Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-04-08T15:04:34Z No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-11T11:10:59Z (GMT) No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-04-11T11:10:59Z (GMT). No. of bitstreams: 2 Tese - Sandra Regina Marcoline Gherardi - 2014.pdf: 1563773 bytes, checksum: 3c2b927eb08946b0b340ab1b4f2018fd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-10-14 / In this work we aimed to evaluate the effect of time and temperature of storage on the quality, chemical composition and functional properties of white and brown eggs from commercial hens aged 50 and 30 weeks, respectively. We conducted two experiments, and in each experiment 240 eggs, white (exp. 1) and brown (Exp. 2) eggs were randomly selected, with an average weight between 55 and 65g stored in natural environment and refrigerated for a period of 28 days. A completely randomized design was adopted in a factorial arrangement with two temperatures (ambient and refrigerated)and five storage periods (1, 7, 14, 21 and 28 days) totaling 10 treatments with six replications and four eggs per experimental unit. The quality parameters evaluated were weight of egg, yolk, albumen and shell, yolk index, Haugh Unit, percentage of eggshell, albumen and yolk and pH of albumen and yolk. In the chemical composition analysis of albumen and yolk, we determined moisture, total lipids, protein, total solids and ash contents. The functional properties studied were foaming, foam stability, emulsion formation, time necessary to destabilize the emulsion and yolk color index. The storage time influenced negatively the parameters for both white and brown eggs with more marked influence on eggs stored under ambient condition. In case of changes, they were more intense in the first seven days of storage regardless of storage condition. The refrigerated conditions improved the stability of foams, decreased the amount of oil required to form emulsion and intensified the yolk color index. Storage under refrigeration guarantees the quality, chemical composition and functional properties of white and brown eggs, during 28-day study, extending the shelf life and hence the use by food industries. / Objetivou-se com este trabalho avaliar a influência do tempo e temperatura de estocagem sobre a qualidade, composição química e propriedades funcionais de ovos brancos e marrons oriundos de aves comerciais com 50 e 30 semanas de idade respectivamente. Foram conduzidos dois experimentos e em cada avaliação, foram selecionados 240 ovos, brancos (Exp. 1) e marrons (Exp. 2), ao acaso, com peso médio entre 55 e 65g sendo armazenados em ambiente natural e sob refrigeração por um período de 28 dias. Foi adotado delineamento inteiramente casualizado, em esquema fatorial, composto de duas temperaturas (ambiente e refrigeração) e cinco períodos de estocagem (1, 7, 14, 21 e 28 dias) totalizando 10 tratamentos com seis repetições e quatro ovos por unidade experimental. Os parâmetros de qualidade avaliados foram peso do ovo, gema, albúmen e casca, índice de gema, Unidade Haugh, porcentagem de casca, albúmen e gema e pH de gema e albúmen. Para determinação da composição química foram determinados os teores de umidade, lipídios totais, proteína bruta, sólidos totais e cinzas tanto de albúmen quanto da gema. As propriedades funcionais estudadas foram capacidade de formação de espuma, estabilidade da espuma formada, capacidade de formação de emulsão, determinação do início da desestabilização da emulsão e índice de coloração da gema. O tempo de armazenamento influenciou negativamente sobre os parâmetros avaliados, tanto para ovos brancos quanto marrons, apresentando influência mais marcante sobre ovos mantidos sob condição ambiente. As modificações quando ocorreram, foram mais intensas nos primeiros dias de estocagem independentemente da condição de armazenagem. A refrigeração melhorou a estabilidade das espumas, diminuiu o volume de óleo necessário para formar emulsão e intensificou o índice de coloração das gemas. O armazenamento sob refrigeração garantiu a manutenção da qualidade, composição química e propriedades funcionais durante os 28 dias do estudo para ovos brancos e marrons, ampliando a vida de prateleira e consequentemente o uso pelas indústrias alimentícias.

