Dissertations / Theses on the topic 'Loquat'

El moteado del níspero, causado por el hongo Fusicladium eriobotryae es la principal enfermedad que afecta al cultivo del níspero, produciendo pérdidas importantes en la cosecha en los años con condiciones climáticas adecuadas. Sin embargo, no existen estudios sobre la epidemiología y el control de esta enfermedad, por lo que éstos constituyen el principal objetivo de la presente Tesis. Para ello, se va ha caracterizar in vitro y en campo la influencia de las principales variables climáticas en el desarrollo de F. eriobotryae, desarrollándose ecuaciones matemáticas que modelicen esta relación. Por otro lado, se va a llevar a cabo un modelo epidemiológico para el moteado del níspero capaz de predecir, en función de las principales variables climáticas el riesgo de infección. Además, se realizará un estudio del control de la enfermedad que comprenderá, por un lado, la evaluación in vitro y en planta de la efectividad de las principales materias activas utilizadas para el control del moteado del níspero, y por otro la evaluación del grado de resistencia de una colección de aislados de F. eriobotryae a DMI y Metil-Tiofanato y la caracterización molecular de la misma.<br>González Domínguez, E. (2014). Biology, epidemiology and control of Fusicladium eriobotryae, causal agent of loquat scab [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/38715<br>TESIS

In loquat, apex of a current shoot changes from vegetative to reproductive stage during summer, i.e. under high temperature conditions. Indeed, just before floral bud differentiation, a decline in the growth rate due to high temperature takes place. The aim of this work is to study the role of this 'summer rest period' on the apex transition from vegetative to reproductive stage. For this purpose 1) sprouting of secondary shoots was promoted at different times, removing the main shoot, before, during and after floral bud differentiation occurred and 2) groups of trees were shifted to a greenhouse under average maximum temperature not exceeding 25 ° C during different periods from June to October. Floral bud differentiation was evaluated. LEAFY (LFY), APETALA (AP1), TERMINAL FLOWERING 1 (TFL1) and FLOWERING LOCUS T (FT1) expression and hormonal content in abscisic acid (ABA), gibberellins (GAs), indoleacetic acid (IAA) and cytokinins (CKs) were analyzed in bud collected during the summer. Results suggest that the date of shoot apex removal determining floral bud differentiation of new shoots, so that the percentage of the new reproductive shoots reduced with the delaying of apex removal. On the other hand, maximum average temperature not exceeding 25 ° C prevented floral bud differentiation. Buds of the trees under indoors conditons displayed lower expression of identity floral genes EjLFY and EjAP1 than buds of trees grown in field. On the contrary, the floral repressor EjTFL1 and EjFT1 gene expressed higher in buds of the trees grown indoors. Time-course of ABA decreased in buds of trees grown in field during studied period while in buds of trees under greenhouse conditions displayed a growing trend. Time-course of GAs, IAA and CKs concentrations did not show remarkable differences between buds of trees growing under field and indoors conditions. Accordingly, 1) secondary shoots emerged from mid- August are unfitness to flower and 2) maximum average tempertature 25±1 °C during the summer prevents floral bud differentiation, enhances ABA biosynthesis, reduces EjLFY and EjAP1 expression and enhance EjTFL1 expression in the apex.<br>El níspero japonés diferencia sus yemas durante el verano, después de un periodo de ralentización del crecimiento vegetativo ligado a las altas temperaturas que se conoce como reposo estival. El objetivo de esta tesis fue estudiar la influencia de la parada estival en la diferenciación floral de esta especie. Para ello se diseñó un experimento en el que se forzó la brotación de brotes anticipados eliminado el ápice principal en diferentes fechas entre julio y septiembre, antes, durante y después de la parada estival. Paralelamente se diseñó otro experimento en el que se cambiaron las condiciones climáticas a grupos de árboles manteniéndolos en un invernadero a una temperatura máxima media de 25 °C durante diferentes periodos de diversa duración. Se evaluó la diferenciación floral y se analizó la expresión de los genes relacionados con la floración LEAFY (LFY), APETALA (AP1), TERMINAL FLOWERING 1 (TFL1) and FLOWERING LOCUS T (FT1) y el contenido hormonal en ácido abscisico (ABA), giberelinas (GAs), ácido indolácetico (AIA) y citoquininas (CKs) en yemas terminales muestreadas a lo largo del verano. Los resultados indican que la fecha de brotación modifica la diferenciación floral de los brotes anticipados siendo el porcentaje de brotes reproductivos inversamente proporcional a la fecha de eliminación del meristemo. Del mismo modo unas condiciones de temperatura máxima no superior a 25 °C impidieron la diferenciación floral. Las yemas de los árboles que estuvieron bajo dichas condiciones mantuvieron unos niveles de expresión de los genes de identidad floral, EjLFY y EjAP1, mucho menor que la de los árboles en condiciones de campo. Por el contrario, la expresión del represor EjTFL1 y del gen EjFT1 fue mayor en los árboles en invernadero. Por otro lado, el contenido endógeno de ABA descendió en los árboles situados en el campo durante el periodo de estudio mientras que en los árboles situados en el invernadero tuvo una evolución ascendente. Las concentraciones de GAs, AIA y CKs no mostraron prácticamente diferencias entre los ápices de los árboles mantenidos en campo y en invernadero. De acuerdo con ello, 1) los brotes anticipados surgidos a partir de mitad de agosto son incapaces de florecer y 2) la ausencia de altas temperaturas del verano promueve la acumulación de ABA, aumenta la expresión del gen represor (EjTFL1) y reduce la expresión de los genes de identidad floral (EjLFY y EjAP1) en yemas de níspero impidiendo su diferenciación floral.<br>El nispro japonés diferència les seus gemmes durant l'estiu, després d'un període d'alentiment del creixement vegetatiu lligat a les altes temperatures que es coneix com repòs estival. L'objectiu d'aquesta Tesi va ser estudiar la influència de la parada estival en la diferenciació floral d'aquesta espècie. Per a això es va dissenyar un experiment en què es va forçar la aparició dels brots anticipats eliminat l'àpex principal en diferents dates entre juliol i setembre, abans, durant i després de l'aturada estival. Paral·lelament es va dissenyar un altre experiment en què es van canviar les condicions climàtiques a grups d'arbres mantenint-los en un hivernacle a una temperatura màxima mitjana de 25 °C durant diferents períodes de diversa durada. Es va avaluar la diferenciació floral i es va analitzar l'expressió dels gens relacionats amb la floració LEAFY (LFY), APETALA (AP1), TERMINAL FLOWERING 1 (TFL1) and FLOWERING LOCUS T (FT1) i el contingut hormonal en àcid abscísic (ABA) , gibberel·lines (GAs), àcid indolacètic (AIA) i citoquinines (CKs) en gemmes terminals mostrejades al llarg de l'estiu. Els resultats indiquen que la data de brotació modifica la diferenciació floral dels brots anticipats i el percentatge de brots reproductius es inversament proporcional a la data d'eliminació del meristema. De la mateixa manera unes condicions de temperatura màxima no superior a 25 ° C varen impedir la diferenciació floral. Les gemmes dels arbres que van estar sota aquestes condicions van mantenir uns nivells d'expressió dels gens d'identitat floral, EjLFY i EjAP1, molt menor que la dels arbres en condicions de camp. Per contra, l'expressió del repressor EjTFL1 i del gen EjFT1 va ser més gran en els arbres en hivernacle. D'altra banda, el contingut endogen d'ABA va baixar en els arbres situats al camp durant el període d'estudi mentre que en els arbres situats a l'hivernacle va tenir una evolució ascendent. Les concentracions de GAs, AIA i CKS no van mostrar pràcticament diferències entre els àpexs dels arbres mantinguts en camp i en hivernacle. D'acord amb això, 1) els brots anticipats sorgits a partir de meitat d'agost són incapaços de florir i 2) l'absència d'altes temperatures de l'estiu promou l'acumulació d'ABA, augmenta l'expressió del gen repressor (EjTFL1) i redueix l'expressió dels gens d'identitat floral (EjLFY i EjAP1) en gemmes de nispro del Japó impedint la seva diferenciació floral.<br>García Lorca, AL. (2017). Temperature regulating floral bud differentiation in loquat (Eriobotrya japonica Lindl.). Hormonal and genetic aspects [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/79873<br>TESIS

Thesis (MScFor (Forest and Wood Science))--University of Stellenbosch, 2005.<br>This study is about marketing of Uapaca kirkiana fruit in Zambia, a fruit that has great economic value especially among the rural and urban poor. It contributes to general food security. In southern Africa, farmers and other stakeholders have identified Uapaca as a priority species for domestication. Current agroforestry initiatives are promoting integration of indigenous trees into farming systems to provide marketable products for income generation. Domestication of trees however, depends on expanding the market demand for tree products. There is considerable evidence that expanding market opportunities is critical for the success of domestication innovations but farmers have been introduced to domestication with little consideration for marketing. The existing market potential can be achieved through sound knowledge on markets and marketing. Information on the marketing of Uapaca fruit is inadequate. This study, therefore, aimed at generating information on the marketing of Uapaca kirkiana fruit, including the basic conditions of demand and supply of the fruit. The main study was conducted in Chipata and Ndola districts in the Eastern and Copperbelt provinces respectively, while fruit pricing was conducted in Lusaka district in Lusaka Province. Questionnaires and participatory research methods were used to collect the data. A total of 37 markets involving 49 collectors, 59 retailers, 189 consumers and 20 government forest workers are included in the study. The study reveals that there is demand for the fresh and secondary products of the fruit and hence substantial fruit trading exists in Zambia. However, the marketing system is characterised by underdevelopment. There is insufficient capacity to satisfy the demand for the fruit and institutional /policy support to Uapaca fruit market expansion. Currently, only basic technology for product development exists. The results suggest a need to address policy and capacity development for expansion of Uapaca kirkiana fruit industry. To improve the Uapaca trade industry, the study proposes developing and scaling up policy strategies, fruit processing sector, research-extension-trader-agribusiness linkages, infrastructure development and knowledge generation for improved understanding of the Uapaca fruit markets.

