Made available in DSpace on 2014-08-20T14:22:08Z (GMT). No. of bitstreams: 1 Tese_Fernando_Jose_Hawerroth.pdf: 6659971 bytes, checksum: e37f7fea29ee58245d356204894a8ce6 (MD5) Previous issue date: 2010-11-23 / The proper balance between vegetative growth and frutification in pome fruit species such as apple and pear, it is essential to increase production efficiency and improving fruit quality. In this sense, control of vegetative growth and frutification increase are needed in the management of these species, which may be obtained by use of growth regulators. The objective of this study was to evaluate the control of vegetative growth and frutification increase of apple and pear orchards in the Southern Brazil climatic conditions in response to the use of growth regulators. For this, three experiments were carried out in this research. In the first experiment, different concentrations of prohexadione calcium were evaluated in 'Imperial Gala' and 'Fuji Suprema' apple trees, grafted on Marubakaido rootstock with M9 interstock, in an orchard located in Fraiburgo/SC. The treatments (0, 165, 330, 495, 660, and 990 g ha-1 prohexadione calcium) were applied during the vegetative growth in 2008/2009 and 2009/2010 growing seasons. The concentrations corresponding to each treatment were applied split into three parts. The first application was done when the shoots of the control treatment showed growth of 10 cm. The second and third applications were made at 30 and 60 days after the first application, respectively. The prohexadione calcium was effective in controlling vegetative growth of apple trees 'Imperial Gala' and 'Fuji Suprema', reducing the total weight and average weight of pruned shoots, as well as the average shoot length, in the Southern Brazil climatic conditions. The use of prohexadione calcium at concentrations ranging from 165 to 330 g ha-1 increased the fruit production of 'Imperial Gala' apples, but high concentrations of this growth regulator tends to reduce the fruit production, especially in 'Fuji Suprema' apples. The reduction of vegetative growth by the use of prohexadione calcium contributed to increase calcium content in fruits of 'Fuji Suprema'. The second experiment was carried out in Pelotas/RS, using 'Hosui' pears grafted on Pyrus calleryana rootstock. Different 8 concentrations of prohexadione calcium were evaluated (0, 275, 550, and 825 g ha- 1), being half the concentration corresponding to each treatment applied when the shoots had between 5 to 10 cm in length, and the remainder applied 30 days after the first application. The use of prohexadione calcium was effective in controlling the vegetative growth of 'Hosui' pears, decreasing the need for winter pruning by reducing the total weight and the number of shoots pruned. The control of vegetative growth by use of prohexadione calcium determined increase the productive capacity of 'Hosui pears, especially at concentrations ranging from 450 to 750 g ha-1. The aim of the third experiment was to evaluate the effectiveness of thidiazuron, gibberellic acid, prohexadione calcium and combination of these substances on frutification increase of 'Shinseiki' asian pears. The following treatments were applied at full bloom: 1. control (no application); 2. thidiazuron (TDZ) 20 mg L-1; 3. gibberellic acid (GA) 20 mg L-1; 4. prohexadione calcium (PCa) 600 mg L-1; 5. PCa 600 mg L-1 + TDZ 20 mg L-1; 6. PCa 600 mg L-1 + AG 20 mg L-1; e 7. AG 20 mg L-1 + TDZ 20 mg L-1. The application thidiazuron at 20 mg L-1, gibberellic acid at 20 mg L-1 and the combination of these substances during the full bloom increased significantly the frutification and the fruit production of 'Shinseiki' pears. The prohexadione calcium, when sprayed at flowering alone or in combination to thidiazuron and gibberellic acid, was not effective to increase the fruit set and fruit production. The use of growth regulators on flowering decreased the number of seeds per fruit. / O adequado balanço entre o desenvolvimento vegetativo e a frutificação em espécies pomáceas, como a macieira e a pereira, é fundamental ao aumento da eficiência produtiva e a melhoria da qualidade dos frutos. Neste sentido, o controle do desenvolvimento vegetativo e o aumento da frutificação são necessários no manejo de tais espécies, podendo ser obtidos pelo uso de fitorreguladores. Objetivou-se com este estudo avaliar o controle do desenvolvimento vegetativo e o aumento da frutificação de macieiras e de pereiras nas condições climáticas do Sul do Brasil em resposta ao uso de fitorreguladores. Para tanto, foram realizados três experimentos. No primeiro experimento, foram avaliadas diferentes concentrações de proexadione cálcio em macieiras Imperial Gala e Fuji Suprema , enxertadas no porta-enxerto Marubakaido com interenxerto de M9, em pomar localizado em Fraiburgo/SC. Os tratamentos (0; 165; 330; 495; 660; e 990 g ha-1 de proexadione cálcio) foram aplicados durante o período de desenvolvimento vegetativo nos ciclos 2008/2009 e 2009/2010. As concentrações respectivas a cada tratamento foram aplicadas parceladamente em três momentos. A primeira aplicação foi realizada quando as brotações do tratamento testemunha apresentavam crescimento de 10 cm. A segunda e a terceira aplicação foram realizadas aos 30 e 60 dias após a primeira aplicação, respectivamente. O proexadione cálcio foi eficiente no controle do desenvolvimento vegetativo de macieiras Imperial Gala e Fuji Suprema , reduzindo a massa total e a massa média de ramos, assim como o comprimento médio dos ramos, nas condições climáticas do Sul do Brasil. O uso de proexadione cálcio em concentrações variando de 165 a 330 g ha-1 aumentou a produção de maçãs Imperial Gala , porém altas concentrações deste fitorregulador tendem a reduzir a produção de frutos por planta, sobretudo em macieiras Fuji Suprema . A redução do desenvolvimento vegetativo pelo uso do proexadione cálcio contribuiu 6 para o aumento dos teores de cálcio em maçãs Fuji Suprema . O segundo experimento foi realizado no município de Pelotas/RS, utilizando pereiras Hosui enxertadas no porta-enxerto Pyrus calleryana. Foram avaliadas diferentes concentrações de proexadione cálcio (0; 275; 550; e 825 g ha-1), sendo metade da concentração respectiva a cada tratamento aplicada quando as brotações apresentavam entre 5 a 10 cm de comprimento, e o restante aplicado 30 dias após a primeira aplicação. O uso de proexadione cálcio foi efetivo no controle do desenvolvimento vegetativo de pereiras Hosui , minimizando a necessidade de poda hibernal pela redução da massa total e do número de ramos podados. O controle do desenvolvimento vegetativo pelo uso de proexadione cálcio determinou aumento da capacidade produtiva de pereiras Hosui , sobretudo em concentrações de 450 a 750 g ha-1. O objetivo do terceiro experimento foi avaliar a efetividade do thidiazuron, ácido giberélico, proexadione cálcio e a combinação destas substâncias no aumento da frutificação de pereiras asiáticas Shinseiki . Os seguintes tratamentos foram aplicados na plena floração: 1. testemunha (sem aplicação); 2. thidiazuron (TDZ) 20 mg L-1; 3. ácido giberélico (AG) 20 mg L-1; 4. proexadione cálcio (PCa) 600 mg L-1; 5. PCa 600 mg L-1 + TDZ 20 mg L-1; 6. PCa 600 mg L-1 + AG 20 mg L-1; e 7. AG 20 mg L-1 + TDZ 20 mg L-1. A aplicação de thidiazuron 20 mg L-1, ácido giberélico 20 mg L-1 e a combinação destas substâncias durante a plena floração aumentaram significativamente a frutificação e a produção de pereiras Shinseiki . O proexadione cálcio, quando aplicado na floração isoladamente ou em combinação ao thidiazuron e ao ácido giberélico, não se mostrou efetivo no aumento da frutificação e na produção de frutos. A utilização dos fitorreguladores na floração diminuiu o número médio de sementes por fruto.

