemical characterization of the respective samples showed that all presented the evaluation parameters in accordance with standards established by the legislation. The results of sensory analysis revealed that the storage time of 180 days in oak barrel was not enough for the occurrence of desired reactions, which influence the sensory characteristics of the beverage, making it nicer and smoother. However, despite short time storage, honey spirit showed good acceptance by the consumers. Most of them expressed acceptance in terms of \"liked slightly\" and \"not liked nor disliked\". Regarding the MDPREF results, these proved that the sample relating to 180 days of storage was preferred by majority of consumers. Regarding the attitude of purchase, the samples stored for 60 days (79.17%) and 180 days (75.83%) showed the best results. Therefore, we conclude that honey can be considered as a viable alternative for wort formulation for the production of spirit, mainly in the period between harvests of sugar cane, contributing to better utilization of the facilities and as alternative source of income for farmers.

Na primeira etapa deste trabalho avaliou-se o desempenho fermentativo de 6 cepas de leveduras comerciais utilizadas na produção de vinhos e cerveja, em mosto de mel 30°Brix suplementado com 0,5 g/L de peptona e 1 g/L de extrato de levedura. Como resposta determinou-se o crescimento das leveduras em número de células e as concentrações de glicose, frutose e etanol por HPLC, que permitiram determinar os parâmetros rendimento, eficiência de fermentação e produtifidade em etanol, bem como a produção de sulfeto de hidrogênio. Assim, selecionou-se a cepa Saccharomyces bayanus - Pasteur Champagne - Red Star, que teve seu comportamento avaliado em mosto de mel suplementado com diferentes nutrientes. Os resultados demonstraram que a suplementação do mosto de mel com sais ou com suplemento comercial Enovit contribuiu para o bom desempenho da referida cepa. Na etapa seguinte, estudou-se a produção de hidromel em escala piloto a 18°C em reatores de polipropileno contendo 130 L de mosto sem suplementação (controle), suplementado com Enovit e suplementado com pedaços de maçãs (10% m/v). Terminada a fermentação, após 60 dias, o hidromel foi devidamente caracterizado físico-quimicamente quanto aos padrões de qualidade e identidade para hidromel estabelecidos pela Legislação Brasileira. Os resultados demonstraram que, com exceção da acidez total, todos os parâmetros avaliados se encontravam em consonância com os referidos padrões. Assim, as formulações estudadas foram adequadas para a obtenção de hidromel. Terminada a fermentação o hidromel foi submetido ao processo de envelhecimento a 20°C, por 216 dias, em tonel de carvalho (50L); galão de plástico (20 L); e frasco de vidro tipo bolha (20L), sendo neste período, caracterizado físico-químicamente, bem como quanto aos teores de compostos fenólicos totais, capacidade antioxidante, presença de aminas bioativas e compostos aromáticos derivados do tonel de carvalho. O hidromel envelhecido em galão de plástico apresentou teores de extrato seco reduzido abaixo do mínimo estabelecido nos padrões da legislação. A utilização de maçãs como suplemento do mosto de mel resultou em hidromel com maior teor de compostos fenólicos e maior capacidade antioxidante. As concentrações de aminas bioativas encontradas não comprometem a qualidade do hidromel, nem oferecem risco à saúde do consumidor. A análise de compostos fenólicos dos hidroméis envelhecidos em tonel de carvalho revelou que o processo de envelhecimento promoveu a extração de furfural, vanilina e ácido gálico, compostos que contribuem para o perfil sensorial da bebida, ficaram abaixo do limiar de percepção sensorial, fato que pode ser alterado pelo aumento do tempo de envelhecimento. Quanto