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The objective of this research was to develop new sample preparation procedures for the isolation of sulfonamides, as well as, to determine the applicability of employing on-line nitrogen selective and mass spectrometric detection methods. The first phase of this research investigated the effect of temperature and pressure on the supercritical fluid extraction (SFE) of sulfonamides from a spiked sand matrix. Temperature effects were either positive or negative with respect to extraction rate and total recovery, depending on the pressure and extraction fluid employed. The second portion of this research compared trifluoromethane (CHF3) and carbon dioxide (CO2) as fluids for the extraction of sulfonamides from spiked non-fat dry milk, beef liver, and egg yolk were found to be more selective using CHF3 than CO2. The polar trifluoromethane improved the extraction efficiency of the polar sulfonamides from the biological matrices and also reduced the amount of co-extractives. The next phase of this research considered the effect of organic modifier and CO2 in the SFE of sulfonamides from chicken liver, beef liver and egg yolk. Methanol, ethanol, acetone, acetonitrile were compared to determine optimum conditions. A SFE method employing 20% acetonitrile modified CO2 yielded quantitative recovery of sulfonamides from chicken liver, but 20% acetone modified CO2 was required to obtain quantitative recovery from beef liver. Either 20% acetone or 20% acetonitrile yielded quantitative recovery from egg yolk. The last phase of this research focused on the evaluation of selective detection methods for sulfonamide analysis. Chemiluminescence nitrogen detection (CLND) parameters were optimized for use with packed column supercritical fluid chromatography (SFC) yielding a minimum detectable quantity (MDQ) of 5 ng of sulfamethazine, on column. Improvements in the detector design decreased the MDQ to 0.5 ng, while, decreasing the column diameter further reduced the MDQ to 125 pg. The second part of this phase evaluated PLC/Atmospheric pressure chemical ionization (APCI) mass spectrometry for the detection of sulfonamides. Sensitivity in selective ion mode was found to be as low as 50 pg on column for sulfamethazine. Supercritical fluid extracts of sulfonamides spiked at 100μg/kg in chicken liver were found to be readily detected by this method. / Ph. D.

Le lait écrémé est utilisé depuis plus d’un demi-siècle comme diluant protecteur des spermatozoïdes de mammifères. Depuis quelques années, il existe une demande grandissante pour des diluants exempts de produits d’origine animale. Toutefois, le mécanisme par lequel le lait protège les spermatozoïdes n’est pas connu, ce qui rend difficile de lui trouver un substitut. Les protéines majeures du plasma séminal de taureau, les protéines « Binder of SPerm » (BSP), sont néfastes lors de la conservation de la semence. Les spermatozoïdes sont en contact avec une grande concentration de protéines BSP qui stimulent une extraction continuelle de cholestérol/phospholipides de leur membrane plasmique. Les lipoprotéines de faible densité (LDL) du jaune d’oeuf, un autre composé utilisé dans les diluants, empêcheraient les protéines BSP de se lier à la membrane des spermatozoïdes de taureaux et de stimuler un efflux des lipides membranaires, ce qui les protégerait durant la conservation. Notre hypothèse était que les protéines du lait protègent les spermatozoïdes durant la conservation en séquestrant les protéines BSP. Premièrement, nous avons démontré par filtration sur gel qu’il y a une interaction entre les protéines BSP bovines et les protéines du lait. Le lait écrémé a été fractionné en trois fractions : F1 (alpha-lactalbumine, bêta-lactoglobuline et caséine kappa), F2 (toutes les protéines du lait) et F3 (sels, sucres et petits peptides). Les protéines BSP1 et BSP5 ont une affinité plus grande pour F1 que BSP3, tandis que toutes les protéines BSP ont une affinité pour F2. Le titrage calorimétrique isotherme a permis de confirmer l’interaction entre les protéines BSP et les protéines du lait. L’association entre la protéine BSP1 bovine et les micelles de caséines est caractérisée par une constante d’affinité (Ka) de 3.5 × 10^5 M-1 et un paramètre stoichiométrique (n) de 4,5 BSP1 