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Publicación a texto completo no autorizada por el autor / Realiza el análisis cualitativo y evalúa el efecto gastroprotector del extracto acuoso de la parte aérea de Oenothera rosea L‟Hér “Chupa sangre”. Se realizó el análisis cualitativo empleando los reactivos de caracterización según Lock de Ugaz y la solución acuosa reconstituida del extracto acuoso seco de la parte aérea de Oenothera rosea L‟Hér “Chupa sangre”. Para evaluar el efecto gastroprotector se emplearon 48 ratas Holtzman que fueron divididas en 6 grupos. Las lesiones fueron inducidas administrando etanol 96°; las dosis de extracto fueron 250 mg/Kg, 500 mg/Kg y 1000 mg/Kg, las cuales fueron comparadas con la dosis de omeprazol de 20 mg/Kg. La solución acuosa reconstituida del extracto acuoso seco de la parte aérea Oenothera rosea L‟Hér “Chupa Sangre” indicó la presencia de compuestos fenólicos como los flavonoides, taninos, quinonas y también azúcares, triterpenoides y esteroides. Se comprobó la eficacia gastroprotectora de la solución acuosa en un modelo de inducción de lesiones y úlceras gástricas por etanol 96° en ratas Holtzman. Según el sistema de Sydney, los resultados se obtuvieron a una escala macroscópica y a una escala microscópica. En ambas escalas se midió la gravedad de la lesión según los grados de severidad y según la región de profundidad de las lesiones o criterios de evaluación. Los ensayos farmacológicos demostraron que los extractos acuosos en las distintas dosis de administración (250 mg; 500 mg y 1000 mg) y en la dosis de omeprazol (20 mg) exhibieron un significativo efecto gastroprotector (p<0,05) respecto al grupo control. Se determinó la no correlación del diagnóstico macroscópico y microscópico para determinar la dosis efectiva. Conclusión: El extracto acuoso de Oenothera rosea L‟ Hér “Chupa sangre” presenta efecto gastroprotector en inducción de lesiones gástrica en ratas Holtzman. / Tesis

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-09-19T17:34:45Z No. of bitstreams: 1 texto completo.pdf: 672894 bytes, checksum: 1244927f20e5eed71f3171916884e265 (MD5) / Made available in DSpace on 2018-09-19T17:34:45Z (GMT). No. of bitstreams: 1 texto completo.pdf: 672894 bytes, checksum: 1244927f20e5eed71f3171916884e265 (MD5) Previous issue date: 2016-02-24 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Botrytis cinerea, o agente etiológico do mofo cinzento, é um patógeno importante em morangueiros. Clonostachys rosea é efetivo no biocontrole dessa doença em diversas culturas, em condições controladas e de campo. O antagonista pode, ainda, promover o crescimento, aumentar a produção e induzir respostas de defesa nas plantas. Objetivando-se investigar os efeitos da aplicação do antagonista na população do patógeno e no metabolismo da planta, estudou-se a interação entre C. rosea, B. cinerea e morangueiros com base na dinâmica populacional dos fungos e no perfil metabólico do hospedeiro. Avaliou-se a colonização foliar de ambos os fungos e a sobrevivência epifítica de C. rosea em folhas, em câmara de crescimento e em cultivos comerciais. O antagonista se estabeleceu e reduziu a colonização por B. cinerea, principalmente quando aplicado semanalmente. A população de C. rosea diminuiu com o tempo após a aplicação, e a intensidade de colonização pelo patógeno foi inversamente proporcional à do antagonista. Em câmara de crescimento, a população epifítica do antagonista reduziu-se substancialmente aos 3 dias da aplicação. Em condições de campo, onde o intervalo mínimo entre a última aplicação e coleta de folhas foi de 7 dias, não foi possível quantificar a população epifítica do antagonista. Comparou-se o perfil metabólico de morangueiros tratados com C. rosea em intervalos de 7, 14 ou 28 dias ao de morangueiros não tratados, e verificou-se que a aplicação semanal alterou o perfil metabólico. Aumentos significativos nos níveis de citrato e succinato, compostos intermediários do ciclo de Krebs, de alguns fitoesteróis, fitoestrogênios, cafeína e piceatanol foram observados. Tais alterações podem estar relacionadas à promoção de crescimento e ativação do sistema de defesa. Concluiu-se que C. rosea afetou a dinâmica populacional de B. cinerea em tecidos assintomáticos e alterou o perfil metabólico de morangueiros. Esses resultados subsidiam estudos subsequentes para elucidar o modo de ação de agentes de biocontrole. / Botrytis cinerea, the causal agent of gray mold, is an important pathogen of strawberry plants. Clonostachys rosea is effective in the biocontrol of gray mold in several crops, in growth chamber or field conditions. The antagonist may also promote growth, increase the production and induce defense responses of plants. Aiming to investigate the effects of antagonist application on pathogen population and plant metabolism, we studied the interaction of C. rosea, B. cinerea and strawberry plants based on the population dynamics of both fungi and metabolic profile of the host. We assessed leaf colonization of both fungi and the epiphytic survival of C. rosea on leaves, under growth chamber and in commercial cultivations. The antagonist established in leaves and reduced the colonization by B. cinerea, mainly when applied weekly. The population of C. rosea decreased over time after the application and tissue colonization by the pathogen was inversely proportional to the colonization by the antagonist. Under growth chamber the antagonist population was substantially reduced 3 days after applied. Under field conditions, which the minimal interval between last application and the collection of leaves was 7 days, it was not possible to quantify the epiphytic population of the antagonist. We compared the metabolic profiles of strawberry plants treated with C. rosea at intervals of 7, 14, or 28 days with the profiles of untreated plants, and found that weekly applications altered the metabolic profiles. There was a significant increase in the levels of citrate and succinate, intermediate compounds in the Krebs cycle, some phytosterols, phytoestrogens, caffeine, and piceatannol. These changes may be related to growth promotion and activation of plant defense system. In conclusion, C. rosea affected the population dynamics of B. cinerea in symptomless tissues and changed the metabolic profile of strawberry plants. These results can support subsequent studies aiming to elucidate the action mode of biocontrol agents.