a intenção de compra, os hidroméis avaliados apresentaram mais de 50% de notas variando entre 3 (tenho dúvidas se compraria) e 5 (certamente compraria), sendo os de melhor aceitação envelhecidos em recipientes de plástico e vidro. A análise sensorial revelou pouca influência do recipiente de envelhecimento sobre os atributos avaliados e a intensão de compra mostrou que os provadores preferiram os hidroméis envelhecidos em recipientes inertes (vidro e plástico). Diante do exposto, conclui-se que a produção de hidromel e seu posterior envelhecimento, por 216 dias, em recipientes inertes ou em tonéis de carvalho recondicionados, resultam em bebidas que apresentam boa aceitação, sendo uma alternativa viável para apicultores diversificarem a sua produção. / In the first stage of this work it was evaluated the fermentation performance of six commercial strains of yeast used in the of wine and beer production in honey wort 30 °Brix supplemented with 0.5 g / L peptone and 1 g / L yeast extract. In response, it was determined the yeast growth by number of cells and concentrations of glucose, fructose and ethanol by HPLC, as well as fermentation parameters as yield, efficiency and productivity in ethanol, and the production of hydrogen sulfide. Saccharomyces bayanus -Pasteur Champagne - Red Star showed better fermentation performance and it was evaluated in honey wort supplemented with different nutrients. The results showed that must supplementation with salts or commercial supplement Enovit contributes to the yeast performance. In the next step, it was studied the production of mead on a pilot scale at 18 ° C in polypropylene reactors containing 130 L of wort without supplementation (control), supplemented with Enovit and supplemented with apple pieces (10% m/v). By the end of fermentation, after 60 days, mead was properly characterized physico-chemically regarding the standards of quality and identity for mead, established by Brazilian legislation. The results showed that, with the exception of total acidity, all parameters were in accordance with the standards. After the fermentation, mead was subjected to aging process at 20 ° C by 216 days in oak barrel (50L) plastic gallon (20 L) and glass bottle (20L), and was characterized physico-chemically, as well as regarding the content of total phenolics, antioxidant capacity, presence of bioactive amines and aromatic compounds derived from the oak barrel. The results showed that the mead aged in plastic gallon presented content of \"reduced dried extract\" below the minimum patterns stated. It was also observed that the use of apples as the must supplement resulted in honey mead with a higher content of phenolic compounds and antioxidant capacity. The concentrations of bioactive amines found do not compromise the quality of mead, neither consumer health. The analysis of phenolic compounds in meads aged in oak barrel revealed that the aging process promoted the extraction of furfural, vanillin, gallic acid, compounds which contribute to the sensory profile of the beverage. Concentrations of these compounds were below the threshold of sensory perception, which can be changed by increasing aging time. Regarding purchase intent, all meads evaluated obtained more than 50% of positive scores, ranging between 3 (I doubt if buy) and 5 (definitely would buy), and meads aged in plastic and glass containers showed better acceptance. Sensory analysis showed little influence of the aging container on the attributes evaluated and the tasters preferred the meads aged in inert containers. So,it is concluded that the production of mead and its subsequent aging in inert containers or reconditioned oak casks, resulted in beverages that have good acceptance and are a viable alternative for beekeepers to diversify their products.