Dans un contexte de changements climatiques, le stress hydrique et la gestion de l'eau sont considérés comme une contrainte importante pour le secteur agricole. Ainsi la sélection pour la tolérance à la sécheresse est devenue un objectif majeur pour de nombreux programmes de sélection. La vulnérabilité à la cavitation est considérée comme un trait d'intérêt pour la sélection à une sécheresse extrême, en particulier pour les plantes ligneuses. Pourtant, l'étendue de sa variabilité et sa relation avec la tolérance à la sécheresse sont mal documentées à l’échelle intra-spécifique, et en particulier dans les espèces cultivées. Dans cette étude, la variabilité génétique de la vulnérabilité à la cavitation a été étudiée sur trois dispositifs expérimentaux différents avec trois arbres d’intérêt agronomique qui sont menacés par le risque de sécheresse: noyer, hévéa et pommier. Une faible ou aucune, variation de la vulnérabilité à la cavitation a été trouvée dans les espèces étudiées ainsi qu'entre deux espèces de noyers malgré les différences de traits précédemment rapportés. Ces résultats suggèrent une canalisation de la résistance à la cavitation dans les organes critiques (branches, tiges). Chez le pommier, le porte-greffe est soupçonné de provoquer de la plus étroite résistance à la cavitation sur le greffon. Par contre, des différences ont bien étés trouvées sur d’autres traits de réponse à la sécheresse comme la régulation stomatique, la chute des feuilles ou encore la vulnérabilité à la cavitation dans le pétiole. Ainsi, la vulnérabilité à la cavitation des organes critiques n’est pas un paramètre pertinent pour la sélection de la tolérance à la sécheresse, et les programmes de sélection sur les espèces étudiées ne semblent pas avoir affecté ce paramètre. / In a context of climatic changes, drought stress and water management are regarded as one of the most important constraints for agricultural sector. Thus the selection for drought tolerance became a main objective for many breeding programs. Vulnerability to cavitation is considered a trait of interest for the selection for extreme drought stress, especially for woody species. However, the extent of its variability and its relation to drought tolerance are poorly documented on intraspecific level, particularly for cultivated species. In this study the genetic variability of vulnerability to cavitation was studied on three different experimental devices with three trees of agronomic interest that are threatened by the risk of drought: walnut, rubber and apple trees. Low or no variation in xylem vulnerability to cavitation was found in the studied species and between two species of walnuts despite differences previously reported features. These results suggested a canalization of cavitation resistance on critical organs (branches and stems). In apple tree, the rootstock was suspected to cause the narrow resistance to cavitation on the scion. On the contrary, differences on other traits in response to drought stress such as stomatal regulation, leaf shedding or vulnerability to cavitation on petiole were found. Therefore, vulnerability to cavitation of critical organs was not a relevant parameter for the selection of drought tolerance and breeding programs on the species studied did not appear to affect this parameter.