Orientador: Antonio Baldo Geraldo Martins<br>Banca: Paulo Sérgio de Souza<br>Banca: Carlos Ruggiero<br>Resumo: O cultivo de nêsperas vem aumentando no Brasil e no mundo, sendo uma importante fonte de renda para pequenas propriedades, no entanto, a produção de mudas pode ser um gargalo de sua expansão. Dessa forma o objetivo do presente trabalho foi avaliar o reflexo do boro na qualidade das mudas de nespereira, bem como o local de seu fornecimento às plantasmães determinando-se o modo de aplicação mais eficiente neste processo. Foram utilizadas quatro doses de boro (0; 2,5; 5,0 e 7,5 mg dm-3 ) e dois locais de aplicação (no solo e foliar). A utilização de boro nas plantas-matrizes melhora o sistema radicular das mudas obtidas por estaquia, sendo a aplicação foliar mais eficiente do que via solo. A aplicação deste micronutriente promove o equilíbrio entre raiz/parte aérea, garantindo a formação de uma muda equilibrada, haja vista que o fornecimento foliar aumenta o volume de raízes da estaca. Portanto o fornecimento de boro às matrizes torna o processo eficaz, reduzindo custo, otimizando tempo e garantindo a qualidade das mudas frutíferas<br>Abstract: The loquat cultivation is increasing in Brazil and in the world and is an important source of income for small farms, however, the production of seedlings can be an obstacle to its expansion. Thus the aim of this study was to evaluate Boron reflection on the quality of loquat seedlings, and the place of supply to plants parent by determining the most effective application method in this process. There were four boron doses (0, 2.5, 5.0 and 7.5 mg dm-3) and two application sites (soil and leaf). The use of boron instock plants improves the root system of plants from cuttings, the most efficient foliar application than the soil. The application of this micronutrient promotes balance between root / shoot, ensuring the formation of a balanced changes, given that the leaf supply increases the volume of the stake roots. Therefore the supply of boron to headquarters makes the process effectively, reducing cost, optimizing time and assuring the quality of fruit seedlings<br>Mestre

A nespereira é uma planta rústica, que prescinde da utilização sistemática de defensivos, o que torna o cultivo atraente, para atender ao público que cresce, dia a dia, à procura de produtos naturais.O trabalho teve por objetivo realizar a caracterização fenológica da nespereira, estabelecendo os estádios fenológicos e o ciclo de produção de 5 cultivares e de uma seleção.Também estudou-se a melhor fase para o ensacamento dos frutos e a intensidade de raleio para a obtenção de frutos de alta qualidade. Foi estabelecida uma escala fenológica contendo sete estádios de desenvolvimento para a nespereira. Os estágios observados foram: 0 Desenvolvimento da Gema; 1 Desenvolvimento das folhas; 3 Desenvolvimento dos Brotos; 5 Emergência da Inflorescência; 6 Florescimento; 7 Desenvolvimento dos frutos; 8 Maturação dos frutos, de acordo com a escala BBCH. Mediante o estudo da escala fenológica das 5 cultivares e da seleção IAC NE3, verificou-se a existência de diferenças no ciclo de produção das cultivares estudadas e também da seleção, possibilitando a observação de três grupos. O primeiro grupo, mais precoce, foi formado pelas cultivares Mizuho e Mizumo, com ciclo médio de 195 dias contados a partir do início do desenvolvimento das gemas até a colheita dos frutos. As cultivares Néctar de Cristal e Centenária formaram um segundo grupo apresentando um ciclo intermediário com uma média de 203 dias. Já a cultivar Mizauto e a seleção IAC NE3 foram as mais tardias, uma vez que o ciclo foi concluído 208 dias, em média, após o início do desenvolvimento das gemas.A intensidade de raleio a ser adotada deve ser de 4 ou 6 frutos por panícula, levando-se em conta a safra esperada; o ensacamento dos frutos deve ser realizado nos estádios 1 (frutos com 1,0 cm de diâmetro) ou 2 (frutos com 1,5 cm de diâmetro).<br>The Loquat is a rustic plant, which renounce the systematic use of defensive, which makes the cultivation attractive, to attend the public that grows, day by day, the demand for natural products. The study aimed to carry out the phenological characterization of loquat, setting the phenological stages and the cycle of production of 5 cultivars and a selection. Also studied is the best stage for the bagging of panicles and thinning intensity for obtaining fruits of high quality. Was established a scale containing seven phenological stages of development for loquat. The stages were observed: 0 - Development of bud, 1 - Development of the leaves, 3 - Development of shoot, 5 - Emergency of Inflorescence, 6 - Blooming; 7 - Development of fruit; 8 - Ripening fruits, according to the scale BBCH. Through the study of phenological scale from 5 cultivars and selection IAC NE3, there are differences in the production cycle of cultivars and the selection, enabling the observation of three groups. The first group, earlier, was formed by cultivars with Mizuho and Mizumo cycle average of 195 days counted from the beginning of the development of buds until the harvest fruit. Cultivars nectar of Cristal and Centenária formed a second group presenting a cycle intermediary with an average of 203 days. The cultivar Mizauto and the select IAC NE3 were the most late, since the cycle was completed 208 days on average after the start of the development of buds. The intensity of thinning to be used must be 4 or 6 fruits per raceme, considering the crop waited and the fruit price in the market. The fruit bagging must be realized in the harvest time 1(fruits with 1,0 cm of diameter) or 2(fruits with 1,5 cm of diameter).

Os experimentos de campo foram realizados em 2002 e 2003 em três pomares no município de Monte Alegre do Sul, SP, representados por uma coleção de linhagens de goiabeiras (janeiro a abril), uma coleção de cultivares de nespereiras (agosto a setembro) e uma coleção de cultivares de pessegueiros (setembro a outubro). Nos ensaios de infestação, foram utilizadas três linhagens de goiaba (Guanabara, L7P28 e 252), duas cultivares de nêspera (Precoce Campinas e a Precoce 264-54) e três cultivares de pêssego (Aurora 2, Dourado 1 e Régis). Para a determinação do período de infestação, aplicou-se o processo de ensacamento e desensacamento quinzenal e semanal da goiaba e nêspera, respectivamente, e apenas o ensacamento semanal no pêssego. Cada experimento iniciou-se com os frutos ainda no início de seu desenvolvimento (frutos verdes). Para o processo de desensacamento, no início dos experimentos foram ensacados 500 e 400 frutos de goiaba e nêspera, respectivamente. Em cada ensaio, desde o início (frutos verdes pequenos) até a completa maturação, quinzenalmente (goiaba) ou semanalmente (nêspera e pêssego) foi ensacada/desensacada uma amostra de 30 frutos, os comprimentos e diâmetro eram mensurados e retirava-se amostras para realização das análises físico-químicas em laboratório. Após o completo amadurecimento, os frutos foram colhidos e levados ao laboratório, onde foram pesados, medidos os diâmetro e comprimento e depois acondicionados individualmente em copos plásticos com areia+vermiculita até aproximadamente a metade e cobertos com “voil” preso com fita elástica. Para o conhecimento da flutuação populacional das moscas, foram instaladas três armadilhas modelo McPhail com torula (atraente alimentar) em cada pomar, de janeiro/2002 a janeiro/2004. Das moscas-das-frutas originadas dos frutos, foram identificadas cinco espécies de tefritídeos e oito espécies de lonqueídeos. Os parasitóides emergidos foram das famílias Braconidae e Figitidae, representados por cinco espécies de cada uma. Ceratitis capitata apresentou comportamento sazonal com picos populacionais durante o segundo semestre. Verificou-se que a população de moscas-das-frutas (Anastrepha spp. e lonqueídeos) se mantém na área dos pomares (goiaba, nêspera e pêssego) durante o ano todo. Os três hospedeiros sofreram o ataque de moscas-das-frutas nos seus primeiros estádios de desenvolvimento, a partir de 2 cm de diâmetro. Ao longo do desenvolvimento, em todos os hospedeiros (goiaba, nêspera e pêssego), a infestação foi ascendente, apresentando as maiores elevações no final do amadurecimento. Os parâmetros físicos (tamanho, firmeza, brix e cor) mostraram-se mais confiáveis para o prognóstico da suscetibilidade do fruto ao ataque das moscas-das-frutas do que os parâmetros químicos (pH, acidez e umidade) para todos os hospedeiros. Os aspectos ecológicos da infestação dos frutos pela moscas-das-frutas e a relação tritrófica (parasitóide/mosca/fruto) foram discutidos.<br>Field experiments were conducted in three orchards in Monte Alegre do Sul, SP, each containing either a collection of strains of peach (September to October), guava (January to April) or loquat (August to September), during 2002 and 2003. Three strains of guava (Guanabara, L7P28 and 252), two of loquat (Precoce Campinas and Precoce 264-54) and three of peach (Aurora 2, Dourado 1 and Régis) were used for infestation assessment. Guava and loquat fruits were bagged and unbagged biweekly and weekly, respectively, for the assessment of the infestation period and only weekly for the peach. In all cases, fruits were bagged at the beginning of their development, while still green. A sample of 30 bagged fruits were unbagged weekly (loquat and peach) or biweekly (guava) until fruits were completely ripe, from a total of 500 (guava) and 400 (loquat) fruits. Fruits were measured and a sample was taken for physicochemical analysis. Fruits were harvested once they were ripe, taken to the laboratory, weighted and measured. They were placed into plastic cups half-filled with sand and covered with a fine fabric to allow for field-collected fly development. Three McPhail traps containing Torula yeast (food attractant) were hung from January/2002 to January/2004 to assess the population fluctuation of fruit flies in each orchard. Five tephritids and eight lonchaeids species emerged from field-collected fruits maintained in the lab. Five species each of braconid and figitid fruit fly parasitoids were also observed emerging from these fruits. Ceratitis capitata showed a sazonal behavior with population density peaking at the second semester. It was also observed that Anastrepha spp. and lonchaeids remained in the orchards throughout the year. All three host fruits larger than 2 cm in diameter were infested by fruit flies at the early stages of development. Fruit infestation increased in all orchards as fruit development occurred with the highest infestation level observed at the end of the ripening. Physical parameters, such as size, firmness, brix and color were more reliable in estimating fruit susceptibility to fruit-flies than the chemical parameters evaluated (pH, acidity and humidity) for all three host fruits. The ecological aspects of the infestation of fruits by fruit flies and the tritrophic interactions (parasitoid/fruit fly/fruit) were discussed.