pour une caséine. L’association entre la protéine BSP1 bovine et l’alpha-lactalbumine (une protéine du sérum principale), est caractérisée par un Ka de 2.4 × 10^5 M-1 et une valeur “n” de 0,8. Ces résultats indiquent que le lait protège les spermatozoïdes bovins en séquestrant les protéines BSP grâce à une interaction protéine : protéine, tandis que le jaune d’oeuf les protège grâce à une interaction protéine : lipoprotéine. Deuxièmement, nous avons démontré par filtration sur gel que les protéines homologues aux BSP bovines retrouvées dans le plasma séminal de porc, d’étalon et de bélier ont une affinité avec les protéines du lait, ce qui suggère que le mécanisme de protection des spermatozoïdes par le lait pourrait être le même chez ces espèces. Troisièmement, nous avons caractérisé l’interaction entre BSP1 bovine et les LDL du jaune d’oeuf qui a un Ka de 3.4 ± 0.4 × 10^6 M-1 et une valeur de « n » de 104 BSP1 pour une particule de LDL, indiquant qu’il existe des différences entre le mécanisme de protection des spermatozoïdes par le lait et le jaune d’oeuf. Nous croyons que les résultats présentés dans cette thèse aideront à créer de nouveaux diluants ne contenant pas de produits d’origine animale afin de cryoconserver les spermatozoïdes des mammifères. / Skim milk is being used as a protective agent for mammalian semen conservation over half a century. Recently, there has been increased interest in developing extenders free of animal products. However, it is difficult to find suitable component in order to replace milk as an extender, because the mechanisms by which milk protect sperm against cooling and freezing damages during the storage is unknown. The Binder of SPerm (BSP) proteins are the major proteins of bull seminal plasma and they are harmful during sperm storage. In fact, sperm would be in contact with a large quantity of BSP proteins that induce a continuous cholesterol and phospholipids efflux from the sperm membrane during storage. When bull sperm is diluted with an extender containing egg yolk, another compound frequently used in extender, the low-density lipoproteins (LDL) present in the egg yolk prevent the binding of the BSP proteins to the sperm membrane, thus, preventing the lipid efflux from the sperm membrane induced by the BSP proteins. Our hypothesis was that milk proteins would protect sperm during storage by binding BSP proteins. First, we demonstrated by gel filtration that bovine BSP proteins could bind the milk proteins. Skim milk was fractionated into three fractions: F1 (alpha-lactalbumin and beta- lactoglobulin, the major whey proteins and kappa-casein), F2 (mainly caseins and all other milk proteins in small amounts) and F3 (salts, sugars and small peptides). Bovine BSP1 and BSP5 have more affinity for F1 as compared to BSP3 and all the BSP proteins have affinity for F2. We confirmed the interaction between bovine BSP proteins and milk proteins by isothermal titration calorimetry. The binding of BSP1 to casein micelles is characterized by an affinity constant (Ka) of 3.5 × 10^5 M-1 and of a stoichiometric parameter for the association (n) of 4.5 BSP1 per casein. The association between BSP1 and alpha-lactalbumin (one of the major whey proteins) is characterized by a Ka of 2.4 × 10^5 M-1 and a “n” value of 0.8. These results support our contention that milk can protect sperm by preventing the BSP proteins’ binding to the sperm membrane attributable to a protein : protein interaction, while egg yolk sperm protection is attributable to a protein : lipoprotein interaction. Second, our studies showed that the homologous BSP proteins found in the boar, stallion and ram seminal plasma can bind the milk proteins. These results indicate that the mechanism of sperm protection by milk in these species should be similar to the one in bovine species. Third, we characterized the interaction between bovine BSP1 protein and LDL from hen’s egg yolk. The binding was characterized by a Ka of 3.4 ± 0.4 × 10^6 M-1 and a « n » value of 104 BSP1 per LDL particle. Our results indicated that there is difference between the mechanism of sperm protection by milk and egg yolk. We believe that the results presented in this thesis may help to create new extenders free of animal product for mammal sperm preservation in liquid or frozen state.