Thesis (MScAgric)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: The objective of this study was to evaluate the use of Musca domestica (common house fly) larvae meal, as protein source, for broiler production. This was done by investigating the nutritive value of house fly larvae meal together with its total tract digestibility, potential toxicity and carcass characteristics of the broilers supplemented with house fly larvae meal. The proximate analysis of house fly larvae meal show that it contained, on a dry matter basis, a gross energy value of 20.10 MJ/kg, 60.38% crude protein, 14.08% crude fat and 10.68% ash and that the house fly pupae contained a gross energy of 20.42 MJ/kg, 76.23% crude protein, 14.39% crude fat and 7.73% ash. House fly pupae meal had the closest match of amino acid profile when compared with the ideal amino acid profile required by broilers and it has arginine relative to the lysine content closer to the ideal amino acid profile than the house fly larvae meal. The essential fatty acid, linoleic acid, was found at levels of 26.25 and 36.27% of the total fats for the house fly larvae and pupae meal respectively. House fly larvae meal supplementation did not induce gizzard erosion or showed toxicity (regarding the gastro intestinal tract, immune system and organ stress) in broilers. Results revealed that house fly pupae meal had higher total tract digestibilities for most nutrients than of the house fly larvae meal. House fly larvae meal had a crude protein total tract digestibility of 69% and that of pupae meal was 79%. Both larvae and pupae meal had high amino acid total tract digestibilities of all the amino acids analysed. The house fly larvae and pupae meal had an apparent metabolizable energy (AME) value of 14.23MJ/kg and 15.15MJ/kg respectively. The larvae meal total tract crude fat and crude fibre digestibilities were 94% and 62% respectively. The pupae meal total tract crude fat and crude fibre digestibilities were 98% and 58% respectively. House fly larvae meal supplementation in a three phase feeding system significantly increased average broiler live weights at slaughter, total feed intake, cumulative feed intake as well as average daily gain (ADG) when compared to commercial maize: soya oil cake meal diet. In direct comparison of larvae inclusion levels with fishmeal in isonitrogenous and isoenergetic diet, no significant differences were observed between a 10% house fly larvae and a 10% fish meal diets regarding performance characteristic. The 25% house fly larvae meal diet yielded significantly better average broiler live weights at slaughter, total feed intake, cumulative feed intake (from the second week until slaughter) as well as average daily gain when compared to the 25% fish meal diet in the growth phases. Carcass characteristics of the 10% larvae, 10% fishmeal and commercial diets were compared. Chicks that received either the 10% house fly larvae meal or 10% fish meal supplementation produced significantly heavier carcasses and breast muscle portions than the chicks that received the commercial maize: soya oil cake meal. No treatment differences were found regarding breast and thigh muscle colour or pH. This study showed that house fly larvae meal can be regarded as a safe protein source that can be used to replace other protein sources and that has the ability to promote broiler performance without having any detrimental effects on carcass characteristics. / AFRIKAANSE OPSOMMING: Die evaluasie van Musca domestica (gewone huisvlieg) larwe meel as ‘n proteien bron vir braaikuiken produksie Die doel van die studie was om die effek van Musca domestica (gewone huisvlieg) larwe meel, as ‘n protein bron, in braaikuikens te evalueer. Dit was gedoen deur die nutrient waarde van huisvlieg larwe meel saam met die totale spysvertering verteerbaarheid, moontlike toksiesiteit en karkas-eienskappe van braai kuikens te evalueer. Laboratoruim analiese toon dat huisvlieg larwe meel 20.10 MJ/kg bruto energie, 60.38% ru- protein, 14.08% ru- vet en 10.68% as bevat en huisvlieg papie meel 20.42 MJ/kg bruto energie, 76.23% ru- protein, 14.39% ru- vet en 7.73% as bevat. Huisvlieg papie meel stem die meeste ooreen met die idiale amino suur profiel soos wat benodig word deur braaikuikens en dit het ‘n arginien tot lisien verhouding wat die meeste ooreenstem met die idiale amino suur profiel in vergelyking met huis vlieg larwe meel. Die essensiele vet suur, linolien suur, was geanaliseer teen vlakke van 26.25- en 36.27% van die totale vette onderskeidelik vir huisvlieg larwe- en papie meel. Huisvlieg larwe meel vervanging het nie spiermaag erosie of enige ander toksiese effekte te veroorsaak nie. Resultate het getoon dat huisvlieg papie meel, in vergelyking met larwe meel, het ‘n hoër totale spysvertering verteerbaarheid vir meeste van die nutrient. Die huisvlieg larwe meel het ‘n totale ru- protein spysvertering verteerbaarheid van 69% en die van papie meel van 79%. Beide larwe en papie meel het hoë amino suur spysvertering verteerbaarheid. Larwe meel en papie meel het skynbare metaboliseerbare energie waardes van 14.23MJ/kg en 15.15%MJ/kg onderskeidelik. Die larwe meel het ‘n ru-vet en ru- vesel spysvertering verteerbaarheid van 94% en 62% onderskeidelik, waar die papies ‘n ru-vet en ru- vesel spysvertering verteerbaarheid van onderskeidelik 98% en 58% het. Huisvlieg larwe meel vervanging in ‘n drie fase voer stelsel het getoon om die gemiddelde braaikuiken lewende gewigte by slag, totale voer iname, sowel as die gemiddelde daaglikse toename te verhoog waneer dit vergelyk word met ‘n kommersiele mielie- soya olie koek dieet. Geen mekwaardige verskille was waargeneem toe die 10% larwe meel dieet direk met die 10% vismeel diet vergelyk was rakende enige produksie einskappe gemeet nie. Die 25% larwe meel dieet het merkwaardig beter gemiddelde braaikuiken lewende gewigte by slag, totale voer iname, sowel as die gemiddelde daaglikse toename getoon wanneer vergelyk word met die 25% vismeel dieet gedurende die verskeie groei fases. Karkas eienskappe van die 10% larwe meel, 10% vismeel en die kommersiele diete was gevergelyk. Kuikens wat 10% larwe meel en 10% vismeel in die diete ontvang het, het swaarder karkasse gelewer met swaarder borsie massas wanneer vergelyk word met die kommersiele mielie- soya olie koek dieet. Geen behandelings verskille was gevind rakende die borsie- en dy spier kleure of pH nie. Die studie toon dat huisvlieg larwe meel as ‘n veillige protein bron kan beskou word, wat gebruik kan word om ander protein bronne te vervang. Huisvlieg larwe meel het ook die vermoë om braaikuiken produksie te verhoog sonder om enige negitiewe effekte rakende die karkas eienskappe te toon nie.