A nêspera (Eriobotrya japonica Lindl.) é uma fruta do tipo pomo, que tem sido cultivada principalmente no estado de São Paulo. Cinco cultivares de nêspera foram selecionadas por se destacarem no sabor e na capacidade produtiva. Foi realizado o estudo das mudanças fisiológicas e físico-químicas dos frutos durante o desenvolvimento e o período pós-colheita. Amostras das cultivares (Centenária, Mizuho, Mizumo, Néctar de Cristal e Mizauto) foram colhidas semanalmente conforme o avanço do estádio de desenvolvimento (na planta), desde a fase verde dos frutos até se tornarem maduros. Os frutos maduros para estudo do período pós-colheita foram colhidos seguindo critérios adotados por produtores, através da cor dos frutos. As determinações físico-químicas realizadas nas nêsperas maduras foram umidade, fibra, ácido ascórbico, açúcares solúveis (glicose, frutose, sacarose e sorbitol), ácidos orgânicos e carotenóides. Durante o período de desenvolvimento das nêsperas, as curvas de respiração decresceram progressivamente, e houve um considerável aumento na produção de etileno, precedendo o aumento na concentração de açúcares e as mudanças de coloração dos frutos (verde a amarela ou alaranjada). A composição de açúcares e ácidos dos frutos maduros foi semelhante a encontrada nos frutos precocemente maduros durante o desenvolvimento na planta. Portanto, a observação visual da cor não foi apropriada para a definição do ponto ideal de colheita. Durante o período póscolheita, as curvas de respiração e de produção de etileno das cultivares tenderam a queda, mostrando comportamento típico de frutos não-climatéricos. O estudo fisiológico e físico-químico das cinco cultivares de nêspera permitiu conhecer melhor o padrão de amadurecimento de cada cultivar, e assim obter informações que auxiliem na definição do ponto ideal de colheita, e que permitam obter frutos de boa qualidade e apropriados para distribuição comercial.<br>Loquat (Eriobotrya japonica Lindl.) fruit is a pome fruit cultivated mainly in the state of São Paulo. Five loquat fruit cultivars were selected to analysis mainly by their high productivity and taste. Physiological and physico-chemical changes were analised during development and ripening fruit. Samples were harvest weekly according to progress in development stage (in the tree), since imature stage (green) until mature stage. Mature stage to harvest was defined by fruit color, like methods\' producer. The physico-chemical parameters analysed were moist value, fiber, ascorbic acid, soluble sugars (glucose, fructose, sucrose and sorbitol), organic acids and carotenoids. During development of loquat fruits, the respiration levels decreased, and show a increase in ethylene levels, which was associed with increase of sugar concentration in the pulp and the changes of fruit color (green to yellow or orange color). According to sugars and acids levels during development (in the tree) and in ripe stage, the composition in ripe fruits was similar to that early mature fruits analyzed during development in the tree. The visual determination of color was not appropriate to define the harvest point of fruits. During postharvest period of the cultivars, the levels in respiration and ethylene production decreased, which is a typical pattern of non-climacteric fruits. The study of physiological and physico-chemical characteristics of loquat cutivars contribute to know the ripening pattern of each cultivar and to auxiliary in the definition of best mature point to harvest and to comercial distribution.

Examensarbetets mål har varit att utforma en artefakt som ska skapa förutsättning för interaktion mellan människor i den offentliga miljön. Intervjuer, observationer och användarstudier har legat till grund för arbetet och litteratur som har stött ledorden vilka har varit användandet, hållbarhet och vardagens estetik. Designprocessen har varvats med ovanstående metoder, formarbete och egna reflektioner i ämnet. Resultatet blev en artefakt som jag har gett namnet Logui vilket betyder tala till, tala om på latin.

Inaugural-Diss.--Theologische Fakultät--Zürich--Universität, 1992.<br>Contient le texte latin avec trad. allemande en regard de la "Disputatio theologica : an haec propositio sit vera in philosophia, 'Verbum caro factum est'" Bibliogr. p. 247-251.

Submitted by MARCIA ROVADOSCHI (marciar@unifra.br) on 2018-08-16T19:56:49Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_LeonardoQuintanaSoaresLopes.pdf: 1473450 bytes, checksum: eb7d00c9d4a740c17c2d7fd0ee98c234 (MD5)<br>Made available in DSpace on 2018-08-16T19:56:49Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_LeonardoQuintanaSoaresLopes.pdf: 1473450 bytes, checksum: eb7d00c9d4a740c17c2d7fd0ee98c234 (MD5) Previous issue date: 2015-03-20<br>The American Foulbrood Disease (AFB) is a fatal larval bee disease. The etiologic agent is the bacteria Paenibacillus larvae, being highly contagious due to the capacity of this microorganism to form spores. The treatment involves incineration of all contaminated materials, leading to high losses. The Glycerol Monolaurate (MLG) is a compound with a known antimicrobial potential, however its use is impaired due to the low solubility in water and the high melting point. In this context, the nanostructuration of some drugs offers advantages like better stability and solubility, increasing effectiveness and reducing toxic effects. The present study aimed to produce, characterize and evaluate the antimicrobial activity, the toxicity in bees and the ecotoxicity of MLG nanoparticles. The nanoparticles produced showed a diameter of 209.9 nm, polidispersion index of 0.044 and zeta potential of -23.4 mV. The study of the stability parameters showed that even after 120 days, the system was stable. The images of transmission electronic microscopy showed the nanometric size and spherical forms. The determination of inhibitory and bactericidal concentration showed that is necessary 142.8 μg/ml of the formulation (MLG nanoparticle) to obtain a bacteriostatic effect and of 285 μg/ml to obtain a bactericidal effect. The assay in bees showed that the free compound has a high toxicity while the nanoparticles showed a low toxic effect, reducing the number of dead bees. When analyzed the ecotoxicity, the springtail test soil results in high toxicity of the free compound, while the nanostructured formulation didn’t influence in mortality either in reproduction of organisms, showing that nanocapsules aren’t toxic to the ecosystem. Concluding, the developed formulation shows positive results in the action to combat AFB beyond not cause damage in bees and in soil, and can be future alternative to the treatment or prevention of AFB.<br>A American Foulbrood Disease (AFB) é uma doença fatal que acomete as abelhas em seu estado larval. Seu agente etiológico é a bactéria Paenibacillus larvae, sendo altamente contagiosa devido à capacidade de esse microrganismo formar esporos. O tratamento atualmente envolve a incineração de todos os materiais contaminados levando a grandes prejuízos. O monolaurato de glicerol (MLG) é um composto com um conhecido potencial antimicrobiano, entretanto, seu uso é prejudicado devido ao seu baixo grau de solubilidade em água e o seu alto ponto de fusão. Neste contexto, a nanoestruturação de fármacos oferece vantagens como aumento de estabilidade em ambientes adversos, melhora as questões de solubilidade, aumentando a eficácia e reduzindo os efeitos tóxicos. O presente estudo teve como objetivo produzir, caracterizar e avaliar a atividade antimicrobiana, toxicidade em abelhas e ecotoxicidade de nanocápsulas contendo MLG. As nanocápsulas produzidas apresentaram um diâmetro médio de 209,9 nm, índice de polidispersão 0,044 e potencial zeta de -23,4 mV. O estudo de parâmetros de estabilidade mostrou que mesmo passados 120 dias, o sistema se manteve estável. As imagens geradas por microscopia eletrônica de transmissão mostraram o tamanho nanométrico além de formas características esféricas. A determinação da concentração inibitória e bactericida mínima mostrou que são necessários 142,8 μg/ml da formulação (Nanocápsulas de MLG) para se obter um efeito bacteriostático e de 285,7 μg/ml para obtenção de um efeito bactericida. O ensaio nas abelhas mostrou que o composto livre possui uma alta toxicidade, enquanto que as nanocápsulas apresentaram um efeito pouco tóxico, reduzindo o número de abelhas mortas. Quando avaliada a ecotoxicidade, o teste no solo com colêmbolos resultou em uma alta toxicidade do composto livre, enquanto que a formulação nanoestruturada não influenciou na mortalidade nem na reprodução dos organismos, mostrando que as nanocápsulas não são tóxicas para o ecossistema. Concluindo, a formulação desenvolvida apresenta resultados positivos em sua ação no combate da AFB além de não causar dano às abelhas e ao solo, podendo ser uma alternativa futura para o tratamento ou prevenção da AFB.

碩士<br>國立中興大學<br>園藝學系<br>83<br>The early-season flower buds of loquat formed during August and September in Southern Taiwan usually fail to set due to relatively poor pollen viability. Hand pollination with viable pollen grains after cold storage produced 15 to 20% fruit set comparable to that under natural condition. Field sampling indicated that pollen yield and rate of germination of loquat increased with the development of inflorescence from September to December. The positive correlation between viability and cold weather led us to predict that the two processes are closely related. Results of temperature regulation experiment showed that the optimal temperature for the development of loquat inflorescence ranged from 18°to 23℃. Temperature above 25℃ were found to be unfavorable. A 4-hour treatment at 34℃ daily for a period of 5 days reduced the highly vital pollen (produced in December with germinating rate of 90%) to the level of viability of these produced in September (30% germinated). Besides, damaged pollen tubes, plasmoptysis were common place among pollens after high-temperature treatment. A measurement of plant tissue temperature especially leaves and flowers in September and October revealed that unshaded floral parts could reach 34℃ midday might have adverse effect on pollen viability. Lowering air temperatkure by shading not only would successfully bring about early-season fruit production ,but also enhanced fruit quality. Data obtained with hand pollination, temperature regulation and early-season shading in this experiment suggest tentatively that air temperature is the limiting factor for the control of pollen viability and successful fruit set of loquat in Taiwan.

碩士<br>國立中興大學<br>園藝學系<br>87<br>Shoot tip culture of the loquat (Eriobotrya japonica Lindl.)was studied, the optimal micropropagation condieions for culture establishment,shoot proliferation and shoot rooting were investigated. Shoot tip were harvested about 5cm in length from summer shoots and shoot apices excised 0.5mm for initial culture.The best grown was achieved when shoot apices cultured on solid medium with 1/2 strength of Mrashige and Skoog (MS) containing BA 1 mg/l,sucrose 30 g/l and agar 7 g/l for 30 days.These explants were transferred to the same component solid medium for further growth. Multiple shoot were induced by MS medium containing BA 4 mg/l, sucrose 30 g/l and agar 8 g/l for 30 days.For elongation of these young shoots,MS medium supplemented with BA 2 mg/l,sucrose 10 g/l and agar 8 g/l or surcose 30 g/l and agar 8 g/l plus kinetin 0.5 mg/l gave a good response. For rooting, the shots grew above 20 mm in hight , were placed on 1/2 MS medium with IBA 10 mg/l , surcose 20 g/l and agar 8 g/l and incubated in darkness for 7 days , then transplanted to an auxin-free 1/2 MS medium for 23 days.

碩士<br>中臺科技大學<br>食品科技研究所<br>100<br>In recent years, changes in diet and lifestyle, making the risk of type 2 diabetes patients increased. If the human body in long-term plasma conditions of high blood sugar, high blood sugar and protein and glucose link easily lead to the formation of non-enzymatic browning is call glycosylation (glycation), will eventually form a higher glycated end product (AGEs). Such AGEs react with the cell receptors lead to in vivo signaling problem or cell damage, and thus make diabetes and related diseases, such as: Diabetic, Retinopathy, Atherosclerosis etc. How to reduce the generation of AGEs and prevent the occurrence of diabetes, or to provide easy and sugar response antioxidants are one of the important topics. Therefore seek natural anti-oxidants to reduce free radicals in body and reduce the development of AGEs also become more important. Loquat in the growth process, the farmers in order to increase the loquat quality, usually flower thinning, fruit thinning, so lots of loquat flowers were discarded each year. In recent years, people healthcare concept gradually increase, there is quite lots of natural plant sold in tea bags, in order to investigate whether the loquat tea also has the effect of health care, this study general composition analysis for loquat flowers, and using alcohol, water / alcohol, 25 ℃, 50 ℃ water extraction loquat flowers antioxidant and anti-glycation substances, anti-oxidation and glycosylated test to assess the loquat flowers extract antioxidant and prevent the diabetic. The loquat flower was processed the general composition analysis of the results show the water content accounted for the most common, followed by the ash content of at least compared to crude protein. Loquat flowers extracted by four different antioxidant test results show that the water /alcohol extract better in total antioxidant, nitric oxide scavenging and DPPH radical clearance rate respectively of 95.5%, 76.5% and 98.1% , its extract phenols and flavonoids content was also more. In the inhibition of linoleic acid oxidation and chelation of ferrous ion is 50 ° C water extract better, 79.1% and 65.9% respectively. Glycosylation due to high blood sugar and protein binding, finally formation irreversible advanced glycation end products. The results showed that with the increase of the protein glycation reaction time, advanced glycation end products (AGEs), the dicarbonyl compounds and the amount of protein carbonyl groups increased more. Loquat flowers extract can effectively reduce the formation of the glycosylation reaction products, inhibition dicarbonyl content of 28.08%, is water / alcohol extract has the best inhibition. Inhibition advanced glycation end products is 25 ℃ water loquat flower extract has 86.87%. The results, loquat flower extract has good resistance to oxidation, in trials of anti-glycated clear dicarbonyl and final glycation products ability, loquat flower extract may have potential prevent diabetes.