Le lait écrémé est utilisé depuis plus d’un demi-siècle comme diluant protecteur des spermatozoïdes de mammifères. Depuis quelques années, il existe une demande grandissante pour des diluants exempts de produits d’origine animale. Toutefois, le mécanisme par lequel le lait protège les spermatozoïdes n’est pas connu, ce qui rend difficile de lui trouver un substitut. Les protéines majeures du plasma séminal de taureau, les protéines « Binder of SPerm » (BSP), sont néfastes lors de la conservation de la semence. Les spermatozoïdes sont en contact avec une grande concentration de protéines BSP qui stimulent une extraction continuelle de cholestérol/phospholipides de leur membrane plasmique. Les lipoprotéines de faible densité (LDL) du jaune d’oeuf, un autre composé utilisé dans les diluants, empêcheraient les protéines BSP de se lier à la membrane des spermatozoïdes de taureaux et de stimuler un efflux des lipides membranaires, ce qui les protégerait durant la conservation. Notre hypothèse était que les protéines du lait protègent les spermatozoïdes durant la conservation en séquestrant les protéines BSP. Premièrement, nous avons démontré par filtration sur gel qu’il y a une interaction entre les protéines BSP bovines et les protéines du lait. Le lait écrémé a été fractionné en trois fractions : F1 (alpha-lactalbumine, bêta-lactoglobuline et caséine kappa), F2 (toutes les protéines du lait) et F3 (sels, sucres et petits peptides). Les protéines BSP1 et BSP5 ont une affinité plus grande pour F1 que BSP3, tandis que toutes les protéines BSP ont une affinité pour F2. Le titrage calorimétrique isotherme a permis de confirmer l’interaction entre les protéines BSP et les protéines du lait. L’association entre la protéine BSP1 bovine et les micelles de caséines est caractérisée par une constante d’affinité (Ka) de 3.5 × 10^5 M-1 et un paramètre stoichiométrique (n) de 4,5 BSP1 pour une caséine. L’association entre la protéine BSP1 bovine et l’alpha-lactalbumine (une protéine du sérum principale), est caractérisée par un Ka de 2.4 × 10^5 M-1 et une valeur “n” de 0,8. Ces résultats indiquent que le lait protège les spermatozoïdes bovins en séquestrant les protéines BSP grâce à une interaction protéine : protéine, tandis que le jaune d’oeuf les protège grâce à une interaction protéine : lipoprotéine. Deuxièmement, nous avons démontré par filtration sur gel que les protéines homologues aux BSP bovines retrouvées dans le plasma séminal de porc, d’étalon et de bélier ont une affinité avec les protéines du lait, ce qui suggère que le mécanisme de protection des spermatozoïdes par le lait pourrait être le même chez ces espèces. Troisièmement, nous avons caractérisé l’interaction entre BSP1 bovine et les LDL du jaune d’oeuf qui a un Ka de 3.4 ± 0.4 × 10^6 M-1 et une valeur de « n » de 104 BSP1 pour une particule de LDL, indiquant qu’il existe des différences entre le mécanisme de protection des spermatozoïdes par le lait et le jaune d’oeuf. Nous croyons que les résultats présentés dans cette thèse aideront à créer de nouveaux diluants ne contenant pas de produits d’origine animale afin de cryoconserver les spermatozoïdes des mammifères. / Skim milk is being used as a protective agent for mammalian semen conservation over half a century. Recently, there has been increased interest in developing extenders free of animal products. However, it is difficult to find suitable component in order to replace milk as an extender, because the mechanisms by which milk protect sperm against cooling and freezing damages during the storage is unknown. The Binder of SPerm (BSP) proteins are the major proteins of bull seminal plasma and they are harmful during sperm storage. In fact, sperm would be in contact with a large quantity of BSP proteins that induce a continuous cholesterol and phospholipids efflux from the sperm membrane during storage. When bull sperm is diluted with an extender containing egg yolk, another compound frequently used in extender, the low-density lipoproteins (LDL) present in the egg yolk prevent the binding of the BSP proteins to the sperm membrane, thus, preventing the lipid efflux from the sperm membrane induced by the BSP proteins. Our hypothesis was that milk proteins would protect sperm during storage by binding BSP proteins. First, we demonstrated by gel filtration that bovine BSP proteins could bind the milk proteins. Skim milk was fractionated into three fractions: F1 (alpha-lactalbumin and beta- lactoglobulin, the major whey proteins and kappa-casein), F2 (mainly caseins and all other milk proteins in small amounts) and F3 (salts, sugars and small peptides). Bovine BSP1 and BSP5 have more affinity for F1 as compared to BSP3 and all the BSP proteins have affinity for F2. We confirmed the interaction between bovine BSP proteins and milk proteins by isothermal titration calorimetry. The binding of BSP1 to casein micelles is characterized by an affinity constant (Ka) of 3.5 × 10^5 M-1 and of a stoichiometric parameter for the association (n) of 4.5 BSP1 per casein. The association between BSP1 and alpha-lactalbumin (one of the major whey proteins) is characterized by a Ka of 2.4 × 10^5 M-1 and a “n” value of 0.8. These results support our contention that milk can protect sperm by preventing the BSP proteins’ binding to the sperm membrane attributable to a protein : protein interaction, while egg yolk sperm protection is attributable to a protein : lipoprotein interaction. Second, our studies showed that the homologous BSP proteins found in the boar, stallion and ram seminal plasma can bind the milk proteins. These results indicate that the mechanism of sperm protection by milk in these species should be similar to the one in bovine species. Third, we characterized the interaction between bovine BSP1 protein and LDL from hen’s egg yolk. The binding was characterized by a Ka of 3.4 ± 0.4 × 10^6 M-1 and a « n » value of 104 BSP1 per LDL particle. Our results indicated that there is difference between the mechanism of sperm protection by milk and egg yolk. We believe that the results presented in this thesis may help to create new extenders free of animal product for mammal sperm preservation in liquid or frozen state.