Thesis (MScAgric (Animal Sciences))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: The objective of this study was to evaluate the use of Musca domestica (common house fly) larvae meal, as protein source, for broiler production. This was done by investigating the nutritive value of house fly larvae meal together with its total tract digestibility, potential toxicity and carcass characteristics of the broilers supplemented with house fly larvae meal. The proximate analysis of house fly larvae meal show that it contained, on a dry matter basis, a gross energy value of 20.10 MJ/kg, 60.38% crude protein, 14.08% crude fat and 10.68% ash and that the house fly pupae contained a gross energy of 20.42 MJ/kg, 76.23% crude protein, 14.39% crude fat and 7.73% ash. House fly pupae meal had the closest match of amino acid profile when compared with the ideal amino acid profile required by broilers and it has arginine relative to the lysine content closer to the ideal amino acid profile than the house fly larvae meal. The essential fatty acid, linoleic acid, was found at levels of 26.25 and 36.27% of the total fats for the house fly larvae and pupae meal respectively. House fly larvae meal supplementation did not induce gizzard erosion or showed toxicity (regarding the gastro intestinal tract, immune system and organ stress) in broilers. Results revealed that house fly pupae meal had higher total tract digestibilities for most nutrients than of the house fly larvae meal. House fly larvae meal had a crude protein total tract digestibility of 69% and that of pupae meal was 79%. Both larvae and pupae meal had high amino acid total tract digestibilities of all the amino acids analysed. The house fly larvae and pupae meal had an apparent metabolizable energy (AME) value of 14.23MJ/kg and 15.15MJ/kg respectively. The larvae meal total tract crude fat and crude fibre digestibilities were 94% and 62% respectively. The pupae meal total tract crude fat and crude fibre digestibilities were 98% and 58% respectively. House fly larvae meal supplementation in a three phase feeding system significantly increased average broiler live weights at slaughter, total feed intake, cumulative feed intake as well as average daily gain (ADG) when compared to commercial maize: soya oil cake meal diet. In direct comparison of larvae inclusion levels with fishmeal in isonitrogenous and isoenergetic diet, no significant differences were observed between a 10% house fly larvae and a 10% fish meal diets regarding performance characteristic. The 25% house fly larvae meal diet yielded significantly better average broiler live weights at slaughter, total feed intake, cumulative feed intake (from the second week until slaughter) as well as average daily gain when compared to the 25% fish meal diet in the growth phases. Carcass characteristics of the 10% larvae, 10% fishmeal and commercial diets were compared. Chicks that received either the 10% house fly larvae meal or 10% fish meal supplementation produced significantly heavier carcasses and breast muscle portions than the chicks that received the commercial maize: soya oil cake meal. No treatment differences were found regarding breast and thigh muscle colour or pH. This study showed that house fly larvae meal can be regarded as a safe protein source that can be used to replace other protein sources and that has the ability to promote broiler performance without having any detrimental effects on carcass characteristics. / AFRIKAANSE OPSOMMING: Die doel van die studie was om die effek van Musca domestica (gewone huisvlieg) larwe meel, as ‘n protein bron, in braaikuikens te evalueer. Dit was gedoen deur die nutrient waarde van huisvlieg larwe meel saam met die totale spysvertering verteerbaarheid, moontlike toksiesiteit en karkas-eienskappe van braai kuikens te evalueer. Laboratoruim analiese toon dat huisvlieg larwe meel 20.10 MJ/kg bruto energie, 60.38% ru- protein, 14.08% ru- vet en 10.68% as bevat en huisvlieg papie meel 20.42 MJ/kg bruto energie, 76.23% ru- protein, 14.39% ru- vet en 7.73% as bevat. Huisvlieg papie meel stem die meeste ooreen met die idiale amino suur profiel soos wat benodig word deur braaikuikens en dit het ‘n arginien tot lisien verhouding wat die meeste ooreenstem met die idiale amino suur profiel in vergelyking met huis vlieg larwe meel. Die essensiele vet suur, linolien suur, was geanaliseer teen vlakke van 26.25- en 36.27% van die totale vette onderskeidelik vir huisvlieg larwe- en papie meel. Huisvlieg larwe meel vervanging het nie spiermaag erosie of enige ander toksiese effekte te veroorsaak nie. Resultate het getoon dat huisvlieg papie meel, in vergelyking met larwe meel, het ‘n hoër totale spysvertering verteerbaarheid vir meeste van die nutrient. Die huisvlieg larwe meel het ‘n totale ru- protein spysvertering verteerbaarheid van 69% en die van papie meel van 79%. Beide larwe en papie meel het hoë amino suur spysvertering verteerbaarheid. Larwe meel en papie meel het skynbare metaboliseerbare energie waardes van 14.23MJ/kg en 15.15%MJ/kg onderskeidelik. Die larwe meel het ‘n ru-vet en ru- vesel spysvertering verteerbaarheid van 94% en 62% onderskeidelik, waar die papies ‘n ru-vet en ru- vesel spysvertering verteerbaarheid van onderskeidelik 98% en 58% het. Huisvlieg larwe meel vervanging in ‘n drie fase voer stelsel het getoon om die gemiddelde braaikuiken lewende gewigte by slag, totale voer iname, sowel as die gemiddelde daaglikse toename te verhoog waneer dit vergelyk word met ‘n kommersiele mielie- soya olie koek dieet. Geen mekwaardige verskille was waargeneem toe die 10% larwe meel dieet direk met die 10% vismeel diet vergelyk was rakende enige produksie einskappe gemeet nie. Die 25% larwe meel dieet het merkwaardig beter gemiddelde braaikuiken lewende gewigte by slag, totale voer iname, sowel as die gemiddelde daaglikse toename getoon wanneer vergelyk word met die 25% vismeel dieet gedurende die verskeie groei fases. Karkas eienskappe van die 10% larwe meel, 10% vismeel en die kommersiele diete was gevergelyk. Kuikens wat 10% larwe meel en 10% vismeel in die diete ontvang het, het swaarder karkasse gelewer met swaarder borsie massas wanneer vergelyk word met die kommersiele mielie- soya olie koek dieet. Geen behandelings verskille was gevind rakende die borsie- en dy spier kleure of pH nie. Die studie toon dat huisvlieg larwe meel as ‘n veillige protein bron kan beskou word, wat gebruik kan word om ander protein bronne te vervang. Huisvlieg larwe meel het ook die vermoë om braaikuiken produksie te verhoog sonder om enige negitiewe effekte rakende die karkas eienskappe te toon nie.