No presente trabalho, primeiramente avaliou-se o desempenho fermentativo de 10 cepas de leveduras isoladas de frutas, de mel centrifugado e de resíduo do processamento de mel após a etapa de sedimentação (borra do mel), em tubo de ensaio contendo mosto constituído de Mel 15 °Brix suplementado, individualmente, com diferentes nutrientes, a saber: extrato de levedura comercial (10,0 g/L), farelo de arroz (20,0 g/L), mistura (1,0 g/L) constituída de farelos de milho, arroz e soja na proporção 5:2:5 e nutriente comercial (D&R) (0,15 g/L). Os parâmetros avaliados coexistiram da determinação do número de células em câmara de Neubauer e das concentrações de açúcares e etanol por HPLC, o que permitiu a determinação dos parâmetros rendimento, eficiência de fermentação e produtividade. Estes resultados sustentaram a seleção do mosto suplementado com extrato de levedura comercial e da melhor cepa de levedura que foram posteriormente avaliados em escala piloto. A cepa selecionada foi a levedura isolada a partir da borra de mel, a qual foi posteriormente identificada como Saccharomyces cerevisiae e nomeada Saccharomyces cerevisiae EEL 2009. A avaliação da produção de aguardente de mel em escala piloto foi realizada nas instalações do Alambique da Associação Rural de Canas, Canas-SP e o destilado obtido armazenado em tonel de carvalho de 200 L por 6 meses. Amostras foram coletadas a cada 30 dias para caracterização físico-química em conformidade com os parâmetros estabelecidos na Legislação Brasileira para aguardente de frutas. Em paralelo, as respectivas amostras foram devidamente avaliadas sensorialmente por 120 provadores não treinados, por meio de testes de aceitação, em escala hedônica, considerando os quesitos aparência, aroma, sabor, corpo e impressão global, além da atitude de compra. Estes resultados foram analisados estatisticamente por ANOVA e teste de Tukey, sendo os resultados de aceitação em relação à impressão global, analisados por meio da análise multivariada o que permitiu traçar o Mapa de Preferência Interno - MDPREF. Os resultados referentes à caracterização físico-química das respectivas amostras demonstraram que todas apresentaram os parâmetros de avaliação em conformidade com os padrões estabelecidos pela legislação. Os resultados da análise sensorial revelaram que o tempo de armazenamento de 180 dias em tonel de carvalho não foi suficiente para a ocorrência de reações desejadas, o que influenciaria nas características sensoriais da bebida, tornado-a mais agradável e suave. No entanto, apesar do pouco tempo de armazenamento, a aguardente de mel apresentou uma boa aceitação por parte dos provadores, cuja maioria manifestou sua aceitação em termos de \"gostei ligeiramente\" e \"não gostei, nem desgostei\". No tocante ao MDPREF, este revelou que a amostra referente a 180 dias de armazenamento foi preferida por um maior número de provadores. Em relação à atitude de compra, as amostras armazenadas por 60 (79,17%) e 180 dias (75,83%) apresentaram os melhores resultados. Diante do exposto, conclui-se que o mel se apresenta como uma alternativa viável para a formulação de mosto para produção de aguardente no período de entre safra de cana-de-açúcar, contribuindo para melhor aproveitamento das instalações e como fonte alternativa de renda para o produtor rural. / In this work, first it was evaluated the fermentation performance of 10 yeast strains isolated from fruits, centrifuged honey and residue of honey processing after the sedimentation (sludge honey) step. This experiment was carried out in test tube containing wort made from honey 15 °Brix supplemented individually with different nutrients, namely: commercial yeast extract (10.0 g/L), rice bran (20.0 g/L), mixture (1.0 g/L) consisting of bran corn, rice and soy in the ratio 5:2:5 and commercial nutrient (D&R) (0.15 g/L). The parameters evaluated included the cell number determination in Neubauer chamber, and sugars and ethanol concentrations by HPLC, which allowed to determine the fermentation parameters as yield, fermentation efficiency and productivity. These results supported the selection of the wort supplemented with commercial yeast extract and the best yeast strain, which were subsequently evaluated in a pilot scale. The selected strain was the yeast isolated from the sludge honey, which was later identified as Saccharomyces cerevisiae and named Saccharomyces cerevisiae EEL 2009. The production evaluation of honey spirit on a pilot scale was conducted at the Canas Rural Association Pilot Plant for cachaça production - Canas-SP and distillate was stored in oak barrel of 200 liters per 6 months. Samples were collected every 30 days for physic-chemical characterization in accordance with the guidelines established in the Brazilian Legislation for fruit spirit. Beyond that, the respective samples were properly sensory evaluated by 120 untrained consumers regarding to acceptance testing employing a hedonic scale, considering characteristics as appearance, aroma, flavor, and overall impression, and the attitude of purchase, as well. These results were statistically analyzes by ANOVA and Tukey`s test and the results of acceptance regarding to the overall impression, were analyzed by multivariate analysis which allowed tracing the Internal Preference Map - MDPREF. The results concerning the physic-chemical characterization of the respective samples showed that all presented the evaluation parameters in accordance with standards established by the legislation. The results of sensory analysis revealed that the storage time of 180 days in oak barrel was not enough for the occurrence of desired reactions, which influence the sensory characteristics of the beverage, making it nicer and smoother. However, despite short time storage, honey spirit showed good acceptance by the consumers. Most of them expressed acceptance in terms of \"liked slightly\" and \"not liked nor disliked\". Regarding the MDPREF results, these proved that the sample relating to 180 days of storage was preferred by majority of consumers. Regarding the attitude of purchase, the samples stored for 60 days (79.17%) and 180 days (75.83%) showed the best results. Therefore, we conclude that honey can be considered as a viable alternative for wort formulation for the production of spirit, mainly in the period between harvests of sugar cane, contributing to better utilization of the facilities and as alternative source of income for farmers.