Abstract Rhodiola rosea is a medicinal plant, mainly used in Asia and Scandinavia. It is characterized as an adaptogen and is reported to have many pharmacological properties, which are ascribed to the glycosides of cinnamyl alcohol and tyrosol. As natural habitats are already overharvested and the cultivation of this plant needs 4–6 years, the production of the pharmacologically important compounds in in vitro cultures could be an alternative. In the work presented here, the production of these glycosides in compact callus aggregate cultures of roseroot was addressed. Biotransformation of exogenously added cinnamylalcohol and tyrosol was studied. Glucosylation of the precursors yielded high amounts of rosin and salidroside and low amounts of rosavin. During the course of this work, four new glycosides of cinnamyl alcohol were found and identified. The optimal concentration of the precursors and the time needed for the biotransformation was also determined. For enhancing the biotransformation rate, glucose was added to the culture medium alongside with sucrose, which doubled the production of cinnamyl alcohol glycosides but did not affect the production of salidroside. A pilot experiment using air-lift bioreactor was performed. A cDNA fragment encoding tyrosine decarboxylase was isolated and described. The expression of this gene was analysed in the leaves and roots of two chemotypes. The results demonstrate the important role of tyrosine decarboxylase in the production of salidroside. The results revealed production of the pharmacologically important glycosides of Rhodiola rosea; however the successful pilot bioreactor experiment remains to be scaled-up. New information was obtained on the biosynthesis of salidroside, which substantiate the metabolic engineering of roseroot.

Evalúa el efecto antiinflamatorio y cicatrizante de la crema farmacéutica a base del extracto etanólico de las hojas de Oenothera rosea “chupasangre” procedente del Departamento de Ancash (Huaraz). Se determina los metabolitos secundarios mediante marcha fitoquímica (flavonoides, alcaloides, taninos, saponinas, fenoles, glucósidos y otros). Además, evalúa el efecto contra la inflamación y su actividad en las cicatrices en 3 grupos poblacionales (contusiones leves, contusiones moderadas y heridas leves cerradas) de 20 a 50 años de edad, de ambos sexos, los cuales se subdividen en grupos experimentales y controles, en el Centro de Salud Ganimedes Disa IV Lima este - MINSA del distrito de San Juan de Lurigancho. Se determina el estado general para un diagnóstico médico; para luego iniciar el uso tópico por medio de controles de observación y medición de la zona afectada hasta su completa recuperación. Los datos son procesados mediante el análisis (ANOVA), Tukey y análisis de varianza, dando como resultado que las cremas al 3 y 5 % muestran buen efecto antiinflamatorio (contusiones leves y contusiones moderadas) y regular efecto cicatrizante (heridas leves cerradas), mientras, que la crema al 1 % no tiene efecto. Además, la crema al 5 % es sometida a estabilidad acelerada a una temperatura de 40 °C durante 90 días teniendo como parámetros los análisis organolépticos (aspecto, color y olor), fisicoquímicos (pH, viscosidad) y carga microbiológica total; obteniendo como resultado una crema estable y que cumple con los criterios de aceptación. / Tesis