Apple scab, the most serious disease of apple is caused by fungal pathogen Venturia inaequalis. Knowledge about the apple response to apple scab attack on the cellular and tissue level is insufficient. For studies of Malus-Venturia interaction on the cellular and tissue level, the establishment of methods for cell structures visualization in apple leaves is necessary. In this work, the experimental plant material grown in vitro and ex vitro was successfully established and the method of apple infection by conidia of V. inaequalis was optimized. Various methods of cell components visualization such as vital staining, in situ immunolocalization, transformation, environmental scanning electron microscopy and confocal microscopy, were tested. Cell structures, such as the cytoskeleton, the cell wall and the cuticle were visualized in apple leaves. Preliminary experiments following specific the changes of cell wall structures induced by V. inaequalis attack were performed. Further, changes of cuticle structure, the first barrier for penetration of pathogen to plant tissues during infection, were observed during the leaf ontogenesis. Powered by TCPDF (www.tcpdf.org)

Esta tese alia dois enfoques principais dentro da Química Farmacêutica. Por um lado, busca explorar uma nova fonte de insumos naturais, cascas de Malus  domestica, com vistas a obtenção dos triterpenos ácidos ursólico (AU) e betulínico (AB); e por outro lado, estuda a relação dos derivados triterpênicos semissintéticos obtidos com a proteína-alvo (PfATP6) destacada atualmente na literatura da terapia da malária com vistas ao planejamento de novos antimaláricos. O primeiro Capítulo inclui o desenvolvimento de um método eficiente, fácil e extremamente rápido onde são combinadas as técnicas de extração por ultrassom (UAE) e análise por Cromatografia Líquida de Alta Eficiência Acoplada a Detector de Espectroscopia de Massas (LC-MS), para identificação e doseamento dos ácidos ursólico (AU) e betulínico (BA) em extratos de cascas frescas de maçã de cinco clones das cultivares Gala e Fuji (“Baigent”, “Fuji Mishima”, “Fuji Suprema”, “Fuji Select” and “Maxi Gala”) oriundas da Região Sul do Brasil. Os parâmetros cromatográficos do método analítico incluem: ionização por eletro spray em modo positivo (ESI+), fluxo de 1,0 mL/min, em modo de eluição isocrático, consistindo de 80% acetonitrila e 20% acetato de amônio 10 mM em pH 6,0 e temperatura ambiente. O método desenvolvido foi validado e mostrou ser seletivo, sensível (LOD e LOQ de 0,087 e 0,266 μg/mL para BA, e 0,398 e 2,117 μg/ mL para UA), coeficiente de regressão linear (r >0.99), preciso, exato e robusto para os analitos de interesse. A otimização do método combinado de UAE com LC-MS permitiu concluir os procedimentos de extração e análise em tempo inferior a 4 h, uma vez que o método não requer a secagem da amostra, etapa que demanda longos tempos de processamento. Este método foi aplicado e forneceu a primeira caracterização fitoquímica dos cinco clones de maçã estudados. Os resultados demonstraram que o método combinado de UAE-LC-MS é adequado às práticas do controle de qualidade. O segundo Capítulo apresenta o estudo da interação dos ligantes semissintéticos derivados dos AU e AB, sintetizados pelo nosso grupo de pesquisa, com a proteína Ca2+-ATPase do Plasmodium falciparum (PfATP6), através do emprego da técnica de Docking Molecular. A PfATP6 é descrita como um importante alvo para novos antimaláricos como Artemisinina (ART), cujo mecanismo de ação inclui, dentre outros, a modulação da homeostasia do cálcio intracelular. Investigações conduziram à hipótese do extravasamento do cálcio, do interior do retículo sarco-endoplasmático como mecanismo plausível para ação dos derivados triterpênicos do AU e AB. Os escores de energia determinados no Docking, de cada um dos nove ligantes (derivados triterpênicos) e quatro compostos de controle (ácidos ursólico e betulínico, artemisinina (ART) e tapsigargina (TPG) foram determinados (análises realizadas em triplicata) e correlacionados com os valores de IC50, para a atividade antimalárica. Os resultados mostraram excelente correlação entre os escores de energia (com a PfATP6) e os valores de IC50, superior a 80% (r > 0,83360). O estudo fornece fortes evidências de que a PfATP6 pode constituir um alvo para os derivados pentacíclicos do estudo, bem como permitiu identificar o perfil conformacional dos ligantes e os principais resíduos do sítio de ligação (SL) da PfATP6 envolvidos nas interações; bem como, contribuiu para a melhor compreensão das propriedades envolvidas na interação do ligante com o receptor e mecanismo de ação. O terceiro Capítulo faz uso de Métodos Clássicos e Quânticos para descrever as mudanças conformacionais da proteína PfATP6. As simulações de Dinâmica Molecular do receptor na forma isolada e complexada com os ligantes foram realizadas durante 10 ns. As conformações finais obtidas para os receptores foram avaliadas em termos RMSD, efeito da presença dos ligantes no sítio de ligação e tipos de interações estabelecidas entre o ligante e o receptor. Os resultados mostraram que as proteínas PfATP6 e SERCA tendem a manter sua conformação nativa e que o modelo utilizado é adequado aos propósitos do estudo. O monitoramento dos resíduos da região citoplasmática das proteínas permitiu evidenciar o efeito alostérico da presença dos ligantes AU e ART no SL, sobre os domínios A e N, da PfATP6. Esse efeito reproduz as conformações E1 e E2, bem estabelecidas para PfATP6, na presença e ausência de Ca2+. As análises de Dinâmica Molecular corroboram os achados do Capítulo II ao evidenciarem o estabelecimento de interações de hidrogênio com os resíduos importantes do SL da PfATP6. Esses resultados fundamentam os indícios de que as bombas de Ca2+-ATPase (SERCA), possam ser de fato, um alvo para os derivados triterpênicos dos AU e AB. / This thesis combines two main focuses within Pharmaceutical Chemistry. On the one hand, it seeks to explore a new source of natural inputs, bark of Malus domestica, in order to obtain ursolic (UA) and betulinic acid (BA) triterpenes. On the other hand, it studies the relation of the semi-synthetic triterpenic derivatives obtained with the target proteins (PfATP6), currently highlighted in the literature on malaria therapy with a view to planning new antimalarial drugs. The first chapter includes the development of an efficient, easy and extremely fast method where ultrasonic extraction techniques (UAE) and high-performance liquid chromatography coupled to mass spectroscopy (LC-MS) are combined for identification and assay of ursolic acid (UA) and betulinic acid (BA) in fresh apple peel extracts from five clones of the Gala and Fuji cultivars (Baigent, Fuji Mishima, Fuji Suprema, Fuji Select and Maxi Gala ") in the Southern Region of Brazil. Chromatographic parameters of the analytical method include: electrospray ionization (ESI +), flow rate of 1.0 mL/min in isocratic elution mode, consisting of 80% acetonitrile and 20% 10 mM ammonium acetate at pH 6.0 and room temperature. The method was validated and proved to be selective, sensitive (LOD and LOQ of 0.087 and 0.266 μg/mL for BA, and 0.398 and 2.117 μg/mL for UA), linear regression coefficient (r> 0.99), accurate, robust for analytes of interest. The optimization of the combined method of UAE with LC-MS allowed to complete the procedures of extraction and analysis in less than 4 h, since the method does not require drying the sample, a stage that demands long processing times. This method was applied and provided the first phytochemical characterization of the five apple clones studied. The results demonstrated that the combined UAE-LC-MS method is suitable for quality control practices. The second chapter presents the study of the interaction of the semi-synthetic ligands derived from the UA and BA, synthesized by our research group, with the Plasmodium falciparum Ca2+-ATPase protein (PfATP6), using the Molecular Docking technique. PfATP6 is described as an important target for new antimalarials such as Artemisinin (ART), whose action mechanism includes, among others, the modulation of intracellular calcium homeostasis. Investigations led to the hypothesis of extravasation of calcium from the interior of the sarco-endoplasmic reticulum as a plausible mechanism for the action of the triterpenic derivatives of UA and BA. The Docking energy scores (binding energy) of each of the nine ligands (triterpenic derivatives) and four control compounds (ursolic and betulinic acids, artemisinin (ART) and tapsigargine (TPG)) with PfATP6, were calculated (analyses performed in triplicate) and correlated with its antimalarial IC50 value. The results showed an excellent correlation between energy scores (with PfATP6) and IC50 values, higher than 80% (r > 0.83360). The study supplies strong evidence that PfATP6 may be a target for the pentacyclic derivatives of the study, and also allowed identifying the conformational profile of the ligands and the main residues of the PfATP6 binding site (BS) involved in the interactions. It further contributed to a better understanding of the properties involved in the interaction of the ligand with the receptor and its action mechanism. The third chapter uses Classical and Quantum Methods to describe the conformational changes of the PfATP6 protein. The Molecular Dynamics simulations of the receptor in the isolated and complexed form with the ligands were performed for 10 ns. The final conformations obtained for the receptors were evaluated in RMSD terms, effect of the presence of ligands at the binding site and types of interactions established between the ligand and the receptor. The results showed that the PfATP6 and SERCA proteins tend to maintain their native conformations and that the model used is adequate for the purposes of the study. The monitoring of the residues of the cytoplasmic region of the proteins allowed evidencing the allosteric effect of the presence of UA and ART ligands in BS, on the A- and N-domains of PfATP6. This effect reproduces the well-established E1 and E2 conformations for PfATP6, in the presence and absence of Ca2+, respectively. Molecular Dynamics analyses corroborate the findings of Chapter II, by showing the possibility of establishing hydrogen interactions with the important residues of PfATP6 BS. These results support the evidence that Ca2+-ATPase (SERCA) pumps may be a target for the triterpenic derivatives of UA and BA.

Orientador: Adriano Sakai Okamoto / Banca: Raphael Lucio Andreatti filho / Banca: Guilherme Augusto Marietto Gonçalves / Resumo: O presente trabalho teve como proposta realizar a caracterização probiótica in vitro de amostras de Lactobacillus spp., provenientes de conteúdo intestinal de perus adultos e saudáveis, através do isolamento e identificação por características morfológicas, moleculares e fisiológicas. Inicialmente foram isoladas 170 amostras, as quais foram avaliadas pelo método de coloração de gram, testes de produção de catalase, hidróxido de potássio, produção de gás através da fermentação da glicose e produção de gás sulfídrico em Triple Sugar Iron (TSI). Após foram pré-selecionadas 74 amostras, que passaram por identificação molecular com a Polymerase Chain Reaction (PCR) e submetidas ao PCR-ARDRA com uso das enzimas Sph I, Nco I, Nhe I, Ssp I, Sfu I, Dra I, Vsp I, Eco RI, Hinc II, Hind III e Avr II. A avaliação da resistência bacteriana ocorreu através dos testes de suco gástrico artificial e sua tolerância a sais biliares, capacidade de adesão à mucosa intestinal, potencial de multiplicação, provas de antagonismo contra Salmonella Heidelberg, produção de peróxido de hidrogênio, antibiograma e avaliação dos genes de resistência antimicrobianos integrons C e submetidas ao sequenciamento genético. Concluiu-se que 11 amostras foram selecionadas, sendo uma de Lactobacillus frumenti, 9 de Lactobacillus reuteri e uma de Lactobacillus johnsonii, todas com resultados nos testes de antagonismo, resistência ao suco gástrico, resistência a sais biliares, hidrofobicidade, potencial de multiplicação, produção de peróxido de hidrogênio e com resistência a mais de 50% dos antimicrobianos testados, porém não apresentaram genes de resistência antimicrobianos na técnica que avalia genes integrons C, baseado nos resultados de todas as análises as amostras selecionadas podem vir a ser administradas in vivo. / Abstract: The aim of the present work was to characterize the in vitro probiotic characterization of Lactobacillus spp. Samples from intestinal contents of adult and healthy turkeys through isolation and identification by morphological, molecular and physiological characteristics. Initially, 170 samples were isolated, which were evaluated by gram staining method, catalase production tests, potassium hydroxide, gas production through glucose fermentation and the production of sulphide gas in Triple Sugar Iron (TSI). After being pre-selected 74 samples, which were submitted to PCR-ARDRA using the enzymes Sph I, Nco I, Nhe I, Ssp I, Sfu I, Dra I, Vsp I, Eco RI, Hinc II, Hind III and Avr II. The evaluation of bacterial resistance occurred through the tests of artificial gastric juice and its tolerance to bile salts, intestinal mucosal adhesion capacity, multiplication potential, Salmonella Heidelberg antagonism tests, hydrogen peroxide production, antibiogram and evaluation of the genes of Antimicrobial resistance and submitted to genetic sequencing. It was concluded that 11 samples were selected, one of Lactobacillus frumenti, 9 of Lactobacillus reuteri and one of Lactobacillus johnsonii, all with results in the tests of antagonism, resistance to gastric juice, resistance to bile salts, hydrophobicity, multiplication potential, Hydrogen peroxide production and resistance to more than 50% of the antimicrobials tested, but did not present antimicrobial resistance genes in the technique that evaluates integron C genes, based on the results of all the analyzes the selected samples can be administered in vivo. / Mestre