Sugar (primarily sucrose) has been a part of the daily diet for literally hundreds of years, but research is now suggesting that sugar intake can be detrimental to our health. In particular, excessive consumption of simple sugars with high glycemic index (GI) values have been shown to cause overeating and weight gain. As well, elevated postprandial hyperglycemia can result after consuming sugars and this has been linked to disease formation and progression, the development of advanced glycation endproducts, inflammation and increased mortality rates. Honey has been recognised as having a number of beneficial health properties, including slower uptake into the bloodstream, a pharmacological action of reducing blood glucose levels and a high level of bioavailable antioxidants, all of which may mean that honey could be less harmful to health than sucrose in the diet. This study was therefore designed to investigate the health benefits of honey in the diet as a replacement for sucrose, using small animal studies. As well, because of the interest in using honey as a replacement for sucrose in sweetened dairy foods, a small number of in vitro investigations were carried out to determine whether honey could retain its bioactive properties when combined with milk/dairy products. Using the in vitro studies, it was shown that the combination of milk with honey had no effect on either the antibacterial or antioxidant capabilities of honey. During the animal feeding studies a number of significant findings were observed. In the earlier work it was shown that honey had a significant effect on protein metabolism when fed for 14 days at a level of 600 g/kg diet (comprising 480 g sugars and 120 g water) compared with animals fed an equivalent amount of sucrose. In this study, honey-fed rats exhibited significantly lower weight gains (p less than 0.001), food intake (p less than 0.05) and nitrogen intakes (p less than 0.05) and significantly higher faecal nitrogen outputs (p less than 0.05) compared with sucrose-fed rats. Animals fed a diet consisting of 480 g/kg of mixed sugars as in honey generally exhibited protein metabolism parameters that were comparable to those of the sucrose-fed rats, suggesting that the effects of honey on protein metabolism were not due solely to its distinctive sugar composition. Furthermore, in another study that specifically investigated the effects of honey on weight regulation, honey (100 g/kg diet) resulted in significantly reduced weight gain after 6 weeks (p less than 0.01) compared with animals fed the same amount of sugars as sucrose, although food intake was not reduced in this study. Percentage weight gains were shown to be comparable between honey-fed rats and those fed a sugar-free diet, suggesting that differences in glycemic control may be partly responsible for the results seen. Fasting lipid profiles and blood glucose levels were also measured in this study, but no significant differences were observed between diet groups. During long-term (12 months) feeding weight gain was again significantly reduced in rats fed honey (p less than 0.05) and a sugar-free diet (p less than 0.01) compared with those fed sucrose, the weights of honey-fed rats and those fed the sugar-free diet being comparable at the end of the study. In addition, blood glucose levels were significantly lower (p less than 0.001), and HDL-cholesterol levels significantly higher (p less than 0.05) in animals fed honey compared with those fed sucrose after 52 weeks, but no differences in these parameters were observed between rats fed sucrose and a sugar-free diet. No other significant differences in lipid profiles were observed. Immunity measures were improved after feeding honey or sucrose for 52 weeks, animals in both of these diet groups having significantly higher levels of neutrophil phagocytosis compared with those fed the sugar-free diet (both p less than 0.0001). In addition, the percentage of leukocytes that were lymphocytes was significantly higher in honey-fed rats at the end of the study. Furthermore, levels of oxidative damage in aortic collagen were significantly reduced in rats fed honey or the sugar-free diet (both p less than 0.05) compared with those fed sucrose after 52 weeks. Full