Made available in DSpace on 2016-06-02T19:06:05Z (GMT). No. of bitstreams: 1 5287.pdf: 3924606 bytes, checksum: 935b5a09df26eb1b41df901a189a6e2a (MD5) Previous issue date: 2013-05-29 / Universidade Federal de Sao Carlos / Geographic information systems with the aid of spatial databases store and manage crisp spatial data (or exact spatial data), whose shapes (boundaries) are well defined and have a precise location in space. However, several spatial data do not have precisely known boundaries or have an uncertain location in space, which are called vague spatial data. The boundaries of a given vague spatial data may shrink or extend, therefore, may have a minimum and maximum extension. Clouds of pollution, deforestation, fire outbreaks, route of an airplane, habitats of plants and animals are examples of vague spatial data. In the literature, there are currently vague spatial data models, such as Egg-Yolk, QMM and VASA. However, according to our knowledge, they focus only on the formal aspect of the model definition. Thus, real or synthetic vague spatial data is not available for use. The main objective of this master thesis is the development of algorithms for the generation of synthetic vague spatial data based on the crisp models of spatial data vague Egg-Yolk, QMM and VASA. It was also implemented a tool, called VagueDataGeneration, to assist in the process of generation such data. For both the algorithms and the tool, the user is able to set the properties related to the data type of model, such as size, shape, volume, complexity, location and spatial distribution. By using the proposed algorithms and the VagueDataGeneration tool, researchers can generate large samples of vague spatial data, enabling new research, such as testing indexes for vague spatial data or evaluating query processing over data warehouses that store vague spatial data. The validation of the vague spatial data generation was conducted using a case study with data from vague rural phenomena. / Sistemas de informação geográfica com o auxílio de bancos de dados espaciais armazenam e gerenciam dados espaciais exatos, cujas formas (fronteiras) são bem definidas e que possuem uma localização exata no espaço. Entretanto, vários dados espaciais reais não possuem os seus limites precisamente conhecidos ou possuem uma localização incerta no espaço, os quais são denominados dados espaciais vagos. Os limites de um dado espacial vago podem encolher ou estender, portanto, podem ter uma extensão mínima e máxima. Nuvens de poluição, desmatamentos, focos de incêndios, rota de um avião, habitats de plantas e de animais são exemplos de dados espaciais vagos. Na literatura, atualmente existem modelos de dados espaciais vagos, tais como Egg-Yolk, QMM e VASA. No entanto, segundo o nosso conhecimento, estes enfocam apenas no aspecto formal da definição do modelo. Com isso, dados espaciais vagos reais ou sintéticos não estão disponíveis para uso. O principal objetivo deste trabalho de mestrado consiste no desenvolvimento de algoritmos para a geração de dados espaciais vagos sintéticos baseados nos modelos exatos de dados espaciais vagos Egg-Yolk, QMM e VASA. Também foi implementada uma ferramenta, chamada VagueDataGeneration, para auxiliar no processo de geração desses dados. Nos algoritmos propostos e na ferramenta desenvolvida, o usuário define as propriedades referentes ao tipo de dado de um modelo, tais como tamanho, formato, volume, complexidade, localização e distribuição espacial dos dados espaciais vagos a serem gerados. Por meio do uso dos algoritmos propostos e da ferramenta VagueDataGeneration, os pesquisadores podem gerar grandes amostras de dados espaciais vagos, possibilitando novas pesquisas, como exemplo, testar índices para dados espaciais vagos ou testar técnicas de processamento de consultas em Data Warehouses que armazenam dados espaciais vagos. A validação da geração de dados espaciais vagos foi efetuada usando um estudo de caso com dados de fenômenos rurais vagos.