碩士<br>國立勤益科技大學<br>企業管理系<br>98<br>With the development of tourism, promotion of industry, and formation of community, the local tourist activities, such as festival, have been growing rapidly than before. Furthermore, the idea to combine festival and local fruit as a main tourist activity in local leisure industry has been widely adopted and promoted for years, such as Grape festival in Da-Tsun, Chan-Hua; Strawberry festival in Da-Hu, Miao-Li; Loquat festival in Taiping, Taichung. Among all these fruit festivals, the Loquat festival is the only annual event that has been ongoing for years. Taiping Loquat Festival has been held continuously for 8 years since 2003. However, not many studies have been conducted in regard of the local government’s policy planning process, traveler’s satisfaction, and the relationship between these two factors. The finding suggested that, despite the fact that the local government of Taiping has intended to provide a variety of subordinate activities accompany the festivals, but the travelers did not find the effort worthwhile and satisfactory. The researcher further analyzed travelers’ satisfactory rate concerning local government’s 4 main political planning goals. The result showed that the travelers were satisfied with the Policy 1 – purchasing quality, Policy 2 - combining governmental and civilian resources, and Policy 4 - a friendly community. Finally, most of the travelers pointed out that related festival activities should be held around the habitat of loquat.

碩士<br>大葉大學<br>食品暨應用生物科技學系<br>107<br>In this study, we used loquat as the main raw material to discuss the optimal extraction conditions of eucalyptus leaves, the preparation of loquat fermented beverages and the addition of eucalyptus leaf extracts to the lyophilized fermented beverages, which were compared with the commercially available sputum fermented beverages, and we have divided into three parts. The first part uses the flavonoid content as an indicator. The results showed that the freeze-dried eucalyptus leaves 40 mesh powder / 95% alcohol at a ratio of 1:10 w/v, 70 ° C, and ultrasonic-assisted extraction for 3 times gave a higher extraction amount of 86.8 mg / g. The second part is the preparation of sputum fermented beverages. After washing the sputum, the sputum is removed, the seeds are removed, and the endometrium is added. The yeast, acetic acid bacteria, and lactic acid bacteria are separately added for the first stage fermentation, and the fermentation broth is coarsely filtered to remove the solid matter. After that, like the fungus, add equal weight (the control group replaces the bacteria with an equal weight of water), and then adjust the sugar content to 65°Brix, and carry out the second stage fermentation to obtain yeast, acetic acid bacteria, lactic acid bacteria, etc. Three kinds of fermented drinks; then these three kinds are compounded into a sputum compound fermented beverage in equal proportions, and a eucalyptus leaf extract is added to improve the functionality. In the third part, five kinds of homemade fermented beverages were compared with the commercially available "loquat fermented drinks" products, and their flavonoids, antioxidants and sensory evaluations were analyzed. The results showed that the total phenolic content, the ability to scavenge DPPH free radicals, the ability to chelate ferrous iron, and the total antioxidant capacity were the highest in the composite fermented beverages with added eucalyptus extract; but the sensory evaluation showed that the acetic acid bacteria group received consumer acceptance. The highest, only the former is slightly bitter, the latter is less functional if properly combined or adjusted to become a potential composite fermented beverage.

碩士<br>元培科技大學<br>生物技術研究所<br>96<br>Loquat, Eriobotrya japonica (Lindley), a member of the Rosaceae family, is originally from Asia. This nutritious fruit have been consumed by the general public for years, and its fruit, leaves and seeds are included in traditional herbal medicine for relieving different symptoms. A previous study showed that the ethanol extract of loquat seeds contained high levels of polyphenolic compounds. Since polyphenols have been shown to exert antioxidant activity, and to modify inflammatory response, cell cycle, signal transduction, etc., the objective of this study was to examine the antioxidant activity and anti-inflammatory effect of loquat seed extract (LSE) on murine macrophage-like RAW264.7 cells treated with lipopolysaccharides (LPS). Results in this study confirmed that LSE contained high levels of polyphenolic compounds and flavonoids, and exhibited strong antioxidant potential s in total antioxidant status and scavenging activities in 1,1-diphenyl-2-picryhydrazl (DPPH) free radical, nitric oxide (NO) and ferric ion. The cytotoxic effect of LSE on macrophage cells was also examined, and it showed that cell proliferation was not influenced as the concentration of LSE was lower than 5 mg/ml. LSE significantly suppressed the synthesis of nitric oxide and pro-inflammatory prostaglandin E2 (PGE2) in LPS-treated cells through inhibitory modulation of nitric oxide synthetase (iNOS) and cyclooxygenase type-2 (COX-2) expression at the translational level, and the suppression was in a dose-dependent manner. Similarly, inhibition also occurred in the synthesis of two inflammatory cytokines, such as tumor necrosis factor  (TNF) and interleukin-6 (IL-6). Besides, LSE significantly reduced 5-desaturation and modulated n-6 polyunsaturated fatty acid (PUFA) metabolism, resulting in cellular dihomo--linolenic acid (DGLA) accumulated, but arachidonic acid (AA) decreased. Taken together, our data suggested that anti-inflammatory effect of LSE might be in part due to its antioxidant potential, inhibitory effect on iNOS and COX-2 expression, or modulation of n-6 PUFA metabolism.

碩士<br>國立中興大學<br>園藝學研究所<br>78<br>The application of GA3 300 ppm on the florescence of loquat did not effect the pollen germanation in vitro viability, but damage the papillate tissure of loquat stigma, resulted in retardation of pollen germination and growth. Distoration of ovule stalk, degratation of embryos sac and abnomal development of ovule were found in the embryo after 7-days of GA3 treatment at full bloom stage. 　　The seedless loquat induced by GA3 300 ppm were treated with BA25 ppm, BA50 ppm, GA3 50 ppm+BA50 ppm or GA3 50 ppm+BA50 ppm twice after 35 days and 71 days of full blooming. Significant increase in fruit weight, fruit length, fruit width, flesh thickness, fruit cell layer and cell size were found in the treatments of GA3 50 ppm+BA 50 ppm and GA3 50 ppm+NAA 50 ppm. The averages fruit weights of these two treatments were 21.56g and 19.96g, respectively. Althoght the fruit weights of seedless fruit were less than that of seed fruits (27.94g), the percentage of flesh were the same. The fruit lengh of these two treatments were 5.25 cm and 5.75 cm, respectively. The fruit width were 0.78 cm and 0.74 cm, respectivity, which are similar to that of seed fruits (0.85 cm) Fruit cell layers of these two treatments are less than that of seeded loquat, but the fruit cell size were larger. The cell division of seed fruit was terminated at 89 days after full blooming while terminated at 98 days in seedless fruits. 　　Increassed in fruit hardiness, flesh percentage, fruit color and total soluble solid of seedless loquat fruit were resulted from treatments of plant growth regulator. The fruit hardiness of loquat kept 2 or 4 fruitles per pedical and treated with GA 50 ppm+NAA 50 ppm, or control were 560 g/cm2, 580 g/cm2, 296 g/cm2, 293 g/cm2, respectivety. GA 50 ppm+NAA 50 ppm traetment increased the percentage of flesh up to 17% and total soluble solids of treated fruit was 9-9.5% higher than seeded fruit. The GA 50 ppm+NAA 50 ppm treated fruit was higher in lightness, hue, saturation than that of the control. The fruits treated with BA 25 ppm had the lowest acid content compared with the control and seeded fruit. 　　Studies on effects of plant growth regulator on the quality of seeded loquat showed that significant increase in the fruit weight, fruit length, flesh percentage, titratable acid, total soluble solids, pH value, and TSS/acidity ratio and decreased in the seed number by the treatment of growth regulators. Loquat fruit treated with BA 50 ppm had lowest fruit seed number. Increase in the fruit weight and fruit length were fuund in the GA3 50 ppm+ BA 50 ppm ppm, in spite of 2, 4, or 6 fruitlet per pedical were kept. The pH value and total soluble solids. were enchanced by spray with GA3 50 ppm+ BA 50 ppm or GA 50 ppm+NAA 50 ppm , GA3 50 ppm+ BA 50 ppm had larger effect on the T.S.S and pH value than other treatments. Overall, growth regulators had a prominent effects on the improvement of the quality and production of seedless and seeded loquat fruit.

碩士<br>淡江大學<br>公共行政學系公共政策碩士班<br>98<br>The main topic of the thesis is using Taiping Loquats Festival in Taichung county as an example, which is based on a viewpoint of elevate local industry festival performance. Introduced different input and output models of Data Envelopment Analysis (DEA) to evaluate the whole performance of the festival in seven years, and to discovered the problems and solutions of the Taiping Loquats Festival, made concrete suggestions of those problems and gave consultations of elevate the quality of the local festival. The discussing questions are as follows: (1)Probing the relative efficiency of Taiping Loquats Festival by comparing the production cost data of Taiping City and Guoshing Township? (2)How to improve the inefficiency unit? (3)Inspecting the problems of Taiping Loquats Festival? Qualitative and quantitative methods are used in this thesis. To understand the extent of Taiping Loquats Festival performance were elevated by analyzing documents, depth interview, and Data Envelopment Analysis. Besides, SWOT analysis is used to discuss the situation and the problems it faced. The results of research are as follows: (1)The per year Decision Making Unit(DMU) of Taiping City had little raise but still decrease of the whole evaluation, the decrease units are 1996、1999、2000、2003、2004、2005、2007、2008 in Taiping City and 1996、1997、2001、2003、2004、2005、2007 in Guoshing Township . (2)After analyzing the external opportunity and threat and internal strength and weakness of Taiping City and Taiping Loquats Festival, producing four models of strategies by TOWS matrix.(3) Taiping Loquats Festival only promote it fame, but doesn’t have any help for farmers.