body DEXA scans were also undertaken in this 12-month study to assess body fat levels and bone mineral composition and density, although they revealed few statistically significant differences. Percentage body fat levels were shown to be nearly 10% lower in honey-fed rats compared with sucrose-fed animals at the end of the study (p less than 0.05), but no other significant differences between diet groups were observed. With one exception, no differences in bone mineral composition or bone mineral density were observed between the three diet groups after 52 weeks. This data agreed with the results generated from two earlier studies that showed that feeding honey short-term (for 6-8 weeks) to rats that were either calcium-deficient or fed a low calcium diet had no effect on bone calcium levels, bone mineral content, bone mineral density or bone breaking parameters. Lastly, long-term feeding of honey to rats had a number of statistically significant effects on anxiety and cognitive performance when assessed using animal maze tasks. Anxiety-like behaviour was significantly reduced in honey-fed rats overall compared with those fed sucrose (p = 0.056) or a sugar free diet (p less than 0.05). Spatial memory was also better in honey fed-rats throughout the 12 month study, these animals not displaying the same degree of age-related spatial memory loss seen in the other two diet groups. No significant differences in recognition memory or learning capability were observed between diet groups after 52 weeks. In conclusion, both short-term and long-term feeding of honey result in a number of health benefits compared with eating similar amounts of sucrose. These include less weight gain, improved immunity, reduced levels of oxidative damage and improved cognitive performance.. These effects of honey are likely to occur through a number of different processes, although the presence of high concentrations of antioxidants and other minor components in honey are likely to be important contributors. Honey may therefore help to improve human heath if it is used as an alternative to sucrose in foods and beverages, although feeding studies in humans are required to assess its efficacy. In addition, more animal studies are needed to assess which features of honey (e.g. fructose content, antioxidant content and bioactivities) are required to achieve optimal effects, and to determine what impact heating and food processing may have on the beneficial health effects of honey.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

The main objective of this study was to investigate if natural honey can be used as a cryoprotecting agent (CP) in vitrification medium to improve the viability of vitrified-warmed bovine oocytes. The first study was conducted to investigate the dehydration capability of natural honey compared with sucrose, and to determine the proper concentration of honey-based medium and the optimum time for sufficiently safe dehydration of bovine oocytes. Matured cumulus-oocyte complexs (COCs) were denuded and introduced individually into different concentrations (0.25, 0.5, 1.0, 1.5 or 2.0 M) of honey and sucrose-based medium followed by rehydration in control media (TCM). Video images were recorded during dehydration and rehydration, and oocyte images were captured at 12 time intervals to calculate oocyte-volume changes during dehydration and rehydration. Results demonstrated that, in honey-based media, the maximum oocyte shrinkage was achieved after 60 sec exposure in 0.25M, 0.5M and 1.0M concentrations; while at higher concentrations 1.5M and 2.0M, the maximum dehydration occurred at 30 and 20 seconds respectively. In sucrose-based medium, the maximum oocyte shrinkage was achieved after 60 sec exposure in 0.25 or 0.5M concentrations. However, at higher concentrations (1M, 1.5M or 2M), the maximum dehydration occurred at 30, 20 and 10 sec. For rehydration, oocytes dehydrated in honey or sucrose-based medium were able to regain their original volume within 60-120 sec. However, oocytes dehydrated in higher concentrations (2M honey, and 1.5M and 2M sucrose) were rehydrated back to their original volume within 20 sec. This study concluded that natural honey and sucrose caused similar cell dehydration. Only