Biological control agents are a promising niche to replace chemical pesticides for treating plant pathogens in agriculture. A potential biocontrol agent is the microparasitic fungi Clonostachys rosea which has the ability to attack various plant pathogens such as other fungi and nematodes. One key feature in the interaction between mycoparasite and prey is degradation of the fungal cell wall where cell wall degrading enzymes are important. One cell wall degrading enzyme is cellobiose dehydrogenase of which it has been found a high number of genes for in C. rosea compared to other mycoparasites. The reason for these many cellobiose dehydrogenase genes being present in C. rosea is what this study aimed to find out. To do so, the different cellobiose dehydrogenase proteins 001, 002, 003 and 004 were successfully expressed in Pichia pastoris. The 003 protein had significantly higher expression levels and were further purified with size exclusion chromatography where some of the resulting purified protein was used to set up a crystallization screen. Unfortunately, no crystals have been formed so far. The enzymatic activity against lactose, cellobiose and laminaribiose of all produced cellobiose dehydrogenase proteins were also analyzed using a 2,6-dichloroindophenol activity assay. The proteins 001 and 002 showed a low activity against lactose and cellobiose whereas the other protein showed no activity for the tested conditions. That these proteins have developed variations in their activities may be one reason for why they are all still existing.

Time-averaging, the mixing of fossils of different ages within a single bed, defines the limit of temporal resolution of the fossil record. Quantitative estimates of this resolution threshold have not been acquired for any group other than mollusks. This study provides the first quantitative estimates of time-averaging for brachiopods, extending our understanding of intrinsic, or group specific controls on this process. Estimates were obtained by direct dating of individual terebratulid brachiopod shells Bouchardia rosea (Mawe) collected from modern surficial shelly accumulations in the Southeast Brazilian Bight (SW Atlantic). Using amino acid racemization dating calibrated with radiocarbon, 82 individual brachiopod shells, collected from four nearshore localities, were dated. The shells vary in age from modern to 3000 years, standard deviation = 680 years. The age distribution is significantly right-skewed (K3=2.48). At 50-year resolution, the temporal completeness is 75% for the last 1000 years and declines to 20% completeness for 1000-2000 yr. BP. Preservational quality (taphonomy) of modern (<50 yr.) shells is statistically indistinguishable from that of older shells, demonstrating that shell taphonomy is not a good predictor of within-assemblage relative age. These results conform to previously published results for mollusks. Therefore, brachiopods can show considerable time-averaging and this time-averaging can be on a scale similar to aragonitic mollusks despite the apparent lack of robustness of calcitic brachiopod shells. This suggests that the brachiopod fossil record can be notably time-averaged, but estimates of this mixing cannot be reliably deciphered from the taphonomic condition of shells. / Master of Science