碩士<br>國立中興大學<br>水土保持學系所<br>105<br>Abstract By discussion on the land investigation analysis and feasibility in regulations, this research analyzed the feasibility of establishing loquat zone in Taiping District, Taichung City, further proposed the specific development and implementation countermeasures for the agriculture in this region and planned the future development direction for loquat zone, so as to meet the demands of overall development in this region and achieve the purpose of caring for farmer&apos;&apos;s rights and interests and natural ecology; meanwhile, evaluation was conducted for the decompilation of protection forest as well as the land management issue and the concrete proposal solution was put forward. According to environment, economy, regulations and land suitability analysis, the establishment of loquat zone will not have the impact on and cause conflict to environment. Based on this, the region suitable for the plantation of loquat only involves the part of decompilation of protection forest. It is suggested that Taichung City Government should file an application according the relevant procedure. In the future, the idea of “conservation of sloping land”, “land-based development” and “industrial sustainability” is planned through local organizational management and the counseling of Taichung City Government and the basic principle of considering farmer&apos;&apos;s rights and interests and livelihood, land resource security, forestland management, environmental security and resource sustainability is followed, so as to promote the loquat zone having ecology, life and production diversity and provide reference for the local residence to create all-around sustainable development in loquat zone.

碩士<br>亞洲大學<br>會計與資訊學系<br>105<br>The farmers’ association plays an important role in not only the history, but the political and social development in Taiwan. After obtaining its membership in the World Trade Organization (WTO) in 2002, agriculture in Taiwan faces international competition, along with the dietary changes domestically, the local agriculture industry changed in order to adapt to fit society’s needs. According to agriculture statistics, Taichung ranked 1st in the production of the loquat fruits, and within the city, Taiping ranked 2nd, just behind Xinshe. It is an important income for Taiping, solidifying the loquat fruit industry as an important factor in the economic development of the district. This research will explore the farmers’ association of Taiping and the methods of production and sales of the loquat fruit, using in-depth interviews to analyze how the farmers’ association instruct local farmers and the development of the association. With this research, one can expect it to improve the continuing development of the association, and the loquat fruit being its most recognizable feature.

碩士<br>國立中興大學<br>植物病理學系所<br>98<br>A new disease, loquat twig blight has occurred in central Taiwan since 2008. Cylindrocladiella isolates Lsc-01 and Lsc-02 and Cylindrocladium isolate Lscy-01 were isolated from diseased loquat twigs. The pathogenicity was confirmed by inoculating loquat seedlings, which showed the same symptoms as diseased plants in the fields. Based on morphological characteristics, ITS and D1/D2 sequences analyses, three isolates were respectively identified as Cylindrocladiella peruviana isolates Lsc-01 and Lsc-02 and Cylindrocladium reteaudii isolate Lscy-01. The optimal temperatures for mycelial growth and conidial germination of Ce. peruviana isolates Lsc-01 and Lsc-02 was at 24-28 ℃, and Cy. reteaudii isolate Lscy-01 also was at 24-28 ℃. The optimal temperature for sporulations of Lsc-01 and Lsc-02 was at 28 ℃, and isolate Lscy-01 was at 16 ℃. Furthermore, conidial suspensions of Ce. peruviana Lsc-01 & Lsc-02 and Cy. reteaudii Lscy-01were respectively sprayed on six fruit crops. The results showed that isolate Lscy-01 could infect seedlings of loquat, guava, plum and wax apple, but Lsc-01 & Lsc-02 could not. Lscy-01 was also able to directly infect fruit without wounds. The fruit rot became more severe when inoculum density of Lscy-01 was increased. After Lscy-01 was inoculated on fruit surface of loquat for 16 hrs, it could form appressorium-like structures around stomata and penetrated directly into the intercellular epidermis. Based upon indoxyl acetate reaction under microscopy observation, it was proved that Lscy-01 was able to produce esterase, but were not Lsc-01 and Lsc-02. Twelve synthetic chemical fungicides were evaluated their effect on suppression of mycelial growth of Lsc-01, Lsc-02 and Lscy-01 on MEA. The results showed that 33.5 % Oxine Copper was very effective in inhibiting compeletly mycelial growth of three isolates at one ppm. Thirty one Chinese medicines were evaluated their effect on suppression mycelial growth of Lsc-01, Lsc-02 and Lscy-01 on MEA. The results showed that 2 ％ Aucklandiae Radix, Scutellariac Radix and Chinensis Galla were effective in inhibiting mycelial growth of three isolates. A wound protectant ointment was formulated with petrolatum and 2 % (w/w) Aucklandiae Radix powder.It was significantly effective in controlling twig blight of loquat when wounded petiole amd twig were treated.

Mestrado em Engenharia Agronómica - Instituto Superior de Agronomia<br>The mode of action of naphthaleneacetic acid (NAA) (20 mg/l) in fruit thinning of loquat was studied through the action of NAA on both female and male gametophyte of ovules and pollen grains cultured in vitro. Additionally, in vivo treatments with NAA to the panicles of different growth stages were carried out. The flowers of these panicles were observed with fluorescence microscopy in order to evaluate the effect of NAA in the ovule’s development and in the pollen grain germination. In vitro results showed that NAA prevents pollen germination, pollen tube growth and ovule’s development from flowers of panicles in the stage 609-700 BBCH. In vivo, NAA also prevents pollen germination and pollen tube growth for 7 days after the treatment. In flowers near the stage 609 BBCH, NAA interrupts the ovule’s development, resulting in the emergence of callose in the chalazal part of the ovule. Ovules in previous or later stages than 605- 700 BBCH are insensitive to NAA, therefore, if this treatment is carried out earlier or later becomes ineffective.---------------------------------Neste trabalho, estudou-se o mecanismo de acção do ácido 1-naftalenoacético (ANA) (20 mg/l) sobre a monda de frutos de nespereira, para tal analisou-se a acção do ANA sobre o gametófito masculino e feminino, procedendo-se à cultura de grãos de pólen e de óvulos in vitro. Paralelamente, foram feitos tratamentos com ANA in vivo, a panículas de diferentes estados fenológicos. As flores destas panículas foram observadas através de microscopia de fluorescência para avaliar o efeito do ANA no desenvolvimento dos óvulos e na germinação de grãos de pólen. Os resultados mostram que o ANA, em condições in vitro, impede a germinação dos grãos de pólen, o crescimento do tubo polínico e o desenvolvimento de óvulos provenientes de flores de panículas no estado 609-700 da escala BBCH. In vivo, o ANA impede a germinação de grãos de pólen e o crescimento do tubo polínico durante 7 dias após o tratamento e nas flores próximas do estado 609 BBCH interrompe o desenvolvimento dos seus óvulos, levando ao aparecimento de calose na calaza do óvulo. Os óvulos em estados anteriores e posteriores a 605-700 BBCH são insensíveis ao tratamento com ANA, consequentemente, se este tratamento for efectuado antecipada ou tardiamente torna-se ineficaz.

碩士<br>國立彰化師範大學<br>地理學系<br>105<br>Taiping District of Taichung City as "Loquat Hometown", but the wholesalers were commissioned to sell loquat in the early, farmers were vulnerable to exploitation, so that profitable meager. Therefore, the Taiping Township Farmers' Association on the one hand through the "Loquat Study Group" to improve the quality of loquat from 1990, on the other hand held loquat evaluation and sales activities to expand direct sales. Then, Taiping City held the first Taiping Loquat Festival in 2003, in one fell swoop started in the name of the Taiping loquat, let the loquat industry from the traditional agriculture to tourism. 　　Based on the theory of "Value Co-Creation", this study analyzes the loquat industry and loquat festival as follows: 1. The best benefit of Taiping Loquat Festival is to increase the unit price and direct sales proportion of loquat in order to maintain the loquat industry; 2. Loquat marketing activities follow Ramaswamy & Gouillart (2010) proposed the four principles, it is fruitful, but the loquat tourist shuttle can not embody the spirit of value co-creation, the effect is not as effective as expected, so value co-creation can conspicuously enhance the festivals and industrial tourism benefits; 3. The major predicament of loquat industry is the production season is too short, if the new varieties are more suitable for Taiwan, and with the provisions of "Spatial Planning Act" Ex-regional Protection forest into agricultural development zones, combined with the Taiping Loquat Festival promotional marketing, Taiping loquat industry will be able to sustainable development.

碩士<br>國立高雄師範大學<br>地理學系<br>101<br>Most of Taiwanese loquats yield from shallow hill regions in the central Taiwan such as Xinshe District, Taiping District and Guoxing Rural Township. The high labor-demanding period of loquat cultivation lasts from the end of October to April next year. However, the scarce population in hill regions can not afford the labor-intensive loquat cultivation. It has become the major issue for loquat farmers to cope with seasonal labor demands and accomplish all the harvests every year. Since loquats are soft and difficult to preserve, another issue drives from the types of selling. Applying semi-structured interviews and field research as the research methods, this study aims to investigate the loquat cultivation in Xinshe, Taiping and Guoxing with the time-geographical approach. The results show the main reason that Xinshe, Taiping and Guoxing have become the main yielding regions of loquats attributes to their perfect cultivation condition. In addition, since loquats are rare and high-priced in the fruit market, the benefits from growing have attracted farmers in the central Taiwan. The first cultivation of loquats began in Tou-Bian-Keng, Taiping, and gradually scattered to the low hills nearby in 1950s. In 1970s, the loquat cultivation had reached its peak, having the maximum yield amount and growing area. The scale of loquat cultivation had reduced in 1980s due to the pests and lower competitiveness, yet the cultivation had also expanded into higher altitudes and formed a phenomenon of “the hill-climbing loquats”. Although some loquats are still cultivated in protection forest nowadays, the government has bizarrely chosen to burn a blind eye and help popularize corps harvested from the over-utilization forest. There are three types of labor allocation as coping with the seasonal demands of loquat harvests. The first type refers to professional cultivation, making loquats the only or high-proportion crop and hiring part-time workers in peak seasons. Multi-crop farming is the second type; it refers to cultivate loquats and other corps with similar proportions and different peak seasons. By planting other crops as the slack seasons of loquats, the farmers can also diversify the risk. The third type of labor allocation refers to hiring no part-time workers. The farmers hold negative attitude toward the loquat cultivation since they are aged and taking the loquat cultivation as an exercise. These farmers are too weak to do heavy work, and they put less attention on loquats but other crops. In order to maximize the selling speed and profits of loquats, the farmers always market their own products for more profits. However, the sustainable management of selling their own loquats also needs to work on customer development and credit accumulation. The reason why Taiping can market more loquats than other two districts attributes to its brand effect of “Loquat Hometown”. On the other hand, some loquat farmers will sell the remaining loquats to the local wholesalers after they tempt to market their own products. There are also some farmers choose to sell the loquats all at once to the local wholesalers when it comes to rich harvests. The loquats are still profitable as sending to the peasant associations for co-cooperative marketing. However, since the loquat famers also have to bear the marketing costs, it is not the most profitable way to sell loquats and the last selling option for farmers. Among the three regions, farmers in Taiping can always find out more profitable selling types than co-cooperative marketing. The co-cooperative marketing business in Xinshe achieves little progress since the farmers tend to put up their fruits to auction in the fruit and vegetable wholesale market. About half of the loquat harvest in Guoxing is sold to the northern Taiwan by the co-cooperative marketing due to the inconvenience of transportation. The northern Taiwan is the prior market of loquat wholesales due to the intensive population and various sorts of consumers with higher income and consuming ability.