Depuis la domestication du pommier, l’homme a progressivement sélectionné des variétés plus performantes, notamment pour la qualité du fruit, la productivité ou la résistance aux pathogènes. Les bases génétiques de ces caractères ont été explorées par cartographie en descendances F1 ne permettant d’explorer qu’une infime partie de la diversité génétique disponible.L’objectif de la thèse portait sur l’analyse des bases génétiques de caractères de qualité du fruit et de résistance du pommier à la tavelure et au feu bactérien dans des collections représentant une diversité plus large. Le génotypage de core collections de variétés anciennes s’est fait à l’aide de deux puces 8k et 480kSNPs ou grâce à du ré-séquençage de gènes. Des traces de différenciation génétique entre pommes à cidre et à couteau ont été identifiées et partiellement reliées à la voie des polyphénols. Après analyse de l’étendue du déséquilibre de liaison à large et fine échelle, une approche de génétique d’association a permis l’identification de régions génomiques associées à la variation de plusieurs caractères de qualité du fruit, dont le haut du groupe de liaison 16 rassemblant l’acidité (locus Ma), la fermeté, la jutosité et l’amertume (gène LAR). Pour la résistance au feu bactérien, une région contenant un homologue du gèneNPR1 (activateur de défenses) a été identifiée.Cette thèse a ainsi permis de préciser la localisation potentielle de QTLs identifiés préalablement par cartographie génétique et d’identifier de nouvelles ressources utiles dans de futurs programmes de sélection assistée par marqueurs. / Since apple domestication, humans have progressively selected improved varieties, especially for traits linked with fruit quality, productivity or resistance to pathogens. The genetic bases underlying these traits have been explored thanks to genetic mapping in F1 segregating populations that only allows the study of a small part of the available genetic diversity. The aim of this work was to analyze the genetic bases of fruit quality and disease resistance against apple scab and fire blight, in collections of old apple varieties representing a much larger diversity. Genotyping of core collections was performed either with arrays of 8k and 480k SNPs or by resequencing of chosen genes. Signs of genetic differentiation were identified between cider and dessert apples and were partially linked to the polyphenols pathway. After studying linkage disequilibrium, both on a large and a small scale, an association genetics approach allowed the identification of genomic regions associated with the variation of several fruit quality traits. Especially, the top of linkage group 16 was found to be linked with acidity (locus Ma), firmness, juiciness and bitterness (LAR gene). Concerning the resistance of apple to fire blight, a region containing a homolog of the NPR1 gene (defense activator) was identified. This thesis allowed the refining of the putative localization of previously identified QTLs and the identification of new genetic resources that could be useful in future selection programs using marker assisted selection.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2017-03-24T15:50:08Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO_GPZ_versãofinal.pdf: 1750560 bytes, checksum: 45861d58aabcf6100696f8b44e555dea (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-03-28T20:23:47Z (GMT) No. of bitstreams: 2 DISSERTAÇÃO_GPZ_versãofinal.pdf: 1750560 bytes, checksum: 45861d58aabcf6100696f8b44e555dea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-03-28T20:23:47Z (GMT). No. of bitstreams: 2 DISSERTAÇÃO_GPZ_versãofinal.pdf: 1750560 bytes, checksum: 45861d58aabcf6100696f8b44e555dea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A demanda por produtos com características semelhantes àquelas das frutas in natura vem sendo uma tendência atual, devido à divulgação dos benefícios trazidos por uma dieta rica em compostos bioativos. Nesse contexto, os sucos ganham espaço no mercado, especialmente quando se amplia a disponibilidade de compostos bioativos. Assim uma das possibilidades é associar aos sucos de maçã pequenos frutos como amora, framboesa e morango, que são ricas fontes de compostos com propriedades antioxidantes, além de fornecer diferenciadas características sensoriais. Com isso, objetivou-se desenvolver suco integral de maçã e suco de maçã com adição de pequenos frutos, avaliando as características físicoquímicas e sensoriais. O trabalho foi realizado em três etapas: 1) desenvolvimento de suco integral de maçã das cultivares Fuji, Gala, Granny Smith e Pink Lady; 2) adição individual de 5, 10, 15 e 20 % amora, framboesa e morango ao suco de maçã ‘Gala’; 3) produção de suco integral de maçã ‘Gala’ com adição individual de 5 % de cada pequeno fruto. Na primeira etapa, observou-se que o suco de maçã ‘Granny Smith’ apresentou maior acidez e maior presença de compostos bioativos. A menor acidez foi evidenciada para o suco da cv. Gala. O suco da cv. Fuji, apresentou maior quantidade de açúcares totais. Em relação à aceitabilidade o suco de maçã ‘Gala’ e ‘Fuji’ tiveram as melhores notas. Com isso, escolheu-se o suco de maçã ‘Gala’ para aplicação de diferentes concentrações de pequenos frutos. Na segunda etapa, podese perceber que as maiores concentrações de pequenos frutos adicionados ao sucos de maçã resultaram em menores notas na avaliação sensorial, demonstrando que a adição de mais do que 10 % de pequenos frutos pode causar depreciação sensorial dos sucos. Na terceira etapa observou que a adição de amora e de morango ao suco de maçã proporcionou o incremento de 18,5 % e 16,2 %, respectivamente, no teor de compostos fenólicos totais. Os principais compostos fenólicos individuais detectados nos sucos de maçã e nos sucos adicionados de pequenos frutos foram ácido cafeico, ácido ferúlico, ácido cumárico e ácido gálico, os quais são importantes antioxidantes. A adição de pequenos frutos proporcionou o incremento de antocianinas ao suco de maçã, a amora forneceu o acréscimo de pelargonidina, cianidinas e keracianina. Quanto aos aspectos sensoriais, a adição de amora e framboesa proporcionaram maior aceitação para o atributo cor, e para o sabor o suco de maçã com morango, o suco de maçã integral e suco de maçã com amora tiveram melhor aceitabilidade. De forma geral, foi observado que a adição de pequenos frutos ao suco de maçã obteve boa aceitação e contribuiu para o aumento do teor de compostos bioativos, sendo uma boa alternativa para o enriquecimento do suco de maçã em compostos potencialmente benéficos à saúde, além de promover maior diversificação de produtos derivados de maçã. / The demand for products with characteristics