oocytes dehydrated in 1M honey-based media reached maximal dehydration after 60 sec and equally regained original volume. Therefore, 1M of honey-based medium is suggested for sufficient and safe oocyte dehydration during vitrification. The second study was conducted to determine in vitro maturation (IVM), in vitro fertilization (IVF) and embryonic development of bovine oocytes vitrified in honey-based vitrifcation media. In Experiment 1, bovine COCs were randomly distributed in control group (non-vitrified; G1), 0.5M sucrose group (second control; G2), and 0.5M, 1M and 1.5M honey groups (G3, G4 and G5 respectively). The COCs were exposed to equilibration solution 1 (VS1) at ~ 22 oC for 5 min and to vitrification solution 2 (VS2) for 1 min, mounted on Cryotops and plunged into LN2. COCs were warmed in TCM and honey/sucrose medium at 38.5oC for 1 min, washed, matured in vitro (IVM), denuded, and immunostained to evaluate maturation. Maturation rate was significantly higher (80.7%) in control group (G1) than in vitrified groups (56, 52, 55 and 51% in G2, G3, G4 and G5, respectively) (P<.0001), whereas there was no significant difference among the vitrified groups (P>0.05). In Experiment 2, bovine COCs distributed in control (not vitrified, G1) and vitrified groups using 1M honey and 0.5M sucrose (G2 and G3 respectively), underwent for IVM, IVF and in vitro culture (IVC) for 9 days. Cleavage rate was significantly higher (P<.0001) in the control group (74%, G1, n=183) than rates of vitrified groups (51% in G2, n=137; and 42% in G3, n=131), whereas no differences among vitrified groups (P=0.0723). Rate of blastocyst formation was significantly higher (34%) in G1 than in the vitrified groups (P<.0001); however, blastocyst formation rates in the honey group were significantly higher (P=0.0026) than in the sucrose group (13% and 3% respectively). Addition of natural honey (1.0M; or 21.7%w/v) in vitrification medium can safely and sufficiently dehydrate bovine oocytes during vitrification procedure. The vitrification of bovine oocytes in 1M honey improved their post-warming maturation abtility and embryonic development.

To enhance the scientific utility and practical application of the honey bee genome and assign the linkage groups to specific chromosomes, I identified chromosomes and characterized the karyotype of the sequenced strain DH4 of the honey bee. The primary analysis of the karyotype and ideogram construction was based on banding and Fluorescence In Situ Hybridization (FISH) for rDNA detection. FISH confirmed two locations for the NOR on telomeric regions of chromosomes 6 and 12 plus an additional less frequent signal on chromosome 1, all three of which were confirmed with silver staining (AgNO3). 4’6-diamidino-2phenylindole (DAPI), and CBanding methods were used to construct the primary ideograms that served as a basis to further identify the chromosomes and locate important structures. The primary map was compared with Giemsa banding, AgNO3-banding, Trypsin banding, and R-banding. The karyotype of the honey bee was established as two metacentric chromosomes (1 and 10), two submetacentric with ribosomal organizer (6 and 12), four submetacentric heterochromatic chromosomes (16, 15, 4 and 13), four euchromatic subtelocentric chromosomes (2, 8, 11 and 14) and four acrocentric chromosomes (3, 5, 7 and 9). In situ nick-translation banding methods were used to verify the heterochromatin distribution. The cytogenetic maps of the honey bee karyotype represented in the ideograms were subsequently used to place 35 mapped BACs (Solignac et. al. 2004) of Solignac’s BAC library. As the BACs hybridized to multiple sites, the mapping was based on strength and frequency of the signals. Location and position of the BACs was compared with those published in the different version of Map Viewer of the NCBI and BeeBase web sites. 10 BACs were confirmed with the last version of Map Viewer V4, 12 BACs were mapped based on high frequency and agreement with the earlier version of Map Viewer. 14 BACs were mapped as confirmed based on moderate frequency of the signal and agreement with the last version of MVV, most of these BACs hits as a secondary signal.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.