La version intégrale de ce mémoire est disponible uniquement pour consultation individuelle à la Bibliothèque de musique de l’Université de Montréal (www.bib.umontreal.ca/MU).<br>Au début du dix-huitième siècle, un nouveau genre opératique en « langue napolitaine » se développe à Naples, la commedeja pe ‘mmuseca (« comédie en musique »). Celui-ci intègre peu à peu dans ses intrigues des personnages s’exprimant en toscan et accompagnés d’une musique inspirée du dramma per musica. Cette intégration de la langue littéraire de la Péninsule et d’un style musical serio est interprétée comme une tentative d’ennoblir le genre et a mené à la théorisation d’une dichotomie entre sphère sérieuse toscane et sphère comique napolitaine. Fondée sur une vision réductrice de la situation linguistique napolitaine, associant le toscan aux nobles et le napolitain au peuple, cette conception ne reflète ni la situation linguistique et politique de l’époque, ni la réalité des livrets et de leur mise en musique, comme ce mémoire se propose de le démontrer. En effet, l’étude du contexte politique et linguistique dans lequel la commedeja s’est développé révèle que le napolitain était la langue usuelle du peuple et de l’aristocratie. De plus, Naples était sous domination étrangère et le toscan était la langue employée par les dominateurs: le napolitain devenait donc un symbole de napolétanéité et du désir d’autonomie des habitants du Royaume de Naples. Dans ce mémoire, le dépouillement d’un corpus de soixante-six livrets datant de 1721 à 1749 et l’analyse de l’opéra Lo frate ‘nnamorato (1732) de Gennaro Antonio Federico et Giovanni Battista Pergolesi suggèrent que l’intégration du toscan dans la commedeja incarne une forme de résistance contre la domination étrangère.<br>The commedeja pe ‘mmuseca, a comic operatic genre in Neapolitan, flourished in early eighteenth-century Naples. Gradually, it began integrating Tuscan-speaking characters into its plots, accompanied by a dramma per musica inspired musical style. This inclusion of the literary language of the Peninsula and of a seria musical style has been interpreted as an attempt to ennoble the genre, and has led to the theorization of a dichotomy between Tuscan serious characters and Neapolitan comic ones. This view of Tuscan is based on the mistaken premise that Neapolitan is the language of the people and Tuscan, that of the aristocracy. I argue that, caused by a misunderstanding of the political and linguistic context of eighteenth-century Naples, it not only overlooks the reality of the music and libretti, but also neglects the political and nationalistic dimensions of the genre’s multilingualism. In fact, Neapolitan was spoken by all Neapolitans, whereas Tuscan was the language of foreigners and of the Austrian ruling power. This leads me to suggest a new interpretation to the commedeja’s multilingualism: accompanied by regional characterization, it is actually a form of local resistance against foreign domination. The analysis of sixty-six libretti, dating from 1721 to 1749, and of Lo frate ‘nnamorato (1732) by Gennaro Antonio Federico and Giovanni Battista Pergolesi, confirm this hypothesis.

Dissertação de Mestrado em Engenharia Zootécnica<br>A loque americana é umas das principais doenças que afeta a criação das abelhas melíferas. Esta doença é altamente virulenta e contagiosa, pois é causada pela bactéria formadora de esporos, Gram-positiva Paenibacllus larvae. Os sinais desta doença são bastante evidentes nos favos da criação, pois após a infeção as larvas passam de uma cor pérola para uma cor castanha escura. Nesta fase, cada larva morta contém aproximadamente 2,5 x 109 esporos que são transmitidos para outras larvas através das abelhas obreiras. O apicultor consegue facilmente observar a criação doente, pois esta apresenta sinais caraterísticos, devendo proceder de imediato à destruição da colmeia afetada, impedindo que ocorra a transmissão de esporos de colónia em colónia e entre apiários. No entanto, a deteção tardia da loque americana baseada apenas em sinais clínicos presentes na colónia, é preocupante visto que o único tratamento possível consiste na destruição da colmeia. Desta forma, o primeiro objetivo deste estudo consistiu em detetar a presença de esporos de Paenibacillus larvae em produtos sem sinais visíveis de contaminação, como o mel e a cera. Posteriormente, foram adicionadas ao estudo amostras de mel tratado com radiação gama, favos com sinais da doença e por fim um suplemento alimentar (pasta de açucar). A metodologia utilizada para a deteção de esporos viáveis do agente patológico presente nas amostras consistiu em técnicas laboratoriais de cultivo em meio sólido suplementado com antibióticos, seguido por testes de identificação, nomeadamente teste da catalase e coloração Gram, e por último confirmação por reação em cadeia da polimerase. Das 75 amostras de mel analisadas, 57% continham esporos viáveis, sendo que a maioria de casos positivos foram detetados nas amostras provenientes dos distritos do Porto, Aveiro, Coimbra, Viseu e Beja. Relativamente à concentração de esporos viáveis por grama de mel verificou-se a existência de dois focus, um na região litoral norte/centro que corresponde ao distrito do Porto, Aveiro e Viseu, e outro em Portalegre, contendo ambos entre 60 a 130 unidades formadoras de colónias. No caso da cera, em 15 amostras analisadas, 20% foram positivas para a deteção de esporos, e continham em média entre 17 a 37 esporos viavéis por grama de cera. No que diz respeito às restantes amostras analisadas, o mel tratado com radiação gama foi negativo para a presença de esporos, sugerindo a eficiência deste tratamento na eliminação de esporos, todas as amostras de favos com sinais de doença foram positivas para a presença de P. larvae, e por último, a pasta de açucar que esteve em contato com uma colmeia infetada foi também positiva para a deteção de esporos viáveis. Esta metodologia mostrou-se eficiente na deteção de esporos viáveis da loque americana nos vários produtos apícolas. Visto que o esporo é altamente resistente a altas temperaturas e a desinfeções químicas, a prevenção e o maneio apícolas são fundamentais no controlo desta patologia. Com esta metodologia é possível avaliar o estado sanitário dos apiários e eventuais produtos apícolas que se pretendam utilizar no maneio das colónias, evitando desta forma a disseminação desta doença.<br>American foulbrood is one of the most important diseases affecting honey bee brood. This disease is highly virulent and contagious, because it is caused by the spore forming Gram-positive bacterium Paenibacillus larvae. The clinical signs of the disease in the brood comb are highly visible because larvae changes from a pearly white to a dark brown color. At this stage, each dead larva has approximately 2.5 x 109 spores, and they are transmitted through nurse bees to other larvae. Beekeepers can easily observe the disease brood, because of its specific signs. They should immediately burn the infected beehive, preventing the spores spreading within and between colonies and apiaries. The late detection of American foulbrood based on clinical signs alone is alarming since the only possible treatment consists of beehive burn. Therefore, the first objective of this study was to detect the presence of Paenibacillus larvae spores in beehive products without any clinical signs of the disease, such as honey and wax. Subsequent to this study, honey treated with gamma radiation was added, combs with clinical disease signs and finally a supplementary feed (candy). The methodologies used for viable spore detection were culture laboratory techniques on solid media supplemented by antibiotics, followed by identification tests, namely catalase test and Gram staining, and finally confirmation tests using polymerase chain reaction. Of 75 honey samples examined, 57% contained viable spores, with majority of positive samples detected in Porto, Aveiro, Coimbra, Viseu e Beja. Relative concentration of viable spores per gram of honey was observed and there are two areas of focus, one in the north/center cost region, corresponding to Porto, Aveiro, and Viseu and another in Portalegre, containing both between 60 to 130 colonies forming units. Of 15 wax samples examined 20% were positive, and the average of colony forming units per gram of wax was between 17 to 37 viable spores. In the case of the remaining samples, the honey treated with gamma radiation was negative for spore presence, suggestion the effectiveness of this treatment for spore elimination, all brood combs samples with clinical signs of disease were positive to P. larvae, and finally, the candy sample that was in contact with infected colonies, was also positive for viable spore detection. The methodology used was shown to efficiently detect American foulbrood viable spores in several beehive products. Since the spore is extremely resistant to high temperatures and to chemical disinfestations, prevention and adequate beekeeping handling were crucial to control this pathology. With the implementation of this methodology, it is possible to determine the sanitary status of apiaries and beehive products, while avoiding the dissemination of the disease.

El objetivo central de este trabajo de tesis fue realizar una evaluación integral de los antibióticos oxitetraciclina, tilosina, tilmicosina y lincomicina para el control de la loque americana. Se puso a punto una técnica para evaluar la sensibilidad in vitro de los cuatro antibióticos contra cepas de Paenibacillus larvae adaptando técnicas propuestas por la NCCLS pero empleando MYPGP como medio basal. Este medio resultó altamente efectivo para determinar la sensibilidad/resistencia de P. larvae frente a los antibióticos probados porque permitió el crecimiento óptimo de esta especie y los controles de calidad se ajustaron a los parámetros propuestos. Se propusieron las siguientes cargas de discos: Tetraciclina: 5 µg; Tilosina: 15 µg; Tilmicosina: 15 µg; Lincomicina: 15 µg y, a partir de las mismas se elaboró un standard para interpretación de los discos: Tetraciclina: sensible > 19 mm de diámetro de halo de inhibición; Intermedio 15-18 mm y resistente ≤ 14 mm; Tilosina: sensible 1 > mm; Intermedio 11-13 mm y resistente ≤ 10mm; Tilmicosina: sensible 14 > mm; Intermedio 11-13 mm y resistente ≤ 10 mm; Lincomicina: sensible 21 > mm; Intermedio 15-20 mm y resistente ≤ 14 mm. Se determinaron los puntos de corte para la especie en estudio con respecto a los valores de Concentración Inhibitoria Mínima (CIM), y se propone el empleo de los siguientes puntos de corte epidemiológicos: Tetraciclina: sensible ≤ 4 µg/ml y resistente > 16 µg/ml; Tilosina: sensible ≤ 2 µg/ml y resistente > 8 µg/ml; Tilmicosina: sensible ≤ 2 µg/ml y resistente > 8 µg/ml; Lincomicina: sensible ≤ 4 µg/ml y resistente > 16 µg/ml. Teniendo en cuenta que ya existen casos de resistencia hacia la oxitetraciclina, el uso de estos puntos de corte epidemiológicos permitirá denotar rápidamente pequeños cambios en la población salvaje de Paenibacillus larvae. Ninguno de los ATB evaluados resultó tóxico para abejas adultas; tanto tilosina, como tilmicosina y lincomicina resultaron virtualmente atóxicos. En el caso de oxitetraciclina los valores de LD50 obtenidos resultaron próximos al límite de compuestos tóxicos, de modo que un mínimo error en la dosificación a campo puede resultar en toxicidad para abejas adultas. Se puso a punto una técnica para determinar la toxicidad larval de los ATB la cual resultó satisfactoria y permitió determinar claramente diferencias estadísticamente significativas. Sólo el tratamiento oxitetraciclina presentó una mortandad larval superior a la normal, lo cual demuestra que resulta tóxico. Tilmicosina, tilosina y lincomicina por el contrario no resultaron tóxicos para las larvas, aunque de los tres, la tilmicosina presentó un mayor porcentaje de toxicidad dentro de los parámetros normales y la lincomicina tuvo los niveles de mortandad más bajos. Finalmente se pudo concluir que todos los antibióticos ensayados en esta tesis resultaron efectivos para el control de la enfermedad a campo a las dosis y en las formas de aplicación probadas. Los resultados de todos los ensayos con tilmicosina para el control de P. larvae, tanto in vitro como a campo y de toxicidad para abejas y larvas, constituyen el primer registro a nivel mundial. En tanto que los resultados de todos los ensayos con lincomicina para el control de P. larvae, tanto in vitro como a campo y de toxicidad para abejas y larvas, son inéditos para la Argentina.