similar to fresh fruits has been a current trend, due to the announcing of the benefits brought by a diet rich in bioactive compounds. In this context, juices gain market, especially when they expand the availability of bioactive compounds. Thus, one of the possibilities is to associate green apple juice with blackberry, raspberry, and strawberry, which are rich sources of compounds with antioxidant properties, and additionally provide unique sensory characteristics. The objective of this study was to develop integral apple juice and apple juice with the addition of small fruits, and to characterize their physicochemical and sensory characteristics. The work was developed in three steps: 1) apple integral juice development using cultivars Fuji, Gala, Granny Smith and Pink Lady; 2) individual addition of 5, 10, 15, and 20% blackberry, raspberry, and strawberry pulp to apple juice ‘Gala’; 3) production of whole apple juice 'Gala' with individuality of 5 % of each small fruit. In the first step, it was observed that 'Granny Smith' juice presented the highest acidity and greatest bioactive content. The lowest acidity was observed for the cv. Gala and the highest sugar content was found in cv. Fuji juice. 'Gala' and 'Fuji' juices had the highest acceptability rates. Therefore 'Gala' was chosen for the application of different concentrations of small fruits. In the second step of the study, it could be noticed that the higher the concentration of small fruits added to the apple juice the lower the sensory evaluation scores, demonstrating that the addition of more than 10 % of small fruits can cause sensorial depreciation of the juices. In the third step, it was observed that the addition of blackberry and strawberry to apple juice contributed to an increase of total phenolic content of 18.5% and 16.2%, respectively. The main individual phenolic compounds detected in apple juices and juices added of small fruits were caffeic acid, ferulic acid, coumaric acid, and gallic acid, important antioxidant compounds. The addition of small fruits contributed to an increase in anthocyanin content in the apple juice; blackberry contributed with increases in pelargonidin, cyanidins, and keracyanin contents. As for the sensory, the addition of blackberry and raspberry contributed to a greater acceptance for the color attribute. Apple juice with strawberry, whole apple juice, and apple juice with blackberry had the best taste acceptability. In general, it was observed that addition of small fruits to apple juice promoted good acceptance and contributed to the increase bioactive compound content. These blends constitute a good alternative for the enrichment of apple juice in health beneficial compounds and promote a greater diversification of apple products.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2017-02-02T13:47:04Z No. of bitstreams: 1 Ana Paula Camboim Leao.pdf: 2376458 bytes, checksum: d62ed4d58dba9fed450d1ac20346cdce (MD5) / Made available in DSpace on 2017-02-02T13:47:04Z (GMT). No. of bitstreams: 1 Ana Paula Camboim Leao.pdf: 2376458 bytes, checksum: d62ed4d58dba9fed450d1ac20346cdce (MD5) Previous issue date: 2016-02-02 / The research was developed through the Consuption, Day By Day and Social Development Pos-Graduation Program and involved public workers from the Federal Rural University of Pernambuco. It sought to answer the following question: How Financial debt strategies impact the lives of this educational administrative staff of public workers, in their ways of consumption and social, professional and family wellness? With that inquiry, it was elaborated as main objective of this work to analyze factors and implications of the financial debt process by payroll loan along with the educational administrative staff of public workers. The specific objectives, on the other hand, were elaborated in order to analyze the factors that motivate this public debt; to analyze products, services and communication strategies that corroborate with the public worker or his/her family’s debt; to map financial debt situations, specifically from payroll loan; and finally, to analyze implications of individual and social nature resulting from the financial debt of these federal public workers. To achieve these goals, a quantitative and qualitative study was done distributing questionnaires among 200 public workers, from witch 90% returned. In a more in-depth approach, 05 intentionally chosen public workers were interviewed about budget imbalance, its causes and consequences, appreciating his/her position in the academic community context. The data treatment was to clash the results from the interviews with the consulted background work. The results showed that expenses related to daily life, allied to recurring payroll loan signings, may contribute negatively to the unbalance of domestic budgets and its repercussions affect psychological, personal and social wellness. / A pesquisa desenvolvida a partir do Programa de Pós-Graduação em Consumo, Cotidiano e Desenvolvimento Social com o(a)s servidore(a)s Técnico- Administrativos da Universidade Federal Rural de Pernambuco buscou responder o seguinte questionamento: como estratégias de endividamento financeiro impactam a vida de servidore(a)s público(a)s Técnico-Administrativos em Educação, em suas práticas de consumo e bem-estar sócio-familiar e profissional? Com essa indagação, elaborou-se como objetivo geral para esta pesquisa analisar fatores e implicações do processo de endividamento financeiro por meio do empréstimo consignado, junto a servidore(a)s federais Técnico-Administrativos em Educação. Os objetivos específicos, por sua vez, foram elaborados com o intuito de analisar fatores que motivam o endividamento este público; analisar produtos, serviços e estratégias comunicacionais que corroboram o endividamento do(a)s servidore(a)s e familiares; mapear situações de endividamento financeiro, especificamente, por meio do empréstimo consignado; por fim, analisar implicações de natureza individual e social decorrentes do endividamento financeiro destes servidore(a)s público(a)s federais. Para o alcance destes objetivos foi realizado um estudo quanti-qualitativo com a distribuição de questionários entre 200 servidore(a)s, no qual se obteve o retorno de 90% destes. Em uma abordagem mais aprofundada, 05 servidore(s) escolhido(a)s intencionalmente foram entrevistado(a)s, acerca do desequilíbrio orçamentário, suas causas e consequências, valorizando a posição dela(e)s no contexto da comunidade acadêmica. O tratamento dos dados processou-se através do confronto dos resultados das entrevistas desenvolvidas com a literatura trabalhada. Os resultados apontam que as despesas relacionadas ao cotidiano, aliadas às contratações recorrentes de empréstimos consignados podem contribuir negativamente no desequilíbrio do orçamento doméstico com repercussões no bem-estar psíquico, pessoal e social.