Made available in DSpace on 2015-03-26T13:08:47Z (GMT). No. of bitstreams: 1 texto completo.pdf: 3042861 bytes, checksum: c35801efccaed5f3893e29c5338bdbb1 (MD5) Previous issue date: 2010-03-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Restinga is an ecosystem associated to Mata Atlântica, in which the species closest to the sea are adapted to adverse climatic and edaphic conditions. Among recent studies, the leaf is the most utilized organ for the understanding of morphoanatomical relationships between individuals and the environment, while works focusing stem and root are yet incipient. Thus, the aim of this work was to characterize the morpho-anatomy of adventitious roots and stems of Blutaparon portulacoides (A. St.-Hil.) Mears (Amaranthaceae), Canavalia rosea (Sw.) Dc. (Fabaceae), Ipomoea pes-caprae (L.) Stweet (Convolvulaceae), Remirea maritima Aubl. (Cyperaceae) e Scaevola plumieri (L.) Vahl. (Goodeniaceae) occurring in a Halophile-Psammophila Reptant formation of the Paulo César Vinha State Park, relating to environment parameters; to describe the secretory structures and to identify the classes of secondary metabolites present in the secretion of leaves of Ipomoea pes-caprae and Ipomoea imperati. Soil was sampled and micro environmental measurements were performed. For anatomical evaluations, roots, stems and leaves of different regions were collected and processed following usual techniques in plant anatomy. The soil had few nutrients and organic matter. The micro environmental data showed that herbaceous plants are susceptible to wind speed conditions, relative humidity and evaporation similar to that of sub shrubby species. Morpho-anatomically, the species presented distinct adaptive strategies,specially stolons or rhizomes, aquiferous tissue, aerenchyma in the roots and starch reserve in both stem and root. In Ipomoea pes-caprae and Ipomoea imperati,mucilaginous trichomes and laticifers were described on leaf lamina, and in I. pescaprae, nectaries on petiole. The morpho-anatomical structures found in stems, roots and leaves of the studied species were outstanding features which allowed the survival and competitiveness of these species in the Non-flooded Herbaceous Formation, once they provide adaptive capacity to high temperature, strong wind, high salinity, lack of nutrients, among others. / Restinga é um ecossistema associado à Mata Atlântica, com as espécies próximas ao mar adaptadas a condições climáticas e edáficas adversas. Dentre os estudos atuais, a folha é o órgão mais utilizado para o entendimento das relações morfoanatômicas dos indivíduos com o ambiente, sendo incipientes os trabalhos sobre caules e raízes. Desta forma, o objetivo foi caracterizar a morfoanatomia de raízes adventícias e caules de Blutaparon portulacoides (A. St.-Hil.) Mears (Amaranthaceae), Canavalia rosea (Sw.) Dc. (Fabaceae), Ipomoea pes-caprae (L.) Stweet (Convolvulaceae), Remirea maritima Aubl. (Cyperaceae) e Scaevola plumieri (L.) Vahl. (Goodeniaceae) ocorrentes na formação Halófila-Psamófila Reptante do Parque Estadual Paulo César Vinha, relacionando com parâmetros ambientais; descrever as estruturas secretoras e identificar as classes de metabólitos secundários presentes no secretado das folhas de Ipomoea pes-caprae e Ipomoea imperati. Foram realizadas coletas de solo e medições microambientais. Para avaliações anatômicas, raízes, caules e folhas de diferentes regiões foram coletados, e processados segundo técnicas usuais em anatomia vegetal. O solo apresentou-se pobre em nutrientes e em matéria orgânica. Os dados microambientais demonstraram que as plantas herbáceas estão sujeitas a velocidade do vento, umidade relativa do ar e evaporação similares às da espécie subarbustiva. Em relação à morfoanatomia, as espécies apresentaram estratégias adaptativas distintas, com destaque para presença de estolões ou rizomas, tecido aquífero e aerênquima nas raízes e reserva de amido tanto no caule quanto na raiz. Em Ipomoea pes-caprae e Ipomoea imperati, foram descritas, na lâmina foliar, tricomas mucilaginosos e laticíferos; e em I. pes-caprae, nectários no pecíolo. As estruturas morfoanatômicas encontradas nos caules, raízes e folhas das espécies estudadas foram características marcantes que possibilitaram a sobrevivência e competitividade dessas espécies na formação Halófila-psamófila Reptante, uma vez que fornecem capacidade adaptativa à alta temperatura, vento forte, alta salinidade, falta de nutrientes, dentre outros.

Laboratory bioassay was used to determine the pathogenicity of nine isolates of fungi of the genera Metarhizium, Beauveria and Clonostachys to the parasitic mite, Varroa destructor. All nine isolates were pathogenic to V. destructor with Metarhizuim anisopliae UAMH 9198, Clonostachys rosea UAMH 9161 and Beauveria bassiana GHA being the most pathogenic within their respective species. Metarhizium anisopliae UAMH 9198 was more lethal to V. destructor than B. bassiana GHA and C. rosea UAMH 9161 with LC50 values of 1.6 x 10(5), 9.6 x 10(6) and 5.4 x 10(6) conidia/mL, respectively. Metarhizium anisopliae and B. bassiana significantly affected brood and adult honey bee survivorship and their immune responses. They were lethal to the bees with LC50s of 3.70 x 10(6) and 2.62 x 10()5 conidia/mL, respectively. The effect of temperature and thymol on conidia germination, production and colony growth of the fungal isolates was determined. Temperature significantly affected conidia germination, production and colony growth (P < 0.05) but thymol did not. Efficacy of the two most promising isolates (M. anisopliae UAMH 9198 and B. bassiana GHA) as potential bio-control agents against V. destructor in hives was evaluated. Fungal inocula were applied as dry formulation, with corn flour as carrier, using dispenser tray and dusting applications. Treatments were either applied alone or in combination with thymol to determine any synergistic effects. All treatments significantly increased mite mortality (P < 0.05), however, the mite control efficacy varied between fungal treatments and application methods. Combined treatments of fungi and thymol caused significantly higher mite mortality than single fungal treatments, which showed control levels of ≤61%. Significant differences in mite mortality were found between the two delivery methods with the differences depending on the fungal isolate. The results suggest that M. anisopliae UAMH 9198 would be a more effective bio-control agent for the management of V. destructor in honey bee colonies than the other isolates tested when dispensed continuously in hives using delivery methods such as a dispenser tray. However, future research is needed to improve delivery methods and investigate the effect of carriers used in the formulation on the efficacy of such entomopathogenic fungi. / Ontario Ministry of Agriculture and Food (OMAF), University of Guelph