Dissertação de mestrado em Bioengenharia<br>A Loque Americana (LA) é uma doença causada pela Paenibacillus larvae, uma bactéria gram-positiva formadora de esporos, que são a forma infetante das primeiras fases larvares das abelhas. Esta doença leva à destruição de colónias inteiras, causando graves perdas financeiras e da população de abelhas, em todo mundo. A restrição à presença de antibióticos no mel imposta pela legislação Europeia, e o aparecimento de resistências aos antibióticos mais usados, dificulta o seu uso no tratamento da LA, tornando urgente desenvolver métodos antimicrobianos alternativos. O uso de endolisinas tem sido considerado uma boa alternativa aos antibióticos, especialmente em bactérias gram-positivas. Neste trabalho avaliou-se o efeito antimicrobiano da endolisina PlyPl23, uma N-acetylmuramoyl-L-alanine amidase derivada do fago phiIBB_Pl23, no controlo de P. larvae. Após clonagem, expressão e purificação da PlyPl23, realizaram-se ensaios in vitro de modo a determinar as condições de reação ótimas e avaliar a sua capacidade antimicrobiana. Todas as estirpes identificadas como P. larvae (71.2 % pertencentes ao genótipo ERIC I e 19 % ao ERIC II) foram sensíveis à PlyPl23 com um maior espetro de ação do que o fago phiIBB_Pl23 (76.2 %). A PlyPl23 apresentou valores de pH ótimo a pH 3 e muito elevado a pH 4 (em média, 56.5 % da atividade apresentada em pH3), que correspondem a níveis de pH encontrados no mel, néctar, pólen e geleia real. Quando suplementada com 200 mM NaCl a pH 7 (pH semelhante ao das larvas, 6.8), apresentou resultados de atividade mais elevados do que sem a adição do sal, (em média, com 0 mM NaCl observou-se 74.6 % da atividade obtida com 200 mM NaCl). Ao contrário, a adição do ião Zn2+ ao tampão de reação não melhorou a atividade. Relativamente à avaliação do efeito antimicrobiano da PlyPl23, verificou-se que uma concentração de 0.2 μM foi suficiente para diminuir a população de P. larvae em cerca de 4 log. Partindo de 104 CFU.ml-1, que se assemelha às concentrações de P. larvae encontradas em larvas infetadas, a enzima reduziu a bactéria para níveis não detetáveis. No que diz respeito aos testes de estabilidade em substâncias importantes a ter em conta para a aplicação in vivo, a enzima, manteve a atividade quando sujeita ao contato com, sacarose, e melhorou o seu efeito antibacteriano após contato com geleia real (aumento médio de 64.9 % de atividade após 30 min) e com homogeneizado de larva (diminuição mais 1 log de atividade após 15 min), relativamente à ação em tampão Tris pH7, 200 mM NaCl. A PlyPl23 é, até à data, a primeira endolisina derivada de um fago de P. larvae a ser reportada, e os resultados obtidos revelaram um elevado potencial para integrar um produto comercial para o controlo da loque americana.<br>The American foulbrood (AFB) is a disease caused by Paenibacillus larvae, a gram-positive spore-forming bacteria. The spores are the infective form, of the first larval stages of honeybees. This disease leads to the destruction of entire colonies, causing serious financial losses and the decline in bee population worldwide. The restriction on the presence of antibiotics in honey imposed by European legislation, and the emergence of bacterial resistance to commonly used antibiotics, hamper its use in the AFB treatment and makes urgent the development of alternative antimicrobial methods. The use of endolysins, bacteriophage-derived enzymes, has been considered a good alternative to antibiotics, particularly in gram-positive bacteria. In this work, the antimicrobial effect of an endolysin, PlyPl23, to control P. larvae was evaluated. PlyPl23, was identified as an N-acetylmuramoyl-L-alanine amidase and derived from phage phiIBB_Pl23. After cloning, expression and purification of the enzyme, in vitro assays were performed to determine the optimal reaction conditions and to evaluate the antimicrobial performance. All the strains identified as P. larvae (71.2 % belonging to genotype ERICI and 19 % to ERIC II) were sensitive to PlyPl23 with a broader spectrum of action than phiIBB_Pl23 phage (76.2 %). PlyPl23 presented the optimum pH at pH 3 and very high activity at pH 4 (56.5 % of the activity observed in pH 3, on average), which correspond to the pH levels found in honey, nectar, pollen and royal jelly. When PlyPl23 acted in Tris pH 7 supplemented with 200 mM NaCl (pH similar to larvae, 6.8), results showed higher activity than without the addition of salt, (on average, with 0 mM NaCl was observed 74.6% of the activity obtained with 200 mM NaCl). In contrast, the addition of Zn2+ to the reaction buffer did not improve the activity. Relatively to the antimicrobial effect of PlyPl23, it was found that a concentration of 0.2 μM was sufficient to decrease the population of P. larvae in about 4 log. Starting from a concentration of 104 CFU.ml-1, which is similar to concentrations found in infected larvae, the enzyme reduced the bacteria to non-detectable levels. Concerning to stability tests when the enzyme was in contact with substances that are important to consider for in vivo applications, it was observed that the enzyme remained active after contact with sucrose, and improved its antibacterial effects when in contact with royal jelly (average increase of 64.9% activity after 60 min) and larvae homogenized (decrease of 1 log of activity after 15 min), comparing with the action in Tris pH 7, 200 mM NaCl. PlyPl23 is, to this date, the first P. larvae phage- derived endolysin that was reported, and the results revealed a high potential of the enzyme to comprise a commercial product for the control of AFB.

La enfermedad más grave de la etapa larval de las abejas (Apis mellifera L.) es la loque americana, causada por la bacteria esporulada Gram (+) Paenibacillus larvae. Es muy contagiosa y posee problemas únicos para su prevención y control debido a que las esporas bacterianas mantienen su capacidad infectiva durante tiempo prolongado y sobreviven bajo condiciones ambientales adversas, no existiendo brotes estacionales ya que se manifiesta en cualquier época del año con la condición que haya cría presente en la colmena. La enfermedad tiene difusión mundial y aparece en la lista de enfermedades de la OIE (Office International des Epizooties - Organización Mundial de la Salud Animal), que incluye enfermedades transmisibles que son consideradas de impacto socio-económico y/o de importancia para la salud pública entre países y que influyen negativamente en el comercio internacional de animales y productos de origen animal. En la Argentina se la detectó por primera vez en 1989 y actualmente está ampliamente diseminada en todas las áreas productoras de miel. Con el objeto de estudiar la diversidad de Paenibacillus larvae se aplicaron procedimientos de microbiología clásica y de biología molecular para caracterizar una colección de 455 cepas del patógeno. El cepario de trabajo se conformó con 106 aislamientos obtenidos de larvas de abejas con síntomas de la enfermedad, con 307 cepas aisladas de mieles contaminadas de distintos orígenes geográficos, 21 cepas provistas por otros laboratorios y 21 cepas de Colecciones Internacionales. Se identificaron 54 aislamientos de restos larvales y/o escamas con síntomas clínicos de la enfermedad y 252 aislamientos de mieles provenientes de distintas localidades de la Argentina. A partir de material de panales con síntomas clínicos remitidos por investigadores de otros países, se obtuvieron 52 aislamientos conformados por 3 aislamientos de Alemania, 5 de Francia, 5 de Italia, 9 de N. Zelanda, 6 de Polonia, 11 de Sudáfrica, 4 de Suecia, 1 de Túnez y 8 de Uruguay. Adicionalmente se aislaron 18 cepas de mieles de Italia, 17 de mieles de EE.UU., 7 de mieles de Francia, 5 de mieles de España, 3 de mieles de Canadá, 2 de mieles de Sudáfrica, 1 de miel de Brasil, 1 de miel de Túnez y 1 de miel de Panamá conformando un total de 55 cepas obtenidas de mieles de otros países productores. Fueron recibidas como cultivo 42 cepas: 2 de Argentina, 2 de Bélgica, 2 de Chile, 2 de Japón, 3 de Inglaterra, 4 de República Checa, 10 de EE.UU. y 13 de origen desconocido. Si bien la colección estuvo integrada por un alto número de cepas provenientes de Argentina (n= 308) el trabajo se realizó también sobre un número considerable de cepas (n=147) de otros orígenes que resultó adecuado para los estudios de diversidad. Adicionalmente, con el fin de incluir controles, se emplearon 5 cepas de Colecciones Internacionales que en la actualidad se clasifican como Paenibacillus larvae pero que pertenecían a la anterior subespecie Paenibacillus larvae subsp. pulvifaciens (1 de EE.UU., 1 de Canadá, 1 de Francia y 2 de origen desconocido). Otras cepas de este grupo proveniente de EE.UU. fueron gentilmente cedidas por el Dr. D. P. Stahly para los estudios con el bacteriófago PPL1c del capítulo III. Se constituyó un segundo cepario con una colección de 282 aislamientos bacterianos de otras especies esporuladas aerobias conformado por 129 aislamientos de Bacillus cereus, 62 de Paenibacillus alvei, 52 de Bacillus megaterium, 8 de Bacillus mycoides, 6 de Bacillus subtilis, 6 de Lysinibacillus sphaericus, 5 de Bacillus thuringiensis, 5 de Brevibacillus laterosporus, 5 de Bacillus circulans, 2 de Bacillus licheniformis, 1 de Bacillus polymyxa y 1 de Bacillus pumilus. Adicionalmente, se emplearon 41 cepas recibidas de Colecciones Internacionales y pertenecientes a distintas especies de los géneros Paenibacillus, Bacillus, Brevibacillus, Lysinibacillus y Virgibacillus. En el Capítulo II se caracterizaron las 455 cepas de Paenibacillus larvae mediante técnicas microbiológicas. El agregado de ácido nalidíxico al medio de cultivo a una concentración final de 9 µg/ml resultó adecuado para la obtención de cultivos puros de Paenibacillus larvae evitando el sobredesarrollo del saprobio Paenibacillus alvei comúnmente presente en las larvas de abejas. Con respecto a los aislamientos provenientes de muestras de mieles, el agregado de la combinación de antibióticos ácido nalidíxico y ácido pipemídico al medio MYPGP permitió obtener colonias de Paenibacillus larvae y mayormente inhibió el desarrollo de otras especies esporuladas presentes en este tipo de muestras, ya que más del 80% de las muestras analizadas contenía más de una especie bacteriana resistente al tratamiento de shock térmico de 80 °C. Independientemente del origen geográfico y/o floral de las mieles analizadas aquí, las especies esporuladas aerobias encontradas con mayor frecuencia han sido Paenibacillus larvae, Paenibacillus alvei, Bacillus cereus, Bacillus megaterium y Bacillus pumilus. Paralelamente y, en menor proporción, se hallaron cepas de Brevibacillus laterosporus, Bacillus mycoides, Bacillus thuringiensis, Bacillus subtilis, Bacillus circulans, Paenibacillus polymyxa, Lysinibacillus sphaericus y Bacillus licheniformis. Mientras que Paenibacillus larvae y Paenibacillus alvei están asociados con enfermedades de las abejas, Bacillus cereus y Bacillus megaterium son especies ubicuas frecuentes en todo tipo de suelos, sus esporas sobreviven la distribución en polvos y aerosoles siendo vehiculizadas desde estos lugares a otros hábitats como las superficies florales. Paralelamente, Bacillus megaterium junto con Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, Bacillus cereus, Bacillus circulans y Paenibacillus alvei son las especies predominantes en los tractos digestivos de larvas, abejas adultas y heces de las mismas. También en este capítulo, se determinó que la configuración superficial de las esporas bacterianas vistas al MEB resultó coincidente con lo descripto por otros autores para cada una de las especies analizadas. En el capítulo III se demostró que el bacteriófago PPL1c resultó una herramienta útil para la identificación y diferenciación rápida de cepas de Paenibacillus larvae, como complemento de otras características fenotípicas. El fago mostró ser altamente específico a nivel de especie encontrándose un 98 % de cepas de Paenibacillus larvae susceptibles al mismo mientras que el resto de las especies de Paenibacillus, Bacillus, Brevibacillus, Lysinibacillus y Virgibacillus resultaron resistentes. En el capítulo IV se estableció que la técnica de rep-PCR resultó adecuada para determinar la variabilidad de las poblaciones de Paenibacillus larvae dónde se halló una alta homogeneidad genética. El empleo de los cebadores BOX y REP reveló la existencia de 4 perfiles de fingerprints a los cuales denominamos A, B, C y D respectivamente, mientras que las cepas de colección pertenecientes al ex. grupo pulvifaciens mostraron el perfil distintivo E. Cuando se emplearon los cebadores ERIC, con excepción de algunas cepas provenientes de Colecciones Internacionales, todos los aislamientos obtenidos de larvas y mieles presentaron un perfil ERIC-I coincidente con lo descripto también por otros investigadores y sólo se halló el perfil ERIC-II en cepas pigmentadas enviadas de la Colección del SAG. Mientras que el perfil ERIC-I es el de mayor distribución a nivel mundial, el perfil ERIC-II ha sido citado en Alemania, Finlandia, Suecia y Austria y, muy recientemente, en muestras de Canadá y Nueva Zelanda. Las cepas pertenecientes al perfil ERIC I estudiadas en esta tesis a su vez se subdividían en alguno de los 4 perfiles BOX (A, B, C o D) y presentaban colonias "típicas" de Paenibacillus larvae no pigmentadas, de color entre blanquecino y grisáceo y formas ligeramente rugosas en agar MYPGP y en agar Columbia suplementado con sangre ovina (Capítulo II). Por otra parte las cepas de colección del grupo ex pulvifaciens, analizadas aquí (n=5) presentaron un perfil ERIC–IV / BOX-E. Si bien en este trabajo no se encontró ningún representante del genotipo ERIC-II, a excepción de los cultivos enviados por el SAG, el mismo pudo haber pasado inadvertido en el análisis de mieles debido a la alta contaminación de las mismas con otras especies esporuladas Gram (+) que hizo imprescindible someter a las muestras a un shock térmico previo al aislamiento en medios semi-selectivos. En el capítulo IV también se concluyó que el gen 16S rRNA es polimórfico entre las especies esporuladas aerobicas comúnmente presentes en la miel, no obstante, no se detectaron polimorfismos a nivel intra-especie en las cepas de Paenibacillus larvae de distintas regiones geográficas cuando se emplearon las endonucleasas (Alu I, Msp I, Hae III, CfoI, Rsa I y Taq I). En cuanto a las comparaciones entre perfiles de rep-PCR y RFLP se observó que los 4 patrones de BOX-PCR observados (A, B, C y D) y los fingerprints ERIC-I y ERIC-II se correlacionaron con el perfil A obtenido por HaeIII, mientras que las cepas de colección que mostraron un perfil BOX-E y un perfil ERIC-IV se correlacionaron con el perfil B obtenido con la endonucleasa HaeIII. Lo mismo ocurrió con la enzima HinfI dónde las cepas pertenecientes a los perfiles ERIC-III y BOX-E mostraron un patrón B y las cepas con perfiles ERIC-I o ERIC-II y BOX-A, BOX-B, BOX-Cy BOX-D no presentaron un sitio de corte con esta endonucleasa. En el capítulo V se seleccionó un subconjunto de aislamientos de Paenibacillus larvae de distintos orígenes geográficos y perfiles de fingerprints para determinar la sensibilidad/ resistencia a tetraciclina y oxitetraciclina mediante las técnicas de CIM y de difusión en agar. La distribución de la sensibilidad por CIMs de la población bacteriana analizada varió entre 0,062 y 128 µg/ml de Tc sugiriendo una distribución bimodal con dos sub-poblaciones que claramente difirieron en su sensibilidad, con 76% de cepas S (CIM entre 0,062 y 2 µg/ml) y 24 % de cepas dentro del rango de la zona I y R. Las cepas I provenían en su mayoría de Argentina y las S cuyos valores de CIM resultaron los más bajos, de países europeos, lo cual es esperable dado que en casi toda la UE está prohibido el uso de antibióticos en apicultura, mientras que en países como EE.UU., Canadá y Argentina, entre otros, es usual el empleo de OTC para el control de loque americana en las colmenas afectadas. Al comparar las CIM obtenidas por E-test con las obtenidas por dilución en agar, en el caso de agar Iso-Sensi Test hubo concordancia de categoría del 100 %, encontrándose la misma proporción de cepas susceptibles y resistentes que por la técnica de dilución en agar; pero al emplear MYPGP hubo concordancia de categoría del 81,81 %. Se concluye entonces que la combinación de las tiras de E-Test con el agar ISO-Sensi Test representa una alternativa válida para determinar las CIM de tetraciclina en cepas de Paenibacillus larvae. Es importante destacar que todas las cepas de Paenibacillus larvae ensayadas mostraron un desarrollo confluente en Iso-Sensi test, el cual podría usarse también como medio alternativo para el desarrollo de las células vegetativas de este patógeno. Se obtuvieron las secuencias completas de los plásmidos pPL373, pPL374 y pPL395. Dichas secuencias se depositaron en el GenBank con los números de acceso KF 433938 para pPL373, KF 536616 para pPL374 y KF440690 para pPL395, provenientes de las cepas PL373, PL374 y PL395, respectivamente que presentaban resistencia a Tc y OTC. Hemos demostrado experimentalmente la transferencia del plásmido pPL374 en la cepa de Bacillus subtilis m351 y del plásmido pPL373 en la cepa de Bacillus subtilis GSY1104 mediante conjugaciones en medio líquido. Al examinar ambas cepas dadoras de Paenibacillus larvae PL373 y PL374 mediante la técnica de lisis in situ se observaron dos plásmidos de aproximadamente 5.000 bp y 8.000 bp, pero sólo el plásmido más pequeño era el que contenía el gen de resistencia a Tc. Luego de efectuar la secuenciación completa de los plásmidos más pequeños obtenidos de las cepas pPL373 y pPL374, respectivamente confirmamos que el gen de resistencia es el tetL y no el tetK como creímos en un principio y que ambos replican por el mecanismo de círculo rodante encontrado en pequeños plásmidos movilizables de alto número de copias presentes en bacterias Gram (+). Todos los experimentos de movilización por conjugación empleando la cepa PL395 que contiene un único plásmido de 5.000 bp denominado pPL395 resultaron infructuosos, no obstante el ADN plasmídico obtenido de la cepa PL395 si pudo transferirse por electroporación a una cepa de Paenibacillus larvae TcS. Los plásmidos pPL373 y pPL374 también pudieron transferirse por electroporación a la misma cepa de Paenibacillus larvae TcS. En todos los casos los plásmidos se mantuvieron en forma estable en sus respectivos aceptores. Los plásmidos más grandes contenidos en las cepas PL373 y PL374, podrían codificar para funciones conjugativas y, al coexistir en la misma cepa bacteriana aportarían a los plásmidos movilizables pPL373 y pPL374 las funciones conjugativas necesarias para su movilización por conjugación; por lo que se justifica estudiarlos en profundidad y secuenciarlos para clarificar los mecanismos de transmisión plasmídica de Paenibacillus larvae en la naturaleza. La presencia de plásmidos con secuencias muy similares aislados de bacterias Gram (+) provenientes de distintas zonas geográficas y de distintos nichos ecológicos (suelo, hábitats marinos, alimentos) de los géneros Bacillus, Paenibacillus, Sporosarcina, Lactobacillus y Bhargavaea nos sugieren que los genes mob presentes en dichos plásmidos pueden estar involucrados en una exitosa THG. El uso extensivo de Tc y OTC para el control de loque americana en algunos países de América pudo haber contribuido al incremento del número de cepas resistentes de Paenibacillus larvae, favoreciendo la transferencia de estos plásmidos entre cepas del mismo patógeno o desde o hacia otras especies de bacterias Gram (+) de los géneros Lactobacillus, Bacillus y Paenibacillus que pertenecen a la microbiota de la colmena (miel, tractos digestivos de abejas adultas y larvas, superficies florales y polen). El uso prolongado de tetraciclina en las colmenas afectadas por loque americana y otras enfermedades bacterianas estaría favoreciendo la ocurrencia de eventos de transferencia genética horizontal como se demostró en cepas del patógeno provenientes de EE.UU.